超高效液相色谱-串联质谱法测定大鼠血浆中淫羊藿素
|
摘要
目的建立超高效液相色谱-串联质谱法测定大鼠血浆中淫羊藿素(ICT)浓度,并用于研究大鼠静脉注射ICT的药动学。方法采用乙腈提取血浆中ICT,以香豆雌酚(CMT)为内标,在BEH C18色谱柱(2.1 mm×50 mm,1.7μm)上以流动相乙腈-水-甲酸铵-甲酸梯度洗脱,流速0.3 ml/min,柱温40℃,分析时间6.5 min;在电喷雾离子化电离源上以多反应监测方式进行负离子检测,用于定量分析的离子反应分别为m/z 367.1→297.1(ICT)和m/z 267.0→211.1(CMT)。大鼠静脉注射10 mg/kg ICT,采集不同时刻血样并测定药物浓度。结果 ICT的线性范围为0.5~20 ng/ml,方法定量下限为0.5 ng/ml。ICT日内和日间精密度(RSD)在8.1%以内,准确度在95.3%~99.3%,提取回收率为90.1%~92.1%,基质效应为97.5%~101.2%。大鼠静脉注射ICT的t1/2为(1.13±0.32)h,表观分布容积为(4.54±0.27)L/kg,清除率为(2.91±0.68)L/(h·kg)。结论该方法速度快、专属性好、准确度高,适用于ICT临床前药动学研究。
Objective To establish an ultra-performance liquid chromatographytandem mass spectrometry( UPLC-MS / MS) method for the quantification of icairitin( ICT),and investigate pharmacokinetics of ICT in rats following a single intravenous administration. Methods ICT and an internal standard coumestrol( CMT) were extracted from rat plasma using acetonitrile and separated on a BEH C18 column( 2. 1 mm × 50 mm,1. 7 μm) using a gradient mobile phase of acetonitrile,water,ammonium formate and formic acid at a flow rate of 0. 3 ml / min. At negative electrospray ionization mode,multiple reaction monitoring of the precursor-product ion transitions of m / z 367. 1→297. 1for ICT,267. 0 →211. 1 for CMT was used for the quantification. Plasma was collected after rats were intravenously administered with ICT at a single dose of 10 mg / kg. Results The linear calibration curve was obtained in a concentration range of 0. 5- 20 ng / ml with a lower limit of quantification of 0. 5 ng / ml. The value of intra- and inter-day precision was less than 8. 1% and accuracy fell in the ranges of 95. 3%- 99. 3%. The recovery ranged from 90. 1% to 92. 1% and the matrix effects from 97. 5% to 101. 2%.After intravenous administration of ICT to rats,t1 /2was( 1. 13 ± 0. 32) h,V was( 4. 54 ±0. 27) L / kg,and CL was( 2. 91 ± 0. 68) L /( h·kg). Conclusion The method was rapid,specific and accurate,suitable for preclinical pharmacokinetics of ICT.
Objective To establish an ultra-performance liquid chromatographytandem mass spectrometry( UPLC-MS / MS) method for the quantification of icairitin( ICT),and investigate pharmacokinetics of ICT in rats following a single intravenous administration. Methods ICT and an internal standard coumestrol( CMT) were extracted from rat plasma using acetonitrile and separated on a BEH C18 column( 2. 1 mm × 50 mm,1. 7 μm) using a gradient mobile phase of acetonitrile,water,ammonium formate and formic acid at a flow rate of 0. 3 ml / min. At negative electrospray ionization mode,multiple reaction monitoring of the precursor-product ion transitions of m / z 367. 1→297. 1for ICT,267. 0 →211. 1 for CMT was used for the quantification. Plasma was collected after rats were intravenously administered with ICT at a single dose of 10 mg / kg. Results The linear calibration curve was obtained in a concentration range of 0. 5- 20 ng / ml with a lower limit of quantification of 0. 5 ng / ml. The value of intra- and inter-day precision was less than 8. 1% and accuracy fell in the ranges of 95. 3%- 99. 3%. The recovery ranged from 90. 1% to 92. 1% and the matrix effects from 97. 5% to 101. 2%.After intravenous administration of ICT to rats,t1 /2was( 1. 13 ± 0. 32) h,V was( 4. 54 ±0. 27) L / kg,and CL was( 2. 91 ± 0. 68) L /( h·kg). Conclusion The method was rapid,specific and accurate,suitable for preclinical pharmacokinetics of ICT.
引文
[1]MA H,HE X,YANG Y,et al.The genus Epimedium:an ethnopharmacological and phytochemical review[J].J Ethnopharmacol,2011,134(3):519-541.
[2]TIONG C T,CHEN C,ZHANG S J,et al.A novel prenylflavone restricts breast cancer cell growth through AhR-mediated destabilization of ERalpha protein[J].Carcinogenesis,2012,33(5):1089-1097.
[3]LAI X,YE Y,SUN C,et al.Icaritin exhibits antiinflammatory effects in the mouse peritoneal macrophages and peritonitis model[J].Int Immunopharmacol,2013,16(1):41-49.
[4]WANG Z,ZHANG X,WANG H,et al.Neuroprotective effects of icaritin against beta amyloid-induced neurotoxicity in primary cultured rat neuronal cells via estrogen-dependent pathway[J].Neuroscience,2007,145(3):911-922.
[5]HUANG J,YUAN L,WANG X,et al.Icaritin and its glycosides enhance osteoblastic,but suppress osteoclastic,differentiation and activity in vitro[J].Life Sci,2007,81(10):832-840.
[6]SHEN P,WONG S P,YONG E L.Sensitive and rapid method to quantify icaritin and desmethylicaritin in human serum using gas chromatography-mass spectrometry[J].J Chromatogr B,2007,857(1):47-52.
[7]SHEN P,WONG S P,LI J,et al.Simple and sensitive liquid chromatography-tandem mass spectrometry assay for simultaneous measurement of five Epimedium prenylflavonoids in rat sera[J].J Chromatogr B,2009,877(1-2):71-78.
[8]CHANG Q,WANG G N,LI Y,et al.Oral absorption and excretion of icaritin,an aglycone and also active metabolite of prenylflavonoids from the Chinese medicine Herba Epimedii in rats[J].Phytomedicine,2012,19(11):1024-1028.
[9]张立.淫羊藿素血药浓度检测方法研究[J].湖北中医杂志,2013,35(12):66-67.
[10]刘海培,孟繁华,郭继芬,等.高效液相色谱-串联质谱法测定大鼠血浆中淫羊藿素[J].药学学报,2009,44(10):1140-1144.
[11]刘海培,孟繁华,郭继芬,等.淫羊藿素在大鼠体内的药动学研究[J].中国药学杂志,2010,45(7):539-543.
[12]WONG S P,SHEN P,LEE L,et al.Pharmacokinetics of prenylflavonoids and correlations with the dynamics of estrogen action in sera following ingestion of a standardized Epimedium extract[J].J Pharm Biomed Anal,2009,50(2):216-223.
[2]TIONG C T,CHEN C,ZHANG S J,et al.A novel prenylflavone restricts breast cancer cell growth through AhR-mediated destabilization of ERalpha protein[J].Carcinogenesis,2012,33(5):1089-1097.
[3]LAI X,YE Y,SUN C,et al.Icaritin exhibits antiinflammatory effects in the mouse peritoneal macrophages and peritonitis model[J].Int Immunopharmacol,2013,16(1):41-49.
[4]WANG Z,ZHANG X,WANG H,et al.Neuroprotective effects of icaritin against beta amyloid-induced neurotoxicity in primary cultured rat neuronal cells via estrogen-dependent pathway[J].Neuroscience,2007,145(3):911-922.
[5]HUANG J,YUAN L,WANG X,et al.Icaritin and its glycosides enhance osteoblastic,but suppress osteoclastic,differentiation and activity in vitro[J].Life Sci,2007,81(10):832-840.
[6]SHEN P,WONG S P,YONG E L.Sensitive and rapid method to quantify icaritin and desmethylicaritin in human serum using gas chromatography-mass spectrometry[J].J Chromatogr B,2007,857(1):47-52.
[7]SHEN P,WONG S P,LI J,et al.Simple and sensitive liquid chromatography-tandem mass spectrometry assay for simultaneous measurement of five Epimedium prenylflavonoids in rat sera[J].J Chromatogr B,2009,877(1-2):71-78.
[8]CHANG Q,WANG G N,LI Y,et al.Oral absorption and excretion of icaritin,an aglycone and also active metabolite of prenylflavonoids from the Chinese medicine Herba Epimedii in rats[J].Phytomedicine,2012,19(11):1024-1028.
[9]张立.淫羊藿素血药浓度检测方法研究[J].湖北中医杂志,2013,35(12):66-67.
[10]刘海培,孟繁华,郭继芬,等.高效液相色谱-串联质谱法测定大鼠血浆中淫羊藿素[J].药学学报,2009,44(10):1140-1144.
[11]刘海培,孟繁华,郭继芬,等.淫羊藿素在大鼠体内的药动学研究[J].中国药学杂志,2010,45(7):539-543.
[12]WONG S P,SHEN P,LEE L,et al.Pharmacokinetics of prenylflavonoids and correlations with the dynamics of estrogen action in sera following ingestion of a standardized Epimedium extract[J].J Pharm Biomed Anal,2009,50(2):216-223.