李丽丽;黄琪;齐佳佳;姚敏;彭代银;
LI Li-li;HUANG Qi;QI Jia-jia;YAO Min;PENG Dai-yin;Department of Pharmacy, Anhui University of Chinese Medicine;Institute of Traditional Chinese Medicine Resources Protection and Development;MOE-Anhui Joint Collaborative Innovation Center for Quality Improvement of Anhui Genuine Chinese Medicinal Materials;

• 1:安徽中医药大学药学院
• 2:中药资源保护与开发研究所
• 3:省部共建安徽道地中药材品质提升协同创新中心

摘要(Abstract)：

风轮菜是治疗子宫异常出血(abnormal uterine bleeding, AUB)常见中药，皂苷类和黄酮类是风轮菜中主要药效活性成分，然而其主要成分的药代动力学行为尚不清晰。该文建立了一种风轮菜总提取物(total extraction of Clinopodium chinense, TEC)中5种成分在AUB大鼠血浆中药物浓度的测定方法并对该分析方法进行了方法学验证。随后，取同1 d受孕SD雌性大鼠，灌胃米非司酮(12.4 mg·kg^(-1))联合米索前列醇(130μg·kg(-1))联合米索前列醇(130μg·kg^(-1)),建立AUB大鼠模型。灌胃给予AUB大鼠TEC后检测大鼠血浆中橙皮苷、柚皮素、芹菜素、柴胡皂苷a及断血流皂苷A的含量。采用DAS 2.0软件计算药动学参数。结果表明，在检测范围内5种化合物线性关系良好。该方法专属性、准确度、精密度、回收率、基质效应和稳定性均满足生物样品的测定要求。灌胃TEC后在AUB大鼠血浆中可检测到5种化合物，橙皮苷、柚皮素、芹菜素、柴胡皂苷a、断血流皂苷A的C_(max)分别为701.6、429.5、860.7、75.1、304.1 ng·mL(-1)),建立AUB大鼠模型。灌胃给予AUB大鼠TEC后检测大鼠血浆中橙皮苷、柚皮素、芹菜素、柴胡皂苷a及断血流皂苷A的含量。采用DAS 2.0软件计算药动学参数。结果表明，在检测范围内5种化合物线性关系良好。该方法专属性、准确度、精密度、回收率、基质效应和稳定性均满足生物样品的测定要求。灌胃TEC后在AUB大鼠血浆中可检测到5种化合物，橙皮苷、柚皮素、芹菜素、柴胡皂苷a、断血流皂苷A的C_(max)分别为701.6、429.5、860.7、75.1、304.1 ng·mL^(-1)。各化合物在体内半衰期较短，消除速率快；表观分布容积提示待测化合物呈广泛组织分布。该方法快速、准确，灵敏，为风轮菜治疗AUB的药效物质基础研究提供了参考。
Clinopodium chinense, a traditional folk medicinal herb, has been used to treat abnormal uterine bleeding(AUB) for many years. Saponins and flavonoids are the main active components in C. chinense. To study the pharmacokine-tics of multiple components from the total extract of C. chinense(TEC), we established a sensitive and rapid method of ultra-perfor-mance liquid chromatography coupled with tandem mass spectrometry(UPLC-MS/MS) for simultaneous determination of five compounds in the plasma of AUB rats. After validation, the AUB model was established with SD female rats which got pregnant on the same day by gavage with mifepristone(12.4 mg·kg(-1)。各化合物在体内半衰期较短，消除速率快；表观分布容积提示待测化合物呈广泛组织分布。该方法快速、准确，灵敏，为风轮菜治疗AUB的药效物质基础研究提供了参考。
Clinopodium chinense, a traditional folk medicinal herb, has been used to treat abnormal uterine bleeding(AUB) for many years. Saponins and flavonoids are the main active components in C. chinense. To study the pharmacokine-tics of multiple components from the total extract of C. chinense(TEC), we established a sensitive and rapid method of ultra-perfor-mance liquid chromatography coupled with tandem mass spectrometry(UPLC-MS/MS) for simultaneous determination of five compounds in the plasma of AUB rats. After validation, the AUB model was established with SD female rats which got pregnant on the same day by gavage with mifepristone(12.4 mg·kg^(-1)) and misoprostol(130 μg·kg(-1)) and misoprostol(130 μg·kg^(-1)). The established method was applied to the detection of hesperidin, naringenin, apigenin, saikosaponin a, and buddlejasaponin Ⅳb in AUB rats after the administration of TEC. The pharmacokinetic parameters were calculated by DAS 2.0. The five compounds showed good linear relationship within the detection range. The specificity, accuracy, precision, recovery, matrix effect, and stability of the method all matched the requirements of biolo-gical sample detection. The above 5 compounds were detected in the plasma of AUB rats after the administration of TEC. The C_(max) va-lues of hesperidin, naringenin, apigenin, saikosaponin a, and clinoposide A were 701.6, 429.5, 860.7, 75.1, and 304.1 ng·mL(-1)). The established method was applied to the detection of hesperidin, naringenin, apigenin, saikosaponin a, and buddlejasaponin Ⅳb in AUB rats after the administration of TEC. The pharmacokinetic parameters were calculated by DAS 2.0. The five compounds showed good linear relationship within the detection range. The specificity, accuracy, precision, recovery, matrix effect, and stability of the method all matched the requirements of biolo-gical sample detection. The above 5 compounds were detected in the plasma of AUB rats after the administration of TEC. The C_(max) va-lues of hesperidin, naringenin, apigenin, saikosaponin a, and clinoposide A were 701.6, 429.5, 860.7, 75.1, and 304.1 ng·mL^(-1), respectively. All the compounds owned short half-life and quick elimination rate in vivo, and the large apparent volume of distribution indicated that they were widely distributed in tissues. Being rapid, accurate, and sensitive, this method is suitable for the pharmacokinetic study of extracts of Chinese herbal medicines and provides a reference for the study of pharmacodynamic material basis of C. chinense in treating AUB.

关键词(KeyWords)： 风轮菜;子宫异常出血;药动学;UPLC-MS/MS;总提取物
Clinopodium chinense;abnormal uterine bleeding;pharmacokinetics;UPLC-MS/MS;total extract

Abstract:

Keywords:

基金项目(Foundation): 安徽省高校自然科学研究重点项目(KJ2021A0539);; 安徽中医药大学自然科学研究重点项目(2020zrzd13);安徽中医药大学人才支持计划项目(2019rcyb005)

作者(Authors): 李丽丽;黄琪;齐佳佳;姚敏;彭代银;
LI Li-li;HUANG Qi;QI Jia-jia;YAO Min;PENG Dai-yin;Department of Pharmacy, Anhui University of Chinese Medicine;Institute of Traditional Chinese Medicine Resources Protection and Development;MOE-Anhui Joint Collaborative Innovation Center for Quality Improvement of Anhui Genuine Chinese Medicinal Materials;

DOI: 10.19540/j.cnki.cjcmm.20220421.702

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