本研究旨在建立同步测定香菇中麦角固醇和维生素 D2的液相色谱 － 串联质谱 法． 超声耦合皂化法制备的样品溶液采用 C8色谱柱处理，以甲醇 － 水作为流动相，270 nm 检测波长、25 ℃柱温、2． 5 mL /min 流速为色谱条件，通过梯度洗脱法分离纯化样品; 采用 电喷雾正离子模式( ESI + ) 电离及多反应( MＲM) 监测检测模式，当气帘气( CUＲ) 、雾化 气( GS1 ) 及辅助气( GS2 ) 压力分别设定为 137． 9、310． 3 及 413． 7 kPa 时，以 5 500 V 离子源 喷射电压( IS) 、500 ℃离子源温度( TEM) 、5 L /min 离子源喷雾流速为质谱条件进行串联 质谱分析． 结果表明: 麦角固醇与维生素 D2色谱峰保留时间分别为 4． 49 min 和 4． 26 min． 经方法学考察，麦角固醇( 0． 40 ～ 3． 60 mg /L) 和维生素 D2 ( 0． 22 ～ 2． 20 mg /L) 的含量与峰 面积有良好的线性关系，麦角固醇和维生素 D2的平均加标回收率范围为 86． 7% ～ 102． 4%， 相对标准偏差为 1． 02% ～ 4． 13% ． 采集的样品采用液 － 质联用法和 HPLC 色谱法同步检 测，实验结果并无显著性差异( P ＞ 0． 05) ，与 HPLC 法色谱峰面积定量相比，液质 － 联用 法具有快速检测时间短、检测限低且更灵敏的优势． 本研究建立的同步检测香菇中麦角固 醇和维生素 D2的含量液相色谱 － 串联质谱法，为香菇食用菌新产品质量标准及产品质量 检测标准的制定奠定了基础，为研究同分异构体的同步检测提供理论依据．

Liquid chromatography coupled with tandem mass spectrometry ( LC-MS /MS) method was established for simultaneous determination of ergosterol and vitamin D2 content in Lentinus edodes． Ergosterol and vitamin D2 were extracted by saponification reflux combined in ultrasonic auxiliary and purified through C8 chromatographic column with optimizing the chromatographic conditions at 25 ℃ of column temperature，270 nm of detection wavelength and 1. 5 mL /min of the flow rate of methanol aqueous solution in the gradient elution methods． The prepared samples were tested by mass spectrometry with electrospray ionization positive ion ( ESI + ) mode and multi-reaction ( MＲM) monitoring mode． Ion source injection voltage ( IS) at 5500V，ionic source temperature ( TEM) at 500℃ and ion source spray velocity at 5 L /min were applied as mass spectrometer conditions when the set pressure of air curtain gas ( CUＲ) ，atomization gas ( GS1 ) and auxiliary gas ( GS2 ) were controlled at 137. 9，310. 3 and 413. 7 kPa， respectively． Ｒesults show that the retention time of ergosterol and vitamin D2 chromatographic peaks are at 4. 49 and 4. 26 min，respectively，the content of ergosterol ( 0. 40 ～ 3. 60 mg /L) and vitamin D2 ( 0. 22 ～ 2. 20 mg /L) within a certain content range of peak area presents a good linear relationship． The average recovery rate of ergosterol and vitamin D2 are within 86. 7% ～ 102. 4% ，and the relative standard deviation are within 1. 02% ～ 4. 13% ，respectively，according to methodological validation． The collection samples were detected by LC-MS / MS established in this experiment and HPLC recommended by the national standard． The detecting results suggest that no significant difference ( P ＞ 0. 05) is found by two methods utilized for samples analysis． Compared with the peak area quantification by HPLC chromatography，LC-MS /MS method has the advantages of shorter detection time，lower detection limit and more sensitive detection． The simultaneous determination of ergosterol and vitamin D2 in Lentinus edodes by liquid chromatography-tandem mass spectrometry established in this study lays a foundation for the establishment of new product quality standards and product quality testing standards for edible fungi，and provides theoretical basis for the simultaneous detection and research of isomers．