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Determination of anticonvulsant drugs and methyl xanthine derivatives in serum by liquid chromatography with direct injection: column-switching method using a new internal-surface reversed-phase silica support as a precolumn

Medicinal plants have been the basis for discovery of various important marketed drugs. Xanthine is one such lead molecule. Xanthines in various forms (caffeine, theophylline, theobromine, etc) are abode in tea, coffee, cocoa, chocolate etc. giving them popular recognition. These compounds are best known for their diverse pharmaceutical applications as cyclic nucleotide phosphodiesterase inhibition, antagonization of adenosine receptor, anti-inflammatory, anti-microbial, anti-oxidant and anti-tumor activities. These properties incentivize to use xanthine as scaffold to develop new derivatives. Chemical synthesis contributes greater diversity in xanthine based derivatisation. With highlighting the existing challenges in chemical synthesis, the present review focuses the probable solution to fill existing lacuna. The review summarizes the available knowledge of xanthine based drugs development along with exploring new xanthine led chemical synthesis path for bringing diversification in xanthine based research. The main objective of this review is to explore the immense potential of xanthine as scaffold in drug development.

A specific and sensitive method for the determination for difluprednate (DFBA) and its metabolite (deacetylated DFBA, DFB) in aqueous humor was developed. DFBA and DFB were initially absorbed on a Pinkerton-type column, then analyzed by high-performance liquid chromatography using a semi-micro column after column switching. Under the optimized conditions, calibration curves for DFBA and DFB showed good linearity over the range of 1.0–50 and 0.5–50 ng/ml, respectively. We applied the method to the analysis of DFBA and DFB in rabbit aqueous humor, and found that DFBA in rabbit aqueous humor 1 h after instillation of 0.002% DFBA ophthalmic emulsion was not detected, but DFB was present at the concentration of 4.3±3.1 ng/ml.

Haloperidol and its two metabolites, reduced haloperidol and 4-(4-chlorophenyl)-4-hydroxypiperidine (CPHP) in human plasma and urine were analyzed by HPLC–MS using a new polymer column (MSpak GF-310), which enabled direct injection of crude biological samples without pretreatment. Recoveries of haloperidol and reduced haloperidol spiked into plasma were 64.4–76.1% and 46.8–50.2%, respectively; those for urine were 87.3–99.4% and 94.2–98.5%, respectively; those of CPHP for both samples were not less than 92.7%. The regression equations for haloperidol, reduced haloperidol and CPHP showed good linearity in the ranges of 10–800, 15–800 and 400–800 ng/ml, respectively, for both plasma and urine. Their detection limits were 5, 10 and 300 ng/ml, respectively, for both samples. Thus, the present method was sensitive enough for detection and determination of high therapeutic and toxic levels for haloperidol and its metabolites present in biological samples.

The measurement of the anti-epileptic drug phenobarbital from serum samples combining immunoassay and size-exclusion chromatography is presented. The immunoreaction is based on the competitive binding of the analyte (unlabelled phenobarbital) and the fluorescent-labelled phenobarbital to anti-phenobarbital antibodies. Mixing of the reagents and the immunoreaction takes place in a flow system. The products are separated on-line on a short gel chromatographic column and the fluorescence intensity of the marker is measured. The calibration curve shows good linearity in the range 5–80 μg/ml, corresponding to therapeutically relevant serum levels. Intra-day precision values are between 7.32 and 9.48%; the accuracy is between 0.97 and 9.43%. Inter-day precision and accuracy measured on 6 different days fall between 5.38 and 10.05% and −8.27 and −4.97%, respectively. The results obtained with the proposed method show a good correlation with those of other methods (radioimmunoassay and fluorescence polarisation immunoassay) already established in clinical laboratories.

Guanidino compounds involved in the urea and guanidine cycles have been found in serum of nephritic patients, and some guanidino compounds have been suspected to be uremic toxins. The simultaneous analysis of naturally occurring metabolites is important for diagnosis of diseases. In this review, liquid chromatographic analysis of natural metabolites of guanidino compounds are described. the information about arginine as a precursor of nitric oxide are included. The reports of pharmaceutical compounds having a guanidino group, peptides containing arginine and aminoglycosides are summarized in Table 1.

Two ion chromatographic methods have been proposed for the simultaneous determination of caffeine, theobromine and theophylline. The separations were based on isocratic elutions and the determinations on ultraviolet absorbance detections at 274 nm. The detection limits (signal-to-noise ratio 3:1) for the three compounds were all below sub-μg ml⁻¹ level. The methods have been successfully applied to the determination of these three compounds in foods and pharmaceutical preparations, and the average recoveries for various samples ranged from 87% to 103%. In addition, the retention mechanisms of these three compounds on various ion-exchange resins were discussed. The results obtained revealed that ion-exchange chromatography is possibly a good alternative to conventional liquid chromatography for the separation and determination of some water-soluble, neutral organic molecules with a certain hydrophobicity.