HPLC-MS/MS 法同时测定Beagle 犬血浆中槲皮素、山萘酚和异鼠李素

中国临床药理学与治疗学 ›› 2008, Vol. 13 ›› Issue (12): 1330-1338.

HPLC-MS/MS 法同时测定Beagle 犬血浆中槲皮素、山萘酚和异鼠李素

黄敏文, 王广基, 李昊, 孙建国

中国药科大学药物代谢动力学研究中心, 南京 210038, 江苏

收稿日期:2008-11-20 修回日期:2008-12-21 发布日期:2020-10-30
基金资助:
This research was supported by Hi-Tech Research and Development Program of China(No2003AA2Z347A), and Key Lab of Drug Metabolism and Pharmacokinetics (BM2001201)

Simultaneous determination of quercetin, kaempferol and isorhamnetin in dog plasma by HPLC-MS/MS

HUANG Min-wen, WANG Guang-ji, LI Hao, SUN Jian-guo

Key Laboratory of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, Nanjing 210038, Jiangsu, China

Received:2008-11-20 Revised:2008-12-21 Published:2020-10-30
Contact: WANG Guang-ji, corresponding author, male, professor, doctoral tutor, research direction:pharmacokinetics. Tel: 025-85391035 E-mail: gjwang@cpu.edu.cn

摘要/Abstract

摘要： 目的建立同时测定犬血浆中槲皮素、山萘酚和异鼠李素浓度的HPLC-MS/MS 方法。方法血浆样品经酸水解后用乙醚提取, 采用选择性反应检测方法测定其血药浓度。仪器为FinniganLC-MS/MS 二级四极杆质谱检测器, 色谱柱为Luna C₁₈ (150 mm ×2.00 mm i.d., 5 μm, Luna,USA);流动相为乙腈-0.1 %甲酸, 梯度洗脱。结果槲皮素、山萘酚和异鼠李素的线性范围均为0.5 ~ 100.0 ng/mL, 三种黄酮的最低定量限均为0.5 ng/mL, 各自日内日间精密度分别小于7.3 %,6.2 %和6.4 %, 回收率分别大于70 %, 66 %和70 %。结论该测定方法经验证符合血浆样品的测定要求, 可以应用于临床前药代动力学研究。

关键词: 槲皮素, 山萘酚, 异鼠李素, HPLC-MS/MS, 药物代谢动力学

Abstract: AIM: A rapid, sensitive and highly selective liquid chromatography-tandem mass spectrometry method was developed and validated for simultaneous determination of quercetin, kaempferol and isorhamnetin in dog plasma.METHODS: Quercetin, kaempferol and isorhamnetin conjugates were hydrolysed chemically.The analytes were extracted from plasma samples by liquid-liquid extraction, separated on a Luna ODS-2 column (150 mm ×2.1 mm I.D., 5 μm particle size), and detected by tandem mass spectrometry with a Finnigan IonSpray ionization interface. The mobile phase consisted of 0.1 % aqueous formic acid (A)and gradient-grade acetonitrile (B).RESULTS: The calibration curves for quercetin, kaempferol and isorhamnetin were linear in concentration ranges of 0.5-100.0 ng/mL in dog plasma.The method has a lower limit of quantification (LLOQ)of 0.5 ng/mL for all the three flavonols.The intra-and inter-day precisions, expressed as the R.S.D., were less than 7.3 %, 6.2 % and 6.4 % for quercetin, kaempferol and isorhamnetin, respectively, and the recovery was more than 70 %, 66 %and 70 %, respectively.The application of this assay was examined in a preliminary pharmacokinetic study of quercetin, kaempferol and isorhamnetin in beagle dogs after oral administration of 6 Ginkgo biloba tablets.CONCLUSION: The present method was suitable for determining quetcetin, kaempferol and isorhamnetin in dog plasma.

Key words: quercetin, kaempferol, isorhamnet, HPLC-MS/MS, pharmacokinetics

中图分类号:

R969.1

黄敏文, 王广基, 李昊, 孙建国. HPLC-MS/MS 法同时测定Beagle 犬血浆中槲皮素、山萘酚和异鼠李素[J]. 中国临床药理学与治疗学, 2008, 13(12): 1330-1338.

HUANG Min-wen, WANG Guang-ji, LI Hao, SUN Jian-guo. Simultaneous determination of quercetin, kaempferol and isorhamnetin in dog plasma by HPLC-MS/MS[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2008, 13(12): 1330-1338.

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