高效液相色谱-串联质谱法快速检测银耳中的米酵菌酸
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摘要
目的建立高效液相色谱-串联质谱法(highperformanceliquidchromatography-tandemmass spectrometry,HPLC-MS/MS)快速检测银耳中的米酵菌酸的方法。方法样品以甲醇提取后,超声振荡,离心分层,上清液用甲醇定容。使用空白基质液配置标准曲线对结果进行校正。在优化后的色谱及质谱条件下,采用(electronsprayionization,ESI)负离子模式进行电离,并通过多反应监测(multi-selectedreactionmonitoring,MRM)模式对目标化合物的定量离子和定性离子进行测定。结果米酵菌酸在0.50～50.00μg/L范围内线性良好,相关系数r为0.9996,3个浓度添加水平的平均回收率范围在80.6%～95.6%之间,相对标准偏差为4.98%～7.21%。方法检出限(limit of detection, LOD)为5.0μg/kg,定量限(limit of quantification, LOQ)为15.0μg/kg。结论该方法操作简便快速,回收率高、灵敏度高、精密度好,适合测定银耳基质中的米酵菌酸。
Objective To establish a method of rapid determination of bongkrekic acid in Tremella by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS). Methods The sample was extracted with methanol, then it was ultrasonically shaken and centrifuged, and the supernatant was adjusted to volume with methanol. The results were calibrated by a blank matrix configuration standard curve. Under optimized chromatographic and mass spectrometric conditions, ionization was performed using an electron spray ionization(ESI) negative ion mode, and quantitative and qualitative ions of the target compound were determined by a multi-selected reaction monitoring(MRM) mode. Results The linearity of bongkrekic acid was good in the range of 0.50～50.00 μg/L, and the correlation coefficient r was 0.9996. The average recovery of 3 concentration levels were 80.6%～95.6%, and the relative standard deviation was 4.98%-7.21%. The method of limit of detection(LOD) was 5.0 μg/kg, and the limit of quantification(LOQ) was 15.0 μg/kg. Conclusion The method is simple and rapid, it has high recovery rate, high sensitivity and good precision, which is suitable for the determination of bongkrekic acid in the Tremella.
Objective To establish a method of rapid determination of bongkrekic acid in Tremella by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS). Methods The sample was extracted with methanol, then it was ultrasonically shaken and centrifuged, and the supernatant was adjusted to volume with methanol. The results were calibrated by a blank matrix configuration standard curve. Under optimized chromatographic and mass spectrometric conditions, ionization was performed using an electron spray ionization(ESI) negative ion mode, and quantitative and qualitative ions of the target compound were determined by a multi-selected reaction monitoring(MRM) mode. Results The linearity of bongkrekic acid was good in the range of 0.50～50.00 μg/L, and the correlation coefficient r was 0.9996. The average recovery of 3 concentration levels were 80.6%～95.6%, and the relative standard deviation was 4.98%-7.21%. The method of limit of detection(LOD) was 5.0 μg/kg, and the limit of quantification(LOQ) was 15.0 μg/kg. Conclusion The method is simple and rapid, it has high recovery rate, high sensitivity and good precision, which is suitable for the determination of bongkrekic acid in the Tremella.
引文
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[10]李红艳,金燕飞,黄海智,等.高效液相色谱-二极管阵列检测器结合固相萃取法快速测定食品中米酵菌酸残留[J].食品科学,2016,37(24):247-251.Li HY,Jin YF,Huang HZ,et al.Fast determination of bongkrekic acid in foods using mixed-mode weak anion exchange solid phase extraction coupled with high performance liquid chromatography with diode array detection(HPLC-DAD)[J].Food Sci,2016,37(24):247-251.
[11]侯佰立.固相萃取-高效液相色谱法快速测定食品中米酵菌酸残留[J].现代食品,2019,(9):169-172.Hou BL.fast determination of bongkrekic acid in foods using solid phase extraction-high performance liquid chromatography[J].Mod Food,2019,(9):169-172.
[12]周霞,陈万勤,傅红雪,等.全自动固相萃取-超高效液相色谱法快速测定食品中米酵菌酸含量[J].安徽农业科学,2018,46(8):159-160,182.Zhou X,Chen WQ,Fu HX,et al.Fast determination of bongkrekic acid in food by auto solid phase extraction-ultra high performance liquid chromatography[J].J Anhui Agric Sci,2018,46(8):159-160,182.
[13]周鹏.超高效液相色谱串联质谱法测定银耳中米酵菌酸[J].食品研究与开发,2015,36(22):123-126.Zhou P.Determination of bongkrekic acid in Tremella fuciformis berk by ultra high performance liquid chromatography-tandem mass spectrometry[J].Food Res Dev,2015,36(22):123-126.
[14]苏肯明,梁达清.液相色谱-串联质谱法测定银耳中的米酵菌酸[J].广东化工,2014,41(16):168-169.Su KM,Liang DQ.Determination of bongkrekic acid in Tremella fuciformis by LC-MS/MS[J].Guangdong Chem Ind,2014,41(16):168-169.
[15]李红娜,袁飞,张辰陽,等.液相色谱和飞行时间质谱联用检测米酵菌酸[J].食品工业,2018,39(7):319-323.Li HN,Yuan F,Zhang CY,et al.The research of detection bongkrekic acid(BA)by LC-ESI-TOF technology[J].Food Ind,2018,39(7):319-323.
[2]刘坤,孟玲芝,王圣博,等.米酵菌酸在食品检测中的研究新进展[J].吉林农业,2018,(17):81.Liu K,Meng LZ,Wang SB,et al.New progress of rice fermentation bacterial acid in food detection[J].Jinlin Agric,2018,(17):81.
[3]Rohm B,Scherlach K,Hertweck C.Biosynthesis of the mitochondrial adenine nucleotide translocase(atpase)inhibitor bongkrekic acid in burkholderia gladioli[J].Organ Biomol Chem,2010,8(7):1520-1522.
[4]Henderson PJF,Lardy HA.Bongkrekic acid-An inhibitor of the adenine nucleotide translocase of mitochondria[J].J Biol Chem,1970,245(6):1319-1326.
[5]宋兴田,吴洪娟,庄宝祥.米酵菌酸中毒小鼠肝和脑组织的形态学改变[J].中国医药导报,2010,7(26):36-38.Song XT,Wu HJ,Zhuang BX.Morphologic changes of intoxicated mice hepatocyte and brain tissues of bongkrekic acid[J].Chin Med Herald,2010,7(26):36-38.
[6]Passam HC,Coleman JOD.The effects of atractyloside and bongkrekic acid on jerusalem artichoke mitochondria in relation to adenine nucleotide translocation[J].J Exper Botan,1975,26(93):536-543.
[7]Ruprecht A,Jonathan J,King MS,et al.The molecular mechanism of transport by the mitochondrial ADP/ATP carrier[J].Cell,2019,176(3):435-447.
[8]NY/T 749-2018绿色食品食用菌[S].NY/T 749-2018 Green food-Edible mushroom[S].
[9]GB 7096-2014食品安全国家标准食用菌及其制品[S].GB 7096-2014 National Food Safety Standard-Edible fungi and their products[S].
[10]李红艳,金燕飞,黄海智,等.高效液相色谱-二极管阵列检测器结合固相萃取法快速测定食品中米酵菌酸残留[J].食品科学,2016,37(24):247-251.Li HY,Jin YF,Huang HZ,et al.Fast determination of bongkrekic acid in foods using mixed-mode weak anion exchange solid phase extraction coupled with high performance liquid chromatography with diode array detection(HPLC-DAD)[J].Food Sci,2016,37(24):247-251.
[11]侯佰立.固相萃取-高效液相色谱法快速测定食品中米酵菌酸残留[J].现代食品,2019,(9):169-172.Hou BL.fast determination of bongkrekic acid in foods using solid phase extraction-high performance liquid chromatography[J].Mod Food,2019,(9):169-172.
[12]周霞,陈万勤,傅红雪,等.全自动固相萃取-超高效液相色谱法快速测定食品中米酵菌酸含量[J].安徽农业科学,2018,46(8):159-160,182.Zhou X,Chen WQ,Fu HX,et al.Fast determination of bongkrekic acid in food by auto solid phase extraction-ultra high performance liquid chromatography[J].J Anhui Agric Sci,2018,46(8):159-160,182.
[13]周鹏.超高效液相色谱串联质谱法测定银耳中米酵菌酸[J].食品研究与开发,2015,36(22):123-126.Zhou P.Determination of bongkrekic acid in Tremella fuciformis berk by ultra high performance liquid chromatography-tandem mass spectrometry[J].Food Res Dev,2015,36(22):123-126.
[14]苏肯明,梁达清.液相色谱-串联质谱法测定银耳中的米酵菌酸[J].广东化工,2014,41(16):168-169.Su KM,Liang DQ.Determination of bongkrekic acid in Tremella fuciformis by LC-MS/MS[J].Guangdong Chem Ind,2014,41(16):168-169.
[15]李红娜,袁飞,张辰陽,等.液相色谱和飞行时间质谱联用检测米酵菌酸[J].食品工业,2018,39(7):319-323.Li HN,Yuan F,Zhang CY,et al.The research of detection bongkrekic acid(BA)by LC-ESI-TOF technology[J].Food Ind,2018,39(7):319-323.