Mass Spectrometric Approaches for the Analysis of Phytosterols in Biological Samples

Plant sterols (phytosterols) are important structural components of plant cellular membranes, and they play a major role during development and metabolism. They have health-associated benefits, especially in lowering blood cholesterol levels. Because of their many health claims, there is a growing interest in their analysis. Although various analytical strategies have been employed in analyzing phytosterols, chromatography linked to mass spectrometry (MS) is superior due to its sensitivity. Furthermore, specificity and selectivity are enhanced by utilizing tandem mass spectrometry (MS/MS). This article reviews the various mass spectrometric strategies used for the analysis of phytosterols. It highlights the applications and limitations associated with each MS strategy in various sample matrixes such as plant, human, animal, food, and dietary supplements. GC-MS was historically the method of choice for analysis; however, the derivatization step rendered it tedious and time-consuming. On the other hand, liquid chromatography coupled to MS (LC-MS) simplifies the analysis. Many ionization techniques have been used, namely, electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI), and atmospheric pressure photoionization (APPI). APCI showed superiority in terms of ion intensity and consistency in ion formation, primarily forming [M + H - H₂O]⁺ ions rather than [M + H]⁺. In addition, matrix assisted laser desorption ionization (MALDI) as well as ambient mass spectrometry such as direct analysis in real time (DART) have also been evaluated.