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,5 μm)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10 μL。结果 在低、中、高浓度(5、10、20 μg·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。
Abstract
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 10 μL. RESULTS At low, medium and high concentrations (5, 10, 20 μg·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
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Key words
Aurantii Fructus Immaturus /
synephrine /
N-methyltyramine /
plasma protein binding rate /
HPLC /
ultrafitration /
free drug concentration
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中图分类号：
R969.1
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脚注
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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)
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