海南崖豆藤提取物HN-1不同种属血浆蛋白结合特性研究

Study on plasma protein binding characteristics of HN-1 isolated from Millettia pachyloba Drake of different species

目的:采用超滤法,研究海南崖豆藤抗炎活性成分6-methoxy-8,8 dimethy-3-(2,4,5-rimethoxypenyl)4H,8H-pyrano[2,3-f]chromen-4-one在大鼠,Beagle犬,食蟹猴和人四个种属中的血浆蛋白结合率。方法:采用超滤法结合液相色谱质谱联用(UFLC-MS/MS)检测方法,对HN-1在大鼠,Beagle犬。食蟹猴和人血浆的蛋白结合率进行测定。结果:在0.1,1,5μg·mL^-1质量浓度FHN-1与大鼠血浆的蛋白结合率分別为(98.57±0.55)%、(96.38±0.55)%、(91.06±0.13)%;与Beagle犬血浆的蛋白结合率分別为(99.44±0.28)%、(96.4±0.88)%、(90.42±0.47)%;与食蟹猴血浆的蛋白结合率分别为(99.73±0.38)%、(95.30±0.34)%、(91.53±1.37)%;与健康人血装的蛋白结合率分别为(99.13±0.07)%、(96.30±0.11)%、(92.06±2.06)%。结论:本研究建立的方法简单准确,特异性强。可用于HN-1的血浆蛋白结合率测定:HN-1与各种属血浆蛋白均为高强度结合(>90%),且在已考察范围内,其血装蛋白结合率无明显种属差异。

Objective:Ultrafiltration was used to study protein binding rate of the anti-inflammatory active ingredient of Millettia pachyloba Drake,6-methoxy-8,8-dimethyl-3-(2,4,5-trimethoxyphenyl)-4H,8H-pyrano[2,3-f]chromen-4-one (HN-1),in rat,Beagle dog,cynomolgus monkey and human.Method:The protein binding rate of HN-1 in rat,Beagle dog,cynomolgus monkey and human plasma was detected by ultrafiltration combined with liquid chromatography-mass spectrometry (UFLC-MS/MS).Result:The protein binding rates of HN-1 in rat plasma were (98.57±0.55)%,(96.38±0.55)%,and (91.06±0.13)%,respectively,at a concentration of 0.1,1,5 μg﹒mL-1.The protein binding rates of Beagle dog plasma were (99.44±0.28)%,(96.94±0.88)%,and (90.42±0.47)%,respectively.The protein binding rates of cynomolgus monkey plasma were (99.73±0.38)%,(95.30±0.34)%,(91.53±1.37)%,respectively.The protein binding rate of healthy human plasma were (99.13±0.07)%,(96.30±0.11)%,(92.06±2.06)%.Conclusion:The method established in this study is simple,accurate and specific,and can be used for the determination of plasma protein binding rate of HN-1.HN-1 is highly bound to various plasma proteins (>90%),and there is no significant species difference in plasma protein binding rate within the range examined.