BACKGROUND:In the present quality control file or technique standards of in vitro fertilization medium, the indicators of the component contents and detection methods have not been clearly defined. To ensure the safety and effectiveness of these products, we should establish the quality standards as early as possible. OBJECTIVE:To establish a method for determining the three main bioactive constituents of in vitro fertilizationmedium including glucose, lactic acid sodium salt, pyruvic acid sodium salt by ultra-high performance liquid chromatography tandem mass spectrometric method (UHPLC-MSMS), and to analyze the content of each constituent. METHODS:The UHPLC-MSMS was used, and UHPLC separation was performed on a SUPELCO Discovery HS F5-3 column (15 cm × 2.1 mm, 3μm) in a gradient elute mode with acetonitrile and water (both containing 0.1%formic acid) as the mobile phase at a flow rate of 0.35 mL/min. The column temperature was 40℃. Mass spectrometry detection was performed with multi-reaction monitoring mode using negative electro spray ionization. RESULTS AND CONCLUSION:The linearity was achieved in the range of 0.1-10μg/mL (r=0.999 8) for glucose, 0.05-5μg/mL (r=0.999 4) for lactic acid sodium salt, and 0.1-10μg/mL (r=0.999 4) for pyruvic acid sodium salt. The recoveries were 96.4%-98.1%with relative standard deviation less than 2.8%. To conclude, the UHPLC-MSMS method is sensitive, rapid, accurate and specific, thus providing a basis for the quality standard study of in vitro fertilization medium.%背景：目前，对于体外生殖培养液的质量控制文件或技术标准中，对成分含量的指标及检测方法未做明确规定，为保证该类产品的安全有效性，应及早建立质量标准。目的：建立采用液相质谱联用法检测体外生殖培养液中葡萄糖、乳酸钠、丙酮酸钠3种指标成分的方法，分析不同用途培养液中3种成分的含量。方法：采用超高效液相色谱三重四级杆质谱联用法，使用SUPELCO Discovery HS F5-3色谱柱(15 cm×2.1 mm，3μm)，流动相为0.1%甲酸水溶液-0.1%甲酸乙腈溶液，梯度洗脱，流速0.35 mL•min-1，柱温40℃；质谱采用电喷雾离子源，负离子模式扫面，扫描方式为多反应监测。结果与讨论：葡萄糖、丙酮酸钠、乳酸钠分别在0.1-10 mg/L(r=0.9998)、0.05-5 mg/L(r=0.9994)、0.1-10 mg/L(r=0.9994)范围线性关系练好；平均加样回收率在96.4%-98.1%之间，相对标准偏差均小于2.8%。说明采用液相质谱联用法检测体外生殖培养液灵敏、快速、准确、专属性强，可为体外生殖培养液的质量标准研究提供依据。
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