LAQV, REQUIMTE, Laboratory of Applied Chemistry, Department of Chemical Sciences, Faculty of Pharmacy, University of Porto, Porto, Portugal.
LAQV, REQUIMTE, Department of Chemistry, University of Aveiro, Aveiro, Portugal.
Crit Rev Anal Chem. 2024;54(8):3038-3065. doi: 10.1080/10408347.2023.2221731. Epub 2023 Jun 19.
Obesity is a disease of epidemic proportions with a concerning increasing trend. Regarded as one of the main sources of energy, lipids can also represent a big part of an unnecessary intake of calories and be, therefore, directly related to the problem of obesity. Pancreatic lipase is an enzyme that is essential in the absorption and digestion of dietary fats and has been explored as an alternative for the reduction of fat absorption and consequent weigh loss. Literature describes a great variability of methodologies and experimental conditions used in research to evaluate the inhibitory activity of compounds against pancreatic lipase. However, in an attempt to choose the best approach, it is necessary to know all the reaction conditions and understand how these can affect the enzymatic assay. The objective of this review is to understand and summarize the methodologies and respective experimental conditions that are mainly used to evaluate pancreatic lipase catalytic activity. 156 studies were included in this work and a detailed description of the most commonly used UV/Vis spectrophotometric and fluorimetric instrumental techniques are presented, including a discussion regarding the differences found in the parameters used in both techniques, namely enzyme, substrate, buffer solutions, kinetics conditions, temperature and pH. This works shows that both UV/Vis spectrophotometry and fluorimetry are useful instrumental techniques for the evaluation of pancreatic lipase catalytic activity, presenting several advantages and limitations, which make the choice of parameters and experimental conditions a crucial decision to obtain the most reliable results.
肥胖是一种流行疾病,且呈上升趋势,令人担忧。脂肪被认为是主要的能量来源之一,但其也可能代表了多余卡路里的摄入,而这与肥胖问题直接相关。胰脂肪酶是一种在吸收和消化膳食脂肪过程中必不可少的酶,它已被探索作为减少脂肪吸收和减轻体重的一种替代方法。文献描述了用于评估化合物对胰脂肪酶抑制活性的研究中存在着大量不同的方法和实验条件。然而,为了选择最佳方法,有必要了解所有的反应条件,并理解这些条件如何影响酶促测定。本综述的目的是了解和总结主要用于评估胰脂肪酶催化活性的方法和相应的实验条件。共纳入了 156 项研究,详细描述了最常用的紫外/可见分光光度法和荧光光度法两种仪器技术,包括对两种技术中所用参数差异的讨论,即酶、底物、缓冲溶液、动力学条件、温度和 pH。本研究表明,紫外/可见分光光度法和荧光光度法都是评估胰脂肪酶催化活性的有用仪器技术,它们具有各自的优点和局限性,因此参数和实验条件的选择是获得最可靠结果的关键决策。