超滤法结合高效液相色谱法测定枳实中两个生物碱在不同种属生物中的血浆蛋白结合率  

Ultrafiltration Combined with HPLC to Determine Plasma Protein Binding Rates of Two Alkaloids in Different Species of Aurantii Fructus Immaturus

作　　者：陈思琦 刘德鸿 黄东 陈伟康 罗跃华 CHEN Si-qi;LIU De-hong;HUANG Dong;CHEN Wei-kang;LUO Yue-hua(Jiangxi Institute for Drug Control,NMPA Key Laboratory for Quality Evaluation of Chinese Patent Medicine,Jiangxi Province Engineering Research Center of Drug and Medical Device Quality,Nanchang 330029,China;School of Pharmacy,Nanchang University,Nanchang 330006,China)

机构地区：[1]江西省药品检验检测研究院,国家药品监督管理局中成药质量评价重点实验室,江西省药品医疗器械质量工程研究中心,南昌330029 [2]南昌大学药学院,南昌330006

出　　处：《中国药学杂志》2023年第15期1410-1415,共6页Chinese Pharmaceutical Journal

基　　金：江西省重点研发计划项目资助(20212BBG73025);江西省药品监督管理局科研项目资助(2021KY44)。

摘　　要：目的建立枳实中辛弗林、N-甲基酪胺血浆蛋白结合率的方法。方法采用超滤法结合高效液相色谱法测定枳实中辛弗林、N-甲基酪胺在SD大鼠和新西兰兔血浆中的蛋白结合率。以Agilent ZORBAX SB-C18(4.6 mm×250 mm,5μm)为色谱柱,甲醇-0.14%庚烷磺酸钠水溶液(60∶40,加磷酸调pH至3.0)为流动相进行等度洗脱,流速为1 mL·min^(-1),柱温为40℃,检测波长为275 nm,进样量为10μL。结果在低、中、高浓度(5、10、20μg·mL^(-1))下,辛弗林的鼠血浆蛋白结合率分别为(14.85±3.18)%、(11.47±1.46)%、(3.70±0.93)%(n=3),兔血浆蛋白结合率分别为(4.28±1.52)%、(5.34±1.45)%、(8.54±2.97)%(n=3);N-甲基酪胺的鼠血浆蛋白结合率分别为(21.27±2.62)%、(9.49±4.58)%、(4.36±0.91)%(n=3),兔血浆蛋白结合率分别为(7.40±8.50)%、(9.85±1.83)%、(15.18±6.11)%(n=3)。结论所建方法操作简单、快速准确,可用于测定枳实中辛弗林、N-甲基酪胺的血浆蛋白结合率。OBJECTIVE To establish a method for determination of plasma protein binding rates of synephrine and N-methyltyramine from Aurantii Fructus Immaturus.METHODS Ultrafiltration method combined with HPLC method was adopted to determine the plasma protein binding rates of synephrine and N-methyltyramine from Aurantii Fructus Immaturus in the plasma of SD rats and New Zealand rabbits.The determination was performed on an Agilent ZORBAX SB-C18 column with mobile phase consisting of methanol-0.14%sodium heptanesulfonate aqueous solution(60∶40,phosphoric acid added to adjust pH to 3.0)at the flow rate of 1 mL·min^(-1).The column temperature was maintained 40℃,and the detection wavelength was set at 275 nm.The sample size was 10μL.RESULTS At low,medium and high concentrations(5,10,20μg·mL^(-1)),the plasma binding rates of synephrine in rats were(14.85±3.18)%,(11.47±1.46)%,and(3.70±0.93)%,respectively(n=3).The plasma binding rates in rabbits were(4.28±1.52)%,(5.34±1.45)%,and(8.54±2.97)%,respectively(n=3).The plasma binding rates of N-methyltyramine in rats were(21.27±2.62)%,(9.49±4.58)%,and(4.36±0.91)%,respectively(n=3).The plasma binding rates in rabbits were(7.40±8.50)%,(9.85±1.83)%,and(15.18±6.11)%,respectively(n=3).CONCLUSION The established method is simple,rapid,and accurate and can be used to determine the plasma protein binding rates of synephrine and N-methyltyramine from Aurantii Fructus Immaturus.

关 键 词：枳实 辛弗林 N-甲基酪胺 血浆蛋白结合率 高效液相色谱法 超滤法 游离药物浓度 

分 类 号：R969.1[医药卫生—药理学]