Pan Ying, Qian Dawei, Liu Pei, Zhang Yang, Zhu Zhenhua, Zhang Li, Duan Jin-Ao
Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, Nanjing 210023, China; Department of Pharmacy, The Second Hospital of Nanjing, Affiliated to Medical School of Southeast University, Nanjing 210003, China.
Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, Nanjing 210023, China; Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Aug 15;1060:221-230. doi: 10.1016/j.jchromb.2017.06.004. Epub 2017 Jun 4.
Xiang-Fu-Si-Wu Decoction (XFSWD) has been used to treat gynecology diseases in clinical practice for hundreds of years in China, especially for primary dysmenorrheal of QiZhi Type. In previous researches, both essential oils (XEO) and non-volatile components (NXEO) extracted from XFSWD showed significant pharmacological activities, but the influence of XEO on NXEO was unclear. The purpose of this paper was to develop a validated analytical method for simultaneous determination of ten components including Vanillic acid, Ferulic acid, Caffeic acid, Gallic acid, Paeoniflorin, Albiflorin, Tetrahydropalmatine, Protopine, Berberine and Tetrahydrocolumbamine, and to compare the pharmacokinetic parameters of these components to illustrate the influences of XEO on NXEO in normal rats. After being extracted by methanol (1:3, v/v), the plasma samples were analyzed with Clarithromycin (IS) and Chloramphenicol (IS) as mixed internal standard (IS). Then the analytes were separated on a ACQUITY UPLC BEH C (100×2.1mm, 1.7μm) column with gradient mobile phase (containing 0.1% formic acid aqueous solution and acetonitrile) at a flow rate of 0.4mL/min. All analytes and mixed IS were performed on an electrospray ionization source (ESI) using multiple-reaction monitoring (MRM) with positive and negative ionization mode. The calibration curves of all the analytes showed good linearity (r >0.99), and the lower limits of quantification (LLOQ) were 0.66-46.88ng/mL. The intra- and inter-day precisions ranged 1.24-12.45% and 2.49-13.24% for all the analytes. The mean extraction recoveries of the analytes were in the range of 73.10-93.71% and the average matrix effects were within 82.39-93.18%. The validated method has been fully applied to compare the pharmacokinetic parameters of ten components in rat plasma after oral administration of NXEO, NXEO+β-cyclodextrin (β-CD), NXEO+XEO, and NXEO+β-XEO (the inclusion of XEO by β-CD). It was found that the area under the concentration curves (AUC) of ten constituents in group NXEO+β-CD increased compared with group NXEO especially Paeoniflorin, Tetrahydropalmatine and Tetrahydrocolumbamine (p <0.05). The maximum concentration (C) and AUC of ten compounds in group NXEO+XEO and NXEO+β-XEO had varying degree of increasing in comparison with group NXEO. The growth of group NXEO+β-XEO ran higher than group NXEO+XEO. As can be seen from above-mentioned results, β-CD could remarkably increase the absorption of Paeoniflorin, Tetrahydropalmatine and Tetrahydrocolumbamine. Both of XEO and β-XEO could improve the absorption and bioavailability of all the ten non-volatile compositions. The promotion effect of β-XEO was stronger than XEO due to the double functions of XEO and β-CD.
香附四物汤（XFSWD）在中国临床实践中用于治疗妇科疾病已有数百年历史，尤其用于治疗气滞型原发性痛经。在以往研究中，从香附四物汤中提取的挥发油（XEO）和非挥发性成分（NXEO）均显示出显著的药理活性，但XEO对NXEO的影响尚不清楚。本文旨在建立一种经过验证的分析方法，用于同时测定香草酸、阿魏酸、咖啡酸、没食子酸、芍药苷、 albiflorin、延胡索乙素、原阿片碱、小檗碱和四氢小檗碱等十种成分，并比较这些成分的药代动力学参数，以阐明XEO对正常大鼠体内NXEO的影响。血浆样品经甲醇（1:3，v/v）提取后，以克拉霉素（内标物）和氯霉素（内标物）作为混合内标物进行分析。然后，在ACQUITY UPLC BEH C（100×2.1mm，1.7μm）色谱柱上，以含0.1%甲酸水溶液和乙腈的梯度流动相，流速为0.4mL/min进行分析物分离。所有分析物和混合内标物均采用电喷雾电离源（ESI），在正离子和负离子模式下进行多反应监测（MRM）。所有分析物的校准曲线均显示出良好的线性（r>0.99），定量下限（LLOQ）为0.66 - 46.88ng/mL。所有分析物的日内和日间精密度范围分别为1.24 - 12.45%和2.49 - 13.24%。分析物的平均提取回收率在73.10 - 93.71%范围内，平均基质效应在82.39 - 93.18%范围内。该经过验证的方法已完全应用于比较口服NXEO、NXEO +β-环糊精（β-CD）、NXEO + XEO和NXEO +β-XEO（β-CD包合XEO）后大鼠血浆中十种成分的药代动力学参数。结果发现，与NXEO组相比，NXEO +β-CD组中十种成分的浓度曲线下面积（AUC）增加，尤其是芍药苷、延胡索乙素和四氢小檗碱（p<0.05）。与NXEO组相比，NXEO + XEO组和NXEO +β-XEO组中十种化合物的最大浓度（C）和AUC有不同程度的增加。NXEO +β-XEO组的增长高于NXEO + XEO组。从上述结果可以看出，β-CD可显著增加芍药苷、延胡索乙素和四氢小檗碱的吸收。XEO和β-XEO均可提高所有十种非挥发性成分的吸收和生物利用度。由于XEO和β-CD的双重作用，β-XEO的促进作用强于XEO。
