Rapid and sensitive hydrophilic interaction chromatography/tandem mass spectrometry method for the determination of glycyl-sarcosine in cell homogenates.
A rapid, selective and sensitive hydrophilic interaction chromatography/tandem mass spectrometry (HILIC/MS/MS) was developed and validated for the determination of glycyl-sarcosine (Gly-Sar) in Caco-2 cell homogenates. After a simple protein precipitation with acetonitrile, the analyte was separated on a HILIC column and detected by a triple quadrupole mass spectrometry equipped with an electrospray ionization (ESI) source. The method was linear among the concentration range of 1-2000 ng/mL for Gly-Sar and the lower limit of quantification (LLOQ) was 1 ng/mL using as l ittle as 50 microL of cell homogenates. The intra-day and inter-day relative standard deviations (RSD) were less than 15% and the relative errors (RE) were all within +/-15%. The validated method was successfully employed in the study of Gly-Sar uptake inhibition in Caco-2 cells by valcytarabine, a potential substrate of the peptide transporter 1 (PEPT1).
Sun
Yongbing
Y
Department of Biopharmaceutics, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang, 110016, China.
Sun
Jin
J
Liu
Jianfang
J
Yin
Shiliang
S
Chen
Ying
Y
Zhang
Peng
P
Pu
Xiaohui
X
Sun
Yinghua
Y
He
Zhonggui
Z
D018938:Caco-2 Cells; D002457:Cell Extracts / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004151:Dipeptides / Q000737:chemistry*; D006801:Humans; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
D002457:Cell Extracts; D004151:Dipeptides; C004194:glycylsarcosine