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胆汁酸与Thio-NAD在3α-羟基类固醇脱氢酶（3α-Ketosteroids，3α-HSD）作用下生成Thio-NADH和3α-酮类固醇，随后3α-酮类固醇与NADH在3α-HSD作用下生成胆酸汁和NAD + 。Thio-NADH在405 nm处具有特征吸收峰，通过吸光值变化即可定量检测总胆酸汁的含量。
Bile acids react with Thio‑NAD in the presence of 3α‑hydroxysteroid dehydrogenase (3α‑HSD) to generate Thio‑NADH and 3α‑ketosteroids. Subsequently, the 3α‑ketosteroids react with NADH in the presence of 3α‑HSD to regenerate bile acids and NAD⁺. Thio‑NADH exhibits a characteristic absorption peak at 405 nm. By measuring the change in absorbance, the total bile acid content can be quantitatively determined.
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