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人血清中总胆固醇的测定

人血清中总胆固醇的测定. [ 实验目的及要求 ] 学会利用氧化酶法测定血清中总胆固醇含量的原理和方法。 [ 实验原理 ] 胆固醇脂 +H2O 胆固醇 + 游离脂肪酸 胆固醇 +O2 ⊿4- 胆甾烯酮 +H2O2 2H2O2+4-AA+4- 氯仿 醌亚胺(红色) +4H2O CEH :胆固醇脂酶 POD :过氧化物酶 CHOD :胆固醇氧化酶 4-AA : 4- 氨基安替吡啉

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人血清中总胆固醇的测定

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  1. 人血清中总胆固醇的测定

  2. [实验目的及要求] • 学会利用氧化酶法测定血清中总胆固醇含量的原理和方法。 • [实验原理] • 胆固醇脂+H2O 胆固醇+游离脂肪酸 • 胆固醇+O2 ⊿4-胆甾烯酮+H2O2 • 2H2O2+4-AA+4-氯仿 醌亚胺(红色)+4H2O • CEH:胆固醇脂酶 POD:过氧化物酶 • CHOD:胆固醇氧化酶 4-AA:4-氨基安替吡啉 • 醌亚胺在500nm有特征吸收,且生成量与样本中的胆固醇含量成正比,可通过测定标准品反应生成的醌亚胺的吸光度值,计算出样本中胆固醇的浓度。

  3. [实验仪器及试剂] • 试管 移液管 UNICO-2000型分光光度计 恒温水浴锅 • 标准血清 样本血清 蒸馏水 酶试剂盒

  4. [实验步骤] • 按下表加入各反应物:

  5. A样本 A标准 胆固醇含量= ×标准血清浓度 • 各管混匀,37℃反应6 min,以空白管校零。

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