基于UHPLC-Q-Exactive Orbitrap-MS快速鉴定补阳还五汤及其在人血浆和尿液中药物化学成分

Rapid Identification of Chemical Constituents for Buyang Huanwu Decoction and Its Chemical Constituents in Human Plasma and Urine by UHPLC-Q-Exactive-Orbitrap-MS

关键词:  补阳还五汤  药效物质基础  血浆  尿液  超高效液相色谱串联四级杆轨道阱质谱  脑卒中

Key words:  Buyang Huanwu decoction  pharmacodynamic material base  plasma  urine  UHPLC-Q-Exactive Orbitrap-MS  stroke

摘要:

目的 应用超高效液相色谱串联四级杆轨道阱质谱（UHPLC-Q-Exactive Orbitrap-MS）技术鉴定补阳还五汤及其在人血浆和尿液中药物化学成分。方法 采集补阳还五汤及其临床患者口服给药后的血浆和尿液样本，样本经80%甲醇水处理后，采用Hyperil Gold C₁₈色谱柱（1.0 mm×50 mm，1.9 μm），流动相为水（A）-乙腈（B）（各含0.1%甲酸和5 mmol·L^-1醋酸铵）进行梯度洗脱，流速0.3 mL·min^-1，柱温40℃。通过对比分析补阳还五汤、给药后人血浆和给药后人尿液样本，同时结合化合物的质谱碎裂规律，表征其制剂、含药血浆和尿液中药物化学成分。结果 在汤剂中共鉴定了55个化合物；在给药后患者的血浆中共鉴定了34个化合物；在尿液中鉴定了23个化合物。这些成分主要是黄酮类、苯酸类、苯酞类和皂苷类化合物。结论 本研究全面阐明了补阳还五汤及其在人血浆和尿液药物化学成分，为揭示补阳还五汤真正治疗脑卒中的药效物质基础提供了参考。

Abstract:

OBJECTIVE To identify Buyang Huanwu decoction and its chemical constituents in human plasma and urine by UHPLC-Q-Exactive Orbitrap-MS. METHODS The plasma and urine samples of patients were collected after oral administration of Buyang Huanwu decoction. The samples were treated by 80% methanol water. The Hyperil Gold C₁₈ column (1.0 mm×50 mm, 1.9 μm) was used. The mobile phase was water(A)-acetonitrile(B)(containing 0.1% formic acid and 5 mmol·L^-1 ammonium acetate) for gradient elution. The flow rate was 0.3 mL·min^-1, and the column temperature was 40℃. Through the comparative analysis of Buyang Huanwu decoction, human plasma after administration and human urine samples after administration, combined with the mass spectrometry fragmentation law of the compound, the medicinal chemical components in its preparation, medicated plasma and urine were characterized. RESULTS Fifty-five compounds were identified in Buyang Huanwu decoction, 34 compounds in plasma and 23 compounds in urine. These components were flavonoids, benzoic acid, phthalide and saponins. CONCLUSION This study comprehensively elucidate the chemical constituents of Buyang Huanwu decoction and its pharmacochemistry in human plasma and urine, which provides a reference for revealing the real material basis of pharmacodynamics in treating stroke.