超滤法结合UPLC-MS/MS法研究树豆酮酸A在不同种属血浆中的血浆蛋白结合率
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篇名: 超滤法结合UPLC-MS/MS法研究树豆酮酸A在不同种属血浆中的血浆蛋白结合率
TITLE:
摘要: 目的:比较树豆酮酸A在不同种属血浆中的血浆蛋白结合率。方法:以超高效液相色谱-串联质谱法(UPLC-MS/MS)为检测手段,采用超滤法测定低、中、高质量浓度(2.5、5、20 μg/mL)的树豆酮酸A分别在大鼠、家兔及人血浆中的血浆蛋白结合率。色谱条件:色谱柱为Waters BEH C18,保护柱为WatersVanGuard BEH C18;流动相为0.01%甲酸溶液(溶剂A)和含0.01%甲酸的乙腈溶液(溶剂B),梯度洗脱;流速为0.15 mL/min;柱温为30 ℃;进样量为2 μL。质谱条件:电喷雾电离源;负离子模式采集;毛细管电压为1.5 kV;锥孔电压为30 V;离子源温度为100 ℃;脱溶剂气温度为400 ℃;锥孔气流量为50 L/h;脱溶剂气流量为800 L/h;扫描范围为质荷比(m/z) 50→1 200。结果:在2.5、5、20 μg/mL质量浓度下,树豆酮酸A在大鼠血浆中的血浆蛋白结合率分别为(75.63±0.90)%、(98.30±0.03)%、(99.42±0.01)%(n=3);在家兔血浆中的血浆蛋白结合率分别为(79.61±1.08)%、(98.48±0.10)%、(99.42±0.03)%(n=3);在健康人血浆中的血浆蛋白结合率分别为(76.74±1.22)%、(97.99±0.11)%、(99.37±0.01)%(n=3),其中在2.5 μg/mL时树豆酮酸A在大鼠和人血浆中的血浆蛋白结合率明显低于在家兔血浆中的血浆蛋白结合率(P<0.05)。结论:5、20 μg/mL的树豆酮酸A在大鼠、家兔及人血浆中的血浆蛋白结合率高于2.5 μg/mL树豆酮酸A;仅在2.5 μg/mL时,树豆酮酸A在不同种属血浆中的血浆蛋白结合率有明显差异;在5、20 μg/mL时,树豆酮酸A在不同种属血浆中的血浆蛋白结合率均无明显差异。
ABSTRACT: OBJECTIVE: To compare plasma protein binding rate of cajanonic acid A with different species of plasma. METHODS:Using UPLC-MS/MS as the detection means. Plasma protein binding rate of low, medium and high concentrations of cajanonic acid A (2.5, 5, 20 μg/mL) with rats, rabbits and human plasma were determined by ultrafiltration method. The chromatographic conditions included that Waters BEH C18 as chromatographic column, WatersVanGuard BEH C18 as guard column, mobile phase consisted of ultrapure water solution containing 0.01% formic acid (solvent A) and acetonitrile solution of 0.01% formic acid (solvent B) gradient elution, at the flow rate of 0.15 mL/min, column temperature of 30 ℃, sample size of 2 μL. Mass spectrum condition included that ESI, negative ion mode acquisition, capillary voltage of 1.5 kV, cone voltage of 30 V, ion source temperature of 100 ℃, desolvent gas temperature of 400 ℃, cone gas flow of 50 L/h, desolvent gas flow of 800 L/h, scanning range of m/z 50→1 200. RESULTS: At the concentration of 2.5, 5 and 20 μg/mL, the plasma protein binding rates of cajanonic acid A were (75.63±0.90)%, (98.30±0.03)% and (99.42±0.01)% in the rats plasma; (79.61±1.08)%, (98.48±0.10)% and (99.42±0.03)% in rabbits plasma (n=3); (76.74±1.22)%, (97.99±0.11)% and (99.37±0.01)% in human plasma (n=3). At the concentration of 2.5 μg/mL, plasma protein binding rates of cajanonic acid A in plasma of rats and human were significantly lower than that in plasma of rabbits (P<0.05). CONCLUSIONS: The plasma protein binding rate of 5,20 μg/mL cajanonic acid A with rats, rabbits and human plasma are higher than that of 2.5 μg/mL cajanonic acid A. There is significant difference in plasma protein binding rate of 2.5 μg/mL cajanonic acid A with different species of plasma,and there is no significant difference in plasma protein binding rate of 5, 20 μg/mL cajanonic acid A with different species of plasma.
期刊: 2019年第30卷第13期
作者: 班玉娟,张丽,陈瑞,朱高峰,王建塔,陈文章,汤磊,黄静
AUTHORS: BAN Yujuan,ZHANG Li,CHEN Rui2,ZHU Gaofeng,WANG Jianta,CHEN Wenzhang,TANG Lei,HUANG Jing
关键字: 树豆酮酸A;血浆蛋白结合率;超滤法;超高效液相色谱-串联质谱法;大鼠;家兔;人
KEYWORDS: Cajanonic acid A; Plasma protein binding rate; Ultrafitration method; UPLC-MS/MS method; Rat; Rabbit; Human
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