Determination of proline-hydroxyproline dipeptide in rat urine by high-performance anion-exchange chromatography coupled with pulsed amperometric detection
- Authors
- Sim, Hee-Jung; Moon, Eunjung; Kim, Sun Yeou; Hong, Seon-Pyo
- Issue Date
- 1-Jul-2013
- Publisher
- ELSEVIER SCIENCE BV
- Keywords
- Anion-exchange chromatography; Pulsed amperometric detection; Barium hydroxide; Proline-hydroxyproline; Hydroxyproline
- Citation
- JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, v.930, pp.70 - 74
- Journal Title
- JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
- Volume
- 930
- Start Page
- 70
- End Page
- 74
- URI
- https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/14431
- DOI
- 10.1016/j.jchromb.2013.04.033
- ISSN
- 1570-0232
- 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 20 min using a mobile phase of 1.2 mM Ba(OH)(2)+ 1.5 mM Ba(OAc)(2). The linear dynamic ranges and their detection limits (S/N = 3) were 1-100 (r(2) = 0.9990-0.9999) and 0.05-0.3 mu M, 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. (C) 2013 Elsevier B.V. All rights reserved,
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