高效液相色谱法测定体外大鼠肠道菌液中大豆苷及其代谢物

张圳; 冯家懿; 果卉; 姚媛; 闻俊; 周婷婷

doi:10.3969/j.issn.1006-0111.2018.04.013

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高效液相色谱法测定体外大鼠肠道菌液中大豆苷及其代谢物

张圳, 冯家懿, 果卉, 姚媛, 闻俊, 周婷婷

文章导航 > 药学实践与服务 > 2018 > 36(4): 347-350

张圳, 冯家懿, 果卉, 姚媛, 闻俊, 周婷婷. 高效液相色谱法测定体外大鼠肠道菌液中大豆苷及其代谢物[J]. 药学实践与服务, 2018, 36(4): 347-350. doi: 10.3969/j.issn.1006-0111.2018.04.013

引用本文:

ZHANG Zhen, FENG Jiayi, GUO Hui, YAO Yuan, WEN Jun, ZHOU Tingting. Simultaneously determination of daidzin and its metabolite in rat intestinal bacteria test solution in vitro by HPLC[J]. Journal of Pharmaceutical Practice and Service, 2018, 36(4): 347-350. doi: 10.3969/j.issn.1006-0111.2018.04.013

Citation:

ZHANG Zhen, FENG Jiayi, GUO Hui, YAO Yuan, WEN Jun, ZHOU Tingting. Simultaneously determination of daidzin and its metabolite in rat intestinal bacteria test solution in vitro by HPLC[J]. Journal of Pharmaceutical Practice and Service, 2018, 36(4): 347-350. doi: 10.3969/j.issn.1006-0111.2018.04.013

高效液相色谱法测定体外大鼠肠道菌液中大豆苷及其代谢物

doi: 10.3969/j.issn.1006-0111.2018.04.013

张圳¹,
冯家懿¹,
果卉¹,
姚媛²,
闻俊¹,
周婷婷^1,

1.
第二军医大学药学院, 上海市药物(中药)代谢产物研究重点实验室, 上海 200433
2.
第二军医大学药学院, 上海市药物(中药)代谢产物研究重点实验室, 上海 200433;安徽中医药大学药学院, 安徽合肥 230012

基金项目: 国家自然科学基金资助项目（81573584，81773862）；第二军医大学本科生创新孵化基地项目（FM2016071）

Simultaneously determination of daidzin and its metabolite in rat intestinal bacteria test solution in vitro by HPLC

1.
School of Pharmacy & Shanghai Key Laboratory for Pharmaceutical Metabolite Research, Second Military Medical University Shanghai 200433, China
2.
School of Pharmacy & Shanghai Key Laboratory for Pharmaceutical Metabolite Research, Second Military Medical University Shanghai 200433, China;School of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, China

摘要: 目的建立HPLC法测定体外大鼠肠道菌液中的大豆苷及其代谢物，研究大鼠肠道菌群对大豆苷的代谢情况。方法采用大鼠离体粪便温孵法，将大豆苷与大鼠肠道菌群培养液进行厌氧温孵。温孵样品采用甲醇沉淀蛋白处理后进行HPLC法分析，色谱条件：色谱柱为Diamonsil C₁₈柱；流动相为0.2%乙酸水溶液与甲醇，梯度洗脱；检测波长为260 nm；柱温为25℃；进样量为10 μl。结果大豆苷及其代谢物大豆苷元分别在98.0~490、57.6~288 μg/ml浓度范围内线性良好，高、中、低3个浓度的相对回收率在96.6%~114.0%之间，相对标准偏差小于9.9%，表明本法的准确度和精密度符合要求。大豆苷在肠道菌液中代谢生成大豆苷元，其消除半衰期为37.6 min。结论所建立的HPLC法适合同时测定体外大鼠肠道菌液中的大豆苷及其代谢物，大豆苷在体外代谢的主要途径为糖苷键断裂转化成对应苷元。
- 肠道菌群 /
- 体外代谢 /
- 大豆苷 /
- 高效液相色谱法
Abstract: Objective To establish an HPLC method for simultaneously determination of daidzin and its metabolite in rat intestinal bacteria test solution in vitro, and study the metabolism of daidzin by rat intestinal bacteria. Methods The incubation of daidzin was carried out at 37℃ with rat intestinal bacteria test solution. The samples were precipitated by methanol followed by a separation with HPLC method. The conditions were as follows:the chromatographic column was Ultimate C₁₈; the mobile phase was 0.2% acetic acid and methanol (gradient elution); the detection wavelength was 260nm, and the column temperature was 25℃. Results Daidzin and daidzein showed good linearity with the range of 98.0~490μg/ml and 57.6~288μg/ml, respectively. The accuracy of daidzin and daidzein was ranged from 96.6%~114.0% and the precision of the two analytes was within 9.9%. Daidzin was transformed to daidzein in the rat intestinal bacteria test solution, and its elimination half-life was 37.6 min. Conclusion The established HPLC method was suitable for the determination of daidzin and its metabolite. Daidzin could be metabolized by rat intestinal bacteria in vitro,and the main metabolic pathway was transforming daidzin into corresponding aglycone through breaking the glucoside bond
- intestinal bacteria /
- metabolism in vitro /
- daidzin /
- HPLC

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高效液相色谱法测定体外大鼠肠道菌液中大豆苷及其代谢物

doi: 10.3969/j.issn.1006-0111.2018.04.013

1. 第二军医大学药学院, 上海市药物(中药)代谢产物研究重点实验室, 上海 200433

2. 第二军医大学药学院, 上海市药物(中药)代谢产物研究重点实验室, 上海 200433;安徽中医药大学药学院, 安徽合肥 230012

基金项目: 国家自然科学基金资助项目（81573584，81773862）；第二军医大学本科生创新孵化基地项目（FM2016071）

收稿日期: 2017-10-02
修回日期: 2018-03-26

关键词:

肠道菌群 /

体外代谢 /

大豆苷 /

高效液相色谱法

摘要: 目的建立HPLC法测定体外大鼠肠道菌液中的大豆苷及其代谢物，研究大鼠肠道菌群对大豆苷的代谢情况。方法采用大鼠离体粪便温孵法，将大豆苷与大鼠肠道菌群培养液进行厌氧温孵。温孵样品采用甲醇沉淀蛋白处理后进行HPLC法分析，色谱条件：色谱柱为Diamonsil C₁₈柱；流动相为0.2%乙酸水溶液与甲醇，梯度洗脱；检测波长为260 nm；柱温为25℃；进样量为10 μl。结果大豆苷及其代谢物大豆苷元分别在98.0~490、57.6~288 μg/ml浓度范围内线性良好，高、中、低3个浓度的相对回收率在96.6%~114.0%之间，相对标准偏差小于9.9%，表明本法的准确度和精密度符合要求。大豆苷在肠道菌液中代谢生成大豆苷元，其消除半衰期为37.6 min。结论所建立的HPLC法适合同时测定体外大鼠肠道菌液中的大豆苷及其代谢物，大豆苷在体外代谢的主要途径为糖苷键断裂转化成对应苷元。

English Abstract

Simultaneously determination of daidzin and its metabolite in rat intestinal bacteria test solution in vitro by HPLC

1. School of Pharmacy & Shanghai Key Laboratory for Pharmaceutical Metabolite Research, Second Military Medical University Shanghai 200433, China

2. School of Pharmacy & Shanghai Key Laboratory for Pharmaceutical Metabolite Research, Second Military Medical University Shanghai 200433, China;School of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, China

Received Date: 2017-10-02
Rev Recd Date: 2018-03-26

Keywords:

Abstract: Objective To establish an HPLC method for simultaneously determination of daidzin and its metabolite in rat intestinal bacteria test solution in vitro, and study the metabolism of daidzin by rat intestinal bacteria. Methods The incubation of daidzin was carried out at 37℃ with rat intestinal bacteria test solution. The samples were precipitated by methanol followed by a separation with HPLC method. The conditions were as follows:the chromatographic column was Ultimate C₁₈; the mobile phase was 0.2% acetic acid and methanol (gradient elution); the detection wavelength was 260nm, and the column temperature was 25℃. Results Daidzin and daidzein showed good linearity with the range of 98.0~490μg/ml and 57.6~288μg/ml, respectively. The accuracy of daidzin and daidzein was ranged from 96.6%~114.0% and the precision of the two analytes was within 9.9%. Daidzin was transformed to daidzein in the rat intestinal bacteria test solution, and its elimination half-life was 37.6 min. Conclusion The established HPLC method was suitable for the determination of daidzin and its metabolite. Daidzin could be metabolized by rat intestinal bacteria in vitro,and the main metabolic pathway was transforming daidzin into corresponding aglycone through breaking the glucoside bond

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高效液相色谱法测定体外大鼠肠道菌液中大豆苷及其代谢物

doi: 10.3969/j.issn.1006-0111.2018.04.013

Simultaneously determination of daidzin and its metabolite in rat intestinal bacteria test solution in vitro by HPLC

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高效液相色谱法测定体外大鼠肠道菌液中大豆苷及其代谢物

doi: 10.3969/j.issn.1006-0111.2018.04.013

1. 第二军医大学药学院, 上海市药物(中药)代谢产物研究重点实验室, 上海 200433 2. 第二军医大学药学院, 上海市药物(中药)代谢产物研究重点实验室, 上海 200433;安徽中医药大学药学院, 安徽 合肥 230012

English Abstract

Simultaneously determination of daidzin and its metabolite in rat intestinal bacteria test solution in vitro by HPLC

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目录

1. 第二军医大学药学院, 上海市药物(中药)代谢产物研究重点实验室, 上海 200433

2. 第二军医大学药学院, 上海市药物(中药)代谢产物研究重点实验室, 上海 200433;安徽中医药大学药学院, 安徽合肥 230012