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目的:对新品红标准试剂的组分进行分离、鉴定,以了解掌握该标准试剂的组分情况,为该批标准试剂的标化提供依据。方法:采用液相色谱-二极管阵列检测器-电喷雾离子化串联质谱法(LC-DAD-ESI-MS/MS)进行分离鉴定。色谱柱为Phenomenx Kinetex C18柱(100 mm!2.1 mm,2.6$m),柱温35℃;流动相A为乙腈,流动相B为0.05 mol·L-1醋酸铵溶液,梯度洗脱(0~30 min,流动相A的比例从5%线性改变至45%),流速为0.2 mL·min-1;二极管阵列检测器的扫描范围为200~700 nm(提取波长:550 nm),质谱的扫描范围为m/z 110~400,裂解电压为200 V。结果:新品红标准试剂(Sigma公司产品)含3种组分,百分比含量分别为71.5%、25.7%和2.8%。根据其色谱保留行为、UV提取图谱、质谱裂解信息,并结合相关的文献报道,将其分别归属为品红Ⅲ(magentaⅢ或new fuchsin(NF),主组分)、品红Ⅱ(magentaⅡ)和品红Ⅰ(magentaⅠ)。薄层色谱结果验证了其组分情况。结论:采用串联色谱技术,利用目标化合物的色谱保留行为、紫外吸收光谱和质谱裂解规律等综合信息可为新品红标准试剂的定性鉴别提供技术依据。
OBJECTIVE: To separate and identify the constituents of new magenta standard reagents in order to know the composition of the standard reagents and provide the basis for the standardization of this batch of standard reagents. Methods: LC-DAD-ESI-MS / MS was used for the identification and identification of the tumor cells by liquid chromatography-diode array detector-electrospray ionization tandem mass spectrometry. The column was Phenomenx Kinetex C18 column (100 mm, 2.1 mm, 2.6 μm) with a column temperature of 35 ° C. The mobile phase A was acetonitrile and the mobile phase B was 0.05 mol·L -1 ammonium acetate solution. 30 min, the mobile phase A ratio changed linearly from 5% to 45%) at a flow rate of 0.2 mL · min-1. The diode array detector was scanned at 200-700 nm (extraction wavelength: 550 nm) The range is m / z 110 ~ 400, the cracking voltage is 200V. Results: The new magenta standard reagent (Sigma product) contained three components with percentages of 71.5%, 25.7% and 2.8%, respectively. According to their chromatographic retention behavior, UV extraction and mass spectrometry cleavage, combined with relevant literature reports, they were assigned to magenta Ⅲ or new fuchsin (NF), magenta Ⅱ and Magenta I. The results of TLC confirmed its composition. Conclusion: The chromatographic retention behavior, UV absorption spectra and mass spectral cleavage laws of target compounds can provide the technical basis for the qualitative identification of new magenta standard reagents by using tandem chromatography.