高效亲和色谱法测定2种中药成分与人血清白蛋白的结合率

2011年4

月Vo.l 29No .4

April 2011

Chi n ese Journal of Chromatography

358~361

研究论文

DOI :10.3724/SP .J .1123.2011.00358

*通讯联系人:张秀莉,副研究员,主要研究天然小分子和多肽的分离分析.Te :l (0411)84379521,E m ai:l zhangx i uli @dicp .ac .cn .基金项目:重大新药创制专项(N o .2009ZX 09313 003)和国家自然科学基金面上项目(No .30801513).收稿日期:2010 12 08

高效亲和色谱法测定2种中药成分与人血清白蛋白的结合率

蔡晓明1

, 张 岩2

, 于 龙1

, 郭志谋1

, 张秀莉1*

, 梁鑫淼

1

(1.中国科学院大连化学物理研究所,中国科学院分离分析重点实验室,辽宁大连116023;

2.Un ivers ity of C alifon ia Irvi ne ,CA 92697 4625,USA )

摘要:采用高效亲和色谱技术(HPAC )对中药成分与人血清白蛋白(HSA )的相互作用进行了研究。首先采用点击化学的方法制备了表面键合有HSA 蛋白的硅胶固定相并装填成亲和色谱柱,根据药物在该色谱柱上与空白硅胶

柱上的保留时间差计算得到药物与蛋白的结合率。利用该方法测得模型化合物华法令与H SA 的结合率与文献中采用超滤法测得的结果基本一致,表明该方法可用于测定药物与HSA 的结合率。在此基础上用该方法测定了葛根素和告依春两种中药成分与HSA 的相对结合率分别为10 26%和10 20%。同时用超滤的方法测定了葛根素与H SA 的结合率为14 25%。结果表明,HPAC 可以作为研究药物与蛋白相互作用的一种简便可行的方法,其测定结果与超滤方法一致。

关键词:高效亲和色谱法;人血清白蛋白;葛根素;告依春;结合率;超滤

中图分类号:O 658 文献标识码:A 文章编号:1000 8713(2011)04 0358 04

Detection of drug hu m an seru m albu m in b ind i ng ratios of t wo

Ch i nese m edici nal ingred ients by h igh perfor m ance

affinity chro matography

CAI Xiao m i n g 1

,Z HANG Yan 2

,YU Long 1

,GUO Zhi m ou 1

,ZHANG X iuli 1*

,LI ANG X in m iao

1

(1.CAS K ey L abora tory of Sepa ra tion Sc ien ce for An a ly tica l Che m istry ,D a lian In stitu te o f

Che m ica l Phy sics,Chin ese Acade m y o f Scie n ces,Da lian 116023,Ch i n a;

2.Un iver sity o f Ca lifon ia Ir vin e,CA 92697 4625,U SA )

A bstract :The i n teraction of t wo Chinese m edici n al i n gredi e nts and hum an seru m al b u m in (HSA )has been investigat ed by high perf or m ance affi n it y chro m atography (HPAC ).HSA bounded silica based stati o nary phase w as prepared based on t he click chem istry strategy ,and packed in a colu m n (nam ed as HSA col u m n).The drug HSA binding rati o was calculated fro m t he diff erence of t he drug s ret enti o n ti m es on the H SA colu m n and silica col u m n (blank col u m n).T he warfari n HSA bindi n g rati o det er m ined by t his m ethod was s i m ilar to t he refer ence reported value by ultrafiltration m et hod .The results i n dicat ed that the new HSA colu m n and t he HPAC m et hod can be used for t he detection of binding ratio of drug and HSA .T he bi n di n g ratios of puerari n and goitri n deter m i n ed by the HPAC m ethod were 10 26%and 10 20%,respecti v e l y .And the binding rati o of puerarin deter m i n ed by ultrafiltrati o n was 14 25%.A ll t hese results showed t hat HPAC is a usef ul m et hod t o i n vestigate the interacti o n bet ween drugs and protein .

K ey words :high perf or m ance affinity chro m atography (HPAC);hu m an seru m albu m i n ;puer ari n ;goitrin ;binding ratio ;ultrafiltration 人血清白蛋白(hu m an seru m albu m in ,HSA)是由585个氨基酸残基组成的一条单肽链,相对分子质量约为66500,是血浆中含量最丰富的重要载

体蛋白[1]

。H SA 上有多个药物结合位点,药物进入

血浆后,首先与血清白蛋白结合,然后再被输送到身体的各个部位产生药效。药物进入体内后的许多重