Development and Validation of an LC/MS/MS Method for Determination of Valproic Acid and Its Metabolite 2-Propyl-4-pentenoic Acid in Monkey Plasma

다국어 초록 (Multilingual Abstract)

A rapid, accurate, and sensitive liquid chromatography/mass spectrometry (MS)/MS method for the quantitative determination of valproic acid (VPA) and its metabolite, 2-propyl-4-pentenoic acid (4-ene VPA), in monkey plasma was developed and validated. The sample extraction was performed using hydrophilic-lipophilic balance cartridge. The analytes were separated on a Kinetex C18 (2.1 mm × 100 mm, 2.6 μm) analytical column under a mobile phase consisting of 10 mM ammonium formate (pH 8.0)/methanol (20/80, v/v) and isocratic flow at 0.15 mL/min.
The tandem mass spectrometer was operated in negative electrospray ionization with selected ion monitoring conditions, 143.0, 141.0,and 121.0 for VPA, 4-ene VPA, and benzoic acid (internal standard), respectively. The linearity of calibration curve ranging from 0.1 to 20 μg/mL was at least 0.9996 (coefficient of correlation, r) for both analytes. Intra- and inter-day precisions for both analytes were lower than 15%, resulting from quality control (QC) samples at concentration of 0.2 (low QC), 1.6 (middle QC),and 16 (high QC) μg/mL except at the lower limit of quantification (LLOQ) (0.1 μg/mL) level, which was less than 20%. The intraand inter-day accuracies were within ±15%. The recoveries were 84.4–90.8% for VPA and 88.2–100.6% for 4-ene VPA. Both analytes were stable throughout short-term temperature, post preparation for 24 h, and three freeze/thaw cycles, validating that this method could be applied to toxicokinetic and pharmacokinetic studies.

참고문헌 (Reference)

1 Davis R, "Valproic aicd. A reappraisal of its pharmacological properties and clinical efficacy in epilepsy" 47 : 332-372, 1994

2 Zimmerman HJ, "Valproate-induced hepatic steatogenesis in rats" 2 : 591-597, 1982

3 Cheng H, "Quantification of valproic acid and its metabolite 2-propyl-4-pentenoic acid in human plasma using HPLC-MS/MS" 850 : 206-212, 2007

4 Ramakrishna NVS, "Liquid chromatography/ electrospray ionization mass spectrometry method for the quantification of valproic acid in human plasma" 19 : 1970-1978, 2005

5 Nakamura M, "Improved procedure for the high-performance liquid chromatographic determination of valproic acid in serum as its phenacyl ester" 310 : 450-454, 1984

6 Vam der Horst FA, "High-performance liquid chromatographic determination of valproic acid in plasma using a micelle-mediated pre-column derivatization" 456 : 191-199, 1988

7 Pucci V, "High sensitivity determination of valproic acid in mouse plasma using semi-automated sample preparation and liquid chromatography with tandem mass spectrometric detection" 19 : 3713-3718, 2005

8 Lucarelli C, "HPLC Method for the Simultaneous Analysis of Valproic Acid and other Common Anticonvulsant Drugs in Human Plasma or Serum" 33 : 37-40, 1992

9 Willmore LJ, "Divalproex and epilepsy" 37 : 43-53, 2003

10 Amini-Shirazi N, "Determination of VPA and its Two Important Metabolites in Iranian Overdosed Patients" 6 : 854-862, 2010

11 Rettenmeier AW, "Biotransformation and pharmacokinetics in the rhesus monkey of 2-npropyl-4-pentenoic acid, a toxic metabolite of valproic acid" 14 : 454-464, 1986

12 Wolf JH, "Automated analysis procedure for valproic acid in blood, serum and brain dialysate by highperformance liquid chromatography with bromomethylmethoxycoumarin as fluorescent label" 487 : 496-502, 1989

13 Granneman GR, "Aspects of the metabolism of valproic acid" 14 : 375-387, 1984

14 Matsuura K, "A simple and rapid determination of valproic acid in human plasma using a nonporous silica column and liquid chromatography with tandem mass spectrometric detection" 22 : 387-393, 2008

15 Jain DS, "A high throughput and selective method for the estimation of valproic acid an antiepileptic drug in human plasma by tandem LC-MS/MS" 72 : 80-88, 2007