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血清5-核苷酸酶(5-NT)的测定,不外乎根据两个原理:①比色测定释放出的磷;②采用一系列有关的酶促反应。后者需多种保存期有限且代价较高的酶试剂。测磷法试剂简单,可作自动化分析,但现行方法须除蛋白,使操作时间延长,结果精密度减低,方法也不十分敏感。作者结合Lawrence法,利用敏感的氯化亚锡/钼反应来测磷,加入十二烷基硫酸钠可不除蛋白,操作简单,可靠,试剂便宜,且较稳定。试剂①巴比妥盐酸缓冲液(pH7.5在37℃时):将巴比妥钠8.25 g、硫酸锰0.238 g加水溶解并补充体至1升,加120 ml 0.2MHCl,必要时调pH至7.73(20℃),贮于4℃。②基质液:称取5-腺苷酸二钠(AMP·二钠·6H_2O)499.2 mg加水至100ml。按1.0 ml 1份分装后,贮于-20℃。③氯化镍(NiCl_2,
Serum 5-nucleotidase (5-NT) determination, nothing more than based on two principles: ① colorimetric determination of released phosphorus; ② using a series of related enzymatic reactions. The latter requires a variety of enzyme reagents with limited shelf life and high cost. Phosphorus reagent is simple and can be used for automated analysis, but the current method to remove protein, so that the operation time is extended, resulting in reduced precision, the method is not very sensitive. The authors combined with Lawrence method, the use of sensitive stannous chloride / molybdenum reaction to phosphorus, sodium dodecyl sulfate can be added without protein, simple, reliable, cheap reagents, and more stable. Reagents ① barbiturates hydrochloric acid buffer (pH7.5 at 37 ℃): 8.25 g of sodium pentobarbital, manganese sulfate 0.238 g dissolved in water and make up to 1 liter, add 120 ml 0.2MHCl, if necessary, adjust the pH to 7.73 (20 ° C) and stored at 4 ° C. ② matrix solution: Weigh 5-adenylate disodium (AMP · disodium · 6H_2O) 499.2 mg water to 100ml. After dispensing 1.0 ml aliquots, store at -20 ° C. ③ nickel chloride (NiCl_2,