UHPLC-MS/MS测定人血浆中4种一线抗癫痫药物浓度及其临床应用

Clinical Application and Determination of Four First-line Antiepileptic Drugs in Human Plasma by UHPLC-MS/MS

关键词:  丙戊酸钠  卡马西平  拉莫三嗪  左乙拉西坦  超高效液相-串联质谱法

Key words:  sodium valproate  carbamazepine  lamotrigine  levetiracetam  UPLC-MS/MS

摘要:

目的 建立对人血浆中丙戊酸钠、卡马西平、拉莫三嗪和左乙拉西坦浓度测定的超高效液相-串联质谱方法，并进行临床实际运用。方法 血浆样本甲醇沉淀蛋白后，采用Waters Acquity UPLC BEH C₁₈色谱柱(100 mm×2.1 mm，1.7 μm)，流动相为0.1%甲酸水-乙腈，梯度洗脱，进样1 µL。选择正负离子同时监测模式分析，卡马西平、左乙拉西坦、拉莫三嗪、丙戊酸钠、替硝唑(内标)MRM通道分别为m/z 236.9→193.9，m/z 171.0→154.2，m/z 256.0→145.0，m/z 143.1→143.1，m/z 248.1→120.9；去簇电压和碰撞电压分别为70 V/21 V，90 V/10 V，90 V/23 V，–50 V/–10 V，90 V/42 V。结果 血浆样本中各代谢物丙戊酸钠(线性范围20~150 µg·mL^-1)、卡马西平(线性范围2~15 µg·mL^-1)、拉莫三嗪(线性范围0.5~15 µg·mL^-1)、左乙拉西坦(线性范围2~60 µg·mL^-1)在该方法下呈良好的线性关系，r²>0.997，血浆样品中各待测物的精密度与准确度试验均符合要求(RSD<6.7%，−12.9%<准确度<10.5%)。共收集77例2021年1月—2021年6月临床癫痫患者服药后血浆进行药物浓度检测分析，丙戊酸钠、卡马西平、拉莫三嗪和左乙拉西坦浓度治疗窗合格率分别是69.57%，70.00%，87.50%和36.11%。结论 本方法适用于临床上大样本量分析，为临床医技部门治疗药物浓度检测提供了一个便捷、可靠、科学的检测手段。

Abstract:

OBJECTIVE To establish a UHPLC-MS/MS method for simultaneous determination of the concentrations of sodium valproate, carbamazepine, lamotrigine and levetiracetam in human plasma and carry out clinical practice. METHODS After methanol precipitation of plasma samples, the Waters Acquity UPLC BEH C₁₈ column(100 mm×2.1 mm, 1.7 μm) was used, the mobile phase was 0.1% formic acid water acetonitrile with gradient elution, and 1 μL was injected. Select the positive and negative ion simultaneous monitoring mode for analysis, the MRM channels of carbamazepine, levetiracetam, lamotrigine, sodium valproate, and tinidazole(internal standard) were m/z 236.9→193.9, m/z 171.0→154.2, m/z 256.0→145.0, m/z 143.1→143.1, m/z 248.1→120.9, respectively. Declustering voltage and collision voltage were 70 V/21 V, 90 V/10 V, 90 V/23 V, -50 V/-10 V, 90 V/42 V, respectively. RESULTS In plasma samples, the metabolites of sodium valproate(linear range 20-150 µg·mL^-1), carbamazepine(linear range 2-15 µg·mL^-1), lamotrigine(linear range 0.5-15 µg·mL^-1), levetiracetam(linear range 2-60 µg·mL^-1) showed a good linear relationship under this method, r²>0.997, the precision and accuracy of each analyte in the plasma sample tested met the requirements(RSD<6.7%, −12.9%<RE<10.5%). A total of 77 patients with clinical epilepsy patients from January 2021 to June 2021 were collected for drug concentration detection and analysis. The concentration of sodium valproate, carbamazepine, lamotrigine and levetiracetam was treated. The treatment window pass rates were 69.57%, 70.00%, 87.50% and 36.11% respectively. CONCLUSION This method is suitable for clinical analysis of large sample sizes, and provides a convenient, reliable and scientific detection method for the detection of therapeutic drug concentration in the clinical medical technology departments.