GC-MS测定莪术中β-榄香烯的血药浓度及大鼠体内药代动力学研究
|
摘要
目的:建立快速灵敏的气质联用(GC-MS)法测定莪术中β-榄香烯在大鼠血浆中血药浓度,并研究β-榄香烯在大鼠体内的药代动力学特征。方法:血浆样品的处理采用正己烷液-液萃取方法,萘为内标。色谱柱:DB-5毛细管柱(30 m×0.25 mm,0.25μm);程序升温:初始60℃,保持1 min,再以30℃·min~(-1)的速度升温至160℃,保持3 min,再以10℃·min~(-1)的速度升温至200℃,保持2 min。采用电子轰击离子源及选择离子扫描模式(SIM);选择监测离子:β-榄香烯m/z 93,内标m/z 128。按β-榄香烯10 mg·kg~(-1)灌胃给药后,测定大鼠血浆中β-榄香烯的浓度,并用DAS 2.0软件计算药代动力学参数。结果:血浆中β-榄香烯质量浓度在0.05～200.0μg·mL~(-1)浓度范围内线性关系良好(r~2=0.999 1),定量下限为0.05μg·mL~(-1);低、中、高3个浓度准确度、日内及日间精密度均小于5%;提取回收率分别为91.7%、105.1%、108.8%;低、中、高3个浓度血浆样品在室温放置12 h,4℃冰箱中放置24 h及反复冻融4次后均能保持稳定。大鼠灌胃莪术提取液后β-榄香烯药-时曲线符合二室模型;主要药代学参数:AUC_(0-t)为(9.83±1.07)μg·L~(-1)·h,t_(1/2α)为(0.47±0.05)h,t_(1/2β)为(1.82±3.11)h,T_(max)为(2.06±0.37)h,C_(max)为(2.61±0.17)μg·mL~(-1)。结论:该方法简便、准确,专属性强,适用于莪术中β-榄香烯在大鼠体内的药代动力学研究。
Objective:To establish a rapid,sensitive method for the determination of plasma concentration ofβ-elemene in Curcuma zedoaria using capillary gas chromatography coupled to mass spectrometry(GC-MS) and to study the pharmacokinetics in rats.Methods:The plasma sample was treated with a liquid-liquid extraction method of n-hexane,and naphthalene was used as an internal standard(IS).The determination was performed on DB-5 column(30 m×0.25 mm,0.25 μm).The initial column temperature was 60 ℃,maintained for 1 min,then raised to 160 ℃ at a rate of 30 ℃·min~(-1),maintained for 3 min,and finally increased to 200 ℃ at a rate of 10 ℃·min~(-1) and maintained for 2 min.EI and single ion monitoring pattern(SIM)were used for ion scanning with m/z 93(β-elemene) and m/z 128(IS).After administration of β-elemene 10 mg·kg~(-1) by oral gavage,the concentration of β-elemene in rat plasma was determined,and the pharmacokinetic parameters were calculated by DAS 2.0 software.Results:There was excellent linearity of the plasma concentration of β-elemene in the range of 0.05-200.0 μg·mL~(-1)(r~2=0.999 1),and the lowest limit of quantification was 0.05 μg·mL~(-1).The accuracy,intraday and inter-day precisions of low,medium and high concentrations were less than 5% and the recovery rates were 91.7%,105.1% and 108.8%,respectively.The plasma samples of low,medium and high concentrations were kept at room temperature for 12 h,and were stable after being placed in refrigerator at 4 ℃ for 24 h and repeatedly frozen and thawed for 4 times.The concentration-time curve of β-elemene of the extract of rats after administration of Curcuma zedoaria by oral gavage was consistent with the two-compartment model.The main pharmacokinetic parameters were as follows:AUC_(0-t) was(9.83±1.07)μg·L~(-1)·h,t_(1/2α) was(0.47±0.05)h,t_(1/2β) wss(1.82±3.11)h,T_(max) was(2.06±0.37)h,and C_(max) was(2.61±0.17)μg·mL~(-1).Conclusion:The established method is simple,accurate,specific and suitable for pharmacokinetic studies of β-elemene in curcuma zedoaria in rats.
Objective:To establish a rapid,sensitive method for the determination of plasma concentration ofβ-elemene in Curcuma zedoaria using capillary gas chromatography coupled to mass spectrometry(GC-MS) and to study the pharmacokinetics in rats.Methods:The plasma sample was treated with a liquid-liquid extraction method of n-hexane,and naphthalene was used as an internal standard(IS).The determination was performed on DB-5 column(30 m×0.25 mm,0.25 μm).The initial column temperature was 60 ℃,maintained for 1 min,then raised to 160 ℃ at a rate of 30 ℃·min~(-1),maintained for 3 min,and finally increased to 200 ℃ at a rate of 10 ℃·min~(-1) and maintained for 2 min.EI and single ion monitoring pattern(SIM)were used for ion scanning with m/z 93(β-elemene) and m/z 128(IS).After administration of β-elemene 10 mg·kg~(-1) by oral gavage,the concentration of β-elemene in rat plasma was determined,and the pharmacokinetic parameters were calculated by DAS 2.0 software.Results:There was excellent linearity of the plasma concentration of β-elemene in the range of 0.05-200.0 μg·mL~(-1)(r~2=0.999 1),and the lowest limit of quantification was 0.05 μg·mL~(-1).The accuracy,intraday and inter-day precisions of low,medium and high concentrations were less than 5% and the recovery rates were 91.7%,105.1% and 108.8%,respectively.The plasma samples of low,medium and high concentrations were kept at room temperature for 12 h,and were stable after being placed in refrigerator at 4 ℃ for 24 h and repeatedly frozen and thawed for 4 times.The concentration-time curve of β-elemene of the extract of rats after administration of Curcuma zedoaria by oral gavage was consistent with the two-compartment model.The main pharmacokinetic parameters were as follows:AUC_(0-t) was(9.83±1.07)μg·L~(-1)·h,t_(1/2α) was(0.47±0.05)h,t_(1/2β) wss(1.82±3.11)h,T_(max) was(2.06±0.37)h,and C_(max) was(2.61±0.17)μg·mL~(-1).Conclusion:The established method is simple,accurate,specific and suitable for pharmacokinetic studies of β-elemene in curcuma zedoaria in rats.
引文
[1]满伟,刘郁山,李文伟,等.莪术油药理研究及临床应用进展[J].时珍国医国药,2000,11(7):663MAN W,LIU YS,LI WW,et al.Pharmological study and clinical application progress of Curcuma zedoaria oil[J].Lishizhen Med Mater Med Res,2000,11(7):663
[2]CHEN CC,CHEN Y,HSI YT,et al.Cytotoxic chemical constituents and anticancer activity of Curcuma zedoaria Rosc.essential oil against non-small cell lung carcinoma cells in vitro and in vivo[J].J Agric Food Chem,2013,61(47):11418
[3]CHEN HW,LEE JY,HUANG JY,et al.Curcumin inhibits lung cancer cell invasion and metastasis through the tumor suppressor HLJ1[J].Cancer Res,2008,68(18):7428
[4]彭炳先,周欣,石京山,等.蓬莪术挥发油及其中3种成分抗肝癌和子宫内膜癌的研究[J].华西药学杂志,2007,22(3):312PENG BX,ZHOU X,SHI JS,et al.Effects of volatile oil and three main components from Curcuma phaeocaulis valetonon liver cancer and endometrial carcinoma cell lines[J].West China J Pharm Sci,2007,22(3):312
[5]王佳丽,王秀,夏泉,等.莪术油中3种倍半萜类化合物对肝癌Hep G2细胞增殖抑制作用的研究[J].中成药,2014,36(7):1535WANG JL,WANG X,XIA Q,et al.The proliferation inhibition effect on hepatoma HepG2 cell of three sesquiterpene components from Curcuma phaeocaulis oil[J].Chin Tradit Pat Med,2014,36(7):1535
[6]LEE JY,LEE YM,CHANG GC,et al.Curcumin induces EGFRdegradation in lung adenocarcinoma and modulates p38 activation in intestine:the versatile adjuvant for gefitinib therapy[J].PLo S One,2011,6(8):e23756
[7]JAVADI S,ROSTAMIZADEH K,HEJAZI J,et al.Curcumin mediated down-regulation ofαVβ3 integrin and up-regulation of pyruvate dehydrogenase kinase 4(PDK4)in Erlotinib resistant SW480 colon cancer cells[J].Phytother Res,2018,32(2):355
[8]DOU H,SHEN R,TAO J,et al.Curcumin suppresses the colon cancer proliferation by inhibiting Wnt/β-catenin pathways via miR-130a[J].Front Pharmacol,2017,8:877
[9]ZOU B,LI Q,ZHAO J,et al.β-Elemene and taxanes synergis tically induce cytotoxicity and inhibit proliferation in ovarian cancer and other tumor cells[J].Anticancer Res,2013,33(3):929
[10]MU L,WANG T,CHEN Y,et al.β-Elemene enhances the efficacy of gefitinib on glioblastoma multiforme cells through the inhibition of the EGFR signaling pathway[J].Int J Oncol,2016,49(4):1427
[11]WANG J,HUANG F,BAI Z,et al.Curcumol inhibits growth and induces apoptosis of colorectal cancer LoVo cell line via IGF-1R and p38 MAPK pathway[J].Mol Sci,2015,16(8):19851
[12]YU X,XU M,LI N,et al.β-elemene inhibits tumor-promoting effect of M2 macrophages in lung cancer[J].Biochem Biophys Res Commun,2017,490(2):514
[13]CHEN Z,SONG Y,CHE J,et al.Validation of a sensitive gas chromatographic-mass spectrometric method for the simultaneous determination ofβ-elemene andβ-elemenal in human plasma[J].J Chromatogr B Analyt Technol Biomed Life Sci,2009,877(4):408
[14]詹琼,周鑫莉,黄若凡,等.β-榄香烯血药浓度测定方法的建立及其药代动力学研究[J].中国药房,2017,28(2):173ZHAN Q,ZHOU XL,HUANG RF,et al.Establishment of determination method for plasma concentration ofβ-elemene and its pharmacokinetic study in human[J].China Pharm,2017,28(2):173
[15]GAN YX,LUO NN,JIANG YP,et al.Simultaneous determination of beta-elemene,curcumol,germacrone and neocurdione in volatile oil of Curcuma phaeocaulis and vinegar products by GC-MS[J].China J Chin Mater Med,2015,40(7):1311
[2]CHEN CC,CHEN Y,HSI YT,et al.Cytotoxic chemical constituents and anticancer activity of Curcuma zedoaria Rosc.essential oil against non-small cell lung carcinoma cells in vitro and in vivo[J].J Agric Food Chem,2013,61(47):11418
[3]CHEN HW,LEE JY,HUANG JY,et al.Curcumin inhibits lung cancer cell invasion and metastasis through the tumor suppressor HLJ1[J].Cancer Res,2008,68(18):7428
[4]彭炳先,周欣,石京山,等.蓬莪术挥发油及其中3种成分抗肝癌和子宫内膜癌的研究[J].华西药学杂志,2007,22(3):312PENG BX,ZHOU X,SHI JS,et al.Effects of volatile oil and three main components from Curcuma phaeocaulis valetonon liver cancer and endometrial carcinoma cell lines[J].West China J Pharm Sci,2007,22(3):312
[5]王佳丽,王秀,夏泉,等.莪术油中3种倍半萜类化合物对肝癌Hep G2细胞增殖抑制作用的研究[J].中成药,2014,36(7):1535WANG JL,WANG X,XIA Q,et al.The proliferation inhibition effect on hepatoma HepG2 cell of three sesquiterpene components from Curcuma phaeocaulis oil[J].Chin Tradit Pat Med,2014,36(7):1535
[6]LEE JY,LEE YM,CHANG GC,et al.Curcumin induces EGFRdegradation in lung adenocarcinoma and modulates p38 activation in intestine:the versatile adjuvant for gefitinib therapy[J].PLo S One,2011,6(8):e23756
[7]JAVADI S,ROSTAMIZADEH K,HEJAZI J,et al.Curcumin mediated down-regulation ofαVβ3 integrin and up-regulation of pyruvate dehydrogenase kinase 4(PDK4)in Erlotinib resistant SW480 colon cancer cells[J].Phytother Res,2018,32(2):355
[8]DOU H,SHEN R,TAO J,et al.Curcumin suppresses the colon cancer proliferation by inhibiting Wnt/β-catenin pathways via miR-130a[J].Front Pharmacol,2017,8:877
[9]ZOU B,LI Q,ZHAO J,et al.β-Elemene and taxanes synergis tically induce cytotoxicity and inhibit proliferation in ovarian cancer and other tumor cells[J].Anticancer Res,2013,33(3):929
[10]MU L,WANG T,CHEN Y,et al.β-Elemene enhances the efficacy of gefitinib on glioblastoma multiforme cells through the inhibition of the EGFR signaling pathway[J].Int J Oncol,2016,49(4):1427
[11]WANG J,HUANG F,BAI Z,et al.Curcumol inhibits growth and induces apoptosis of colorectal cancer LoVo cell line via IGF-1R and p38 MAPK pathway[J].Mol Sci,2015,16(8):19851
[12]YU X,XU M,LI N,et al.β-elemene inhibits tumor-promoting effect of M2 macrophages in lung cancer[J].Biochem Biophys Res Commun,2017,490(2):514
[13]CHEN Z,SONG Y,CHE J,et al.Validation of a sensitive gas chromatographic-mass spectrometric method for the simultaneous determination ofβ-elemene andβ-elemenal in human plasma[J].J Chromatogr B Analyt Technol Biomed Life Sci,2009,877(4):408
[14]詹琼,周鑫莉,黄若凡,等.β-榄香烯血药浓度测定方法的建立及其药代动力学研究[J].中国药房,2017,28(2):173ZHAN Q,ZHOU XL,HUANG RF,et al.Establishment of determination method for plasma concentration ofβ-elemene and its pharmacokinetic study in human[J].China Pharm,2017,28(2):173
[15]GAN YX,LUO NN,JIANG YP,et al.Simultaneous determination of beta-elemene,curcumol,germacrone and neocurdione in volatile oil of Curcuma phaeocaulis and vinegar products by GC-MS[J].China J Chin Mater Med,2015,40(7):1311