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Archive > Volume 10 Issue 2 > 2018,10(2):205-213. DOI:10.1016/j.chmed.2018.03.006 Prev Next

基于腦微透析聯(lián)合LC-MS/MS技術(shù)的交泰丸在正常與失眠大鼠的腦內(nèi)藥動學(xué)特性比較研究

Original articles

Comparative Brain Pharmacokinetic Study of Jiaotai Pills in Normal and Insomnic Rats using Brain Microdialysis Combinated with LC-MS/MS

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    摘要:
    目的：比較交泰丸口服給藥后，其主要活性成分（小檗堿、巴馬汀、黃連堿、表小檗堿和藥根堿）在正常和失眠大鼠的腦內(nèi)藥動學(xué)特性。 方法：運(yùn)用液相色譜-串聯(lián)質(zhì)譜結(jié)合腦微透析方法對五種原小檗堿型生物堿進(jìn)行測定。于大鼠的大腦海馬體內(nèi)植入腦微透析探針，交泰丸提取物灌胃給藥后每30 min 收集一管透析液，持續(xù)10 h。采用BDS Hypersil C18色譜柱進(jìn)行分離，流動相為乙腈-水（含5 mmol 乙酸銨并調(diào)pH至5.0），時長4 min。檢測方式為多反應(yīng)檢測模式，MS/MS條件分別為小檗堿m/z 336.0-320.1，巴馬汀m/z 352.0-336.1，藥根堿m/z 338.0-322.1，表小檗堿m/z 336.0-320.1，黃連堿m/z 320.0-292.1，四氫巴馬?。▋?nèi)標(biāo)）m/z 356.4-192.1。 結(jié)果：建立的LC-MS/MS方法的線性、準(zhǔn)確度、精密度、穩(wěn)定性和基質(zhì)效應(yīng)均符合要求，定量下限為0.05 ng•mL-1。五種原小檗堿型生物堿能迅速分布于腦內(nèi)，其在正常組和模型組的腦內(nèi)藥動學(xué)行為具有顯著性差異，主要表現(xiàn)在AUC和半衰期。 結(jié)論：本實驗有效地使用了LC-MS/MS結(jié)合腦微透析的方法對交泰丸中的五種原小檗堿型生物堿進(jìn)行腦內(nèi)藥動學(xué)研究。結(jié)果顯示，這五種原小檗堿型生物堿能直接作用于海馬體，且其在失眠大鼠中的吸收顯著高于正常大鼠。

    Abstract:
    Objective To compare the brain pharmacokinetics of five protoberberine-type alkaloids (i.e. berberine, palmatine, coptisine, epiberberine, and jatrorrhizine), which were the main bioactive constituents of Jiaotai Pills (JTP), in normal and insomnic rats orally administrated with the JTP. Methods The detection was conducted by a fully validated liquid chromatography-tandem mass spectrometry combinated with brain microdialysis method. Brain microdialysis probes were inserted into the hippocampus of rats. JTP extracts were administrated intragastrically and then brain microdialysates were collected at 30 min time intervals for 10 h. The separation of the five protoberberine-type alkaloids was carried out on a BDS Hypersil C18 column using a mobile phase consisting of acetonitrile and water (containing 5 mmol ammonium acetate adjusted to pH 5.0) within 4 min. The quanti?cation was performed by multiple reaction monitoring with the transitions of m/z 336.0-320.1 for berberine, m/z 352.0-336.1 for palmatine, m/z 338.0-322.1 for jatrorrhizine, m/z 336.0-320.1 for epiberberine, m/z 320.0-292.1 for coptisine and m/z 356.4-192.1 for IS. Results The lower limit of quanti?cation for five protoberberine-type alkaloids was 0.05 ng/mL. Linearity, accuracy, precision, stability and matrix effect of five analytes were all satisfactory. Five protoberberine-type alkaloids were quickly distributed in the brain. Moreover, signi?cant differences in the principal pharmacokinetic parameters such as AUC and T1/2 of the analytes were observed between two groups. Conclusion The LC-MS/MS method combinated with microdialysis is useful in the brain pharmacokinetic study of five protoberberine-type alkaloids. The results indicated that the rates of analytes absorption in insomnic rats were signi?cantly higher than those in normal rats. Besides, the protoberberine-type alkaloids could bring a direct effect on the neuron in the hippocampus.

    關(guān)鍵詞:
    腦內(nèi)藥動學(xué);失眠大鼠;交泰丸;LC-MS/MS;微透析;原小檗堿型生物堿

    Keywords:


    Project Supported:
    This work was supported by the National Natural Science Foundation of China (No. 81473319 and No. 81473540), the Key Projects of Guangdong Natural Science Foundation (No. 2017A030311022), the Guangdong Science and Technology Program (No. 2015A030401031, 2014A020209026), the Guangdong Natural Science Foundation (No. 2015A030313123), the Zhongshan Science and Technology Program (No. 2016C1015), and the Science Program for Overseas Scholar of Guangzhou University of Chinese Medicine (Torch Program) (No. XH20170111).

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