The estimation of serum acid phosphatase using α-naphthyl phosphate as substrate: observations on the use of Fast Red TR salt in the assay
1982; Elsevier BV; Volume: 126; Issue: 3 Linguagem: Inglês
10.1016/0009-8981(82)90304-7
ISSN1873-3492
AutoresJonathan D. Cooper, D C Turnell, C P Price,
Tópico(s)Molecular Sensors and Ion Detection
ResumoThe estimation of acid phosphatase (EC 3.1.3.2) in serum by reaction rate analysis, utilising α-naphthyl phosphate as substrate and a diazonium salt (Fast Red TR) for colour development is examined. In the absence of substrate the occurrence of a pseudo catalytic activity is shown to be due to the hydrolysis of the Fast Red TR with the subsequent production of 4-chloro-2-methylphenol. The 4-chloro-2-methylphenol reacts with the Fast Red TR to form an azo compound which absorbs at a wavelength of 340 nm. The production of 4-chloro-2-methylphenol during the reaction is first order with respect to the Fast Red TR concentration, is temperature dependent and is accelerated in the presence of human albumin or Triton X-100. Monitoring the reaction at a wavelength of 585 nm eliminates the need for substrate independent blank correction. The final reaction conditions are summarised, together with a comparison of the method with that of the Kind and King [9] procedure.
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