iMed.ULisboa, Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal.
LAQV-REQUIMTE, Department of Chemistry, NOVA School of Science and Technology, NOVA University, Caparica, Portugal.
J Sci Food Agric. 2024 Nov;104(14):8561-8565. doi: 10.1002/jsfa.13683. Epub 2024 Jun 21.
Green tomato extracts, an agro-food industry waste, are rich in the glycoalkaloid tomatine, which presents activity against several diseases. High-performance liquid chromatography (HPLC) with ultraviolet (UV) detection is one of the most used techniques for quantification of bioactive compounds. The aim of this study was to optimize and validate a selective HPLC method with diode array detector (DAD) for the quantitative analysis of tomatine extracted from green tomatoes by subcritical water.
Chromatographic runs were performed on a InertSustain Phenyl (250 mm × 4.6 mm, 5 μm) analytical column, at a wavelength of 205 nm. A concentration range of 50-580 μg mL was used. The validation process was performed considering the linearity, precision, trueness, limit of detection (LOD) and limit of quantitation (LOQ) of the method. The selected mobile phase composed of acetonitrile and a solution of 20 mmol L potassium dihydrogen phosphate (KHPO) pH 3, resulted in suitable retention times and a standard calibration curve with adequate linearity (R = 0.9999). The method trueness was evaluated by the recovery assay, obtaining a mean recovery of 105% and the precisions were 1.4% and 0.9% (percentage relative standard deviation, RSD%) for the tomatine standard and extract samples, respectively. The inter-day variability was 2.7-9.0% (RSD%) for the standards and 6.9% (RSD%) for extract. The LOD and the LOQ of the method were determined at 8.0 and 24.1 μg mL, respectively.
The herein described method was successfully used for the quantification of tomatine in a tomato-derived extract. Furthermore, the method constitutes a simple and rapid analytical approach able to be used as a routine protocol. © 2024 Society of Chemical Industry.
绿番茄提取物是农业食品工业的一种废弃物，富含糖苷生物碱龙葵碱，对多种疾病具有活性。高效液相色谱（HPLC）与紫外（UV）检测是定量分析生物活性化合物最常用的技术之一。本研究旨在优化和验证一种选择性 HPLC 方法，该方法采用二极管阵列检测器（DAD），用于从绿番茄中用亚临界水提取的龙葵碱进行定量分析。
在 InertSustain Phenyl（250 mm×4.6 mm，5 μm）分析柱上进行色谱运行，检测波长为 205 nm。使用 50-580 μg mL 的浓度范围。验证过程考虑了方法的线性、精密度、准确度、检测限（LOD）和定量限（LOQ）。选择的流动相由乙腈和 20 mmol L 磷酸二氢钾（KHPO）pH 3 溶液组成，得到了合适的保留时间和标准校准曲线具有良好的线性（R = 0.9999）。通过回收试验评估了方法的准确度，得到了 105%的平均回收率，标准和提取物样品的精密度分别为 1.4%和 0.9%（相对标准偏差，RSD%）。标准的日间变异性为 2.7-9.0%（RSD%），提取物的为 6.9%（RSD%）。方法的 LOD 和 LOQ 分别为 8.0 和 24.1 μg mL。
本文描述的方法成功地用于番茄衍生提取物中龙葵碱的定量分析。此外，该方法构成了一种简单快速的分析方法，可作为常规方案使用。 © 2024 化学工业协会。
