RP-HPLC法测定大鼠血浆中丹参酮IIA浓度及其药代动力学研究

RP2HPLC??????????? IIA??????????? ??? , ??? , ??? 3 , ???????? (???????、???????? ????? , ?? 100050) ?? : ?? 　??????????????????? IIA?????。?? 　??????2???? ,? HPLC?????。???? YMC C18 ( 5μm, ID 310 mm ?150 mm) ;??????2?2??? (74?26?1) ;?? 013 mL?min - 1 ;???? 270 nm;??? 42??? 。?? 　????? 0105 mg?L - 1?6140 mg?L - 1 ,??????? 0105 mg?L - 1 。?、?、? 3?????????????? 9819 % ,10211 %? 10014 %。??、????? (RSD) ??? 5 %。?? 　?????、??、?? ,?????? IIA????????????????。 ??? : ??????? ; ??? IIA; ???? ; ????? ????? : R917. 101 　　　????? : A　　　???? : 0513 - 4870(2003)05 - 0368 - 03 Deter mination of tanshinone IIA in rat plasma and the phar macokinet ics by RP2HPLC method QIAO Jin2ping , HOU Pei2ling , LI Ya2wei 3 , ABLIZ Zeper ( Institute of Materia Medica , Chinese Academy of Medical Sciences and Peking Union Medical College , Beijing 100050 , China) Abstract : Aim 　To develop a sensitive and rapid HPLC method for the determination of tanshinone IIA (TS) in rat plasma and to study its pharmacokinetics in rats. Methods 　TS and 42chlorodiphenyl (internal standard) were extracted from plasma with ethyl acetate. After li quid2li quid extraction , the sample was analyzed by HPLC with YMC C18 column (5μm , 150 mm ?310 mm ID) . The mobile phase consisted of acetontrile2 water2acetic acid (74?26?1) at the flow rate of 013 mL?min - 1 , the UV detection wave length was 270 nm. Result s　The calibration curve was linear ( r = 01998 1) in the range from 0105 to 6140 mg?L - 1 . The lowest detectable concentration was 0105 mg?L - 1 . The recoveries at the concentration of 0105 , 1160 and 6140 mg?L - 1 were 9819 % , 10211 % and 10014 % , respectively. The inter2 and intra2day RSDs were all less than 5 %. Conclusion 　Thi s method is proved to be rapid , preci se and reliable enough to be applied to the pharmacokinetics studies of TS in rat s after a single dose of 15 mg?kg - 1 by oral admini stration. Key wor ds : HPLC; tanshinone IIA ; plasma drug concentration ; pharmacokinetics ???? 2 2 ???? ???????????? (3 6 ) 3 ???? 　T 6 63 65 , F x 6 63 5 , 2 z @ 　　??? IIA (tanshinone IIA ,TS)??????? ?? ( Salvia miltiorrhiza Bunge) ????????? ?????? ,????????? ,?????? ?? ,??? ,??? ,?????? ,?????? ??[ 1] 。???? 1995 ?? TS ???????? ???????????[2 ] ,?????????2 ?????[3 ] 、????2???[ 4 ] 、?????? ?[ 5 ]? SFE2CGC ?[ 6] ??????? TS ???。 ?? RP2HPLC??? TS??????????? ??至?????。????????????? ?? (RP2HPLC)??????? TS ??? ,??? ? TS ????????????????? ,?? ????????。 ??? II ( II ) ??? 2??? ( 2 y )?????? ?863? ???? Acta Pharmaceutica Sinica 2003 ,38(5) :368 - 370 ? : 2002 07 22. : 99708 1 . el : 8 - 10 - 1 218 a : 8 - 10 - 0177 7 E mail : eper imm. ac . cn A tanshinone A 4 4 chlorodiphen l : ????? ?? 　?? Jasco ????????? (?? PU2980 ? ,UV2975 ???) ; SRI MODEL 203 Peak Simple Chromatography Data System;??????? ; ?????。 ????? 　TS ??? (?? > 99 %) ?? DELTA??????????? ;42??? (?? , ???) ,?????? ;?????????? , Merck?? ;??? ,???? ,??? ,???? ? ;?????。 ???? 　SD ?? , ?,?? (210 ?20) g ,?? ????????????。 ???? 　??? : YMC C18 ? ( 5μm , ID 310 mm ?150 mm) ;??? :??2?2??? (74?26?1) , ?? :013 mL?min - 1 ,???? 270 nm。 ?????? 　????? 012 mL ,???? ?? (0105 g?L - 1 42???????) 30μL ,?? , ?????? 3 mL ,???? 10 min ,?? 10 min (3 000 r?min - 1 ) ,??????? ,?40 ????? ?? ,????? 200μL ???? ,?? 1 min ,?? ????? 20μL ??[ 7] 。 ?? 1 　???? ???????? ,????、??????? ??????? 1 ?? ,?????? ,????? ????? ,??????? 11168 min , TS ??? ??? 13153 min。 2 　???? ???????012 mL ,?? TS ?????? 50μL ,?????????0105 ,011 ,012 ,014 ,018 , 1160 ,3120 ? 6140 mg?L - 1????? ,????? ??“??????”????。???????? ? ( C)???? ,????????????? ( Y) ???? ,????? Y = 1 6 1 , = 1 ,??????? 1 5 ?L ( SΠN = 3) 。 A : Blank plasma sample ; B : Plasma sample spiked with 3120 mg?L - 1 t anshinone IIA (1) and internal standard (2) ; C : Plasma sample 0117 h after an oral administration of TS (15 mg?kg - 1) to a SD rat ; IS: 42 chlorodiphenyl Figure 1 　HPLC Chromatograms of tanshinone IIA ( TS) and internal standard ( IS) 3　??????? ???????? TS ??? 0105 , 1160 ? 6140 mg?L - 1????? ,?“??????”??? ??? ,?????? 5???? ,??????? ??? 9819 % ,10211 %? 10014 %。? 1 d ??? ???? ,?? 3 ?????????? RSD ?? ?312 % ,214 %? 116 % ( n = 5) 。??????? ??? 5 d ???? RSD ??? 417 % ,316 %? 219 %( n =5) 。 4　????? ??????? TS ??????????? ?[ 8 ] 。????????2???? 3 ???? ,? ? TS ???????? (????? 10 %??) ; 4 ?? ,TS??????至???? 3 ?? ;??? ????????至?????? 24 h。 5　TS ???????????? SD?? 10 ? ,??????? 2 ? ,?? 5 ? ,????????????? ,?????? 5 ??? ,?? 1 ??????????? 0108 , 0125 ,015 ,110 ,310 ,610 ,1010 ,1610 ? 2410 h ;? 2 ? ?????????? 0 ,0117 ,0133 ,0167 ,210 ,410 , 810 ,1210 ? 2010 h。????? 12 h ,????? ?。???280 ???????????? TS ?? ? ,ig 15 mg?kg - 1 ,?????????????? ?? 500μL ,?????????? ,????? ? ,??????“??????”???? ,??? ???? TS ???????2???? ,??? ? 。 ?963????? :RP2HPLC??????????? IIA??????????? ? 0 021 4 C - 0 000 771 r 0 998 1 0 0 mg - 1 ig 2 Figure 2 　Mean plasma concentration2time curve of tanshinone IIA after single oral dose of 15 mg?kg - 1 in rats 　　??????? 3P87 ?????????? , ??????、??、???? ,????2???? ? po ?????? ,??????????? 1。 Ta ble 1 　Pha rmacokinetic parameter s of tanshinone IIA aft er single oral dose of 15 mg?kg - 1 in rats Parameter Values Standard error (SE) 　　A Πmg?L - 1 30182 2157E +01 　　B Πmg?L - 1 3187 1180E +0 　　αΠh - 1 1127 3118E - 01 　　βΠh - 1 0119 7199E - 02 　　T1Π2αΠh 0155 　　T1Π2βΠh 3163 　　T1Π2 kaΠh 0141 　　kaΠh - 1 1169 3117E - 01 　　TmaxΠh 0185 　　CmaxΠmg?L - 1 5157 　　VdΠmL 1134 　　AUC Πmg?h?L - 1 24109 ?? ?????? RP2HPLC ????? IIA??? ?????????? ,???????????。 ??????? ,???????。???? ,?? ??????????、??????????? ??? ,??????????????????。 ???????? ,?? ig??? IIA? ,?? ????? 0155 h ,??????? 3163 h??? ?????。????? IIA ????????? ????。 Refer ences : [ 1] Handbook of the Composit ion and Pharmacology of Common Chinese Drugs (???????????) [Z] . Beijing: China Medico2Pharmaceutical Science and Technology Publishing House , 1994. 598. [ 2] The State Pharmacopoeia Committee of the People’s Republic of China . Pharmacopoeia of the People’s Republic of China (? ?? ?? ?? ?? ) [ S ] . Guangzhou: Guangdong Science and Technology Press , 1995. 62 - 63. [ 3] Shun WJ , Meng HB , Gao H. Determination of tanshinone IIA in Salvia milt iorrhiza Bunge by solid phase extraction2HPLC [J ] . Chin J Pharm Anal (??????) , 1996 ,16 (3) : 199 - 200. [ 4] Liu WJ , Wang BQ , Li XF. Determination of tanshinone IIA by TLC2fluorescence chromatography [ J ] . Northwest Pharm J (?????? ) , 1996 ,11 (6) :224 - 226. [ 5] Hu SY. Research of determination of tanshinone IIA by ultrasonic extraction2HPLC [J ]. Chin J Mod Appl Pharm (? ???????) , 1993 ,10(4) :18 - 20. [ 6] Chen B , Zhu M , Xing WJ , et al . SFE2CGC Determination of Tanshinone IIA in Salvia miltiorrhiza Bunge [ J ] . Acta Pharm Sin (????) , 2001 ,36(1) :55 - 57. [ 7] Xue M , Cui Y, Wang HQ , et al . Pharmcokinetics of cryptotanshinone and its metabolite in pigs [J ] . Acta Pharm Sin (????) , 1999 ,34(2) :81 - 84. [ 8] Du Y, Chen XY, Yang HY, et al . Determination of wogonin in rat plasma by liquid chromatography2tandem mass spectrometry [J ] . Acta Pharm Sin (????) , 2002 ,37 (5) :362 - 366. ?073? ???? Acta Pharmaceutica Sinica 2003 ,38(5) :368 - 370 ?