Application of a UPLC–MS/MS method to the protein binding study of TM-2 in rat, human and beagle dog plasma

Liu, Hui; Wu, Panpan; Yang, Mingjing; Men, Lei; Lin, Hongli; Zhao, Yunli; Tang, Xing

doi:10.1016/j.jpha.2015.08.001

Cited by 12 publications

(3 citation statements)

References 12 publications

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“…The samples were processed as described above and analyzed by UPLC‐MS/MS. The plasma protein bound of R ‐/ S ‐HFBA was calculated using the following formula

Bound (%) = (D_{t} - D_{f}) / D_{t} \times 100 %

where D t is the total drug concentration in the dialysis bag and D f is the free form drug concentration in PBS buffer.…”

Section: Methodsmentioning

confidence: 99%

Determination of protein binding for novel 2‐(2‐hydroxypropanamido)‐5‐trifluoromethyl benzoic acid enantiomers to rats, dogs, and humans plasma by UPLC‐MS/MS

et al. 2020

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R-/S-2-(2-hydroxypropanamido)-5-trifluoromethyl benzoic acid (R-/S-HFBA) is a novel COX inhibitor with remarkable anti-inflammatory and antiplatelet aggregation activities, but no gastrointestinal toxicity. In our previous study, the different pharmacokinetic profiles of the two enantiomers in rats were observed after administration of R-HFBA and S-HFBA. Stereoselective protein binding of the two enantiomers may be a reason for the different pharmacokinetic behaviors. In this study, we developed and validated an UPLC-MS/MS method for determining stereoselective binding of HFBA enantiomers to rat, dog, and human plasma in vitro. Chromatographic separation was achieved by gradient elution with a flow rate of 0.4 mL/min. MS/MS detection was operated in positive electrospray using multiple reaction monitoring (MRM) mode. The method was proved to be linear over the concentration range of 0.005 to 10 μg/mL with a lower limit of quantification of 0.005 μg/mL. The developed method was successfully employed to the plasma protein binding study of HFBA enantiomers. Equilibrium dialysis method was applied to assess drug-plasma protein interactions.The results showed that the enantiomers were both extensively bound to three species plasma and protein binding of R-/S-HFBA was concentration dependent.R-HFBA and S-HFBA showed significant species difference among rat, dog, and human plasma and stereoselective plasma protein binding. K E Y W O R D Splasma protein binding, R-/S-2-(2-hydroxypropanamido)-5-trifluoromethyl benzoic acid, species difference, stereoselectivity, UPLC-MS/MS

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“…The samples were processed as described above and analyzed by UPLC‐MS/MS. The plasma protein bound of R ‐/ S ‐HFBA was calculated using the following formula

Bound (%) = (D_{t} - D_{f}) / D_{t} \times 100 %

where D t is the total drug concentration in the dialysis bag and D f is the free form drug concentration in PBS buffer.…”

Section: Methodsmentioning

confidence: 99%

Determination of protein binding for novel 2‐(2‐hydroxypropanamido)‐5‐trifluoromethyl benzoic acid enantiomers to rats, dogs, and humans plasma by UPLC‐MS/MS

et al. 2020

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Section: Introductionmentioning

confidence: 99%

“…Before any preclinical pharmacological study can be conducted, it is necessary to develop a bioanalytical method and characterize the pharmacokinetic properties for a drug candidate. 14,15 To date, there has been no report on quantification methodology to be used for MFD or on any investigation of its pharmacokinetics. In the present study, an efficient and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the assay of MFD in monkey plasma.…”

Section: Introductionmentioning

confidence: 99%

LC−MS/MS assay for the determination of a novel anti‐fibrotic candidate mefunidone in monkey plasma and its application to a pharmacokinetics study

Han

Wen

Fan³

et al. 2019

Drug Testing and Analysis

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Mefunidone (MFD) is a promising anti‐fibrotic candidate molecule with greater anti‐fibrotic activity than pirfenidone (PFD). However, there has been no report on the methodology used for the quantification of MFD or on any investigation of its pharmacokinetics. In this study, an efficient and reliable liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed to assay MFD in monkey plasma. This assay method was validated and applied to a pharmacokinetics study in monkeys. The lower limit of quantification of this assay was 0.1 μg·mL−1, and the linear calibration curve was acquired with R2 > 0.99 between 0.1 and 60 μg·mL−1. The intra‐day and inter‐day precision were evaluated with coefficient of variations of 1.5%–5.8%, whereas the mean accuracy ranged from 91.7% to 106.9%. A negligible matrix effect and good recovery were obtained using this assay, with average extraction recoveries of MFD and the internal standard (IS) in the range of 85.5%–124.8% and 84.1%–94.0%, respectively. The precision of the absolute matrix effect of MFD and the IS was 1.2–3.0% and 1.2–7.3%, respectively. The samples were stable under all experimental conditions. Linear pharmacokinetics were observed for MFD in monkeys, where the exposures of MFD increased proportionally with increasing MFD doses at the range of 10–90 mg·kg−1. Moderate elimination of MFD from the body was observed, with t1/2 of 5–7 h, and the elimination rate of MFD was stable during multiple dosing. In conclusion, this method provides an reliable analytical approach for quantification of MFD in plasma and was successfully applied to a pharmacokinetics study in monkeys.

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Enantioseparation of N‐acetyl‐glutamine enantiomers by LC–MS/MS and its application to a plasma protein binding study

Gao

Xue

Zhang

et al. 2019

Biomedical Chromatography

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A novel chiral method was developed and validated to determine N‐acetyl‐glutamine (NAG) enantiomers by liquid chromatography–tandem mass spectrometry (LC–MS/MS). Enantioseparation was achieved on a Chiralpak QD‐AX column (150 × 4.6 mm i.d., 5 μm) using methanol–water (50 mm ammonium formate, pH 4.3; 70:30, v/v) at a flow rate of 500 μL/min. The detection was operated with an electrospray ionization source interface in positive mode. The ion transition for NAG enantiomers was m/z 189.0 → 130.0. The retention time of N‐acetyl‐l‐glutamine and N‐acetyl‐d‐glutamine were 15.2 and 17.0 min, respectively. Calibration curves were linear over the range of 0.02–20 μg/mL with r > 0.99. The deviation of accuracy and the coefficient of variation of within‐run and between‐run precision were within 10% for both enantiomers, except for the lower limit of quantification (20 ng/mL), where they deviated <15%. The recovery was >88% and no obvious matrix effect was observed. This method was successfully applied to investigate the plasma protein binding of NAG enantiomers in rats. The results showed that the plasma protein binding of NAG enantiomers was stereoselective. The assay method also exhibited good application prospects for the clinical monitoring of free drugs in plasma.

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Application of a UPLC–MS/MS method to the protein binding study of TM-2 in rat, human and beagle dog plasma

Cited by 12 publications

References 12 publications

Determination of protein binding for novel 2‐(2‐hydroxypropanamido)‐5‐trifluoromethyl benzoic acid enantiomers to rats, dogs, and humans plasma by UPLC‐MS/MS

Determination of protein binding for novel 2‐(2‐hydroxypropanamido)‐5‐trifluoromethyl benzoic acid enantiomers to rats, dogs, and humans plasma by UPLC‐MS/MS

LC−MS/MS assay for the determination of a novel anti‐fibrotic candidate mefunidone in monkey plasma and its application to a pharmacokinetics study

Enantioseparation of N‐acetyl‐glutamine enantiomers by LC–MS/MS and its application to a plasma protein binding study

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