Development of a Fluorescent Labeling Reagent for Determination of Proline and Hydroxyproline in Myeloma Patient Plasma by HPLC-FLD

作者：Teng, Zhenyuan; Lv, Hongying; Zhang, Yaoyao; Zhang, Yi; Guan, Jun^*; Wang, Chengyin

来源：Journal of Chromatographic Science, 2016, 54(10): 1743-1751.

DOI：10.1093/chromsci/bmw132

• PERFORMANCE LIQUID-CHROMATOGRAPHY
• TERMINAL PROLYL DIPEPTIDES
• AMINO-ACIDS
• PRECOLUMN DERIVATIZATION
• MASS-SPECTROMETRY
• URINARY HYDROXYPROLINE
• BIOLOGICAL-FLUIDS
• BIOGENIC-AMINES
• CELL-CULTURES
• GC-MS

A fluorescent derivatization reagent, 3,6-dimethoxy-9-phenyl-9H-carbazole-1-sulfonyl chloride (DPCS-Cl), previously designed, synthesized and developed for amino acids labeling, is here employed. It was used as a pre-column derivatization reagent for the determination of proline (Pro) and hydroxyproline (Hyp) with high-performance liquid chromatography. Both of the analytes were labeled with DPCS-Cl at 60 degrees C for 30 min. The optimized concentration of DPCS-Cl was 25 mu gmL(-1) and the molar ratio of analytes to DPCS-Cl was 1:6. Excitation wavelength of 318 nm and emission wavelength of 440 nm were selected for the fluorescence detection. For Pro and Hyp, the linear correlation coefficients were all >0.999; the linear ranges of calibration curve were all 5.0 nmol L-1 to 5.0 mu mol L-1; the detection limits were 20.0 and 10.0 fmol, respectively; the intraday and inter-day relative standard deviations were 2.6-3.3% and 3.5-4.6%, respectively. This reagent was applied to the determination of Pro and Hyp in plasma of myeloma patients with its merits of high efficiency of derivatization, stability of the derivatives and high sensitivity.

• 出版日期2016-11
• 单位扬州大学