LC-MS/MS assay for the simultaneous quantitation of thromboxane B2 and prostaglandin E2 to evaluate cyclooxygenase inhibition in human whole blood

Journal of Applied Bioanalysis

卷:6

Heather E. Murrey¹  Yifan Shi¹  Kay Ahn¹  Naidong Weng¹  Shefali Patel¹ 

[1] Janssen R&D, Springhouse, PA, USA;

关键词: cox inhibitor;    lc-ms/ms;    lipids;    txb2;    pge2;    whole blood assay;   

DOI  :  10.17145/jab.20.014

来源: DOAJ

【 摘 要 】

OBJECTIVES: A high-throughput LC-MS method for TXB2 and PGE2 was developed for human whole blood assay for COX inhibition.METHODS: A surrogate analyte approach was used for the quantitation of TXB2 and PGE2 by LC-MS. Fifty microliters plasma was processed using solid-phase extraction. TXB2-d4 and PGE2-d4 were used as surrogate analytes. The calibration curves were established for TXB2 from 0.1 to 500 ng/mL and for PGE2 from 0.05 to 500 ng/mL. TXB1 was used as internal standard.RESULTS: The response factor and parallelism between surrogate and authentic analyte were verified. Heparinized whole blood assay for COX inhibition was optimized for sample pretreatment, stimulant concentration and incubation time.CONCLUSION: The LC-MS assay was successfully used to analyze inhibitory activity of four commercially available COX inhibitors. The presented method offers a sensitive, high throughput and low-cost alternative to ELISA for human whole blood assay for COX inhibition.

【 授权许可】

Unknown