title: Determination of proline-hydroxyproline dipeptide in rat urine by high-performance anion-exchange chromatography coupled with pulsed amperometric detection.
abstract: A method for determination of proline-hydroxyproline dipeptide (PHP) was developed using high-performance anion-exchange chromatography-pulsed amperometric detection (HPAEC-PAD). This method resulted in good separation of proline (Pro), hydroxyproline (Hyp), and PHP within 20min using a mobile phase of 1.2mM Ba(OH)2+1.5mM Ba(OAc)2. The linear dynamic ranges and their detection limits (S/N=3) were 1-100 (r(2)=0.9990-0.9999) and 0.05-0.3μM, respectively. Mean recoveries were 91.6-121.3% and 92.2-110.3% for intra- and inter-day assays, respectively. Our HPAEC-PAD method showed clear differences in the corrected PHP levels measured in urine samples from two groups of rats, sham-operated and ovariectomized, without the need for prior acid hydrolysis or sample derivatization.
authors.affiliation: Department of Oriental Pharmaceutical Sciences, Kyung Hee East-West Pharmaceutical Research Institute, College of Pharmacy, Kyung Hee University, Hoegi-dong, Dongdaemoon-gu, Seoul 130-701, Republic of Korea.
authors.forename: Hee-Jung
authors.initials: HJ
authors.lastname: Sim
authors.forename: Eunjung
authors.initials: E
authors.lastname: Moon
authors.forename: Sun Yeou
authors.initials: SY
authors.lastname: Kim
authors.forename: Seon-Pyo
authors.initials: SP
authors.lastname: Hong
mesh_terms: D000818:Animals; D000838:Anions / Q000737:chemistry; D017609:Barium Compounds / Q000737:chemistry; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004151:Dipeptides / Q000737:chemistry / Q000652:urine*; D055664:Electrochemical Techniques / Q000379:methods*; D005260:Female; D057230:Limit of Detection; D010052:Ovariectomy; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D013696:Temperature
chemical_list: D000838:Anions; D017609:Barium Compounds; D004151:Dipeptides; C533879:prolyl-4-hydroxyproline; C012766:barium hydroxide