| 摘要: |
| 目的:建立并验证一种简便快捷的超高效液相色谱(UPLC) 测定癫痫患儿血浆中氯巴占的药物浓度方法。方法:采用
Waters ACQUITY UPLC BEH C18 色谱柱(100 mm×2. 1 mm×1. 7 μm)。流动相为乙腈-10 mmol/ L 磷酸二氢铵溶液(体积比45 ∶ 55,
甲酸调至pH 4. 0),流速0. 15 mL/ min,进样量2 μL,检测波长230 nm,柱温30 ℃。分别精密吸取血浆样品100 μL 和蛋白沉淀
剂200 μL 涡旋混合后以12 000 r/ min 离心10 min,将上清液过滤(0. 22 μm 针头式过滤器)后进样测定。结果:氯巴占的保留时
间为2. 8 min。氯巴占的血药浓度在10~600 ng/ mL 范围内呈现出良好的线性关系(Y =21 922X+143 806,r =0. 999 8)。检测下
限为10 ng/ mL,定量下限为30 ng/ mL。该方法的定量下限( LLOQ,30 ng/ mL)、低浓度质控( LQC,90 ng/ mL)、中浓度质控
(MQC,240 ng/ mL)和高浓度质控(HQC,480 ng/ mL)4 个质控浓度的日内、日间精密度良好,相对标准偏差(RSD) <15. 0%,提取
回收率为91. 18%~99. 92%,稳定性良好( RSD<15. 0%)。32 例癫痫患儿服用氯巴占10 ~ 12 h 后氯巴占平均血药浓度为
52. 10 ng/ mL,治疗有效率为65. 62%,不良反应发生率为31. 25%。年龄>3~6 岁组患儿的氯巴占血药浓度高于年龄>14~18 岁
组(P<0. 05);体质量指数(BMI)18. 5~<24. 0 kg/ m2 组患儿的氯巴占血药浓度高于BMI≥24. 0 kg/ m2 组(P<0. 05)。氯巴占的稳
态血药浓度参考区间为33. 80~339. 50 ng/ mL。结论:本研究建立的UPLC 法能在癫痫患儿在使用氯巴占过程中实时准确快速
地检测血药浓度。 |
| 关键词: 超高效液相色谱法 癫痫 氯巴占 血药浓度 |
| DOI：doi:10.13407/j.cnki.jpp.1672-108X.2025.11.003 |
|
| 基金项目:基金项目:新疆维吾尔自治区第三批“天山英才”培养计划———青年托举人才项目,编号2024TSYCQNTJ0046。 |
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| Determination of Clobazam Blood Concentration in Children with Epilepsy by Ultra-High PerformanceLiquid Chromatography and Its Clinical Application |
| Zhao Ting1, Zhang Huilan1, Sun Yan2, Li Hongjian1, Yu Luhai1 |
| (1. People’ s Hospital of Xinjiang Uygur Autonomous
Region, Urumqi 830000, China; 2. Children’s Hospital of Xinjiang Uygur Autonomous Region, Urumqi 830000, China) |
| Abstract: |
| Objective: To establish and validate a simple and rapid ultra-high performance liquid chromatography (UPLC) method for
the determination of clobazam blood concentration in children with epilepsy. Methods: The chromatographic separation was performed
on the Waters ACQUITY UPLC BEH C18 column (100 mm×2. 1 mm×1. 7 μm) with a mobile phase consisting of acetonitrile and 10
mmol/ L ammonium dihydrogen phosphate solution (45 ∶ 55, adjusted to pH 4. 0 with formic acid). The flow rate was 0. 15 mL/ min,
the injection volume was 2 μL, the detection wavelength was set at 230 nm, and the column temperature was maintained at 30 ℃.
Totally 100 μL plasma sample and 200 μL protein precipitant were vortex mixed and centrifuged at 12 000 r/ min for 10 min. The
supernatant was filtered through a 0. 22 μm syringe filter and determined by injection. Results: The retention time of clobazam was 2. 8
min. The method demonstrated good linearity over the concentration range of 10 to 600 ng/ mL (Y =21 922X+143 806, r =0. 999,8).
The detection limit of the method was 10 ng/ mL, and the quantitative limit was 30 ng/ mL. The lower limit of quantification (LLOQ, 30
ng/ mL), low concentration quality control (LQC, 90 ng/ mL), medium concentration quality control (MQC, 240 ng/ mL) and high
concentration quality control (HQC, 480 ng/ mL) of the method had good intra- and inter-day precision (relative standard deviation,
RSD<15. 0%), the extraction recovery was from 91. 18% to 99. 92%, and the stability was good (RSD<15. 0%). The average blood
concentration of clobazam in 32 cases of children with epilepsy measured 10 to 12 hours after administration was 52. 10 ng/ mL, the
efficacy rate of clobazam treatment was 65. 62%, and the incidence of adverse drug reactions was 31. 25%. The blood concentration of
clobazam in children aged from 3 to 6 years was significantly higher than those aged from 14 to 18 years (P < 0. 05). The blood
concentration of clobazam in children with body mass index (BMI) of 18. 5 to <24. 0 kg/ m2 was higher than that in children with BMI of
⩾24. 0 kg/ m2(P<0. 05). The reference range for the steady-state blood concentration of clobazam was from 33. 80 to 339. 50 ng/ mL.
Conclusion: The UPLC method established in this study can enable real-time, accurate, and rapid detection of the blood concentration
of children with epilepsy during the clobazam treatment. |
| Key words: ultra-high performance liquid chromatography epilepsy clobazam blood concentration |
