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Objective To analyze the correlation of valproic acid and its metabolites (2-propyl-4-pentenoic acid, 3-hydroxy valproic acid,5-hydroxy valproic acid) with liver injury reference index. Methods 328 plasma samples from epilepsy patients were collected and divided into two groups（123 samples from patients with abnormal liver function, experimental group; 205 samples from patients with normal liver function, control group）.The plasma concentrations of valproic acid and its metabolites in the two groups were determined by LC-MS/MS method and the diagnostic value of the concentrations to liver disfunction was analyzed by ROC curve. Results The mean plasma concentration of valproic acid and its three metabolites in the patients with abnormal liver function was higher than that in the control group with was statistically difference（P<0.05）.The concentration of valproic acid and its metabolites could be used as a reference for the diagnosis of liver injury,5-hydroxy valproic acid had better diagnostic value than valproic acid. Conclusion The metabolites of valproic acid were associated with hepatotoxicity, which could be used as a diagnostic index of liver injury and could be a reference for clinical safe application of valproic acid.
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Objective:To investigate the correlation of the concentrations of valproic acid (VPA) and 2-propyl-4-pentenoic acid (4-ene-VPA) with their adverse reactions,and to guide the clinical safety and rational use of VPA.Methods:We collected 254 epilepsy outpatients who took long-term use of sodium valproate oral solution single or combined with other antiepileptic drugs from Xiangya Hospital.The plasma concentrations of VPA and 4-ene-VPA in patients were determined by gas chromatography-mass spectrometry (GC-MS).The double variable correlation analysis was performed to analyze the effect of plasma 4-ene-VPA and VPA concentrations on adverse reactions.Results:The correlations between the PLT level and the dosage ofVPA (P＜0.01 and P＜0.05,respectively),and the RBC level and the concentration of VPA (All P＜0.01) were significant negatively.The concentrations of 4-ene-VPA,VPA,ALT,and AST in the polytherapy group were much higher than those in the monotherapy group (All P＜0.05).In the monotherapy group,the ALT and AST levels in patients younger than or equal to 2 years old were significantly higher than those over 2 years old (P＜0.001).In the polytherapy group,the levels of AST,WBC,and PLT in patients younger than or equal to 2 years old were higher than those over 2 years old (P＜0.05).The levels of AST did not show positive correlation with the concentrations of 4-ene-VPA and VPA (r=0.031,r=0.035,all P＞0.05),and the levels of ALT also did not show positive correlation with the concentrations of 4-ene-VPA and VPA (r=-0.064,r=-0.089,all P＞0.05).Conclusion:VPA may affect blood routine indexes.Age and combination therapy with the non-enzyme-induced anti-epileptic drugs are risk factors for VPA-related liver dysfunctions and renal impairment.The determination of VPA and 4-ene VPA is not a suitable tool for early warning of the VPA-induced liver dysfunction.
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Objective To confirm the hepatotoxicity of valproic acid (VPA ) and its metabolites (2-propyl-4-pentenoic acid ,3-hydroxy valproic acid ,5-hydroxy valproic acid) on human liver cells .Methods Cells were divided into control group and VPA-treated group .The control group was conventionally cultured while the VPA-treated group was treated with valproic acid and its metabolites . The rate of cell proliferation was assayed by CCK 8 protocol . The mRNA levels of CYP1A1 , CYP1A2 ,PCNA ,Bax and Bcl-2 were measured by real time PCR .The correlated protein levels were measured by Western Blotting .The activity of LDH ,AST and ALT were also detected .Results Compared to the control group ,with the increases of concentrations and reaction time of VPA and its metabolites ,the proliferation rate of L02-cell was reduced ,the mRNA and protein levels of CYP1A1 ,CYP1A2 ,and Bax was increased ,the mRNA and protein level of PCNA and Bcl-2 was decreased , AST ,ALT ,and LDH were also elevated in the treated group .Conclusion Valproic acid and its metabolites were positively re-lated to hepatotoxicity .
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OBJECTIVE: To establish a method for simultaneous determination of valproic acid(VPA) and 2-propyls-pente-noic acid(4-ene-VPA) in human plasma by gas chromatography-mass spectrometry (GC-MS) to provide a means for monitoring the plasma concentration of VPA and early warning the valproic acid induced liver toxicity. METHODS: The analytes as TMS derivatives by BSTFA +1% TMCS were separated and identified on HP-5MS column(30 m × 0.25 μm, 0.25 mm) by temperature-programming. Split injection mode was used. The analytes spiked in plasma were extracted with ethyl acetate after acidification with NaH2PO4 buffer solution(pH 5) and then determined and identified using selected ion monitoring(SIM) mode and scan mode, respectively. The internal standard method was used for the determination. RESULTS: VPA and 4-ene-VPA were well separated on their SIM chromatograms and also on the total ion current(TIC) chromatograms. The calibration curves for VPA and 4-ene-VPA showed excellent linearity(r > 0.999). Extraction recoveries were higher than 96%. Intra-day and inter-day RSDs were all lower than 2.49% and 4.63%, the lower limit of quantitation is 5.21 and 5.25 ng · mL-1, respectively. The detection limits were 1.6 and 1.5 ng · mL-1, respectively. CONCLUSION: An optimized analytical method for simultaneous analysis of VPA and 4-ene-VPA in plasma are developed using GC-MS. With good accuracy and precision, high extraction recoveries, it is suitable as a means for monitoring the plasma concentration of VPA and early warning the valproil acid induced liver toxicity. In this paper, it is the first time of simultaneously determination VPA and 4-ene-VPA among Chinese people by GC-MS for providing basis for VPA clinical application.