摘要
对一种新的 HPLC 方法进行了优化和验证,用于测定土耳其特有物种Asparagus lycicus植物中的天冬酰胺,并对该植物进行了植物化学分析。对于天冬酰胺分析,在甲酸 (FA): 甲酸铵 (AF) 缓冲液作为流动相的情况下,使用 C18 反相柱使用水: 甲醇混合物 (95:5, v/v) 获得最佳分离效果。在这种新方法中,天冬酰胺的保留时间(0.61 分钟)非常短。研究了线性、准确度、精密度、检测限 (LOD)、定量限 (LOQ) 和选择性参数,以验证优化方法。通过这种方法,天冬酰胺分析获得了令人满意的回收率 (100.10 ± 2.86%),LOD 和 LOQ 值确定为 0.28 mg L-1和0.92mg L -1,分别。该方法的冻融稳定性相当高。基质效应实验结果表明,芦笋植物基质对天冬酰胺的定量分析没有显着影响。在地方性芦笋 lycicus植物的植物化学表征中,天冬酰胺含量(6.14 mg g -1植物)、总酚(13.22 mg GAE g -1植物)和黄酮类化合物(3.24 mg CE g -1植物)含量、酚类和黄酮类成分( 28.8 毫克迷迭香酸、109.2 毫克阿魏酸、531.2 毫克绿原酸、2.8 毫克邻香豆酸、17.6 毫克p-香豆酸和 6.0 mg 芥子酸/kg 植物)、总抗氧化活性(13.2 mg mg -1 DPPH)、维生素 C 含量(234.6 mg kg -1植物)、有机酸含量如苹果酸(2.9 mg kg -1植物) )、糖成分(每公斤植物 25.15 g 果糖和 18.25 g 葡萄糖)、总花青素含量(5.15 mg kg -1植物)、总油含量(9.65%)和脂肪酸成分(油酸(55.36%)和亚油酸)酸 (32.01%) 和棕榈酸 (7.61%))。这项研究的结果表明,地方性芦笋 lycicus植物可能被认为是具有抗氧化和生物活性成分的有前途的天然来源,在制药和食品工业的潜在应用中。
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Optimization and Validation of a New HPLC Method for the Determination of Asparagine Active Ingredient in Asparagus lycicus and Phytochemical Characterization of Endemic Asparagus lycicus Specie
Abstract
A new HPLC method was optimized and validated for the determination of asparagine in Asparagus lycicus plant, an endemic specie found in Turkey, and the phytochemical analysis of this plant was carried out. For asparagine analysis, optimum separation was obtained with water:methanol mixture (95:5, v/v) in the presence of formic acid (FA):ammonium formate (AF) buffer as mobile phase by using a C18 reversed phase column. In this new method, retention time (0.61 min) of asparagine was very short. The linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), and selectivity parameters were studied for the validation of optimized method. By this method, satisfactory recovery (100.10 ± 2.86%) was obtained for asparagine analysis and LOD and LOQ values were determined to be 0.28 mg L−1 and 0.92 mg L−1, respectively. Freeze-thaw stability of the method is quite high. The results of matrix effect experiments showed that the matrix of Asparagus lycicus plant does not significantly affect the quantification analysis of asparagine. In the phytochemical characterization of endemic Asparagus lycicus plant, asparagine content (6.14 mg g-1 plant), total phenolic (13.22 mg GAE g-1 plant) and flavonoid (3.24 mg CE g-1 plant) contents, phenolic and flavonoid components (28.8 mg rosmarinic acid, 109.2 mg ferulic acid, 531.2 mg chlorogenic acid, 2.8 mg o-coumaric acid, 17.6 mg p-coumaric acid and 6.0 mg sinapic acid per kg plant), total antioxidant activity (13.2 mg mg-1 DPPH), vitamin C content (234.6 mg kg-1 plant), organic acid content as malic acid (2.9 mg kg-1 plant), sugar components (25.15 g fructose and 18.25 g glucose per kg plant), total anthocyanin content (5.15 mg kg-1 plant), total oil content (9.65%) and fatty acid components (oleic acid (55.36%), and linoleic acid (32.01%) and palmitic acid (7.61%)) were determined. The results of this study exhibited that endemic Asparagus lycicus plant might be considered as a promising natural source with antioxidant and bioactive constituents in the potential applications of pharmaceutical and food industries.