气相色谱-燃烧炉-同位素比质谱法测定奶粉中 孕酮δ13C值

Determination of δ13C of progesterone in milk powder by gas chromatography-combustion-isotope ratio mass spectrometry

中文关键词:  孕酮  奶粉  气相色谱-燃烧炉-同位素比质谱法

英文关键词:progesterone  milk powder  gas chromatography-combustion-isotope ratio mass spectrometry

中文摘要:

      目的 基于高效液相色谱纯化分离, 应用气相色谱-燃烧炉-同位素比质谱法(gas chromatography-combustion-isotope ratio mass spectrometry, GC-C-IRMS)建立奶粉中测定孕酮激素碳同位素比值的方法, 以气相色谱-质谱法提供定量支持。方法 样品经水溶解和乙腈提取, 经乙腈饱和的正己烷除脂肪和过C18固相萃取柱净化, 经高效液相色谱纯化、浓缩后进行GC-C-IRMS检测。结果 外源性孕酮δ13C平均值为-32.297‰, 内源性孕酮δ13C平均值为-21.387‰, 方法回收率为52.9%, 检出限为22.7 μg/kg, 相对标准偏差小于5%(n=6)。结论 内源性和外源性孕酮δ13C值有显著性差异, 本方法可以区分内外源性孕酮。但由于方法局限性, 只能检测孕酮大于22.7 μg/kg的样品。

英文摘要:

      Objective Based on the separation and purification of high performance liquid chromatography (HPLC), gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) was established for determining the stable carbon isotope ratio of progesterone in milk powder, and gas chromatograph mass spectrometry (GC-MS) was established for determining the residues of progesterone in milk powder. Methods The samples were dissolved in water and extracted with acetonitrile. Then the lipid substances were removed by liquid-liquid extraction with acetonitrile saturated n-hexane. The purification was carried out on a C18 solid phase extraction (SPE) column. The extracts were purified and concentrated by HPLC and detected by GC-C-IRMS. Results Average δ13C value of endogenous progesterone was -32.297‰ and average δ13C value of exogenous progesterone was -21.387‰. The recovery was 52.9%, the limit of detection was 22.7 μg/kg and the relative standard deviations (RSDs) were less than 5% (n=6). Conclusion There is a significant difference in δ13C value between endogenous and exogenous progesterone, and the method can distinguish endogenous and exogenous progesterone. However, only when the residues of progesterone in powdered milk are more than 22.7 μg/kg, it can be detected by GC-C-IRMS.KEY WORDS: progesterone; powdered milk; Gas chromatograph mass spectrometer; Gas Chromatography-Combustion-Isotope Ratio Mass Spectrometry