UPLC-MS/MS法测定都梁丸中37个成分在大鼠体内的药代动力学

doi:10.5246/jcps.2023.07.047

摘要/Abstract

摘要：

都梁丸是中药经典方剂, 由白芷和川芎组成。为了研究都梁丸的药效物质基础, 我们采用Diamonsil C18柱、乙腈和0.05%甲酸水溶液为流动相、电喷雾电离源和多重反应监测模式, 建立了同时测定白芷、川芎中37种成分的UPLC-MS/MS法并进行了方法学验证。同时, 我们采用该方法测定了口服都梁丸提取物后的大鼠体内药代动力学, 发现这37种成分均能迅速吸收入血, 大部分化合物在24小时内被完全清除, 白芷香豆素的口服吸收效果优于川芎内酯类化合物, 都梁丸中白芷香豆素的清除速度明显慢于白芷单一给药。该结果为都梁丸的药效物质基础和白芷-川芎配伍作用机制研究提供了依据。

关键词: 都梁丸, 白芷, 川芎, UPLC-MS/MS, 药代动力学

Abstract:

Duliang Pill (DLP), consisting of Baizhi and Chuanxiong, is the classical formula of traditional Chinese medicine. In order to investigate the substance basis of DLP, we developed a highly sensitive and reliable UPLC-MS/MS method capable of simultaneously detecting 37 constituents from both Baizhi and Chuanxiong. To achieve chromatographic separation, we utilized a Diamonsil C₁₈ column along with a mobile phase consisting of acetonitrile and 0.05% formic acid in water. MS analysis was conducted in the multiple reaction monitoring mode, utilizing electrospray ionization in either the positive or negative mode, depending on the compound being analyzed. The method was validated for specificity, linearity, sensitivity, precision, accuracy, recovery, matrix effect, and stability, and found to be satisfactory. The validated method was then applied to a pharmacokinetic study in rat plasma following oral administration of DLP extract. Our findings revealed that all compounds were rapidly absorbed from the gastrointestinal tract into the plasma, with most being eliminated completely within 24 h. Notably, coumarins derived from Baizhi exhibited superior oral absorption effects compared to ligustilides from Chuanxiong. Furthermore, we observed that the elimination of coumarins from Baizhi within DLP was slower compared to their elimination in single Baizhi. These results provided valuable insights for the overall study of active substances within the DLP formula and further research into the interaction mechanisms of Baizhi-Chuanxiong compatibility.

Key words: Duliang Pill, Baizhi, Chuanxiong, UPLC-MS/MS, Pharmacokinetics

Supporting:

杨雁芳, 韦玮, 张雷, 徐嵬, 杨秀伟. UPLC-MS/MS法测定都梁丸中37个成分在大鼠体内的药代动力学[J]. 中国药学(英文版), 2023, 32(7): 560-573.

Yanfang Yang, Wei Wei, Lei Zhang, Wei Xu, Xiuwei Yang. Simultaneous quantification of 37 constituents in Duliang Pill by UPLC-MS/MS: Application to the pharmacokinetic studies in rat plasma[J]. Journal of Chinese Pharmaceutical Sciences, 2023, 32(7): 560-573.

图/表 6

Figure 1. Chemical structures of primary components from DLP. (1) Osthenol, (2) dehydrogeijerin, (3) 7-demethylsuberosin, (4) psoralen, (5) bergapten, (6) xanthotoxol, (7) xanthotoxin, (8) 5-hydroxy-8-methoxypsoralen, (9) isopimpinellin, (10) isodemethylfuropinarine, (11) alloimperatorin, (12) alloisoimperatorin, (13) imperatorin, (14) phellopterin, (15) isoimperatorin, (16) heraclenol, (17) byakangelicin, (18) heracol, (19) isooxypeucedanin, (20) oxypeucedanin hydrate, (21) oxypeucedanin, (22) oxypeucedanin ethanolate, (23) oxypeucedanin methanolate, (24) isogosferol, (25) 2??R-neobyakangelicol, (26) byakangelicol, (27) isobyakangelicol, (28) (2?S,3?R)-3?-hydroxymarmesinin, (29) senkyunolide I, (30) senkyunolide H, (31) senkyunolide G, (32) senkyunolide A, (33) Z-ligustilide, (34) 3-butylidenephthalide, (35) neocnidilide, (36) cnidilide, and (37) levistolide A.

Table 1. MW, precursor/product ion, and mass parameters of the 37 analytes and daidzein (IS).

Figure 2. MRM results of the analytes (A. blank plama; B. blank plama spiked with analytes and IS; C. plasma sample at 2 h after oral administration of DLP).

Table 2. Intra- and inter-day precisions and accuracies, recoveries, stabilities, and matrix effects of the 37 analytes in rat plasma (mean ± SD, n = 6).

Figure 3. Mean concentration-time curves of the 37 compounds in rat plasma after oral administration of DLP at a single dose of 20 g/kg (n = 5).

Table 3. Pharmacokinetics parameters of the 37 analytes in rat plasma (mean ± SD, n = 5).

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