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Simultaneous determination of human plasma protein binding of bioactive favonoids in Polygonum orientale by equilibrium dialysis combined with UPLC–MS/MS

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Journal of Pharmaceutical Analysis 2013年第5期3卷 376-381页

作者： Yong Huang Hui Chen Feng He Zhi-Rong Zhang Lin Zheng Yue Liu Yan-Yu Lan Shang-Gao Liao Yong-Jun Li Yong-Lin Wang Provincial Key Laboratory of Pharmaceutics in Guizhou Province School of Pharmacy Guiyang Medical University

A simple and selective ultra performance liquid chromatography--electrospray ionization tandem mass spectrometry （UPLC-ESI-MS/MS） assay was developed for the determination of the human plasma protein binding of four bioactive ftavonoids （such as orientin and vitexin） in Polygonum orientale. Protein precipitation was used for sample preparation. equilibrium dialysis technique was applied to determine the plasma protein binding under physiological conditions. The separation was achieved through a Waters C i s column with a mobile phase composed of 0.1% formic acid in acetonitrile and 0.1% aqueous formic acid using step gradient elution at a flow rate of 0.35 mL/min. A Waters ACQUITYTM TQD system was operated under the multiple reaction monitoring （MRM） mode of positive electrospray ionization. All of the recovery, precision, accuracy and stability of the method met the requirements. Good correlations （r 〉 0.99） of the four compounds were found, which suggested that these compounds can be simultaneously determined with acceptable accuracy. Results showed that the plasma protein bindings of the four bioactive flavonoids were in the range of 74-89% over the six concentrations studied. The binding parameters containing protein binding affinity, protein binding dissociation constant, and protein binding site were studied. The maximum ability to bind with protein was also determined in the assay in order to understand the drug-protein binding of each compound better.

关键词： Polygonum orientale Plasma protein binding Bioactive compounds equilibrium dialysis UPLC–ESI-MS/MS

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Simultaneous determination of LY3214996,abemaciclib,and M2 and M20 metabolites in human plasma,cerebrospinal fluid,and brain tumor by LC-MS/MS

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Journal of Pharmaceutical Analysis 2022年第4期12卷 601-609页

作者： Tigran Margaryan Mackenna Elliott Nader Sanai Artak Tovmasyan Ivy Brain Tumor Center Barrow Neurological InstitutePhoenixAZ85013USA

A sensitive and rapid liquid chromatography tandem mass spectrometry(LC-MS/MS)method was established for the quantification of total and unbound concentrations of LY3214996,an extracellular signal-regulated kinase inhibitor;abemaciclib,a cyclin-dependent kinase 4/6 inhibitor;and abemaciclib active metabolites,M2 and M20,in human plasma,brain tumor,and cerebrospinal fluid samples.The method was validated over a concentration range of 0.2e500 nM within a total run time of 3.8 min using isocratic elution on a Kinetex^(TM) F_(5) column.Detection was performed on a Sciex QTRAP 6500t mass spectrometer employing multiple reaction monitoring mode under positive electrospray ionization.The intra-and inter-batch accuracy as well as the precision of the method for all matrices was within ±20% and ≤20% at the lower limit of quantification,and within ±15% and ≤15% for other quality control levels for all analytes.The unbound fractions of drugs and metabolites in spiked and patient samples were determined using an optimized equilibrium dialysis.The validated method was successfully applied in a phase 0/2 clinical trial to assess the central nervous system penetration of LY3214996 and abemaciclib.

关键词： LC-MS/MS LY3214996 Abemaciclib Brain tumor penetration equilibrium dialysis

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Binding equilibrium study between Mn( Ⅱ ) and HSA or BSA

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Chinese Journal of Chemistry 2000年第1期18卷 35-41页

作者：梁宏涂楚桥张宏志沈星灿周永洽申泮文 Research Institute of Bioinorganic Chemistry Department of Chemistry and Chemical Engineering Guangxi Normal University Guilin Guangxi 541004 China Department of Chemistry Nankai University Tianjin 300071 China

The binding of Mn( Ⅱ ) to human serum albumin (HSA) or bovine serum albumin (BSA) has been studied by equilibrium dialysis at physiological pH (7. 43). The Scatchard analysis indicates that there are 1.8 and 1.9 strong binding sites of Mn( Ⅱ ) in HSA and BSA, respectively. The successive stability constants which are reported for the first time are obtained by non-linear least-squares methods fitting Bjerrum formula. For both Mn( Ⅱ )-HSA and Mn( Ⅱ )-BSA systems, the order of magnitude of K1 was found to be 104. The analyses of Hill plots and free energy coupling show that the positive cooperative effect was found in both Mn( Ⅱ )-HSA and Mn( Ⅱ )-BSA systems . The results of Mn ( Ⅱ ) competing with Cu ( Ⅱ ) 、 Zn(Ⅱ)、Cd( Ⅱ) or Ca( Ⅱ ) to bind to HSA or BSA further support the conjecture that there are two strong binding sites of Mn( Ⅱ) in both HSA and BSA. One is most probably located at the tripeptide segment of N- terminal sequence of HSA and BSA molecules involving four groups composed of n

关键词： Mn (Ⅱ)-serum albumin equilibrium dialysis successive stability constants positive cooperative effect coordination analysis

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Binding equilibrium Studies Between Co^(2+) and HSA or BSA

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Chemical Research in Chinese Universities 2000年第3期16卷 276-279页

作者： BIAN He dong LIANG Hong TU Chu qiao ZHANG Hong zhi SHEN Pan wen Research Institute of Bioinorganic Chemistry Department of Chemistry and Chemical EngineeringGuangxi Normal UniversityGuilin 541004P.R.China Department of Chemistry Nankai UniversityTianjinP.R.China

The interaction of HSA and BSA with Co2+ and a carboxyl atom of Asp is investigated. The number of binding sites and the cooperation among the binding sites are analyzed. According to the equilibrium dialysis results and the study of competition between Co2+ and Cu2+, Ca2+ or Zn2+ to be bound to HSA or BSA, it is suggested that there are three strong binding sites of in both HSA and BSA.

关键词： Co 2+ serum albumin equilibrium dialysis Coordination analysis Positive cooperative effect

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