The method optimization and validation for the determination of asparagine active ingredient in Asparagus lycicus (Likya asparagus) specie by high-performance liquid chromatography (HPLC) method and phytochemical characterization of Asparagus lycicus plant

In this study, the method optimization and validation of the asparagine in Asparagus lycicus plant which is an endemic specie of Turkey, were realized with high performance liquid chromatography (HPLC) method. Also, the phytochemical characterization of the plant was obtained by determining the total phenolic and flavonoid content, phenolic and flavonoid components, total antioxidant activity, vitamin C, organic acid components, sugar components, total anthocyanin content, total oil and fatty oil components. The optimum method conditions were determined as water:methanol:formic acid mixture (95:5:0.1 by volume) for mobile phase, as 0,3 mL/min for flow rate, as 35 °C for column temperature and as 3 µL for injection volume by studying different mobile phase, flow rate, column temperature and injection volume trials. The linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ) and selectivity parameters were obtained for performing the method validation. LOD and LOQ values were calculated as 0.28 mg/L and 0.92 mg/L, respectively. The biochemical content analyses were made to determine the phytochemical characterization of Asparagus lycicus plant. The asparagine content was determined as 6.14 mg/g plant. The total phenolic and total flavanoid contents were found to be 13.22 mg GAE/g plant and 3.24 mg CE/g plant, respectively. The amount of rosmarinic acid, ferulic acid, chlorogenic acid, o-coumaric acid, p-coumaric acid and sinapic acid were detected as 28.8, 109.2, 531.2, 2.8, 17.6 and 6.0 mg/kg plant, respectively in the phenolic and flavanoid component analyses. The total antioxidant activity was found to be 13.2 mg/mg DPPH. Vitamin C content was obtained as 234.6 mg/kg plant. The amount of malic acid was determined as 2.9 mg/kg plant in the organic acid component analysis. The quantity of fructose and glucose were obtained as 25.15 g/kg plant and 18.25 g/kg plant, respectively. The total anthocyanin content was determined as 5.15 mg/kg plant. In the oil analysis, the total oil content was found to be 9.65%. The main fatty acid components were detected as oleic acid (55.36%), linoleic acid (32.01%) and palmitic acid (7.61%).