期刊
CHEMICAL & PHARMACEUTICAL BULLETIN
卷 52, 期 5, 页码 552-555出版社
PHARMACEUTICAL SOC JAPAN
DOI: 10.1248/cpb.52.552
关键词
p-nitrophenol; cathodic detection; HPLC; electrochemical detector; phosphatase; p-nitrophenyl phosphate
Utilizing a commercially available helium-purging device and PEEK tubes for all tubing, especially for connection between the mobile phase and pump, high performance liquid chromatography with an electrochemical detector (ECD/HPLC) at the cathodic mode is a simple and precise method for the determination of p-nitrophenol (NP). Studies with cyclic and hydrodynamic voltammetry indicated that 25% aqueous MeOH containing 0.1% (v/v) CF3CO2H and -0.8V vs. Ag/AgCl are the best mobile phase and detection potential for cathodic ECD/HPLC. With the present system, the limits of detection and determination were 0.2 and 0.25 mum, respectively, and up to 50 mum, a linear calibration curve was afforded. Within-day precisions for the analysis of 5 and 50 mum NP were 0.8 and 0.7% (n = 6), respectively, and between-day precisions (n = 6) for these samples were 3.5 and 2.2%, respectively. Compared with the commonly used Bessey-Lowry-Brock method, cathodic ECD/HPLC was useful for the assay of acid phosphatase in urine samples with p-nitrophenyl phosphate disodium salt as a substrate.
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