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Quantification of peptides in human synovial fluid using liquid chromatography–tandem mass spectrometry

Author：Araceli Garcia-Ac, Sung VoDuy, Sébastien Sauvé, Florina Moldovan, V. Gaëlle Roullin, Xavier Banquy   《Talanta》

ABSTRACTS
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.

Graphical abstract


KEY WORDS
Quantitative bioanalysis; Tandem mass spectrometry; Peptide quantification; B1 receptor; Endothelin receptor antagonist; Synovial fluid; Knee osteoarthritis; Distribution coefficient
SCREENSHOT

RELATED PRODUCTS
 BQ123, Cyclo(-D-Trp-D-Asp-L-Pro-D-Val-L-Leu), 95.51% pure was purchased from China peptides Co.
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https://www.sciencedirect.com/science/article/pii/S003991401830345X
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