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  • Column liquid chromatography  (90)
  • Ultrastructure  (33)
  • Zea
  • Springer  (124)
  • American Institute of Physics
  • Cell Press
  • International Union of Crystallography (IUCr)
  • Oxford University Press
  • 2005-2009
  • 1985-1989  (124)
  • 1945-1949
  • 2005
  • 1988  (124)
  • 1978
  • 1946
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  • 2005-2009
  • 1985-1989  (124)
  • 1945-1949
  • 1975-1979  (117)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Sexual plant reproduction 1 (1988), S. 208-216 
    ISSN: 1432-2145
    Keywords: Megagametophyte ; Synergids ; Brassica campestris ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Brassica campestris, both synergids of the ovule degenerate before the arrival of the pollen tube. Synergid degeneration does not depend on pollination. At the non-degenerated stage, the synergids are completely filled with a complexly organized cytoplasm containing numerous mitochondria with many cristae, a large number of dictyosomes with many associated vesicles, and a very extensive rough endoplasmic reticulum. The degenerative changes that occur in the cytoplasm of the synergids are characterized by a loss of visibility of the membranes of the endoplasmic reticulum and the simultaneous formation of dense deposits on the surrounding membranes of the mitochondria. Locally, the plasma membranes of the synergids disappear, and some ground plasma of the synergids penetrates into the space between the plasma membranes of the egg cell and the central cell.
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  • 2
    ISSN: 1432-2285
    Keywords: Nuclear crystalloid inclusions ; Olea europaea ; Cytochemistry ; Ultrastructure ; Glycoprotein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The nuclei of mesophyll cells of olive trees contain numerous sizeable crystalloid inclusions. Cytochemical examination using epoxy resin-embedded, semithin-sectioned tissue indicated the presence of proteins and oligoor polysaccharides in these inclusions. Their electron microscopical analysis revealed a crystalline substructure consisting of intersected subunits of high order. The spacing of the lattice fibrils and the angles of intersection were determined and used to establish a model of the unit cell of crystallization. It is suggested that the nuclear crystalloids of olive trees consist of glycoprotein molecules. They differ from the intranuclear crystalloids observed in other species predominantly in the high density of their subunit arrangement.
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  • 3
    ISSN: 1432-0878
    Keywords: Basement membrane ; Lipids ; Ultrastructure ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Developing rat incisors were treated with malachite green-aldehyde fixative solution (MGA), which retains and stains lipids. We observed positive staining occurring as dots in the basement membrane. Most of these dots (2–3.5 nm in diameter) were grouped in the lamina densa but some were also present in the lamina lucida and the lamina fibroreticularis. These data provide evidence for the existence of lipids in the dental basement membrane and suggest that they are distributed together with the various groups of proteins so far detected.
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  • 4
    ISSN: 1432-0878
    Keywords: Adrenergic nerves ; Steroidogenic cells ; Embryonic ovary ; β 2Adrenoceptors ; Cytochemistry ; Ultrastructure ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The present study investigates the innervation of the embryonic chick ovary with regard to (i) development and compartmentalization of catecholaminergic nerves, and (ii) presence of adrenoceptors on steroidogenic target cells of catecholaminergic nerve terminals. Catecholaminergic nerve fibers visualized by glyoxylic acid-induced histofluorescence first appeared at embryonic day (E) 13. From E15 through E21 the density of fluorescent aminergic nerves increased markedly in parallel with the concentration of catecholamines and numbers of nerve bundles and single axons seen at the electron-microscopic level. Catecholaminergic nerves were confined to the ovarian medulla and closely associated with interstitial cells. Nerve terminals approached interstitial cells up to a distance of 20 nm and, in their majority, exhibited uptake of the false adrenergic transmitter 5-hydroxydopamine. Although adrenaline amounted to 14% of the total catecholamine content at E21, adrenaline immunoreactivity was only detected in adrenal chromaffin cells, but not in nerve fibers or cell bodies within the ovary. Interstitial cells structurally matured between E15 and E21 as documented by an increase of smooth endoplasmic reticulum and tubular mitochondria. Monoclonal antibodies mAB 120 and BRK 2 raised against avian β 1 and mammalian β 2-adrenergic receptors revealed the presence of β 2-adrenoceptor-like immunoreactivity on the surface of interstitial cells, but not on any other cell type. The results are consistent with the notion of a dense adrenergic innervation of the embryonic chick ovarian medulla and its steroidogenic interstitial cells, and suggest the chick ovary as an excellent model for elucidating the functional role of a neural input to steroidogenic cells during development.
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  • 5
    ISSN: 1432-0878
    Keywords: Smooth muscle ; Intermediate filaments ; Dual network ; Ultrastructure ; Ascidian ; Halocynthia roretzi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An extensive network of intermediate filaments that interconnected cytoplasmic dense bodies and connected the dense bodies to the cell surface was revealed in double-fixed, tannic acid-stained preparations of ascidian smooth muscle. The filament network ran through spaces in the continuous network of myofibrils, connecting them longitudinally, obliquely and transversely to form an intimately associated, dual network. In their transverse passage, the intermediate filaments ran across myofibrils along I-zones exclusively, interconnecting successive dense bodies. The pattern of attachment of intermediate filaments to dense bodies was predominantly “one-sided.” The filaments, which themselves were not incorporated into the contractile apparatus, remained folded or unfolded between myofibrils and between sarcomere-like structures in synchrony with the contraction-relaxation cycles. These results suggest that the intermediate filaments mechanically maintain the organization and arrangement of myofibrils via an intimate association with the myofibrils in the regions of the dense bodies, in such a way that the filaments do not impede muscle function. Based on these observations, a new model for the network of intermediate filaments in smooth muscle cells is proposed.
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  • 6
    ISSN: 1432-0878
    Keywords: Tentacles ; Ultrastructure ; Adhesion ; Leptosynapta spp (Holotheorie) ; Echinodermata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The tentacles of the apodous holothurian Genus Leptosynapta have been studied by use of transmission and scanning electron microscopy. The gross anatomy, water vascular system, fibre systems and ectoneural nerve ring are described. A fuzzy coat of attenuated filaments covers the surface of the tentacle, broken only by secretory ducts. A cuticle underlies the fuzzy coat. Bacteria are common in the subcuticular space. Fixation without osmium gives poor preservation of the surface coats. The epidermis consists of a single layer of columnar cells consisting of Type-1, Type-2, support, goblet and uniciliated cells. Type-1 cells secrete electron-dense material and appear to be homologous to adhesive cells of the tentacles of other holothurians. The support cells contain large, granular vesicles not found in other holothurians. Goblet cells contain flocculent mucus and have an apical cilium. Goblet cells are not found in other holothurian tentacles and may function to lubricate and wrap adhering particles to aid their ingestion. The uniciliated cells are rare, poorly developed and the cilium does not extend past the cuticle. The ultrastructure of the tentacles is discussed in relation to those of other holothurians.
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  • 7
    ISSN: 1432-0878
    Keywords: Lung ; Ultrastructure ; Marsupial ; Newborn ; Surfactant ; Dasyurus hallucatus, Trichosurus vulpecula
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The lungs of newborn northern native cats, Dasyurus hallucatus and newborn brushtail possums, Trichosurus vulpecula were examined by both light and electron microscopy. The native cat has a birth weight of 18 mg after a gestation of about 21 days, whereas the brushtail possum weights 200 mg at birth and has a gestation period of 17.5 days. The lungs of the native cat are two large respiratory sacs, with a respiratory lining of squamous cells and surfactant-secreting cells. The capillaries are located within the connective tissue just below this respiratory epithelium. The visceral covering of the lung is formed by squamous cells. The lungs of the possum are composed of numerous large respiratory sacs which are separated by connective tissue septa in which the capillaries are located. The sacs, as in other species, are lined with squamous cells and surfactant secreting cells. It is proposed that the structure of the lung of the newborn marsupial is related more to the size of the newborn rather than to the length of the gestation period.
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  • 8
    ISSN: 1432-0878
    Keywords: Implantation ; Embryo ; Ultrastructure ; Vasculature ; Rat (Sprague-Dawley) ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Preimplantation-stage embryos were transferred to the anterior eye chamber of recipient rats and guinea-pigs. After implantation had occurred the influence of the embryo on the iris vasculature was examined ultrastructurally. In both species, the earliest effect of embryonic implantation was an increased stromal oedema. Under increasing embryonic influence the vascular endothelial cells showed an increased number of projections into the vascular lumen, while in the rat, endothelial projections were also found pushing back into the basement membrane. In the rat, the endothelium became very irregular in thickness prior to complete disintegration and loss during more advanced stages of implantation. Rat embryonic trophoblast was found invading iris vasculature, particularly in areas where the iridial endothelium was partially or completely missing. Other cells in the iris, including the stroma, appeared to be less affected. In the guinea-pig, however, trophoblast cells appeared to be capable of invading the vasculature by displacing endothelial cells that still appeared morphologically normal.
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  • 9
    ISSN: 1432-0878
    Keywords: Lateral-line sense organs ; Mechanoreceptors ; Supporting cells ; Ultrastructure ; Pleurodeles waltlii (Urodela)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructural and histochemical features of the accessory cells of the neuromast of the salamander P. waltlii have been examined. Three types of accessory cells, supporting, mantle, and basal, were found, but only the first 2 are considered in this article. Supporting cells characterized by a highly dilated endoplasmic reticulum occur among and surrounding sensory cells. Mantle cells, morphologically different from the supporting cells, surround the remainder of the neuromast. Both types of accessory cells exhibit histochemically different secretory materials. Our morphological results suggest that both accessory cells contribute to the formation of cupular material.
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  • 10
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    Cell & tissue research 254 (1988), S. 247-249 
    ISSN: 1432-0878
    Keywords: Pinealocyte ; Light exposure ; Ultrastructure ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Although it is generally known that light strongly influences N-acetyltransferase activity and melatonin production in the pineal gland, little information is available concerning morphological changes following light exposure. As exposure of rats to a short light pulse at night rapidly depresses melatonin synthesis, we decided to determine whether this experimental condition produces rapid changes in the pinealocyte organelles. A 30-min light pulse at night (six hours after lights out) provoked rapid changes in the relative volumes of some pinealocyte organelles. The volume fractions of mitochondria, Golgi apparatus and lipid droplets, and the numbers of dense-core vesicles and “synaptic” ribbons decreased, whereas the volume fraction of lysosomes increased. There were no differences in the volumes of granular endoplasmic reticulum and vacuoles containing flocculent material in those animals exposed to light compared with control animals. These results indicate that a short light pulse at night causes ultrastructural changes that can be interpreted as morphological features of diminished activity of pinealocytes.
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  • 11
    ISSN: 1432-0878
    Keywords: X-organ ; Sinus gland ; Ultrastructure ; Cyclic activity ; Siriella armata (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of the medulla interna-medulla externa X-organ (MI-ME Xo)-sinus gland (SG) complex in the eyestalk of Siriella armata is described during the normal and the experimentally inhibited molt cycle. In the normal SG, four types of neurosecretory axon terminals, each containing distinguishable neurosecretory granules, can be described. Thus, type-2 granules are synthesized by G1 neurons forming the MI-ME Xo. The cell bodies and axonal endings of these cells in the sinus gland have been examined at the following molt stages: intermolt (stage C4), premolt (D0 and D2), and postmolt (A1, A2 and B). Changes in ultrastructure of the G1 cells have been monitored and correlated to inhibitions of the molt-and reproductive cycle produced by electrocauterization of the MI-ME Xo. The results obtained suggest that the neurosecretion from the G1 cells exerts a positive influence on molt and brood preparation. The occurrence of a distal group of G1 cells whose axons terminate at a different site from the SG suggests that the neural factors of the MI-ME Xo are diverse and control different physiological activities.
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  • 12
    ISSN: 1432-0878
    Keywords: Neuropeptide Y (NPY) ; Penis erection ; Perivascular nerve fibers immunocytochemistry ; Ultrastructure ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neuropeptide Y 1–36 (IR-NPY) immunoreactive nerve-fiber processes have been observed in tunicae of veins and arteries and in smooth muscles of the human penis taken at autopsy or during surgery by use of light-and electron-microscopic immunohistochemical techniques. Numerous IR-NPY nerve fibers were mostly concentrated in the inner part of the adventitia close to the media of the arterial and venous vessels and among the intracavernous smooth muscle cells. IR-NPY nerve fibers were less abundant in veins than in arteries. Positive somata were not observed in the penises. At the ultrastructural level, IR-NPY were localized exclusively in large, dense granules of nerve terminals by means of the postembedding immunogold technique. In the deep dorsal vein, IR-NPY nerve fibers were also located in the media formed by an outer circular and an inner longitudinal layer. In the intracavernous and dorsal arteries, they showed the highest density in the inner part of the adventitia. In the corpora cavernosa and in the corpus spongiosum, IR-NPY nerve processes were intermingled between the smooth-muscle fibers around the sinusoid spaces. IR-NPY nerve fibers were present in the cavernous nerves close to the central arteries. The urethra did not show any IR-NPY-positive nerve fibers. This peculiar distribution of IR-NPY nerve fibers suggested that they could participate in regulating arterial and venous blood flow and intracavernous smooth-muscle tone. NPY may therefore be of importance in some of the mechanisms of penile erection especially during detumescence.
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  • 13
    ISSN: 1432-0878
    Keywords: Thymus ; Cultured thymic fragments ; Epithelial cells ; Microenvironment ; Ultrastructure ; Rat (nude)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cultured thymic fragments correspond to the thymic microenvironment depleted of lymphocytes and dendritic cells. When these fragments are implanted under the kidney capsule of congenitally athymic rats, lymphocytes and dendritic cells of host origin enter the graft and induce thymus-dependent immunity in the recipient. This paper describes the ultrastructure of the fragments and the changes that occur during the restoration of normal thymic architecture. At the end of the culture period of 6–9 days and in the early stages after implantation, the grafts consist of keratin-containing epithelial cells of unusual morphology that can be labelled with antibodies raised against the epithelium of the mid/deep cortex and the subcapsule/medulla. Normal thymic architecture develops, including nerves and blood vessels, as lymphocytes populate the environment, and by 4–6 weeks the epithelial cells are the same phenotypically and ultrastructurally as those found in normal rat thymus. However, some areas without lymphocytes still contain the atypical epithelial cells seen before implantation. Large multinucleated giant cells are also present with a few associated epithelial cells of subcapsular/medullary phenotype. In conclusion, the cultured thymic fragments contain a hitherto unknown precursor epithelial cell with an atypical ultrastructure and phenotype that is not seen in normal development.
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  • 14
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    Cell & tissue research 252 (1988), S. 581-587 
    ISSN: 1432-0878
    Keywords: Normal resting breast ; Ultrastructure ; Mitosis ; Cytokinesis ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The parenchyma of the normal “resting” human breast was examined by electron microscopy to characterize the cells undergoing mitosis and the mechanism by which the normal tissue architecture is maintained during this process. In this study of 112 mitotic cells, it was found that the mitotic cells were luminally positioned, polarised epithelial cells with no evidence of myoepithelial cell division. Ultrastructurally, the nuclear and cytoplasmic changes were consistent with previous reports of mitosis in other tissues. However, unlike all previous reports, two specific orientations of the nuclear spindle and thus the planes of cytokinesis were observed. In a few cases the spindle formed parallel to the lumen and division resulted in two luminally positioned daughter cells. However, in the majority of mitotic cells the spindle was approximately at right angles to the lumen and this orientation resulted in a luminally and a basally positioned daughter cell. It is proposed that the abnormally positioned basal daughter cell could develop into a myoepithelial cell or undergo deletion (apoptosis). Thus the two orientations of mitosis may explain the mechanism by which the epithelial and myoepithelial cell populations were maintained by a single progenitor cell without disrupting the integrity of the tissue architecture.
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  • 15
    ISSN: 1432-0878
    Keywords: Ovary ; Steroid-secreting cells ; Ultrastructure ; Chorionic gonadotropin ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructural characteristics of the ovarian medulla of the newly hatched white leghorn chick were studied in control animals and compared with chicks that were treated with human chorionic gonadotropin during embryonic development. The ovarian medulla was mainly occupied by epithelial cells which formed cords or islets surrounded by a basal lamina. Within this epithelial compartment, steroidogenic cells, poorly differentiated cells and a lacunary system could be recognized. When chicks were treated with human chorionic gonadotropin, steroidogenic cells became discernible; there was an increment in the amount of cytoplasm and the area of mitochondria. Poorly differentiated cells exhibited signs of stimulation, and transitional images suggested the transformation of these cells into steroidogenic cells. The epithelial cells of the lacunar system also displayed stimulated cytoplasmic organelles. Evidence was supplied suggesting that relatively undifferentiated cells persist in the ovarian medulla until hatching and can develop into steroidogenic cells under gonadotropic stimulation.
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  • 16
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    Cell & tissue research 253 (1988), S. 137-143 
    ISSN: 1432-0878
    Keywords: Endostyle ; Ultrastructure ; Immunocytochemistry ; 5-Hydroxytryptamine ; Granules ; Ciona intestinalis ; Corella parallelogramma, (Tunicata) ; Ascidia mentula (Tunicata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The cellular and subcellular distribution of serotonin (5-hydroxytryptamine, 5-HT) in the endostyle of three species of ascidians, Ciona intestinalis, Corella parallelogramma, Ascidia mentula, was studied by light-(immunoperoxidase) and electron-microscopic (immunogold) immunocytochemistry. At the light-microscopic level 5-HT-like immunoreactivity (5-HT-LI) was exclusively found in cells located in the lateral portion of the endostyle, between zone 7, known to have iodinating capacity, and zone 8, which consists of ciliated cells. At the electron-microscopic level, the 5-HT-immunoreactive cells were found to correspond to cells containing polymorphous, dense granules, 100–300 nm in diameter. The granules were located in the supranuclear cytoplasm facing the endostyle lumen as well as in the infranuclear cytoplasm facing the extracellular space. Quantification showed that the 5-HT-LI was considerably higher (13–67 times) in cytoplasmic areas containing granules as compared to areas devoid of granules. Most, but not all, of the 5-HT-LI was associated with the dense core of the granules. In conclusion, serotonin-containing cells are located in the peripheral portion of the endostyle, between zones 7 and 8. Serotonin is stored in cytoplasmic granules that are present both in the apical and basal cytoplasm. This suggests the possibility that the cells are bipolar and secrete serotonin both in a basal direction to the extracellular space, and in an apical direction to the pharyngeal lumen.
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  • 17
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    Cell & tissue research 251 (1988), S. 87-93 
    ISSN: 1432-0878
    Keywords: Paneth cells ; Ultrastructure ; Morphometry ; Facultative anaerobes ; Germ-free mice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Germ-free mice were given live or heat-killed facultative anaerobes, and the ultrastructure of ileal Paneth cells was quantitatively examined with special reference to secretory granules showing a bipartite substructure (central core and peripheral halo). After administering live or heatkilled bacteria, there was a decrease in the area occupied by the cores of secretory granules in Paneth cells, and exocytosed core material was observed in the crypt lumen. There were no changes in the area occupied by the halo of secretory granules. None of the examined Paneth cells phagocytosed bacteria. It is concluded that certain bacteria may affect the secretion of antibacterial agents contained in the secretory granules of Paneth cells.
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  • 18
    ISSN: 1432-0878
    Keywords: Endothelium ; Swim bladder ; Capillaries ; Vesicles ; Ultrastructure ; Cryofixation ; Anguilla rostrata (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Several recent studies comparing chemically fixed and cryofixed endothelium have indicated that glutaraldehyde fixation may result in increases in the population of “vesicles” in the cytoplasm. Other reports based on ultrathin serial-section reconstruction of chemically fixed endothelium have revealed that the vesicular system is comprised of interconnected membranous compartments, which are ultimately continuous with either cell surface but do not extend across the endothelial cell. In this study, we have investigated the three-dimensional organization of the vesicular system in directly frozen, freeze-substituted capillaries of the rete mirabile from the swim bladder of the eel, specifically using the same block of embedded capillaries in which frozen capillaries had previously been found to contain less “vesicles” than chemically fixed capillaries. The results show that essentially all vesicles remain inter-connected with each other and are part of two separate sets of invaginations from the luminal and abluminal cell surface like in chemically fixed tissue. Any increase in vesicle number resulting from glutaraldehyde fixation does not affect the overall three-dimensional organization of the vesicular system in these endothelial cells.
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  • 19
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    Cell & tissue research 252 (1988), S. 123-132 
    ISSN: 1432-0878
    Keywords: Uterine epithelium ; Cell culture ; Proteins ; Ultrastructure ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Morphological and biochemical changes occurring in rabbit endometrial epithelial cells when placed in culture were investigated. Cells were examined by scanning- and transmission electron microscopy and freeze-fracture. Morphologically, cultured cells are shorter and broader than the columnar epithelial cells in vivo, but retain their polarity as indicated by the presence of apical microvilli and a well-developed junctional belt. To study changes in biochemical function, proteins synthesized by cells in primary culture were analyzed by two-dimensional gel electrophoresis. Proteins were labeled during a 24-h incubation with 35S-methionine and gels examined by fluorography. The pattern of proteins changed after cells had been in culture for 48 h. On day 3 new proteins were synthesized and several protein species labeled during days 1 or 2 of culture, including uteroglobin, no longer appeared. On days 3–8 of culture the protein patterns were similar. Addition of progesterone, estradiol, prolactin, or combinations of these hormones to the culture medium for 24–144 h failed to elicit consistent changes in the pattern of labeled proteins established after 3 days of culture. Minor differences in protein patterns among unrelated cultures appear to have been derived from the original cells of the culture. These results indicate that after 48 h in primary culture, cells grown in vitro resemble endometrial epithelial cells morphologically, but no longer reflect functionally the character of epithelial cells in the uterus.
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  • 20
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    Cell & tissue research 252 (1988), S. 157-163 
    ISSN: 1432-0878
    Keywords: Frog skin culture ; Desmogenesis ; Interdigitations ; Lamellipodia ; Ultrastructure ; Rana esculenta
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Small trypsinized explants from ventral skin of frogs (Rana esculenta) were maintained in culture for 4 days during which a newly formed epithelium differentiated along the cut edges of the dermis. During the first 6 h adjacent cells produced numerous interdigitating lamellipodia. After 2 days, epithelial polarity was restored by the formation of zonulae occludentes and the epithelial cells were joined by a few small newly formed desmosomes and by numerous interdigitations. Bipartite junctional complexes consisting of a zonula occludens, followed by a series of typical desmosomes, and characteristic of adult frog epidermis were formed only after 4 days. When cultured in the presence of an inhibitor of protein synthesis (cycloheximide) the trypsinized epidermis no longer formed desmosomes. Therefore pools of one or more crucial desmosomal proteins must be very low or non-existent. However, cycloheximide did not prevent the formation of cell contact specializations, consisting of a highly developed system of complex lamellar interdigitations, between adjacent cells.
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  • 21
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    Cell & tissue research 251 (1988), S. 243-248 
    ISSN: 1432-0878
    Keywords: Thoracic duct ; Smooth muscle cell ; Ultrastructure ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The three-dimensional cytoarchitecture and ultrastructure of the smooth muscle cells in the wall of the rat thoracic duct were investigated by scanning and transmission electron microscopy. The muscle layer basically consists of a single layer of circularly arranged cells. The smooth muscle cell is fusiform or ribbon-like in shape, as in veins or venules with a similar or smaller diameter. Connections by spinous processes are observed between adjacent muscle cells along their length. Spot-like membrane contacts frequently occur in areas where facing membranes are closely apposed. These are thought to be gap junctions and may be responsible for electrical coupling and mechanical attachment. Large invaginations arranged regularly in rows on the surface of the smooth muscle cells can be observed. These invaginations are closely associated with a flattened sarcoplasmic reticulum, and caveolae tend to open into the invaginations.
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  • 22
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    Cell & tissue research 253 (1988), S. 397-402 
    ISSN: 1432-0878
    Keywords: Paneth cells ; Ultrastructure ; Morphometry ; Atropine ; Germ-free/Ex-germ-free ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Previous studies have shown that the secretory products of Paneth cells contain antibacterial agents (lysozyme, IgA) that are affected by the bacterial milieu in the intestine. To investigate whether Paneth-cell secretion is controlled via cholinergic mechanisms, the ultrastructure of Paneth cells was studied in four animal groups: (1) germfree (GF) control mice (Jcl: ICR [GN], male, 13 weeks old), (2) GF mice injected subcutaneously with atropine sulfate (200 mg/kg body weight, dissolved in physiological saline 20 mg/ml), (3) ex-GF mice inoculated with feces from specific-pathogen-free (SPF) mice, and (4) ex-GF mice injected with atropine and inoculated with feces from SPF mice. In ex-GF mice inoculated with feces, 70–90% of the Paneth cells showed fewer secretory granules than those from GF mice (p〈0.01). Approximately 30% of the Paneth cells had a large vacuole (3–10 μm diameter) in the apical cytoplasm. Exocytosed electron-dense material from secretory granules was observed in a few crypt lumens. In ex-GF mice inoculated with feces and given atropine, about 90% of the Paneth cells contained numerous secretory granules, like those in GF control mice, but vacuolated Paneth cells and exocytotic figures were rare; thus the secretion of Paneth cells was blocked by atropine. It is therefore possible that the bacterial milieu in the intestine affects the secretory activity of Paneth cells via cholinergic mechanisms.
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  • 23
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    Cell & tissue research 251 (1988), S. 189-195 
    ISSN: 1432-0878
    Keywords: Lymphocytes ; Epithelium ; Mast cells ; Granular leukocytes ; Basophils ; Ultrastructure ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Examination, by light and electron microscopy, of the morphology and the staining properties of intraepithelial lymphocytes from the intestine of the chicken revealed a population of lymphoid cells, of which a proportion (up to 20%) is granulated. The majority of cells were immunoreactive with anti-T cell serum and can therefore be considered to be related to T-lymphocytes, but they did not proliferate when cultured with phytohaemagglutinin. The granulated cells were identical to those previously designated globule-containing leukocytes, but were distinct from mast cells in their morphology, staining reactions and the stability of the granules in different fixatives and buffers.
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  • 24
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    Cell & tissue research 251 (1988), S. 205-214 
    ISSN: 1432-0878
    Keywords: Kupffer cells ; Granulocytes ; Ultrastructure ; Lipopolysaccharide ; Liver ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The fine structure of Kupffer cells has been studied at various times after an intravenous injection of lipopolysaccharide of Salmonella abortus equii. The most prominent effects were: an increase in the number and dimensions of phagocytic vacuoles (often containing ingested LPS and neutrophilic granulocytes); mitochondrial damage, including disintegration of the matrix and cristae; an increase in the amount of dilated, lucent rough endoplasmic reticulum; presence of fat droplets in the cytoplasm. Five days after injection of lipopolysaccharide, the Kupffer cells had resumed their normal ultrastructure. Several minutes after injection of lipopolysaccharide, platelets adhered to the Kupffer and endothelial cells. Between one and six hours, neutrophilic granulocytes accumulated in the liver sinusoids. The resulting obstruction of the hepatic microcirculation most probably affected cellular ultrastructure by ischaemia. At three days, the number of Kupffer cells was doubled, and increased further at later time intervals.
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  • 25
    ISSN: 1432-0878
    Keywords: Chromatophores ; Color change ; Ultrastructure ; Melanophores ; Macrophages ; Xanthophores ; Cichlasoma citrinellum (Teleostei: Cichlidae)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We describe the histological basis of color metamorphosis in the polychromatic Midas cichlid, Cichlasoma citrinellum. Eight percent of the individuals in a natural population transform from gray with black markings to orange, simultaneously losing their ability to adjust coloration in response to background and social context. This trait is inherited. Light- and electron microscopy revealed that this transformation is a two-step process. First, the melanophores die, then macrophage-like cells remove the debris. As a result of this initial process, the underlying xanthophores become visible, producing the orange coloration. A similar process may occur in individuals that further transform to white, or go directly from gray to white.
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  • 26
    ISSN: 1615-6102
    Keywords: Cytoskeleton ; Plasma membrane ; Microtubules ; Ultrastructure ; Freeze-fracture ; Flagellates ; Proteromonas lacertae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The anterior half of the cell surface of the parasitic flagellateProteromonas lacertae is corrugated while the posterior half is covered by hair-like appendages, called somatonemes. In the anterior part, the cortical microtubules are lined by a zig-zag shaped microfibril. Here, these two structures seem to be separated from the plasma membrane. In the posterior half of the cell the somatonemes, analogous to the mastigonemes of chrysophytes, are anchored to the cortical microtubules by paired small deposits of dense material. This was clearly demonstrated by Triton X 100 treatment which solubilized the plasma membrane but left the somatonemes attached to the cortical microtubules. Freeze-fracture images revealed the alignment of clustered intramembrane particles on the P-face of the plasma membrane which correspond to the attachment sites of the somatonemes, seen as dots in thin sections. The ER-derived membrane-associated somatonemes are probably linked to the cortical microtubules by anchoring proteins which are part of the plasma membrane.
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  • 27
    ISSN: 1615-6102
    Keywords: Heat shock ; Soybean root ; Ultrastructure
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    Topics: Biology
    Notes: Summary Soybean seedlings (Glycine max) were incubated in narrow temperature regimes to study the effects of heat shock on cell structures. The incubation temperatures used were as follows: 1. 28 °C (2h); 2. 40 °C (2h); 3. 45 °C (2h); 4. 40 °C (2h)→45 °C (2h); 5. 47. 5 °C (10 min); 6. 40 °C (2h)→47. 5 °C (10 min). Both optical and electron micrographs were taken of the different tissues of root meristems as they responded to heat shock. Cells of roots heated to 45 °C (2h) or 47.5 °C (10 min) with lethal treatment showed drastic heat injuries:e.g., membrane damage, coagulated plasmolysis, protoplasmic contraction, and leakage of cell content. Nucleolar segregation occurred in cells treated at both lethal and supraoptimal temperatures. Seedlings preincubated at 40 °C (2 h) became thermo-tolerant to lethal temperature treatment of 45 °C (2 h) or 47.5 °C (10 min), by protecting the plasmalemma, mitochondria, plastids and nuclei from heat damage. Without preincubation, however, these structures were destroyed.
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  • 28
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    Protoplasma 146 (1988), S. 65-71 
    ISSN: 1615-6102
    Keywords: Apiaceae ; Eryngium campestre ; Metabolism ; Pollen grain ; P-particle ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The metabolism of P-particles (polysaccharide particles) was investigated in mature pollen grains ofEryngium campestre L. Numerous P-particles, originating from dictyosome activity, are found to be accumulated near the apertures, followed by mitochondria. A single layer of ER profiles seems to prevent the fusion of the P-particles with the intine. Instead of this, they fuse with each other forming nonmembrane-bounded polysaccharide-aggregates, which subsequently change their granulated structure to an amorphous. Mitochondria together with small vesicles are involved in the conversion-process. The so formed wall precursors pass through the ER and fuse into the intine.
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  • 29
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    Chromatographia 25 (1988), S. 31-36 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Radioactivity detector ; Solid scintillator ; Fibre-packed flow cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A new type of flow-cell radiation detector for use in liquid chromatography which is packed with aligned scintillator fibers is described. A primary advantage of the fiber packed cell is that light generated by the scintillator is absorbed to a much less extent by the fibers than by the powder scintillator used in conventional flow cells. A detection efficiency of 55% has been obtained for carbon-14 using 0.1-mm diameter hand-pulled glass fibers. Computer modeling has shown that even better results can be obtained by using smaller and more uniform diameter fibers which will allow better packing. The fiber cell also demonstrates back pressures which are a factor of 50 less than the conventional cell and much less susceptibility to absorption of compounds because of its lower surface area.
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  • 30
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    Chromatographia 25 (1988), S. 854-860 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polystyrene ; Gradient elution ; Turbidimetry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Polystyrene samples of narrow molecular-weight distribution have been eluted according to their molecular weight from columns packed with bare silica Si50, phenyl, or C18 bonded phase by gradients of methanol and tetrahydrofuran (THF) or ofiso-octane and THF. Among the six combinations investigated,iso-octane/THF with a silica column formed a proper normal-phase system whereas methanol/THF with a C18 column formed a proper reversed-phase system. The combinations of C18 column andiso-octane/THF or of Si50 column and methanol/THF gradient did not correspond to the approved polarity rules in high-performance liquid chromatography but were nevertheless effective in separating polystyrene mixtures by molecular weight. Methanol andiso-octane are nonsolvents for polystyrene whereas THF is a solvent. The solubility of polystyrene as a function of molecular weight and concentration was determined by means of turbidimetric titration of solutions in THF with the nonsolvents used in the gradients. The solubility and elution characteristics were almost identical on C18 columns or in methanol/THF combinations. The elution from phenyl bonded phase and Si50 columns usingiso-octane/THF gradients required more THF than the solubility experiments. Information is also given on the occurrence of multimodal elution patterns.
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  • 31
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polycyclic aromatic compounds ; Alkylated PAC ; Retention characteristics ; Normal phase
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    Topics: Chemistry and Pharmacology
    Notes: Summary Retention characteristics of series of polymethyl and mono-n-alkyl derivatives of benzene and pyrene, and also of parent polycyclic aromatic hydrocarbons (PAH), were studied using silica and aminopropyl- and cyanopropyl-modified silica. Differences in the selectivities for the studied compound groups were found between the three phases. Deviations from linear behaviour in plots of log (k′)vs. carbon number were observed for the methyl series. These are explained in terms of differences in π-electron delocalisation within the aromatic ring systems. Further, the effect of methyl substitution on selectivity decreased with an increasing number of aromatic rings. Results were obtained which indicated that the primary adsorption site in a cyano column used in normal phase mode, at least for PAH molecules, is the cyano group.
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  • 32
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    Chromatographia 25 (1988), S. 43-50 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed phase ; Void volume ; Adsorption isotherms
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    Topics: Chemistry and Pharmacology
    Notes: Summary In reversed-phase liquid chromatography with n-alkyl bonded silica, the dead volume (V0) of the column is theoretically indeterminate owing to adsorption of organic modifier on n-alkyl chains and of water on silanol groups. With binary mobile phases, retention volumes of the mobile phase components and of their deuterated species are relaeed to the adsorption isotherms and V0 by equations which can be solved with some assumptions on the adsorbed layer composition. Methanol-water and acetonitrile-water systems are studied. As the experimental excess isotherm shows a linear part in the concentration range 50–80% in organic modifier, the hypothesis of an adsorbed layer of constant composition in this range is possible. When increasing the water content of the mobile phase, adsorption of water occurs up to saturation of silanol groups. Then the assumption of a constant water content for a mobile phase having more than 50% of water is applied. With the hypothesis of a constant adsorbed content of organic modifier when the eluent has more than 80% of organic modifier, V0 and the absolute isotherms are calculated over the entire range of mobile phase composition. Experimental retention behavior of the mobile phase components are totally explained by these V0 determinations. The retention times of commonly used V0 markers are compared with V0 values. It is shown that, when buffering the eluent, no visible effect on the distribution equilibrium is observed, so that injection of concentrated potassium nitrate is a convenient method to measure V0. With a few solutes with are UV detectable it is possible to measure V0 whatever the mobile phase composition in methanol-water and acetonitrile-water systems.
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  • 33
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Stationary phase solubility parameters ; Cyano-silica ; Amino-silica ; Diol-silica
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    Topics: Chemistry and Pharmacology
    Notes: Summary Extended solubility parameters have been determined for aminopropyl, cyanopropyl and 1,2-dihydroxypropyl propyl ether (diol) normal bonded-phase HPLC columns. Parameters were calculated from both the retention data of solutes (partition model) and empirically determined solvent strengths (adsorption-displacement model). The use of solvent strengths to calculate solubility parameters for these silica-based bonded stationary phases appears to be superior, since this technique avoids many of the problems that arise from their inherent heterogeneity. Normalized solubility parameters were also used to position these columns on a stationary phase selectivity triangle. The amino and cyano phases appear in regions of the tringle expected based on the properties of the pure liquids, but an ether linkage in the diol apparently neutralizes some of the acidity expected from this phase.
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  • 34
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Macrolide antibiotic azithromycin ; Intermediates of azithromycin ; Optimization of separation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary HPLC is the most convenient method for the assay of the azithromycin (10-dihydro-10-deoxo-11-methyl-11-azaerythromycin A), a semisynthetic macrolide antibiotic and its reaction intermediates. Data concerning the effects of pH, temperature and column type are presented. The analytical procedures enabling the reliable assay of azithromycin and its intermediates, as well as other impurities are defined. Use of a reversed-phase octadecyl column, a pH of 9.3–9.5, and isocratic mode at ambient temperature are the best conditions of analysis.
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  • 35
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Metabolic disorders ; Xanthine and 2,8-dihydroxyadenine ; Urine analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary 2,8-Dihydroxyadeninuria and xanthinuria are inborn enzyme disorders which must be under lifelong therapeutic control. Quantitative determination of 2,8-DHA and xanthine using reversed phase HPLC was performed. A new application for the separation of purines shortens analysis-time. Examples of the determination of 2,8-DHA and xanthine before and under certain therapies are shown. Long term monitoring of the patients offers the possibility of a reliable prophylaxis against stone recurrence.
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  • 36
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Indirect detection of non-electrolytes ; Simulation of induced peaks ; RP systems
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    Topics: Chemistry and Pharmacology
    Notes: Summary Induced peaks observed in indirect detection of nonelectrolytes in high-performance liquid chromatography are simulated. The equation representing the signal intensity of the induced peaks is derived, and it is verified by the experimental results. The peak area is proportional to (K b ′ +1) k a ′ /| a ′ —k b ′ |, where k a ′ ad k b ′ are the capacity factors of the visualization agent and the analyte, respectively.
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  • 37
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    Chromatographia 25 (1988), S. 162-166 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion chromatography ; Analysis of metal ions ; Post column derivatisation ; Amperometric detection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A method for determination of metal ions subsequent to separation by ion chromatography has previously been developed. The method is based on Indirect Amperometric Detection whereby the decrease in the oxidation current, due to a dithiocarbamate ligand added post column, is monitored. Upon elution from the chromatographic column the metal ions are complexed by the ligand. As the complexes formed are electroinactive at the applied potential, the background current decreases according to the metal ion concentration. The method developed in this work involves addition to the reagent of Zn(II) as a auxiliary metal ion to displace the analyte metals from the chromatographic eluent ligand complexes after separation. Sodium bis(2-hydroxy-ethyl) dithiocarbamate was used as the postcolumn derivatising reagent. The addition of Zn(II) to the reagent causes some unforeseen behaviour in the chromatographic system.
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  • 38
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    Chromatographia 25 (1988), S. 199-204 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Column switching ; Catecholamines ; Urine ; Serum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary We developed an automated, two-column HPLC-method that can be used routinely to quantify the catecholamines norepinephrine and epinephrine in body fluids. The method is based on a new, laboratory-prepared SEC-HPAC column material, on the application of a microprocessor-controlled column-switching technique, and on the use of an integrated reaction system for postcolumn derivatization and trihydroxyindol-fluorescence monitoring. It allows the direct injection (upto 500 μl) of an appropriate biological fluid and distinguishes on its integrated sample-processing mode, its practicability, its chemoselectivity and-specivity, its detection limit (2 pg), its within- and between-run precision and its speed of analysis.
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  • 39
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Pheophytinatonicke(II) ; Chlorophyll ; Pheophytin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary HPLC determination of pheophytinatonicke(II) (Pheo-Ni) prepared by the replacement of magnesium(II) in chlorophyll with nicke(II) is described. The good separation of PheoNi was obtained by using chemically bonded C18 as the stationary phase and acetone-methanol (50∶50, vol/vol) as the mobile phase. Conventional spectrophotometric method was also used for the determination of PheoNi. For the synthetic samples prepared by mixing (pheophytinato a) nicke(II) [(Pheo-a) Ni] and (pheophytinato b) nicke(II) [(Pheo-b) Ni], analytical values obtained by the spectrophotometric method were very high compared to those obtained by HPLC. In the proposed HPLC method, (Pheo-a) Ni and (Pheo-b). Ni could be determined in the concentration range of 0.028–30μg/ml and 0.038–30μg/ml with relative standard deviations (n=10) of 3.1% and 0.8%, respectively.
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  • 40
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Flavonoid compounds ; Structure-retention relationships ; Reversed-phase systems
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Reversed-phase HPLC retention behaviour of different flavonoid compounds in different columns and with a range of methanol-buffer eluents was examined. Repeatibility and reproducibility of logk′ and Δlogk′ values were analyzed. Sharp group contributions, expressed as Δlogk′, were observed. These did not depend significantly on acid modifier type, column packing characteristics (C8 and C18) and dimensions (5,7 and 10μm) or the percentage of methanol in the mobile phase. A list of twelve group contributions is presented which is in agreement with literature data.
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  • 41
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    Chromatographia 25 (1988), S. 443-446 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Large scale separations ; Dissolution of silica ; Metallic contaminants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Possible contamination of purified product by the silica matrix is of considerable importance when considering process scale high-performance liquid chromatography (HPLC) as a separation methodology. The stability of silica supports in normal phase, reversed phase and affinity chromatography modes has been examined with organic solvents and water. Both inorganic and organic components were found, contributed by the support, and the chromatographic and distillation equipment. In all cases, based on assumed values of production, they were found to contribute less than 0.01% of impurities. In critical cases crystallisation or dialysis may be considered as a last stage of purification, with re-chromatography to maximise yields.
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  • 42
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Fast atom bombardment mass spectrometry ; Nonionic detergents
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Microcolumn liquid chromatography (LC) combined with fast-atom-bombardment mass spectrometry (FABMS) was applied to the analysis of nonionic detergents. The porous silver filter attached to the top of the capillary fused-silica tubing worked as the interface for coupling of micro LC and FABMS.
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  • 43
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    Chromatographia 25 (1988), S. 577-581 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Process scale ; Examples of separations
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    Topics: Chemistry and Pharmacology
    Notes: Summary The flexibility of process scale high performance liquid chromatography is demonstrated by three examples of reversed-phase separations (a) a standard purification (b) isolation of trace compounds, and (c) recovery from crystallisation mother liquors. It was shown that a material, thought to be acceptably pure, contained a toxic impurity, and a remaining impurity, a previously unseen component of potential therapeutic interest. Favourable cost data were indicated for example (3).
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  • 44
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    Chromatographia 25 (1988), S. 717-720 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed-phase chromtography ; Antifungla agent ; Pharmacokinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A high-performance liqid chromatographic method is reported for the measurement of miconazole in systemic, fungal infectious patients. Pharmacokinetic data are presented for a single patient receiving miconazole therapy. Sample preparation involves protein precipitation by acetonitrile (1:1, vol/vol). Analyses are carried out on a reversed-phae chromatographic system using octadecylsilane stationary phase: a mobile phase consisting of 0.05 M acetate buffer (pH 7.4)acetonitrile (20:80, vol/vol) is used to elaute miconazole is quantified on the basis of ultraviolet absorption at 220 nm. The precision of the method ranged from 3.21% at 0.5 mg/L to 0.85% at 2.0 mg/L. The limit of quantification was established as 0.1 mg/L. Interference from other drugs that are co-adimistered such as amphotericin B, 5-fluorocytosine of ketoconazole and most other comonly encountered drugs was not observed.
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  • 45
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Fractional factorial experimental designs ; Ruggedness test ; Method validation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Recently much attention has been directed to the validation of analytical methods. The level of validation will depend on the method application. If a method is developed for general use, for instance by different analysts, instruments or laboratories, then the effect of these changes need evaluation. This is normally referred to as ruggedness testing, where the limitations of the method to changes in the specified conditions are examined. This paper examines the use of Plackett-Burman fractional factorial experiment designs for ruggedness testing in high performance liquid chromatography (HPLC). The ability of these designs to identify confounding effects is applied to the stability indicating analysis of Salbutamol and its major degradation product, AH4045. The results show that confounding effects, proving extremely pertinent to HPLC, can be identified. The assay proved rugged to most tested changes although not to the use of column packing material from different manufacturers.
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  • 46
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Alkyl-bonded phases ; Retention behaviour ; Ligand density ; n-alkyl chain length
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Two series of bonded phases were synthesized employing LiChrospher Si 100, 10 μm and n-alkyldimethylmonochlorosilanes as silanizing reagents. In series A the n-alkyl chain length, n, of the bonded phase was varied between 1 and 20 at a constant ligand density of 3.5±0.2μmol·m−2. In series B the ligand density, d, was gradually changed from 0 to 4.1μmol·m−2 on the C1, C4, C6, C8 and C18 bonded phases, respectively. The capacity factors of benzoic acid esters and anilines as solutes were found to increase linearly with the n-alkyl chain length of packings at constant eluent composition (series A) up to a so-called critical chain length, ncrit, where the capacity factor remained constant. ncrit was in the range from 11 to 14. The same pattern was observed when plotting the solute capacity factor against the ligand density of the packing at constant n and constant eluent composition (series B). The critical ligand density, αcrit, varied between 2.3 and 3.2 μmol·m−2 depending on n and the solute. Furthermore, solute retention was slightly higher on RP packings with an even number of carbon atoms in the alkyl chain than on those with an odd number. These findings are consistent with the results of Berendsen and de Galan (J. Chromatogr., 196, 21 (1980)), Dill (J. Phys. Chem., 91, 1980 (1987)) and Simpson and Lau (to be published). The observed phenomena reflect the dynamic structure of RP silicas which are related to the mobility of solvated n-alkyl chains. Due to the lack of a precise conformational analysis of the surface of RP silicas, a semiquantitative model was applied to interprete the described dependencies.
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  • 47
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    Protoplasma 147 (1988), S. 55-63 
    ISSN: 1615-6102
    Keywords: Generative cell ; Sperm cell ; Treetomato ; Ultrastructure ; Vegetative nucleus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The structure of the generative cell and its association with the vegetative nucleus in the pollen tube ofCyphomandra betacea Sendt. were observed with the electron microscope. The generative cell, bounded by its own plasma membrane and the inner plasma membrane of the vegetative cell, possesses the cytoplasmic extension which lies within the embayments of a vegetative nucleus. The generative cell contains the normal complement of organelles and, especially, microtubules which cluster into several groups adjacent to the plasma membrane, oriented along the longitudinal axis of the cell. In the pollen tube reaching the lower end of the style aftersemivivo pollination, both of the sperm cells are elongated and polyribosomes and microtubules are the outstanding feature in the cytoplasm. The two sperm cells are connected by a common transverse cell wall, while cytoplasmic channels exist in both the periplasm of the two sperm cells and the transverse wall. The leading sperm cell (Svn) is closely associated with the vegetative nucleus. Thus the present study demonstrates the existence of the male germ unit in the pollen tube ofC. betacea. The possible cytoplasmic continuity between the sperm cells and between the gametes and vegetative cell is considered.
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  • 48
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    Protoplasma 146 (1988), S. 166-173 
    ISSN: 1615-6102
    Keywords: Entomophthorales ; Entomophthora muscae ; Ultrastructure ; Cytochemistry ; Protoplast ; in vitro growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Entomophthora muscae (C.) Fres. can be grownin vitro as protoplasts. Light and electron microscopical studies of thein vitro developed protoplasts have demonstrated the absence of an organized wall over the protoplasmic Con A-positive membrane at all stages of growth. The cytological organization is typical of the Entomophthorales with condensed chromatin in the interphase nuclei and small eccentric metaphase spindles. Long strands of endoplasmic reticulum, microubules and vesicles surrounding the plasmalemma may be involved in maintaining the precise shape ofE. muscae protoplast. Starvation of the fungus induces the formation of hyphal bodies after deposition of Con A- and WGA-positive wall material at the plasmalemma surface.
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  • 49
    ISSN: 1615-6102
    Keywords: Aphanochaete ; Astral microtubules ; Immunofluorescence ; Phycoplast ; Spindle breakdown ; 3-D reconstruction ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The spatial and temporal organization of the microtubular cytoskeleton at the transitional stage of mitosis and cytokinesis has been studied in the chaetophoralean green algaAphanochaete magna using indirect immunofluorescence light microscopy and transmission electron microscopic analysis of serial sections including computer-aided three-dimensional reconstruction. At late mitosis, elaborate asterlike microtubule systems including bundles interconnecting both centriolar regions are present. These systems disappear a the onset of interzonal spindle disintegration. The incipient phycoplast consists of a star-shaped microtubule assemblage projecting from the intact interzonal spindle. It develops strongly at the time of spindle disintegration, later on it becomes compressed by daughter nuclei movement. Cell plate formation is associated with a two-dimensional phycoplast. Phycoplast microtubules remain for a while associated with the completed cross wall but finally they depolymerize. The general occurrence of astral microtubule systems (includingA. magna) is evaluated. The subsequent developmental stages of the phycoplast, formation, maturation and depolymerization, are discussed.
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    Protoplasma 146 (1988), S. 143-149 
    ISSN: 1615-6102
    Keywords: Hyphal tip ; Ultrastructure ; Freeze substitution ; Sclerotium rolfsii
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The hyphal tip ofSclerotium rolfsii was examined after fixation by freeze substitution. The Spitzenkörper consisted of a dense mass of apical vesicles and microvesicles surrounding a vesicle-free zone. Linear arrangements of microvesicles were occasionally observed within the Spitzenkörper. Abundant microfilaments were seen within the Spitzenkörper region, often in close association with apical vesicles and microvesicles. Microtubules passed through the Spitzenkörper and terminated at the plasmalemma at the extreme hyphal apex. Filasomes were mostly observed within the apical region and were in close proximity to the plasmalemma. Rough ER, mitochondria, microtubules, and vacuoles were abundant in the subapical region and were usually oriented parallel to the long axis of the hypha. Ribosomes were aligned on the outer surfaces of mitochondria. Golgi body equivalents were observed throughout the subapical region and appeared as inflated cisternae of varying shapes and electron opacities. Relationships to other basidiomycetous hyphal tip cells are discussed.
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    Environmental biology of fishes 22 (1988), S. 69-77 
    ISSN: 1573-5133
    Keywords: Acid ; Metals ; Ultrastructure ; Olfaction ; Electrical response ; Pollution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Synopsis The effects of acid ((H2SO4) and aluminum AIKSO4 in acidified water on rainbow trout, Salmo gairdneri, olfactory organ were examined using scanning electron microscopy and electrophysiology. Exposure to pH 4.7 resulted in an increase in the number of mucus droplets over parts of the olfactory epithelium, primarily along the ridges of the secondary folds. The addition of aluminum (5.0, 9.5, 20.0 µmol · 1−1) at pH 4.7 resulted in loss of receptor cell cilia, irregularly shaped olfactory knobs, clumped microvilli and swellings on microridge cells. Electrical responses recorded from the olfactory nerve in response to the amino acid L-serine were similar to controls in fish exposed to acidified water. When fish were exposed to acidified water and aluminum the response was depressed. These morphological and electrophysiological changes could be used to indicate metal-induced stress in fish from natural ecosystems.
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  • 52
    ISSN: 1573-5044
    Keywords: maize ; Zea ; amino acid analogs ; selection ; tissue culture ; plant regeneration
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    Topics: Biology
    Notes: Abstract Tissues resistant to lethal levels of equimolar L-lysine plus L-threonine (LT), 5-methyl-DL-tryptophan (5MT, a tryptophan analog), or S-2-aminoethyl-L-cysteine (AEC, a lysine analog) were selected from maize callus capable of plant regeneration (H99 and W77-R3019 genotypes). Resistance to LT resulted from resistant calli having a 19 times greater level of free threonine than wild type tissues. The resistance was expressed in roots of whole plants; threonine levels were two to nine times greater in leaves and kernels of resistant plants than in wild type plants. Slightly greater levels of isoleucine, lysine and methionine were also noted, particularly in the kernel. Genetic studies with individual resistant plants did not always produce inheritance ratios typical of simple Mendelian inheritance, but by the third generation after plant regeneration a trend towards homozygosity was apparent and the data suggests that LT resistance is inherited as a single dominant nuclear gene. Resistance to 5MT resulted from resistant calli having a 133 to 161 times greater level of free tryptophan than wild type tissues. Also, phenylalanine was 22 to 30 times as great and histidine, tyrosine and valine were about two times as great as in wild type tissues. Resistance was expressed in roots of whole plants, and tryptophan levels were at least 2000 times greater in resistant than in wild type plants. Phenylalanine was also 32 times greater. All regenerant plants resistant to 5MT were both male and female sterile. Resistance to AFC was caused by decreased AEC uptake by the callus tissue and was not due to increased levels of free lysine. Plants were not regenerated from this callus.
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  • 53
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    Chromatographia 25 (1988), S. 265-271 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Chiral phases ; Polymer coating ; Quinine substituted polysiloxanes
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    Topics: Chemistry and Pharmacology
    Notes: Summary Stationary phases for chiral separations have been synthesized by chemical modification of porous small particle silica using new procedures of fixation of the chiral moiety. So called pre-polymers of the methylpolysiloxane polysiloxane type are immobilized on silica surfaces by different procedures. These pre-polymers are substituted by chiral groups. Their synthesis is done externally i.e. not in-situ on the support surface. The immobilization on the silica surface is achieved by crosslinking and/or by chemical bonding. Anchor groups within the pre-polymer (e.g. SiH) as well as on the silica surface (SiOH) give rise to the chemical bonding therewith. Chiral phases with quinine as the chiral moiety were obtained which show high separation efficiency as well as chemical stability, in addition to the enantiomeric selectivity required for the separation of certain types of aliphatic and aromatic alcohols.
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  • 54
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    Chromatographia 25 (1988), S. 288-294 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Mobile phase volume ; Ionic solutes ; Eluent electrolyte
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    Topics: Chemistry and Pharmacology
    Notes: Summary A new method for the determination of the mobile phase volume (V m) in liquid chromatography is presented based on the model regarding the retention of ionic solutes in the presence of eluent electrolytes. TheV m value can be determined by measuring the retention volumes of two ions that have the same charge in two eluent electrolyte systems. Compared with the methods using isotopically labelled eluent components or inorganic salts asV m markers, the method presented is proved to give more reasonableV m values for both normal and reversed-phase liquid chromatography. As well as in binary mixed solvent systems, theV m values in single solvent systems can be determined by this method.
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  • 55
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    Chromatographia 25 (1988), S. 339-342 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Gradient elution ; Prediction of retention
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    Topics: Chemistry and Pharmacology
    Notes: Summary A general equation for the final retention of a solute chromatographed under conditions of stepwise gradient elution has been derived. The elution process and the distances travelled by solutes as a function of eluent volume were simulated by computer for the optimization of stepwise gradient prorams from isocratic HPLC data. The validity of the equations was experimentally veritied.
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  • 56
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    Chromatographia 25 (1988), S. 272-278 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polyaromatic hydrocarbons ; Molecular parameters ; Retention prediction
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    Topics: Chemistry and Pharmacology
    Notes: Summary The present research studies the possibility of using the correlation dependence between molecular parameters of unsubstituted polyaromatic hydrocarbons (PAH) and their retention in reversed-phase liquid chromatography to optimize the conditions for the separation and identification of unknown peaks on the chromatograms of multicomponent mixtures. A linear correlation equation, that takes the number and environment of the carbon atom in the PAH molecule into account as well as the differences in the specific interactions of isomeric molecules with polar eluent, has been proposed. The adequacy of the proposed PAH retention model was verified by comparing the calculated retention values with the experimental data. The possibility of identifying unsubstituted PAH according to the number of carbon atoms of various types and according to the values of the molecules lengths (calculated on the basis of the retention of these substances under different eluent compositions) was exemplified by various chromatographic systems (reversed phase-eluent-PAH molecules).
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  • 57
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    Chromatographia 25 (1988), S. 327-331 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Olefin group selectivity ; Methylene group selectivity ; Non-aqueous reversed-phase
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    Topics: Chemistry and Pharmacology
    Notes: Summary The effects of mobile phase composition upon olefin group selectivity (the ratio of the retention factor of a n-alkane to 1-olefin of equal carbon number) has been examined for non-aqueous reversed-phase liquid chromatorphy. Under time-normalized conditions, large variations in olefin group selectivities were noted as the mobile phase constitutents were changes. However, methylene group selectivities were found to be insensitive to the nature of the mobile phase under these conditions. Mobile phases containing alcohols demonstrated low olefin group selectivities compared to those containing acetonitrile as weak solvent. The results of this study explain variations previously observed in the LC separation of olive oil triglycerides that differ in the number of methylene groups and double bonds.
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  • 58
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Cyano bonded phases ; Residual silanol groups ; Diethyldithiocarbamates
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    Topics: Chemistry and Pharmacology
    Notes: Summary When a cyano bonded phase is used together with a nonaqueous eluent, it is universal that the silanol groups which remain at the surface of the silica gel after bonding affect the retention of solutes. With solutes containing such atoms as N, S and O, hydrogen bond may form between the solute and the residual silanol group, leading to dual retention mechanism. Based on the understanding of retention mechanism, methods were developed to separate metal-diethyldithiocarbamate (DDTC) chelates and crown ethers on cyano bonded phase, the mobile phase being a nonaqueous solvent containing triethylamine (TEA).
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  • 59
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    Chromatographia 25 (1988), S. 409-412 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Separation of proteins ; Biological activity after separation
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    Topics: Chemistry and Pharmacology
    Notes: Summary The proteins of the large ribosomal subunit fromEscherichia coli have been separated by size-exclusion, ion-exchange and reversed-phase high-performance-liquid chromatography (HPLC) using various buffer systems. The biological activity of the isolated proteins was tested via their ability to assemble into active 50S subunits (total reconstitution). The activity of the reconstituted subunits was measured with poly(U)-dependent poly-(Phe) synthesis. Reversed-phase HPLC techniques yielded active proteins (80–100%) by application of 2-propanol or acetonitrile. Proteins prepared by size-exclusion chromatography employing ammonium acetate as buffer also gave highly active proteins (70%). On the other hand, separation of the proteins on ion-exchange columns, using urea containing buffers, resulted in reduced activity (up to 50%).
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  • 60
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Phospholipids ; Amniotic fluid ; Fetal lung maturity
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    Topics: Chemistry and Pharmacology
    Notes: Summary A new quantitative analytical method for the determination of phospholipids in amniotic fluid by high performance liquid chromatography (HPLC) is described. In addition to the main compounds, phosphatidylcholine (lecithin) and sphingomyelin, the so-called minor phospholipids, phosphatidylglycerol, phosphatidylinositol and phosphatidylethanolamine can also be determined. Separation is achieved using a guard-column of Lichrosorb Si 60 and an analytical column of Lichrosorb DIOL. Acetonitrile/water is used as mobile phase at an elevated temperature. By determining the recovery rates, the within-run and the between-run precision, it was shown that sufficient accuracy and precision could be achieved for all the parameters examined. The method is highly sensitive, the detection limit for sphingomyelin is 0.2 μg and 0.1 μg for all the other components. A single determination of 5 phospholipids in an amniotic fluid sample takes about two hours. By performing simultaneous extractions it is possible to analyse 5 samples per day.
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  • 61
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed-phase diphenyl and naphtylethyl bonded phases ; Retention/molecular polarizability relationships ; Peropyrene aromatics
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    Topics: Chemistry and Pharmacology
    Notes: Summary The retention behavior of 15 peropyrene-type polycyclic aromatic hydrocarbons was investigated on various bonded stationary phases in reversed-phase liquid chromatography. On diphenyl and naphthylethyl bonded phases, high correlations were obtained between the molecular polarizability of solutes and their retention. However, very low or no correlations were found on various octadecyl bonded phases. These facts are discussed by using the electrostatic interaction concept between the solutes and the stationary phase. We conclude that these observations are due to two reasons: the difference in the degree of planarity of polycyclic aromatic hydrocarbons and the high ability of planarity recognition of octadecyl bonded phases.
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  • 62
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    Chromatographia 25 (1988), S. 504-506 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Porous polymeric sorbent ; Solid-phase extraction ; Chlorophenols
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    Topics: Chemistry and Pharmacology
    Notes: Summary A new porous polymer obtained by the copolymerization of two cross-linking agents: 1,4-di(methacryloyloxymethyl) naphthalene and divinylbenzene was used for the off-line preconcentration of chlorophenols from water solutions by solid-phase extraction. In order to establish its applicability to preconcentration the recoveries and breakthrough volumes of phenol, 2-; 3-chlorophenols; 2,3-; 2,4-dichlorophenols and 2,4,6-trichlorophenols were studied. The recoveries of the studied compounds were compared with those obtained for chemically bonded phases containing hexyl and octadecyl groups.
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  • 63
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    Chromatographia 25 (1988), S. 515-519 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Retention/structure relationships ; Retention prediction ; Substituted phenols
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    Topics: Chemistry and Pharmacology
    Notes: Summary A retention prediction system (RPS) for substituted phenols in reversed-phase HPLC using a ternary mobile phase was investigated. The RPS was used to predict the retention times of the phenols, and then evaluated by comparing measured and predicted retention data. Excellent agreement between both values was obtained. In addition, the retention mechanism of the phenols was investigated by means of a correlation analysis relating the parameters of the RPS to the physicochemical properties of the phenols. The three properties investigated (partition coefficient, hydrophobic substituent constant and Hammett's constant) were used to describe quantitatively the structure-retention relationship. Significant correlating equations between these descriptors and the retention data were obtained.
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  • 64
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; NAD analogs ; NAD glycohydrolase
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    Topics: Chemistry and Pharmacology
    Notes: Summary Pig brain NAD glycohydrolase immobilized on Affi-Gel 10 or nylon 6 was used for the conversion of NAD into 3-acetylpyridine adenine dinucleotide (APAD) or 3-aminopyridine adenine dinucleotide (AAD). A reversed-phase chromatographic system consisting of a C18 Resolve column and phosphate buffer (pH 6.2)-methanol as the mobile phase was used to monitor the production of APAD and AAD.
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  • 65
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    Chromatographia 25 (1988), S. 603-608 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion interaction reagent HPLC ; Separation of amines
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    Topics: Chemistry and Pharmacology
    Notes: Summary Ion interaction reagent RP-HPLC has been employed for the separation of some typical aliphatic and aromatic amines. The effects on retention of the alkyl chain length of the eluent, and of the eluent flow-rate have been studied. The use of solutions of hexylaminium-, octylaminium-and decylaminium-salicylate has been tested and compared, employing both conductometric and spectrophotometric detection. On the basis of the results obtained, the optimal experimental conditions can be chosen for each separation. Aliphatic and aromatic mono and diamines can be separated and detected, with an average sensitivity of the order of 40 ng without any pretreatment or derivatization. The retention data obtained for amines, compared with those obtained with the same ion interaction reagents for anions, help in the interpretation of the mechanism involved in the technique.
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  • 66
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    Chromatographia 25 (1988), S. 609-612 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Gas chromatography ; Triglyceride composition of grapes ; Identification system
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    Topics: Chemistry and Pharmacology
    Notes: Summary The triglyceride composition of the Tempranillo grape (Vitis vinifera) has been examined by a combination of HPLC and GLC. To identify the triglycerides, equations were applied relating log k′ with the molecular variables: equivalent carbon number, chain length and number of double bonds in each of the fatty acids in the glycerides. Ten triglycerides were found, the principal ones being trilinolein (35.75%), dilinoleyl-olein (21.03%) and dilinoleyl-palmitin (17.02%).
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  • 67
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Collagenase ; Enzyme activity
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    Topics: Chemistry and Pharmacology
    Notes: Summary The activity of bacterial collagenase Clostridiopeptidase A was estimated using a labelled synthetic peptide, 4-phenylazobenzyloxycarbonyl-L-Pro-L-Leu-Gly-L-Pro-D-Arg, as substrate. The N-protected dipeptide obtained after enzymatic hydrolysis of Leu-Gly peptide bond was quantified by reversed-phase, high-performance liquid chromatography using 4-phenylazobenzyloxycarbonyl-L-Pro-L-Phe as internal standard. The time dependence of the appearance of the hydrolysis product and the dependence of rates of hydrolysis on collagenase concentration were linear. Kinetic parameters for collagenase were determined to test the suitability of the described procedure.
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  • 68
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Racemic resolution ; Effect of temperature
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    Topics: Chemistry and Pharmacology
    Notes: Summary The effect of temperature on the resolution of racemates was investigated with respect to the resolution of racemic mixtures of aminoacid esters and 2,2,2-trifluoro-1-(9 anthryl) ethanol. Generally the capacity factor (k′) was found to increase with decreasing temperature. The values of the difference in the standard change of molar Gibbs energy, (ΔΔG), between the more retained and the less retained enantiomer do not appear to vary in a particular fashion. The sign of the enthalpic (ΔΔH) and the entropic, (ΔΔS) quantities for all the mixtures also depend upon the nature of the solute mixture.
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  • 69
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    Chromatographia 25 (1988), S. 791-796 
    ISSN: 1612-1112
    Keywords: Gel permeation ; Column liquid chromatography ; Column switching ; Additives in cellulose acetate
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    Topics: Chemistry and Pharmacology
    Notes: Summary The capability of elution-elution multi-dimensional liquid chromatography was investigated. A “column scaling” approach was evaluated for the quantification of low-molecular-weight additives in cellulose acetate. A small-bore (1-mm i.d.) gel-permeation column was used to separate the higher-molecular-weight polymer from the lower-molecular-weight components. Once separated these additives were transferred to a C18 reversed-phase column via a switching valve. The reversed-phase system successfully separated and quantified individual additives. Analysis time for an ultraviolet inhibitor, Tinuvin®P, in cellulose acetate, including re-equilibration, was approximately 30 minutes. Both accuracy and precision were good. Precision over a three day period was about 1.5%.
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  • 70
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    Chromatographia 25 (1988), S. 811-814 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Separation of trimedlure isomers ; Medfly ; Ceratitis capitata
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    Topics: Chemistry and Pharmacology
    Notes: Summary Semi-preparative high-performance liquid chromatographic procedures on 5 μm silica were developed for the isolation of gram quantities of eight trimedlure isomers (trans: A, B1, B2, and C;cis: V, W, X, and Y) for comparative biological evaluation and NMR studies. Isolations were made from an eight-component 2∶1cis: trans-trimedlure mixture, a four-componentcis-trimedlure mixture, trimedlure-B2:X and trimedlure-C:W epimerization merization mixtures, a trimedlure-B1:Y:B2 mixture, and a trimedlure-A concentrate.
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  • 71
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    Chromatographia 26 (1988), S. 133-138 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Electrochemical detection ; Flavonoids ; Metal-complexes
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    Topics: Chemistry and Pharmacology
    Notes: Summary The electrochemical and chromatographic behaviour of flavonoid standards and of flavonoids extracted from food (green tea, black tea and onions) is investigated with respect to metalbinding properties. It is shown that metals such as iron, copper or aluminium are complexed by flavonoids, preferrably by those having an aromatic o-dihydroxy structure. This is confirmed by cyclic voltammetry on HPLC fractions (stopped flow) and by AAS measurement of metals. As the complexing sites of flavonoids are closely related to electrochemical properties, this is used for an indirect detection of metal species at low oxidation potentials. For iron species in particular a sensitive and selective detection is possible. For copper reductive detection can also be used.
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  • 72
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Electrochemical detection ; Morphine ; Polymeric column
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    Topics: Chemistry and Pharmacology
    Notes: Summary A simple, sensitive, accurate and reliable method for morphine determination in biological samples has been developed. It uses reverse-phase HPLC on a polymeric column with an eluent (0.05 mol/L dibasic sodium phosphate: acetonitrile, 85∶15) at pH 9.5, allowing both the suppression of ionization of morphine amine and the promotion of oxidation of the phenolic group. Amperometric detection at mild oxidizing potential (350 mV) proves very selective, and, therefore, only a simple and rapid one-step liquid-liquid sample preparation is required. Under these conditions “clean” chromatograms are obtained even with complex biological matrices such as cadaveric blood, urine and hair. Minimum detectable amount of morphine is about 200 pg injected. A calibration line with a correlation coefficient of 0.99996 has been produced over the range 7.5–250 ng/mL. Precision results comparable to other HPLC methods.
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  • 73
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Electrochemical detector ; Phenolics ; Lignocellulosic materials
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    Topics: Chemistry and Pharmacology
    Notes: Summary The application of HPLC with an electrochemical detector for the determination of phenolics in lignocellulosic materials is reported. The separation of phenolic acids and aldehydes (gallic acid, p-hydroxybenzoic acid, vanillic acid, p-coumaric acid, syringic acid, ferulic acid, vanillin, syringaldehyde and p-hydroxybenzaldehyde) on two different columns (reversed phase C6 and styrene-divinylbenzene PLRP-S) is shown. Chromatograms of phenolic compounds in neutral, basic and oxidative extracts of wheat straw treated with NaOH and white rot fungusStropharia rugosoannulata are reported along with quantitative results.
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  • 74
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    Chromatographia 26 (1988), S. 234-236 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Preparative-scale separations ; Teicoplanin ; Structure determination
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    Topics: Chemistry and Pharmacology
    Notes: Summary Teicoplanin, a lipoglycopeptide antibiotic active against gram-positive bacteria, is produced as a complex mixture consisting of six major components and four chemically related minor cmmponents. Preparative HPLC was used to isolate small amounts of the pure minor components, in order to determine their structures. In the present paper the isolation procedures are presented, as well as the analytical HPLC conditions. The retention times of the minor components with respect to those of the more abundant compounds, permittedus to make hypotheses on their structures, which were then confirmed by NMR and FAB-MS investigations.
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  • 75
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Phase-transfer catalysis ; On-line derivatization ; Phenolic steroids
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    Topics: Chemistry and Pharmacology
    Notes: Summary Commercially available equipment from two manufacturers served to set up an automated system for the precolumn phase-transfer-catalyzed dansylation of phenolic steroids, using ethynyl estradiol (EE) and estradiol (E) as model compounds. Using different mixing techniques, the on-line determination of EE and E in 200μl untreated urine samples was achieved at a rate of 3–8 analyses per hour. Detection limits were calculated to be 3–5 ng/ml. Calibration curves in urine were linear over two orders of magnitude with r=0.999 (n=5) for EE and r=0.999 (n=6) for E. The repeatability of the determination of EE in urine (1μg/ml) was 3.9% (RSD; n=20) and of E (1.5μl/ml) 3.8% (RDS; n=10). The use of plasma instead of urine in the on-line procedures was not possible due to rapid formation of emulsions, but E and EE were determined in 100μl plasma samples using a mild off-line mixing procedure in 10min. Detection limits were calculated to be ca 10ng/ml. A reaction detector, based on a solvent-segmented system, was developed for the on-line post-column dansylation of phenols and was coupled with a reversed-phase LC system. The highly selective system showed excellent linearity over at least two orders of magnitude with r=0.9999 (n=6) for both phenol and 2,5-dimethylphenol. The reproducibility was good with RSD values of around 2%. Detection limits for loop injections from standard solutions were calculated to be between 4 and 11ng.
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  • 76
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Enantiomeric purity ; Chiral HPLC ; Oxyphenonium ; Post-column extraction detection
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    Topics: Chemistry and Pharmacology
    Notes: Summary A method is described for the determination of the enantiomeric purity (enantiomeric excess) of the anticholinergic drug oxyphenonium. The method for this quaternary ammonium compound is based on the direct HPLC analysis with a chiral stationary phase. Two kinds of α1-acid glycoprotein-bonded phases were used. For the detection a post-column extraction with fluorescence detection of the ion-pair counter ion dimethoxyantracene sulphonate was used.
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    Chromatographia 26 (1988), S. 297-299 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Electrochemical detection ; 2-Mercaptobenzothiazole ; River water
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    Topics: Chemistry and Pharmacology
    Notes: Summary A method is proposed for the determination of 2-mercaptobenzothiazole in river waters using multi-electrode electrochemical detection HPLC. 2-mercaptobenzothiazole determination is unsatisfactory by gas chromatography as it degrades readily on the column. Multielectrode electrochemical detection HPLC combines sensitivity and the ability to ‘screen out’ other electrochemically active species. The development work leading up to the proposed method is discussed. The method has a limit of detection of 0.798μgl−1 2-mercaptobenzothiazole and a total standard deviation of 2.06μgl−1 2-mercaptobenzothiazole at a concentration of 7.97μgl−1 2-mercaptobenzothiazole in river water.
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    Chromatographia 26 (1988), S. 369-371 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Urinary trypsin inhibitors
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    Topics: Chemistry and Pharmacology
    Notes: Summary Human urine contains a family of trypsin inhibitors. Procedures for their purification and characterization involve laborious techniques and the conclusions are different in the identification of the separated compounds. We report results obtained by applying our RP-HPLC method to some procedures adopted by different authors.
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  • 79
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Determination of S-Carboxymethyl-L-cysteine ; Precolumn derivatization ; HPLC on-line clean-up
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    Topics: Chemistry and Pharmacology
    Notes: Summary In order to follow levels of S-Carboxymethyl-L-cysteine in biological fluids for a period as long as three half-lives after drug administration during pharmacokinetic studies, an improved method for its determination had to be developed. Like the previous one, this method uses a protein precipitation step followed by an O-Phthalaldehyde derivatization step and then an HPLC on-line clean-up. This latter was obtained by means of a switching valve system, including a Nucleosil CN 5 μm (3 cm × 4.6 mm i.d.) precolumn and a Spherisorb ODS 5 μm (15 cm×4.6 mm i.d.) analytical column. The sensitivity limit was improved to 0.1 μg/ml in plasma samples and 0.2 μg/ml in urine samples. This method was applied in studies comparing single (0.75 g) and repeated (0.75 g tid) oral administration of the drug to 30 elderly patients and 20 healthy volunteers. Results showed that the half-life was 40% longer in elderly patients than in healthy volunteers, and that area under the plasma concentration versus time curve (AUC) values in elderly patients were twice those obtained with young subjects.
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    Chromatographia 25 (1988), S. 895-898 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ozone ; Formaldehyde ; Annular denuder ; Air monitoring
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A denuder sampling method combined with HPLC analysis for the simultaneous determination of formaldehyde and ozone in ambient air is described. It is based on the reactions of CH2O and O3 with 2,4-dinitrophenylhydrazine (DNPH) and 4-allyl-2-methoxyphenol (eugenol)_respectively, both acting as coatings of two annular denuders connected in series. Formaldehyde released from the ozonolysis of eugenol is quantitatively collected on a third downstream DNPH-coated denuder. The two DNPH denuders are then extracted and analyzed as hydrazone derivative by HPLC with UV absorbance detection. The stoichiometric factor of the eugenol-ozone reaction was found to be 2.0±0.1 moles of O3 per mole of CH2O. The limits of detection are 0.8μgm−3 CH2O and 3μgm−3 O3 for 100l air sampled, corresponding to 1-h sampling at 1.7l min−1.
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  • 81
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Silica modifiers ; Aromatic hydrocarbons
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The retention parameters of benzene, naphthalene; anthracene and phenanthrene on untreated silica gel were determined in the presence of four modifiers: 1-methyl-2-pyrrolidone (MP), 4-formyl morpholine (FM), γ-butyrolactone (BL) and sulfolane (SF). Better column performance was found in the case of FM and MP only.
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  • 82
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    Chromatographia 25 (1988), S. 870-874 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Gel-permeation ; Pectinase activity ; Aspergillus Niger andRhizopus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A rapid liquid chromatographic (HPLC) method for the determination of pectinolytic activity of enzymes produced by Aspergillus Niger and Rhizopus species is reported. Compared with more conventional methods, HPLC is more reliable and has a much improved maximum sensitivity. The limit of quantitation of galacturonic acid is 0.1μg.
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  • 83
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    Chromatographia 25 (1988), S. 887-890 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Thiopental ; Pharmacokinetics ; Serum and plasma samples
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A high-performance liquid chromatographic method for the analysis of thiopental in 100μl of human serum or plasma is described. The procedure involves protein precipitation with acetonitrile. The supernatant is directly injected into a chromatograph containing a reversed-phase CLC-ODS (Shimadzu) column. A 50∶50 (v/v) mixture of water-acetonitrile, at a flow-rate of 1.0ml/min is used as the mobile phase. Detection is carried out ata wavelength of 280nm. Total analysis time per sample is 10min. The assay was found to be linear in the range of 0.1 to 120μg/ml. Reproducibility was good, with intra-assay coefficients of variation from 1.780 to 3.208% and inter-assay coefficients of variation from 3.241 to 4.860%. The absolute recoveries were 97.4 to 101,4%. Other drugs were tested for potential interference with the assay, but none was found.
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  • 84
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Chiral separations ; Chymotrypsin on silica ; Amino acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A new HPLC stationary phase was synthesized by the covalent immobilization of α-chymotrypsin on silica. This increased the stability of the enzyme, without decreasing its activity. The initial chromatographic studies show that this phase can be used for chiral separations of enantiomeric solutes. The stereochemical resolutions of amino acids and amino acid derivatives are reported.
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  • 85
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    Chromatographia 25 (1988), S. 1034-1038 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Determination of propafenone ; 5-OH propafenone ; Reversed-phase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary This paper presents a method for the simultaneous detection of propafenone and 5-hydroxypropafenone (5-OH propafenone) using HPLC. The method is sensitive, selective, and reproducible. The chromatographic separation is based on a 25×0.4 cm 5 μm ODS column, a mobil phase of 0.1 M potassium phosphate (K2HPO4) buffer (pH 2.5) and acetonitrile (63∶37), and UV absorbance detection. There is excellent intra- and interassay variation. Retention times of the internal standard, propafenone, and 5-OH propafenone are 4.3, 6.0 and 2.9 minutes respectively. This method shows linearity over the therapeutic range for propafenone and 5-OH propafenone (0.15 to 3.0 μg/ml and 0.075 to 1.5 μg/ml respectively). No interference has been found from other commonly administered drugs, including several antiarrhythmic drugs, at therapeutic levels.
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  • 86
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Soyabean leaf extracts ; Kaempferol glycosides ; Kaempferol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary An HPLC method is described for the determination of kaempferol glycosides in soybean leaf extracts. The method is rapid and can provide qualitative and relative quantitative results for 9 kaempferol glycosides. The flavonol glycosides are extracted from soybean leaves by shaking the samples in 50% methanol; the extracts are filtered, evaporated to dryness and reconstituted in methanol before further purification through a C-18 Sep-Pak column. The samples are injected onto a C-18 HPLC column, separated by gradient elution with a 1% phosphoric acid: methanol mixture and detected on a UV/VIS diode-array detector. Flavonols were monitored at 265 and 348 nm and spectra from 180 to 400 nm were stored and used as an aid in identification. Relative quantities of the kaempferol glycosides among soybean cultivars were calculated from their proportion of peak area in the chromatograms. Total kaempferol concentration of the extracts was calculated after acid hydrolysis of the kaempferol glycosides to the aglycone and comparison of peak areas to kaempferol standards.
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  • 87
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Gel permeation ; Human serum IgG ; Fluorescent immune complex ; Fluoresence detection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A high-performance liquid gel-permeation chromatographic method is described for the determination of human serum immunoglobulin G (IgG) by separating the fluorescent immuno complex from the free fluorescence-labeled antibody. Fluorescence-labeled antibody used in this study was fluorescein isothiocyanate (FITC)-labeled Fab fragment goat anti-human IgG (anti-IgG Fab). Immuno complexes and antibody of different molecular sizes can be separated. FITC-labeled anti-IgG Fab was added to the serum and the mixture is passed through the column. An immuno complex separates as well-delineated peak in the column void volume, and was measured by the fluorescence of the column eluate (Ex=490nm, Em=520nm). The total analysis time for a serum sample was approximately 15min. The minimum detection limit was 25 mg/dl. The relative standard deviation was below 2% (peak area). The results of the HPL-GPC analysis correlate well with those obtained by laser nephelometric assay (r=0.992).
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  • 88
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    Chromatographia 25 (1988), S. 1067-1071 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Retention behaviour of 1,n-halo(alkylthio)alkanes ; Effect of the halogen atom ; Effect of the number of methylene groups
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The RPLC retention of 1,n-halo(alkylthio)alkanes on an ODS phase with methanol/water (95%/5%) has been investigated. Replacement of a methyl group in a dialkyl sulphide R-S-R by a halogen atom X, to yield the title compounds X-(CH2)n-S-R, generally results in a retention decrease. This retention loss results either from the halogen atom proper, or from its immediate vicinity, or from a reduced retention contribution of the alkyl units. The methylene groups (n) between halogen and sulphur produce somewhat greater retention than the methylene groups in the alkyl groups R.
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  • 89
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Aliphatic amines ; Separation factors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary An analytical method is presented for the identification and separation of aliphatic monoamines, diamines and polyamines. Dansyl chloride as the derivatizing agent and fluorimetric detection were employed. The behaviour of different column packings and eluent compositions was tested and compared. The interaction mechanisms of the dansylated compounds with stationary and mobile phases are discussed on the basis of the analysis of the capacity factors evaluated in the homologous series of monoamines CH3(CH2)nNH2, with n varying up to 9 and diamines NH2(CH2)nNH2, with n varying up to 12. An application of this method to a real sample is presented.
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  • 90
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Detection, chiroptical, polarimetric, circular dichroic ; Enantiomers ; Sorbents, optically active
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Polarimetric and/or circular dichroic detection of enantiomers after liquid chromatography on optically active sorberts has been used for the following purposes: determination of enantiomeric purity in spite of peak overlap, investigation of enantiomerization during chromatography, monitoring of racemizations and recording of circular dichrograms without preparative enrichment of enantiomers. Novel examples for all applications are give. The general prospects for chiroptical detection are discussed.
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  • 91
    ISSN: 1612-1112
    Keywords: Size-exclusion chromatography ; Column liquid chromatography ; Peptides ; Hydrolysates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Gel filtration (size-exclusion) and high-performance liquid chromatography have been used to separate peptic peptides from haemoglobin hydrolysate. Elution profiles on Sephadex G-25 displayed nine fractions with molecular weights lower than 6500 daltons. Each fraction was analysed for total amino acid content and showed less than 1% free amino acids. Reversed phase HPLC, using ammonium acetate buffer and acetonitrile as solvent, was applied to each fraction in order to obtain pure peptide peaks. The importance of acquiring a better knowledge of such an hydrolysate is discussed. Various potential applications of this type of hydrolysate, some of them already being undertaken, are envisaged.
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  • 92
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Charge transfer mechanisms ; Aromatic esters ; Structure/retention relationships
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The separation of aromatic esters has been studied on acceptor bonded phases. It is likely that donor-acceptor complex (DAC) formation occurs between the acceptor phase and the electrons of the carbonyl groups. The influence of substituents attached to the aromatic rings and the role of the alkyl chains of phthalates on retention are discussed.
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  • 93
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Gradient elution ; Prediction of retention ; Aldehydes and amines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A general chromatographic model has been set up starting from a set of equations based on the concept of the velocity of a solute along the column. The composition of the mobile phase is taken into account solely as a numerical factor entering into suitable equations and totally independent of the chemical-properties of the constituents. A few isocratic experimental runs are necessary as input data, and subsequently a small amount of computational effort is sufficient to make predictions of retention times under gradient elution conditions for solutes of whatever chemical structure. The prediction errors are dependent on the steepness of the linear gradient chosen but are, in any case, acceptably low.
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  • 94
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    Chromatographia 25 (1988), S. 437-442 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Large scale separations ; Optimisation procedures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Scale up of analytical High-Performance Liquid Chromatography (HPLC) to the process scale offers promise as a preferred separation method for many high value fine chemicals, both for development purposes and continuous production. It's early adoption is curtailed by the widely held perception that preparative HPLC is a low efficiency high cost technique of limited application. This view has largely beeen generated by the generalised use of large particle size, low quality angular silica supports, and difficulties with packing methodologies. A systematic study of a new silica, in new axially compressed equipment using Plackett-Burman statistics, has demonstrated that very high efficiences, leading to high capacities, can be attained.
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  • 95
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Racemic resolution ; Aminoacid esters ; Mobile phase composition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The work in this paper demonstrates the effect of the mobile phase composition on interactions involving racemic esters of aminoacids and a chiral stationary phase derived from N-formylisoleucine. Aprotic solvents have been employed to test the resolution of these racemates. An increase in the alcohol concentration of the mobile phase results in the deterioration of selectivity. These solvents seem to favour high separation factors. The solvent 1,1,2-trichlorotri fluoroethane is a suitable mobile phase modifier for normal phase liquid chromatographic resolution of enantiomers of aminoacid esters.
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  • 96
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    Chromatographia 25 (1988), S. 697-700 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Micro columns ; Fluorometric detection ; Packed flow cell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A packed flow cell was used for fluorometric detection in micro high-performance liquid chromatography. The flow cell consisted of fused-silica tubing packed with the same material as the separation column. A focusing effect of the stationary phase on the signal intensity was observed, leading to an improvement of the mass detection limit, as achieved by on-column detection.
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  • 97
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    Chromatographia 25 (1988), S. 815-816 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; On-column detection ; Fused silica microcolumn ; Fibre optics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Modification of the UV absorbance detector for LC is described. It is based on a simple combination of a commercial photometer with optical fibres to conduct the light. The performance of the detector is illustrated by the analysis of polyaromatic hydrocarbons on a packed fused-silica column. The minimum detectable concentration for naphthalene was measured as 6·10−6M.
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  • 98
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Inductive effects ; Resonance effects ; Methylquinoline derivatives
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Hydrophobicity of methylquinolines was compared with their retention data measured on two stationary phases by reversed-phase HPLC, and with the experimental partition coefficients (log P). The influence of the substituent position on the hydrophobicity is discussed and a two-parameter correlation equation is proposed permitting the prediction of the log k′ values with help of the inductive and resonance effects.
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  • 99
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion-pair chromatography ; Valve switching ; Fluorouracil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Fluorouracil has been analyzed with a reversed-phase ion-pair liquid chromatographic system applying PRP-1 as the stationary phase. A valve switching technique was applied for the separation of the analyte and some other chemotherapeutic agents. The described procedure gives a rapid screening for the analyte in urine and plasma samples without interferences from late eluting solutes. Biological samples are pretreated by means of a liquid-liquid extraction resulting in a recovery of about 52% for plasma and urine with minimum detectable amounts of about 5ng/ml. The sensitivity and the selectivity of the procedure allows the monitoring of fluorouracil in body fluids and subsequent pharmaco-kinetic studies of the fluoropyrimidine.
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  • 100
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    Chromatographia 26 (1988), S. 120-124 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Chlorophylls ; Pheophytins ; Fluorimetric detection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A HPLC method for the separation and estimation of chlorophylls and pheophytins in fresh and frozen peas was assessed with the aim of following the colour changes of the products during storage under frozen conditions. The method involves the acetone extraction of the pigments, followed by HPLC on RP18 with isocratic elution by acetone: ethanol:water, (70:17:13), detecting and estimating the separated compounds by fluorimetry. The mobile phase composition was selected by monitoring the separation efficiency of several mixtures of solvents by HPLC. The reliability and the accuracy of the method were checked.
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