Quantification of peptides in human synovial fluid using liquid chromatography–tandem mass spectrometry
Article [Accepted Manuscript]
Under embargo until: 2020-04-06
Is part ofTalanta ; vol. 186, p.124-132
A method to explore the stability of two anti-inflammatory peptides in human synovial fluid (HSF) has been developed and validated using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The two peptides are BQ123 Cyclo(-D-Trp-D-Asp-L-Pro-D-Val-L-Leu, Mw = 610.7) and R-954 (AcOrn[Oic2, (αMe)Phe5, DβNal7, Ile8]desArg9-bradykinin, Mw = 1194.4). Human synovial fluid samples were analyzed after a protein precipitation step with acetonitrile and dilution with mobile phase. DMSO was used as anti-adsorptive agent. We used an octyl silane column with formic acid (0.1%, v/v) in water as the aqueous mobile phase and acetonitrile isopropanol-formic acid (20:80, 0.1 v/v) as the organic mobile phase and 0.7 mL/min flow rate. The peptides CY-771 and pepstatin A were used as internal standards. Selective detection was performed by tandem mass spectrometry with a heated electrospray source (HESI), operated in positive ionization mode and in selected reaction monitoring acquisition (SRM). The method limit of quantification (injection volume = 10 µL) was 0.17 ng and 1.2 ng, corresponding to 28 and 102 nmol L−1 for BQ123 and R-954 respectively in human synovial fluid. Calibration curves obtained using matrix-matched calibration standards and internal standard were linear from 20 to 1000 nmol L−1. Precision values (%R.S.D.) were ≤ 14% in the entire linear range. Accuracy measured at a low and a high concentration level ranged from 93.1% to 102%. The recoveries (at 800 nmol L−1) were 96.4% for BQ123 and 102.0% for R-954. The method was successfully applied to follow the degradation kinetics of both peptides in human synovial fluid from arthritic patients during 72 h.
Quantification of peptides in human synovial fluid using liquid chromatography–tandem mass spectrometry Araceli Garcia-Ac, Sung Vo Duy, Sébastien Sauvé, Florina Moldovan •V Gaëlle Roullin, •Xavier Banquy Talanta 186:124-132