气质联用法测定莪术油多效应成分及其在大鼠体内的整合药动学研究

Multiple Effective Components Determination in Essential Oil from Curcuma phaeocaulis by GC-MS and Its Integrated Pharmacokinetic Study in Rats

目的:建立测定大鼠血浆中莪术油多效应成分质量浓度的方法,并进行整合药动学研究。方法:16只大鼠单次灌胃莪术油提取液1.0 g/kg(按生药量计),分别于给药后0、0.17、0.5、1、2、2.5、3、4、6、8、10、12、24 h自眼眶取血300～400μL,采用气质联用法测定大鼠血浆中α-蒎烯、1,8-桉叶油素、龙脑、β-榄香烯、莪术醇、吉马酮、莪术二酮的质量浓度。色谱柱为DB-5毛细管柱,载气为氦气,进样口温度为270℃,柱温采用程序升温,流速为1.2 mL/min,分流比为20∶1,进样量为1μL;离子源为电喷雾离子源,以选择反应监测模式进行正离子扫描,扫描范围为m/z 20～500。采用DAS 2.0软件计算上述各效应成分的药动学参数,并以其AUC0-∞在AUC0-∞总和中所占的比例自定义权重系数,估算莪术油多效应成分在大鼠体内的整合药动学参数。结果:α-蒎烯、1,8-桉叶油素、龙脑、β-榄香烯、莪术醇、吉马酮、莪术二酮血药浓度的线性范围分别为2.71～173.54、7.76～496.88、3.37～215.72、21.68～1 387.50、40.21～2 573.44、24.84～3 179.69、47.78～3 057.81 ng/mL(r>0.99),定量下限分别为2.71、7.76、3.37、21.68、40.21、24.84、47.78 ng/mL,精密度、准确度、基质效应等均符合生物样品定量分析的相关要求。α-蒎烯、1,8-桉叶油素、龙脑、β-榄香烯、莪术醇、吉马酮、莪术二酮的cmax分别为(34.72±9.97)、(99.86±5.54)、(16.10±3.37)、(248.98±86.19)、(673.75±104.15)、(2 353.64±637.83)、(2 420.04±708.51)ng/mL;tmax分别为(2.33±0.29)、(0.67±0.29)、(1.33±0.58)、(1.83±0.76)、(0.83±0.29)、(0.89±0.18)、(1.17±0.76)h;t1/2分别为(8.64±1.46)、(8.98±1.63)、(12.43±2.88)、(19.86±4.05)、(15.63±5.50)、(14.17±4.13)、(7.14±0.67)h;AUC0-t分别为(189.78±89.10)、(454.74±82.43)、(100.55±8.27)、(1 067.37±216.55)、(3 154.16±405.94)、(16 501.24±663.88)、(12 524.92±3 222.10)ng·h/mL;AUC0-∞分别为(229.57±93.50)、(524.32±81.67)、(146.28±10.74)、(2 092.70±416.18)、(5 388.65±661.86)、(28 198.87±4 102.62)、(14 139.35±3 109.19)ng·h/mL。整合药动学参数cmax为1 880.94 ng/mL,tmax为0.50 h,t1/2为11.22 h,AUC0-t为13 050.89 ng·h/mL,AUC0-∞为19 015.21 ng·h/mL。结论:该方法可用于大鼠血浆中莪术油多效应成分血药浓度的检测;整合后的莪术油药动学参数与单一效应成分的差异较大,可为表征其整体药动学特征提供参考。

OBJECTIVE:To develop the determination method for plasma concentration of effective components in essential oil from Curcuma phaeocaulis,and to study its integrated pharmacokinetics.METHODS:Sixteen rats were given the extract of essential oil from C.phaeocaulis 1.0 g/kg(by crude drug)intragastrically;blood samples 300-400μL from orbit were collected 0,0.17,0.5,1,2,2.5,3,4,6,8,10,12,24 h after medication.The plasma concentration ofα-pinene,1,8-cineole,borneol,β-elemene,curcumol,germacrone and curdione in rats were determined by GC-MS.The determination was performed on DB-5 capillary column,using helium as carrier gas,at the flow rate of 1.2 mL/min.The injector temperature was 270℃,by temperature programming,and split ratio was 20∶1.The sample size was 1μL.The ion source was electrospray ion source.The selective reaction monitoring mode was used for the positive ion scanning in the range of m/z 20-500.Pharmacokinetic parameters of above effective components were calculated by using DAS 2.0 software.The weight coefficients were customized according to the proportion of AUC0-∞in the sum of AUC0-∞.The integrated pharmacokinetic parameters of multiple effective components in essential oil from C.phaeocaulis were calculated.RESULTS:The linear range ofα-pinene,1,8-cineole,borneol,β-elemene,curcumol,germacrone,and curdione were 2.71-173.54,7.76-496.88,3.37-215.72,21.68-1 387.50,40.21-2 573.44,24.84-3 179.69,47.78-3 057.81 ng/mL,respectively(r>0.99).The lower limits of quantitation were 2.71,7.76,3.37,21.68,40.21,24.84,47.78 ng/mL,respectively.The precision,accuracy and matrix effects were in line with related requirements of quantitative analysis of biological samples.The pharmacokinetic parameters ofα-pinene,1,8-cineole,borneol,β-elemene,curcumol,germacrone,and curdione were as follows that cmax were(34.72±9.97),(99.86±5.54),(16.10±3.37),(248.98±86.19),(673.75±104.15),(2 353.64±637.83),(2 420.04±708.51)ng/mL;tmax were(2.33±0.29),(0.67±0.29),(1.33±0.58),(1.83±0.76),(0.83±0.29),(0.89±0.18),(1.17±0.76)h;t1/2 were(8.64±1.46),(8.98±1.63),(12.43±2.88),(19.86±4.05),(15.63±5.50),(14.17±4.13),(7.14±0.67)h;AUC0-t were(189.78±89.10),(454.74±82.43),(100.55±8.27),(1 067.37±216.55),(3 154.16±405.94),(16 501.24±663.88),(12 524.92±3 222.10)ng·h/mL;AUC0-∞were(229.57±93.50),(524.32±81.67),(146.28±10.74),(2 092.70±416.18),(5 388.65±661.86),(28 198.87±4 102.62),(14 139.35±3 109.19)ng·h/mL,respectively.After integration,the pharmacokinetic parameters were as follows that cmax was 1 880.94 ng/mL;tmax was 0.50 h;t1/2 was 11.22 h;AUC0-t was 13 050.89 ng·h/mL;AUC0-∞was 19 015.21 ng·h/mL.CONCLUSIONS:The method can be used for the detection of plasma concentration of effective components in rats;pharmacokinetic parameters of essential oil from C.phaeocaulis after integration are greatly different from single effective component,which can provide reference for characterization of its overall pharmacokinetics.