Simultaneous Determination of Lamotrigine, Carbamazepine and Sodium Valproate in Plasma of Patients with Epilepsy by HPLC-MS/MS
YU Si-yuana, CHEN Ke-qiangb, FENG Zhi-pinga
a. Department of Pharmacy, b. Department of Neurology, Jiangmen Central Hospital, Affiliated Jiangmen Hospital of Sun Yat-San University, Jiangmen 529030, China
Abstract：OBJECTIVE To establish an HPLC-MS/MS method for the determination of lamotrigine, carbamazepine and sodium valproate in the plasma and apply it for the determination of the plasma drug concentration in patients with epilepsy. METHODS The plasma procedure involved a single-step protein precipitation by acetonitrile, then chromatographed on a ZORBAX Eclipse XDB-C18 (4.6 mm×50 mm, 1.8 μm) using a mobile phase composed of methanol-5 mmol·L-1 ammonium formate with gradient elution was selected. The flow rate was 0.4 mL·min-1, with loratadine as internal standard. The ions were monitored by positive and negative ion switching scanning technology, using electrospray source ion. The multi-reaction monitoring mode (MRM) was used to quantitatively analyze the ionization pairs as m/z 256.0→210.9 (lamotrigine), m/z 237.0→194.0 (carbamazepine), m/z 143.0→143.0 (sodium valproate) and m/z 383.0→337.0 (loratadine). RESULTS Lamotrigine and carbamazepine had good linear relationships in the concentration ranges of 0.5~20.0 μg·mL-1, and the lower limits of quantification was 0.5 μg·mL-1. Sodium valproate also had good linear relationship in the concentration ranges of 32~160 μg·mL-1, and the lower limits of quantification was 32 μg·mL-1. The calibration curves of lamotrigine and carbamazepine were linear over the concentration range of 0.5~20.0 μg·mL-1, the lower limit of quantitation was 0.5 μg·mL-1. The calibration curves of sodium valproate was linear over the concentration range of 32~160 μg·mL-1, the lower limit of quantitation was 32 μg·mL-1. The intra-day and inter-day RSD were less than 15%. The average extraction recoveries of three analytes were more than 85%. The stability was good and is not affected by matrix effect. CONCLUSION This method is simple, rapid and accurate, and is suitable for routine clinical monitoring of the blood concentration of lamotrigine, carbamazepine and sodium valproate.
BEGHI E, GIUSSANI G, SANDER J W. The natural history and prognosis of epilepsy[J]. Epilept Disord. 2015, 17(3): 243-253.
[2]
ELECTROENCEPHALOGRAM AND EPILEPSY CHAPTER, CHINESE SOCIETY OF NEUROLOGY. Expert consensus opinion in treatment of epilepsy[J]. Chin J Neurol(中华神经科杂志), 2011, 44(1): 56-65.
[3]
PATSALOS P N. Drug interactions with the newer antiepileptic drugs (AEDs)-Part 1: Pharmacokinetic and pharmacodynamic interactions between AEDs[J]. Clin Pharmacokinet, 2013, 52(11): 927-966.
[4]
ZHU X, ZHANG T, TIAN M, et al. Comparison of carbamazepine serum concentrations determined by high performance liquid chromatography and fluorescence polarization immunoassay [J]. Chin J Clin Pharmacol(中国临床药理学杂志), 2017, 37(1): 292-295.
[5]
LI Z Y, YAN C L, YAN R, et al. Analytical performance of the Abbott Architect i2000 tacrolimus assay in Chinese patients after renal transplantation[J].J Transplant Proc, 2010, 42(10): 4534-4537.
[6]
XU S S, ZHANG T, ZHOU J K, et al. Determination of concentrations of lamotrigine, oxcarbazepine and its active metabolite in human serum by HPLC method[J]. Chin J Clin Pharm(中国临床药理学杂志), 2015, 31(12): 1176-1179.
[7]
EL HMA, WADA M, IKEDA R, et al. Validation of an LC-MS/MS method for the determination of propofol, midazolam, and carbamazepine in rat plasma: application to monitor their concentrations following co-administration[J]. Biol Pharm Bull, 2015, 38(8): 1250-1253.
[8]
SHEN C J, CHEN X G, WANG Y, et al. Modified high performance liquid chromatography to determine the concentration of sodium valproate in human serum[J]. Acta Acade Med Wannan(皖南医学院学报), 2016, 43(1): 18-21.
[9]
LI W, PENG M, YAO D W. Comparison of on-line column extraction HPLC and pre-column derivatization HPLC method for determination of valproic acid in plasma[J]. Chin Pharm J(中国药学杂志), 2018, 53(20): 1785-1788.
[10]
SHEN Q, WANG Y G, ZENG H, et al. Determination of lamotrigine, phenobarbital, oxcarbazepine and its active metabolite monhydroxycarbazepin concentrations in human serum by RP-HPLC[J]. J China Pharm(中国药房), 2016, 27(11): 1493-1496.
[11]
YANG X, LI L, HE L J, et al. Determination of lamotrigine, olanzapine and quetiapine in human plasma by ultra-performance liquid chromatography-tandem mass spectrometry and its application in therapeutic drug monitoring[J]. Chin Pharm J(中国药学杂志), 2020, 55(1): 44-51.
[12]
CHEN K V, LUO L P, XIE W Y, et al. Simultaneous determination of lamotrigine and carbamazepine in human serum by LC-MS/MS[J]. Eval Anal Drug Use Hosp China(中国医院用药评价与分析), 2012, 12(12): 1108-1110.
[13]
WANG L J, ZHOU C H, WANG J X, et al. Simultaneous determination of lamotrigine and sodium valproate in plasma of patients with epilepsy by HPLC-MS/MS [J]. Chin J Clin Pharm(中国临床药理学杂志), 2019, 35(19): 2390-2394.
[14]
CHEN X, ZHU B, YANG Y, et al. Simultaneous determination of lamotrigine and carbamazepine in human plasma by UPLC-MS/MS[J]. Pract Pharm Clin Remed(实用药物与临床), 2019, 22(10): 1083-1086.
2021, Vol. 56 
