Investigation of Rat Plasma Protein Binding Characteristics of Anemoside B4 by High-throughput Equilibrium Dialysis Combined with UPLC-MS/MS Technique

JIA Jia; OUYANG Hui; HE Ming-yu; HE Ming-zhen; QIU Jia-ming; FENG Yu-lin; YANG Shi-lin

doi:10.13422/j.cnki.syfjx.2017180071

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Investigation of Rat Plasma Protein Binding Characteristics of Anemoside B4 by High-throughput Equilibrium Dialysis Combined with UPLC-MS/MS Technique

更新时间：2024-01-04

- Investigation of Rat Plasma Protein Binding Characteristics of Anemoside B4 by High-throughput Equilibrium Dialysis Combined with UPLC-MS/MS Technique
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 23, Issue 18, Pages: 71-74(2017)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.2017180071
  CLC： R285.5
- Published：2017
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JIA Jia, OUYANG Hui, HE Ming-yu, et al. Investigation of Rat Plasma Protein Binding Characteristics of Anemoside B4 by High-throughput Equilibrium Dialysis Combined with UPLC-MS/MS Technique[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(18): 71-74.
DOI：

JIA Jia, OUYANG Hui, HE Ming-yu, et al. Investigation of Rat Plasma Protein Binding Characteristics of Anemoside B4 by High-throughput Equilibrium Dialysis Combined with UPLC-MS/MS Technique[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(18): 71-74. DOI： 10.13422/j.cnki.syfjx.2017180071.

摘要

目的：建立血浆中白头翁皂苷B4的分析方法，测定白头翁皂苷B4大鼠血浆蛋白结合率。方法：采用96通道高通量平衡透析系统（HTD 96b）进行透析，利用UPLC-MS/MS测定透析内外液中白头翁皂苷B4的浓度，研究白头翁皂苷B4在大鼠血浆中的血浆蛋白结合率，使用Waters XTerra MS C18色谱柱（2.1 mm×50 mm，5 μm），流动相0.1%甲酸水溶液-乙腈梯度洗脱，流速0.7 mL · min-1，柱温40℃，进样量5 μL。结果：白头翁皂苷B4在5~2 000 ng · L-1线性关系良好，其精密度RSD及准确度均<8.0%，重复性RSD<9.0%，稳定性的RSD均<15%；提取回收率和基质效应均在80%~115%。白头翁皂苷B4在低、中、高（6，12，24 mg · L-1）3个质量浓度下大鼠血浆蛋白结合率分别为（95.32±0.37）%，（94.32±0.63）%，（88.64±0.37）%。结论：白头翁皂苷B4与大鼠血浆具有较强的蛋白结合率，且结合率不具有质量浓度依赖性。

Abstract

Objective: To establish an analytical method for investigating the protein binding rate of anemoside B4 in rats plasma.Method: High-throughput equilibrium dialysis was employed to dialysis

an UPLC-MS/MS for determination of anemoside B4 both in the inner and outer dialysate was established and then the protein-binding rate with rats plasma was calculated

Waters XTerra MS C18 column (2.1 mm×50 mm

5 μm) was employed with mobile phase of acetonitrile-0.1% formic acid aqueous solution for gradient elution.Result: The linearity of anemoside B4 was in the range of 5-2 000 ng · L-1.The RSDs of precision

accuracy

repeatability and stability of anemoside B4 were all less than 15%

and its extraction recovery and matrix effect were in 80%-115%.The protein binding rates of anemoside B4 in high

middle and low concentrations(6

24 mg · L-1) were (95.32±0.37)%

(94.32±0.63)% and (88.64±0.37)%

respectively.Conclusion: Anemoside B4 has a higher binding intension with plasma protein

which has on concentration-dependent manner.

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