Key Laboratory of Basic and Application Research of Beiyao, Ministry of Education, Heilongjiang University of Chinese Medicine, Harbin 150040, China.
Department of Pharmaceutical Analysis and Analytical Chemistry, College of Pharmacy, Harbin Medical University, Harbin 150086, China.
Molecules. 2022 Aug 24;27(17):5412. doi: 10.3390/molecules27175412.
A selective and rapid ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established and validated for the determination of ziyuglycoside I, 3,19-dihydroxyurs-12-en-28-oic-acid 28--d-glucopyranosyl ester, and pomolic acid in rats after the oral administration of ziyuglycoside I, 3,19-dihydroxyurs-12-en-28-oic-acid 28--d-glucopyranosyl ester, pomolic acid, and L. extract. The separation was carried out on an ACQUITY UPLCHSS T column (2.1 mm × 100 mm, 1.8 μm), using methanol and 5 mmol/L ammonium acetate water as the mobile phase. The three compounds were quantified using the multiple reaction monitoring mode with the electrospray ion source in both the positive and negative mode. Liquid-liquid extraction was applied to the plasma sample preparation. Bifendate was selected as the internal standard. The intra-day and inter-day precision and the accuracy of the method were all within receivable ranges. The lower limit of quantification of ziyuglycoside I, 3,19-dihydroxyurs-12-en-28-oic-acid 28--d-glucopyranosyl ester, and pomolic acid were 6.50, 5.75, and 2.63 ng/mL, respectively. The extraction recoveries of analytes in rat plasma ranged from 83 to 94%. The three components could be rapidly absorbed into the blood (, 1.4-1.6 h) both in the single-administration group or extract group, but the first peak of PA occurred at 0.5 h and the second peak at 4-5 h in the extract. Three compounds were eliminated relatively slowly (, 7.3-11 h). The research was to establish a rapid, sensible, and sensitive UHPLC-MS/MS method using the multi-ion mode for multi-channel simultaneous mensuration pharmacokinetics parameters of three compounds in rats after oral administration of extract. This study found, for the first time, differences in the pharmacokinetic parameters of the three compounds in the monomer compounds and extract administration, which preliminarily revealed the transformation and metabolism of the three compounds in vivo.
建立并验证了一种选择性和快速的超高效液相色谱-串联质谱（UHPLC-MS/MS）方法，用于测定齐墩果酸苷 I、3,19-二羟基乌苏-12-烯-28-酸 28-O-β-D-吡喃葡萄糖酯和熊果酸在大鼠口服齐墩果酸苷 I、3,19-二羟基乌苏-12-烯-28-酸 28-O-β-D-吡喃葡萄糖酯、熊果酸和 L.提取物后的体内药代动力学参数。分离在 ACQUITY UPLCHSS T 柱（2.1mm×100mm，1.8μm）上进行，使用甲醇和 5mmol/L 乙酸铵水作为流动相。采用正、负离子多反应监测模式对三种化合物进行定量，电喷雾离子源。采用液-液萃取法进行血浆样品制备。双飞蓟宾被选为内标。该方法的日内和日间精密度及准确度均在可接受范围内。齐墩果酸苷 I、3,19-二羟基乌苏-12-烯-28-酸 28-O-β-D-吡喃葡萄糖酯和熊果酸的定量下限分别为 6.50、5.75 和 2.63ng/mL。分析物在大鼠血浆中的提取回收率为 83%至 94%。在单剂量组或提取物组中，三种成分均能迅速被血液吸收（1.4-1.6h），但提取物组中 PA 的第一个峰值出现在 0.5h，第二个峰值出现在 4-5h。三种化合物的消除相对较慢（7.3-11h）。该研究旨在建立一种快速、灵敏、灵敏的 UHPLC-MS/MS 方法，采用多离子模式，对大鼠口服提取物后三种化合物的多通道同时测定药代动力学参数。本研究首次发现单体化合物和提取物给药后三种化合物药代动力学参数的差异，初步揭示了三种化合物在体内的转化和代谢。
