己糖激酶（Hexokinase, HK）能够催化葡萄糖和ATP生成6-磷酸葡萄糖，6-磷酸葡萄糖脱氢酶（Glucose-6-Phosphate Dehydrogenase, G6PDH）进一步催化6-磷酸葡萄糖和NADP+生成6-磷酸葡萄糖酸和NADPH，NADPH在340 nm处具有特征吸收峰，通过吸光值变化即可定量检测ATP的含量。
标准物质： ATP①试剂四和试剂六配制后有效期较短，为便于实验安排，均附赠一支作为备用，每支均可满足至少100个样本的检测用量；
②提取过程应严格在冰浴条件下进行；
③测定过程中试剂四应用液、试剂六应用液和待测样本应置于冰上放置，以免变性和失活；
④提取过程中第一次离心后的上清液若为浑浊属于正常现象，不影响检测结果；
⑤提取液出现晶体析出属于正常现象，50-60℃水浴加热至完全溶解，冷却至室温后使用即可；
⑥准确在10 s和190 s处完成吸光值测定，以确保实验结果的准确性和重复性；若使用96孔UV板测定应使用多道移液器且分批进行测定，以确保组间反应时间一致；
⑦若A2测定超出标准吸光值线性范围：高于最高值建议将待测样本使用蒸馏水适当稀释后再进行测定，低于最低值建议制备更高浓度的样本后再进行测定，计算时相应修改；
⑧若吸光值超过3，建议检查96孔板是否为96孔UV板，96孔板会因材质问题造成吸光值接近设备量程；
⑨提取液中含有蛋白沉淀组分，待测样本不能直接用于蛋白含量测定，若使用蛋白浓度计算ATP含量，则需要另取样本使用PBS或生理盐水按照相同步骤制备为待测样本，再进行蛋白浓度测定；
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