护肾(Ⅲ)号胶囊中三七皂苷R1和人参皂苷Rg1在大鼠体内药代动力学研究(1)
摘要：目的 建立超高液相色谱串联质谱(UHPLC-MS/MS)测定护肾(Ⅲ)号胶囊给药后大鼠血浆中三七皂苷R1、人参皂苷Rg1含量的方法，并对其药代动力学特征进行研究。方法 SD大鼠灌胃护肾(Ⅲ)号胶囊2 g/kg后按时间点取血，以地高辛为内标，采用UHPLC-MS/MS测定给药不同时点血浆中三七皂苷R1和人参皂苷Rg1浓度，并用DAS2.0软件计算药代动力学参数。结果 三七皂苷R1和人参皂苷Rg1的线性关系良好(r≥0.997 5)；日内、日间精密度RSD均小于13.1%，准确度、回收率及稳定性符合生物样品分析要求。大鼠体内三七皂苷R1药代动力学参数：T1/2(7.86±1.69)h，Tmax(4.00±1.04)h，Cmax(0.23±0.05)mg/L；人参皂苷Rg1药代动力学参数：T1/2(4.58±0.95)h，Tmax(6.00±0.00)h，Cmax(0.32±0.03)mg/L。结论 本研究建立的UHPLC-MS/MS分析方法灵敏、准确可靠，适用于护肾(Ⅲ)号胶囊的药代动力学研究。
关键词：护肾(Ⅲ)号胶囊；超高液相色谱串联质谱；药代动力学
DOI：10.3969/j.issn.1005-5304.2018.07.017
中图分类号：R285.5 文献标识码：A 文章编号：1005-5304(2018)07-0071-04
Abstract： Objective To establish a UHPLC-MS/MS method for the determination of notoginsenoside R1 and ginsenoside Rg1 in rat plasma after oral administration of Hushen (Ⅲ) Capsules； To study its pharmacokinetics. Methods SD rats were given Hushen (Ⅲ) Capsules 2 g/kg and then the blood was collected at time point. Digoxin was used as internal standard. The drug concentrations of notoginsenoside R1 and ginsenoside Rg1 in plasma were determined by UHPLC-MS/MS. And the relative pharmacokinetic parameters were calculated by using DAS2.0 software. Results The linear relationship of notoginsenoside R1 and ginsenoside Rg1 was good (r≥0.997 5). The RSD of intra-day and inter-day precision were less than 13.1%. The results of accuracy， recovery and stability met the requirements for the biology sample analysis. For notoginsenoside R1， the pharmacokinetics parameters T1/2， Tmax， Cmax in the plasma were (7.86±1.69)h， (4.00±1.04)h， (0.23±0.05)mg/L. For ginsenoside Rg1， the pharmacokinetics parameters T1/2， Tmax， Cmax in plasma were (4.58±0.95)h， (6.00±0.00)h， (0.32±0.03)mg/L. Conclusion The established UHPLC-MS/MS method is sensitive， accurate and reliable， and is suitable for the pharmacokinetic study of Hushen (Ⅲ) Capsules.
Keywords： Hushen (Ⅲ) Capsules； UHPLC-MS/MS； pharmacokinetics
目前，血尿形成机制尚未完全阐明，通常认为与免疫炎症反应导致肾小球基底膜受损有关，西医治疗往往效果不佳，迁延难愈。肾性血尿属中医学“血证”范畴，中医治疗血尿积累了丰富的临床经验，并取得了较好的疗效[1]。护肾(Ⅲ)号胶囊是张家港市中医医院复方制剂，由白茅根、三七、大黄炭组成，具有清热解毒、滋肾降火功效，临床常用于治疗血尿為主的各种肾病。三七作为方中臣药，具有良好的止血、活血化瘀、消肿止痛作用，辅助君药白茅根治疗血尿诸症。三七总皂苷为三七的主要有效成分，具有抗炎、增强免疫功能的作用。已有研究采用HPLC对护肾(Ⅲ)号胶囊的有效成分进行了含量测定[2]。本研究采用高效、灵敏的超高液相色谱串联质谱(UHPLC-MS/MS)技术对护肾(Ⅲ)号胶囊中三七皂苷R1、人参皂苷Rg1在血浆中的含量进行分析，并对其在大鼠血浆中的药动学特征进行研究，为进一步开展护肾(Ⅲ)号胶囊临床应用研究，明确其作用机制奠定基础。
1 仪器、试药与动物
Agilent 1290超高效液相色谱仪；Agilent 6430 LC-MS三重四级杆质谱仪，配有电喷雾化离子源(ESI)，MassHunter色谱工作站；AE240电子天平(上海梅特勒-托利多有限公司)；MICRO-17R冷冻离心机(美国Thermo公司)；WH-2微型旋涡混合仪(上海沪西分析仪器厂)；Drict-Q5超纯水机(法国Millipore公司)。, 百拇医药(钱伟 冯星月 陈晓峰 王建平 蒋志涛 余辉)
关键词：护肾(Ⅲ)号胶囊；超高液相色谱串联质谱；药代动力学
DOI：10.3969/j.issn.1005-5304.2018.07.017
中图分类号：R285.5 文献标识码：A 文章编号：1005-5304(2018)07-0071-04
Abstract： Objective To establish a UHPLC-MS/MS method for the determination of notoginsenoside R1 and ginsenoside Rg1 in rat plasma after oral administration of Hushen (Ⅲ) Capsules； To study its pharmacokinetics. Methods SD rats were given Hushen (Ⅲ) Capsules 2 g/kg and then the blood was collected at time point. Digoxin was used as internal standard. The drug concentrations of notoginsenoside R1 and ginsenoside Rg1 in plasma were determined by UHPLC-MS/MS. And the relative pharmacokinetic parameters were calculated by using DAS2.0 software. Results The linear relationship of notoginsenoside R1 and ginsenoside Rg1 was good (r≥0.997 5). The RSD of intra-day and inter-day precision were less than 13.1%. The results of accuracy， recovery and stability met the requirements for the biology sample analysis. For notoginsenoside R1， the pharmacokinetics parameters T1/2， Tmax， Cmax in the plasma were (7.86±1.69)h， (4.00±1.04)h， (0.23±0.05)mg/L. For ginsenoside Rg1， the pharmacokinetics parameters T1/2， Tmax， Cmax in plasma were (4.58±0.95)h， (6.00±0.00)h， (0.32±0.03)mg/L. Conclusion The established UHPLC-MS/MS method is sensitive， accurate and reliable， and is suitable for the pharmacokinetic study of Hushen (Ⅲ) Capsules.
Keywords： Hushen (Ⅲ) Capsules； UHPLC-MS/MS； pharmacokinetics
目前，血尿形成机制尚未完全阐明，通常认为与免疫炎症反应导致肾小球基底膜受损有关，西医治疗往往效果不佳，迁延难愈。肾性血尿属中医学“血证”范畴，中医治疗血尿积累了丰富的临床经验，并取得了较好的疗效[1]。护肾(Ⅲ)号胶囊是张家港市中医医院复方制剂，由白茅根、三七、大黄炭组成，具有清热解毒、滋肾降火功效，临床常用于治疗血尿為主的各种肾病。三七作为方中臣药，具有良好的止血、活血化瘀、消肿止痛作用，辅助君药白茅根治疗血尿诸症。三七总皂苷为三七的主要有效成分，具有抗炎、增强免疫功能的作用。已有研究采用HPLC对护肾(Ⅲ)号胶囊的有效成分进行了含量测定[2]。本研究采用高效、灵敏的超高液相色谱串联质谱(UHPLC-MS/MS)技术对护肾(Ⅲ)号胶囊中三七皂苷R1、人参皂苷Rg1在血浆中的含量进行分析，并对其在大鼠血浆中的药动学特征进行研究，为进一步开展护肾(Ⅲ)号胶囊临床应用研究，明确其作用机制奠定基础。
1 仪器、试药与动物
Agilent 1290超高效液相色谱仪；Agilent 6430 LC-MS三重四级杆质谱仪，配有电喷雾化离子源(ESI)，MassHunter色谱工作站；AE240电子天平(上海梅特勒-托利多有限公司)；MICRO-17R冷冻离心机(美国Thermo公司)；WH-2微型旋涡混合仪(上海沪西分析仪器厂)；Drict-Q5超纯水机(法国Millipore公司)。, 百拇医药(钱伟 冯星月 陈晓峰 王建平 蒋志涛 余辉)