Bradley R T, Manowitz P
Department of Psychiatry, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway 08854-5635.
Anal Biochem. 1988 Aug 15;173(1):33-8. doi: 10.1016/0003-2697(88)90154-6.
A highly sensitive assay for arylsulfatase A was developed using high-performance liquid chromatography with electrochemical detection. The retention time of the enzymatic product, p-nitrocatechol, on a reverse phase column was approximately 2.0 min. The assay was able to detect 0.43 pmol p-nitrocatechol in a 20-microliter ethanol extract of the reaction mixture. The coefficients of variation for seven determinations of the intra- and interassays were 9 and 8%, respectively. The levels of arylsulfatase A activity in human saliva samples and leukocyte and platelet lysates determined by this assay were within 6 and 11%, respectively, of the levels determined by a spectrophotometric assay. The high-performance liquid chromatography assay may have utility in measuring arylsulfatase A activity in biological samples with low activity or specific activity or in samples with compounds which interfere with the spectrophotometric assay.
采用高效液相色谱-电化学检测法开发了一种高灵敏度的芳基硫酸酯酶A检测方法。酶促产物对硝基儿茶酚在反相柱上的保留时间约为2.0分钟。该检测方法能够在反应混合物的20微升乙醇提取物中检测到0.43皮摩尔的对硝基儿茶酚。批内和批间七次测定的变异系数分别为9%和8%。通过该检测方法测定的人类唾液样本、白细胞和血小板裂解物中芳基硫酸酯酶A的活性水平分别在分光光度法测定水平的6%和11%以内。高效液相色谱检测法可能有助于测量低活性或比活性的生物样品中或含有干扰分光光度法测定的化合物的样品中的芳基硫酸酯酶A活性。
