Abstract
A new method has been developed for the quantitation of methylated blood proteins in persons exposed to methylating agents. Using different derivatizing agents, 9-fluorenylmethyl chloroformate was the most suitable with regard to both sensitivity and peak separation after subsequent HPLC. After optimization of the chromatographic conditions, two groups who worked with methyl bromide and a control group of non exposed persons were compared with regard to their adduct rates (S-methylcysteine) in the two blood proteins albumin and globin. Albumin and globin were isolated from whole blood samples. After total hydrolysis of the proteins with hydrochloric acid, an amino acid analysis was performed using HPLC with precolumn fluorescent derivatization. This method enabled a quantitation of methylated cysteine in the fmol range. Background levels of S-methylcysteine (15 nmol/g protein) in both albumin and globin were found in non-exposed control persons. Levels were up to 10 fold higher in exposed fumigators working with the methylating agent methyl bromide.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Kramers PGN, Voogd CE, Knaap AGAC, Heijden CA van der (1985) Mutat Res 155:41–47
Danse LHJC, Velsen FL van, Heijden CA van der (1984) Toxicol Appl Pharmacol 72:262–271
Bolt HM, Gansewendt B (1993) CRC Crit Rev Toxicol 23:237–253
Neumann HG (1984) Arch Toxicol 56:1–6
Ehrenberg L, Hiesche D, Osterman-Golkar S, Wennberg I (1974) Mutat Res 24:83–103
Peters T (1970) Adv Clin Chem 13:37–111
Osterman-Golkar S, Ehrenberg L, Segerbäck D, Hällström I (1976) Mutat Res 34:1–10
Allenmark S (1990) Tetrahedron Lett 31:1455–1458
Travis J, Bowen J, Tewksbury D, Johnson D, Pannell R (1976) Biochem J 157:301–306
Anson ML, Mirsky AE (1930) J Gen Physiol 13:469–476
Müller A, Jorritsma U, Gansewendt B (1992) Naunyn-Schmiedeberg's Arch Pharmacol [Suppl] 345:R42 abstr. 167
Hirs CHW, Stein WH, Moore S (1954) J Biol Chem 211:941–950
Lin JK, Wang CH (1980) Clin Chem 26/5:579–583
Hill DW, Walters FH, Wilson TD, Stuart JD (1979) Anal Chem 51:1338–1341
Betner I, Földi P (1988) LC-GC 6:832–840
Betner I, Földi P (1986) Chromatographia 22:381–387
Bailey E, Connors TA, Farmer PB, Gorf SM, Rickard J (1981) Cancer Res 41:2514–2517
Törnqvist M, Osterman-Golkar S, Kautiainen A, Näslund M, Calleman CJ, Ehrenberg L (1988) Mutat Res 204:521–529
Hallier E, Langhof T, Dannappel D, Leutbecher M, Schröder K, Goergens HW, Müller A, Bolt HM (1993) Arch Toxicol 67:173–178
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Müller, A.M.F., Hallier, E., Westphal, G. et al. Determination of methylated globin and albumin for biomonitoring of exposure to methylating agents using HPLC with precolumn fluorescent derivatization. Fresenius J Anal Chem 350, 712–715 (1994). https://doi.org/10.1007/BF00323669
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00323669