title: Liquid chromatography-tandem mass spectrometry to determine the stability of collagen pentapeptide (KTTKS) in rat skin.
abstract: The objective of this study was to develop a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine the stability of collagen pentapeptide (KTTKS), which is a subfragment of collagen and has been proved to promote the extracellular release of collagen in skin fibroblast, in rat skin. The chromatographic condition was optimized on an Acclaim C-18 column (2.1 mm × 150 mm, 3 μm) under isocratic elution using a mobile phase consisting of deionized water and acetonitrile (87:13, v/v) mixture containing 5mM pentafluoropropionic acid as an ion-pairing reagent. The quantitation of KTTKS was performed on a triple quadrupole mass spectrometer in multiple reaction monitoring mode. The calibration curve showed good linearity in the concentration range of 0.05-10.0 μg/mL (r(2)>0.999). The intra- and inter-day precisions were 0.8-6.5% and 2.4-5.8%, respectively, and the intra- and inter-day accuracies were 96.3-102.7% and 92.8-98.5%, respectively. The developed LC-MS/MS method was successfully applied to investigate the degradation rate and sites of KTTKS in rat skin homogenate. KTTKS was found to be very susceptible to the peptide bond cleavage by aminopeptidases present in the skin.
authors.affiliation: College of Pharmacy, Kyungsung University, Nam-ku, Busan, Republic of Korea.
authors.forename: Eun Ji
authors.initials: EJ
authors.lastname: Park
authors.forename: Myung Sun
authors.initials: MS
authors.lastname: Kim
authors.forename: Yun Lim
authors.initials: YL
authors.lastname: Choi
authors.forename: Young-Hee
authors.initials: YH
authors.lastname: Shin
authors.forename: Hye Suk
authors.initials: HS
authors.lastname: Lee
authors.forename: Dong Hee
authors.initials: DH
authors.lastname: Na
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D003094:Collagen / Q000032:analysis / Q000737:chemistry*; D004355:Drug Stability; D007700:Kinetics; D016014:Linear Models; D009842:Oligopeptides / Q000737:chemistry*; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012867:Skin / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D009842:Oligopeptides; C525923:lysyl-threonyl-threonyl-lysyl-serine; D003094:Collagen