Determination of Methyl p-Hydroxy-benzoate, the Metabolite of the Candidate Vaginal Contraceptive Drug 4߰-Carbomethoxyphenyl 4-Guanidinobenzoate Mesylate, in Rabbit Plasma: Application of a rapid and sensitive liquid chromatography-tandem mass spectrometry method

Abstract

A rapid, sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the determination of methyl p-hydroxybenzoate (CAS 99-76-3), the metabolite of the candidate vaginal contraceptive drug 4′-carbomethoxyphenyl 4-guanidinobenzoate mesylate (4′-CMGB) in rabbit plasma. The samples were prepared by liquid-liquid extraction with ethyl acetate after spiking with the internal standard, ethyl p-hydroxybenzoate (PHBA-Et). Chromatographic separation was achieved on a C18 column (4.6 mm × 150 mm, 5 µm) with a mobile phase composition of methanol-water (55:45, v/v) delivered at 1.0 ml/min. LC-MS/MS analysis was carried out on a LC-triple-quadrupole mass spectrometer using atmospheric pressure chemical ionization (APCI) and negative multiple reaction monitoring (MRM). Ions monitored were m/z 151 → 92 for PHBA-Me and m/z 165 → 92 for PHBA-Et with argon at a pressure of 0.8 Pa and collision energy of 32 eV for collision-induced dissociation (CID). The assay was linear over the range of 2.0–100 ng/ml for PHBA-Me. The average recovery was 103.9%. The established method has been applied in a pharmacokinetic study of 4′-CMGB gel in rabbit.

Key words

4′-Carbomethoxyphenyl 4-guanidinobenzoate mesylate, LC-MS/MS, metabolite, pharmacokinetics - CAS 99-76-3 - Contraceptives - Methyl p-hydroxybenzoate

• Literature

• 1 Lv JG, Zhao YN, Zhou YJ, Zhu J, Ji YZ, Zhang WN et al. inventors. China patent CN 1438217A.. Aug 27 2003
• 2 Kaminski JM, Bauer L, Mack SR, Anderson RA, Waller DP, Zaneveld LJ. Synthesis and inhibition of human acrosin and trypsin and acute toxicity of aryl 4-guanidinobenzo-ates.. J Med Chem 1986; 29 (4) 514-9
• 3 Zhao YN, Ji YZ, Sheng Y. Antifertility effects of intraviginal application of acrosin inhibitor KF-950 in rabbit.. Reprod Contracep 2000; 20 (5) 294-6
• 4 Zhao YN, Ji YZ, Lv JG, Yi B, Sheng Y, Jiang D. Effects of acrosin inhibitor 4-guanidinobenzoate on human sperm function.. Acad J Sec Mil Med Univ 2000; 21 (12) 1149-51
• 5 Zhan Y, Zhao YN, Li ZL, Li L, Song Q. Effect of acrosin inhibitor KF-950 on acrosin activity and acrosome of human sperm.. Chin. J. Andro 2002; 8 (2) 92-4
• 6 Zhao YN, Sheng Y, Ji YZ, Sha JY, Liu YH, Lv JG et al. Acute toxic and mutagenic effects of the acrosin inhibitor KF-950.. Carcinogen Teratogen Mutagen 2000; 12 (3) 162-4
• 7 Kaminski JM, Nuzzo NA, Bauer L, Waller DP, Zaneveld LJ. Vaginal contraceptive activity of aryl 4-guanidinobenzoates (acrosin inhibitors) in rabbits.. Contraception 1985; 32 (2) 183-9
• 8 Kaminski JM, Smith D, Reid DS, Kennedy W, Jeyendran RS, Zaneveld LJ. Effect of aryl 4-guanidinobenzoates on the acrosin activity of human spermatozoa.. Biol Reprod 1987; 36 (5) 1170-6
• 9 Chvapil M, Eskelson CD, Stiffel V, Owen JA, Droegemueller W. Studies on Nonoxynol-9. II. Intravaginal absorption, distribution, metabolism and excretion in rats and rabbits.. Contraception 1980; 22 (3) 325-39
• 10 Minami Y, Iida K, Tajima H. Absorption of a vaginal contraceptive, nonoxynol (polyoxyethylene nonylphenyl ether) and its metabolism to nonylphenol in female rabbits.. Yakugaku Zassh 2000; 120 (3) 298-303
• 11 Walter BA, Agha BJ, Digenis GA. Disposition of [14C]non-oxynol-9 after intravenous or vaginal administration to female Sprague-Dawley rats.. Toxicol Appl Pharmacol 1988; 96 (2) 258-68
• 12 Tsukagoshi S. Pharmacokinetics studies of nafamostat mesilate (FUT), a synthetic protease inhibitor, which has been used for the treatments of DIC and acute pancreatitis, and as an anticoagulant in extracorporeal circulation.. Gan To Kagaku Ryoho 2000; 27 (5) 767-74
• 13 Midgley I, Hood AJ, Proctor P, Chasseaud LF, Irons SR, Cheng KN et al. Metabolic fate of 14C-camostat mesylate in man, rat and dog after intravenous administration.. Xenobiotica 1994; 24 (l) 79-92
• 14 Göke B, Stöckmann F, Müller R, Lankisch PG, Creutzfeldt W. Effect of a specific serine protease inhibitor on the rat pancreas: systemic administration of camostate and exocrine pancreatic secretion.. Digestion 1984; 30 (3) 171-8
• 15 Beckh K, Göke B, Müller R, Arnold R. Elimination of the low-molecular weight proteinase inhibitor camostate (FOY 305) and its degradation products by the rat liver.. Res Exp Med (Berl) 1987; 187 (6) 401-6