通过酸性和碱性提取液分别提取样品中NAD+和NADH，NADH可通过PMS的递氢作用，还原氧化型噻唑蓝（MTT）生成甲瓒，产物在570 nm处具有特征吸收峰；NAD+可被乙醇脱氢酶还原为NADH，进一步采用MTT还原法生成甲瓒，通过吸光值变化即可定量检测辅酶Ⅰ NAD (H) 的含量。
标准物质： NAD+ & NADH①试剂三和试剂四配制后有效期较短，为便于试验安排，各附赠一瓶作为备用，每瓶均可满足至少25个样本的测定；
②试剂五配制后未使用完试剂应置于4℃保存，严禁-20℃保存，使用过程中试剂五应用液需置于冰上放置；
③2 µmol/mL NAD+标准液和2 µmol/mL NADH标准液配制后4℃可保存1周，严禁-20℃保存；
④实验过程中吸取上清液时应避免吸入沉淀物而影响结果的准确性；
⑤反应时间对实验结果有较大影响建议精确控制，测定样本较多时建议分批进行测定，以避免组内反应时间不一致对结果造成影响；
⑥反应体系中试剂不能按比例配制为混合液使用，必须按反应体系顺序依次加入；
⑦若A测定大于1.0，建议将待测样本适当稀释后再进行测定，计算时相应修改；
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