Laboratory of Mass Spectrometry, University of Camerino, School of Pharmacy, Via S. Agostino, 62032 Camerino, Italy.
Laboratory of Pharmaceutical and Biotechnological Sciences, University of Camerino, School of Pharmacy, Via S. Agostino, 62032 Camerino, Italy.
Molecules. 2020 Jul 3;25(13):3047. doi: 10.3390/molecules25133047.
In recent decades, mass spectrometry techniques, particularly when combined with separation methods such as high-performance liquid chromatography, have become increasingly important in pharmaceutical, bio-analytical, environmental, and food science applications because they afford high selectivity and sensitivity. However, mass spectrometry has limitations due to the matrix effects (ME), which can be particularly marked in complex mixes, when the analyte co-elutes together with other molecules, altering analysis results quantitatively. This may be detrimental during method validation, negatively affecting reproducibility, linearity, selectivity, accuracy, and sensitivity. Starting from literature and own experience, this review intends to provide a simple guideline for selecting the best operative conditions to overcome matrix effects in LC-MS techniques, to obtain the best result in the shortest time. The proposed methodology can be of benefit in different sectors, such as pharmaceutical, bio-analytical, environmental, and food sciences. Depending on the required sensitivity, analysts may minimize or compensate for ME. When sensitivity is crucial, analysis must try to minimize ME by adjusting MS parameters, chromatographic conditions, or optimizing clean-up. On the contrary, to compensate for ME analysts should have recourse to calibration approaches depending on the availability of blank matrix. When blank matrices are available, calibration can occur through isotope labeled internal standards and matrix matched calibration standards; conversely, when blank matrices are not available, calibration can be performed through isotope labeled internal standards, background subtraction, or surrogate matrices. In any case, an adjusting of MS parameters, chromatographic conditions, or a clean-up are necessary.
近几十年来，质谱技术，特别是与高效液相色谱等分离方法结合使用时，在药物、生物分析、环境和食品科学应用中变得越来越重要，因为它们具有高选择性和灵敏度。然而，由于基质效应（ME），质谱技术存在局限性，在复杂混合物中，当分析物与其他分子一起共洗脱时，ME 会特别明显，从而定量改变分析结果。这在方法验证期间可能是有害的，会对重现性、线性、选择性、准确性和灵敏度产生负面影响。本文从文献和自身经验出发，旨在为 LC-MS 技术中克服基质效应提供一种简单的操作条件选择指南，以便在最短的时间内获得最佳结果。该方法可以应用于不同领域，如药物、生物分析、环境和食品科学。根据所需的灵敏度，分析人员可以最小化或补偿 ME。在灵敏度至关重要的情况下，必须通过调整 MS 参数、色谱条件或优化净化来尽量减少 ME。相反，为了补偿 ME，分析人员应该根据空白基质的可用性来使用校准方法。当有空白基质时，可以通过同位素标记内标和基质匹配校准标准进行校准；相反，当没有空白基质时，可以通过同位素标记内标、背景扣除或替代基质进行校准。在任何情况下，都需要调整 MS 参数、色谱条件或进行净化。
