Lin Yuan-yuan, Wang Yin-ping, Lu Hao-yang, Guo Xiu-cai, Liu Xia, Wu Chuan-bin, Xu Yue-hong
School of Pharmaceutical Science, Sun Yat-sen University, Guangzhou 510006, China.
The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
J Ethnopharmacol. 2014 Apr 28;153(2):454-60. doi: 10.1016/j.jep.2014.03.009. Epub 2014 Mar 12.
Acupoint application of cold asthma recipe (CAR) was a traditional Chinese medicine (TCM) method, widely used as an alternative medicine for clinical prevention of the common winter diseases of asthma and bronchitis. Tetrahydropalmatine (THP) was a main active ingredient of CAR extract. The aim of this study is to compare plasma pharmacokinetics and lung distribution of THP between Feishu (FS) acupoint (BL 13) and Non-Feishu (NFS) acupoint application of CAR extract by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).
The extract of CAR was topically administrated in FS and NFS acupoint of rats for plasma pharmacokinetics, and topically administrated in FS and NFS acupoint of mice for lung distribution. The plasma and lung homogenates were pretreated by protein precipitation with acetonitrile. Chromatographic separation was performed on an ACQUITY UPLC BEH C18 column with a mobile phase consisted of 0.1% formic acid in acetonitrile and 0.1% formic acid in water. The detection was accomplished by multiple-reaction monitoring (MRM) scanning in the positive electrospray ionization (ESI(+)) mode. All pharmacokinetic parameters were estimated by non-compartmental analysis.
A sensitive, accurate and precise UPLC-MS/MS method was successfully established for determination of THP in 100 μL plasma and lung homogenate. The lower limit of quantification (LLOQ) of THP was 0.05 ng/mL and 0.072 ng/mL, respectively. The pharmacokinetic results manifested that THP was absorbed and eliminated slowly in plasma. Additionally, it was found that there was significantly higher amount of THP absorbed into blood and lung after FS acupoint application compared to NFS acupoint application.
Both of the rat plasma pharmacokinetics and mice lung distribution of THP could support that FS acupoint application of CAR extract has greater advantages of absorption into the blood circulation and distribution in target tissue over NFS acupoint application. The results might be helpful in providing a rational explanation for why the TCM chose the acupoint application and elucidating the underlying mechanism of this treatment.
寒喘方（CAR）穴位贴敷是一种传统中医方法，作为临床预防哮喘和支气管炎等常见冬季疾病的替代药物被广泛应用。延胡索乙素（THP）是CAR提取物的主要活性成分。本研究旨在通过超高效液相色谱-串联质谱法（UPLC-MS/MS）比较CAR提取物在肺俞穴（BL 13）和非肺俞穴应用后THP的血浆药代动力学和肺分布。
将CAR提取物局部应用于大鼠的肺俞穴和非肺俞穴以进行血浆药代动力学研究，并局部应用于小鼠的肺俞穴和非肺俞穴以进行肺分布研究。血浆和肺匀浆用乙腈进行蛋白沉淀预处理。采用ACQUITY UPLC BEH C18色谱柱进行色谱分离，流动相由乙腈中的0.1%甲酸和水中的0.1%甲酸组成。通过正电喷雾电离（ESI(+)）模式下的多反应监测（MRM）扫描进行检测。所有药代动力学参数均通过非房室分析进行估算。
成功建立了一种灵敏、准确且精密的UPLC-MS/MS方法，用于测定100μL血浆和肺匀浆中的THP。THP的定量下限（LLOQ）分别为0.05 ng/mL和0.072 ng/mL。药代动力学结果表明，THP在血浆中吸收和消除缓慢。此外，发现与非肺俞穴应用相比，肺俞穴应用后进入血液和肺的THP量显著更高。
THP在大鼠血浆中的药代动力学和在小鼠肺中的分布均表明，与非肺俞穴应用相比，CAR提取物肺俞穴应用在血液循环吸收和靶组织分布方面具有更大优势。这些结果可能有助于合理解释中医为何选择穴位贴敷并阐明这种治疗方法的潜在机制。
