Determination of Total Hydroxyproline and Proline in Human Serum and Urine by HPLC with Fluorescence Detection
| Autor: | Yuuko Date, Yasuto Tsuruta, Kazuya Kohashi, Hirofumi Inoue, Hironori Yoshitomi |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Adult
Male Pharmacology Detection limit chemistry.chemical_classification Chromatography Proline Imino acid Pharmaceutical Science General Medicine Urine High-performance liquid chromatography Fluorescence Hydrolysate Hydroxyproline chemistry.chemical_compound chemistry Humans Female Quantitative analysis (chemistry) Chromatography High Pressure Liquid Aged |
| Zdroj: | Biological and Pharmaceutical Bulletin. 19:163-166 |
| ISSN: | 1347-5215 0918-6158 |
| Popis: | A method to measure total hydroxyproline (Hyp) and proline (Pro) in human serum and urine by HPLC was developed. Hyp and Pro in acid hydrolysates of serum and urine were derivatized with 4-(5,6-dimethoxy-2-phthalimidinyl)phenysulfonyl chloride after treatment with o-phthaladehyde and cleanup on Bond Elut C18 column. The derivatives of imino acids were separated on a reversed phase column by gradient elution with acetonitrile and phosphate buffer (1 mmol/l, pH 7) and detected by fluorescence measurement at 315 nm (excitation) and 385 nm (emission). Detection limits for both Hyp and Pro were 10 fmol per injection. The within-day and day-to-day relative standard deviations for Hyp and Pro in serum and urine were less than 3.19%. The recoveries of Hyp and Pro added to serum and urine were about 100%. The present method was applied to determine total Hyp and Pro in serum and urine from normal subjects and patients with chronic renal failure. |
| Databáze: | OpenAIRE |
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