高效液相色谱法测定尿中咖啡因主要代谢物浓度
HPLC determination of caffeine metabolites in urine
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摘要: 本文建立了测定尿中咖啡因5种主要代谢物(AFMU、1X、1U、17X、17U)浓度的高效波相色谱法。尿样用硫酸铵饱和后加氯仿及异丙醇混合液(85:15)提取2次,空气流吹干,蒸馏水溶解进样。以Shim PackC18为固定相,甲醇-动腈-0.05%醋酸(10:1:89)为流动相,扑热息痛为内标,流速为1.2ml/min,在280um处定量检测。本法精确稳定可靠。用此方法测定了120名健康志愿者口服咖啡后的尿样,对人体内N-乙酰化转移酶和CYP1A2酶活性作了初步分析。Abstract: A HPLC method was developed for determination of caffeine metabolites in urine(AFMU, 1X, 1U, 17X, 17U).The samplb added ammonium sulfate was extracted twice by a mixture of chloroform-isopropanol(85:15 vol/vol)and evaporated to dryness with a gentle stream ofair below)25℃.HPLC analysis was carried out on Shim Pack C18 column with methanol-ace-tonitrile-0.05% acetic acid (10:1:89 vol/vol) as the mobile phase and UV detection at 280nm.The assay procedure has been applied to determining urinary metabolites excreted after oral caffeine in a healthy sample(n = 120).Caffeine metabolite ratios were used to assess the poymorphicN-acetyltransferase and CYP1A2 activities.
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Key words:
- Caffeine /
- HPLC /
- N-acetyltransferase /
- CYP1A2
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