Simultaneous Extraction Optimization and Analysis of Flavonoids from the Flowers of Tabernaemontana heyneana by High Performance Liquid Chromatography Coupled to Diode Array Detector and Electron Spray Ionization/Mass Spectrometry
Flavonoids are exploited as antioxidants, antimicrobial, antithrombogenic, antiviral, and antihypercholesterolemic agents. Normally, conventional extraction techniques like soxhlet or shake flask methods provide low yield of flavonoids with structural loss, and thereby, these techniques may be considered as inefficient. In this regard, an attempt was made to optimize the flavonoid extraction using orthogonal design of experiment and subsequent structural elucidation by high-performance liquid chromatography-diode array detector-electron spray ionization/mass spectrometry (HPLC-DAD-ESI/MS) techniques. The shake flask method of flavonoid extraction was observed to provide a yield of ？(mg/g tissue). With the two different solvents, namely, ethanol and ethyl acetate, tried for the extraction optimization of flavonoid, ethanol (80.1？mg/g tissue) has been proved better than ethyl acetate (20.5？mg/g tissue). The optimal conditions of the extraction of flavonoid were found to be 85°C, 3 hours with a material ratio of 1？:？20, 75% ethanol, and 1 cycle of extraction. About seven different phenolics like robinin, quercetin, rutin, sinapoyl-hexoside, dicaffeic acid, and two unknown compounds were identified for the first time in the flowers of T. heyneana. The study has also concluded that L16 orthogonal design of experiment is an effective method for the extraction of flavonoid than the shake flask method. 1. Introduction Herbal or medicinal plant products, in various forms, have been used to treat different illness for many hundreds of years across the world. About 70–80% of the world population, particularly in the developing countries, rely on nonconventional medicine in their primary healthcare [1]. India has a rich flora that is widely distributed throughout the country, and a large number of Indian medicinal plants are attributed with various pharmacological activities, because of diversified class of phytochemicals, but still, the efficacy of these plants are yet to be scientifically documented [2]. In general, phytochemical constituents are essential for the survival and proper functioning of plants. They provide protection against herbivores, microorganisms, and competitors, regulate growth (e.g., delaying seed germination until an appropriate time), and control pollination, fertilization, and rhizosphere environment [3]. The main secondary metabolite present in plants includes lignins, flavonoid, phenols, alkaloids, amino acid derivatives, organic acids, terpenoids, steroids, and sugar derivatives. Among different phytochemicals, flavonoid exerts a wide
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