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      Microwave-Assisted Extraction of Cannabinoids in Hemp Nut Using Response Surface Methodology: Optimization and Comparative Study

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          Abstract

          Hemp nut is commonly incorporated into several food preparations; however, most countries set regulations for hemp products according to their cannabinoid content. In this study, we have developed an efficient microwave-assisted extraction (MAE) method for cannabinoids (i.e., Δ9-tetrahydrocannabinol, cannabidiol, and cannabinol) in hemp nut. Optimization of the MAE procedure was conducted through single factor experiments and response surface methodology (RSM). A comparative study was also conducted to determine the differences in the extraction yields and morphology of hemp nut between MAE and reference extraction methods, namely heat reflux extraction (HRE), Soxhlet extraction (SE), supercritical fluid extraction (SFE), and ultrasound-assisted extraction (UAE). Among the independent variables in RSM, the temperature was the most significant parameter. The optimal conditions of MAE were as follows: extraction solvent of methanol, microwave power of 375 W, temperature of 109 °C, and extraction time of 30 min. Compared with reference extraction methods, MAE achieved the highest extraction yields of total cannabinoids in hemp nut (6.09 μg/g for MAE; 4.15 μg/g for HRE; 5.81 μg/g for SE; 3.61 μg/g for SFE; 3.73 μg/g for UAE) with the least solvent consumption and shortest time. Morphological observations showed that substantial cell rupturing occurred in the microstructure of hemp nut after MAE, indicating enhanced dissolution of the target compounds during the extraction process. The MAE method is thus a rapid, economic, and environmentally friendly extraction method that is both effective and practical for industrial applications.

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          Most cited references26

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          Ultrasonically assisted extraction (UAE) and microwave assisted extraction (MAE) of functional compounds from plant materials

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            Analytical-scale microwave-assisted extraction.

            Microwave-assisted extraction (MAE) is a process of using microwave energy to heat solvents in contact with a sample in order to partition analytes from the sample matrix into the solvent. The ability to rapidly heat the sample solvent mixture is inherent to MAE and the main advantage of this technique. By using closed vessels the extraction can be performed at elevated temperatures accelerating the mass transfer of target compounds from the sample matrix. A typical extraction procedure takes 15-30 min and uses small solvent volumes in the range of 10-30 ml. These volumes are about 10 times smaller than volumes used by conventional extraction techniques. In addition, sample throughput is increased as several samples can be extracted simultaneously. In most cases recoveries of analytes and reproducibility are improved compared to conventional techniques, as shown in several applications. This review gives a brief theoretical background of microwave heating and the basic principles of using microwave energy for extraction. It also attempts to summarize all studies performed on closed-vessel MAE until now. The influences of parameters such as solvent choice, solvent volume, temperature, time and matrix characteristics (including water content) are discussed.
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              Development of a new extraction technique and HPLC method for the analysis of non-psychoactive cannabinoids in fibre-type Cannabis sativa L. (hemp)

              The present work was aimed at the development and validation of a new, efficient and reliable technique for the analysis of the main non-psychoactive cannabinoids in fibre-type Cannabis sativa L. (hemp) inflorescences belonging to different varieties. This study was designed to identify samples with a high content of bioactive compounds, with a view to underscoring the importance of quality control in derived products as well. Different extraction methods, including dynamic maceration (DM), ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE) and supercritical-fluid extraction (SFE) were applied and compared in order to obtain a high yield of the target analytes from hemp. Dynamic maceration for 45min with ethanol (EtOH) at room temperature proved to be the most suitable technique for the extraction of cannabinoids in hemp samples. The analysis of the target analytes in hemp extracts was carried out by developing a new reversed-phase high-performance liquid chromatography (HPLC) method coupled with diode array (UV/DAD) and electrospray ionization-mass spectrometry (ESI-MS) detection, by using an ion trap mass analyser. An Ascentis Express C18 column (150mm×3.0mm I.D., 2.7μm) was selected for the HPLC analysis, with a mobile phase composed of 0.1% formic acid in both water and acetonitrile, under gradient elution. The application of the fused-core technology allowed us to obtain a significant improvement of the HPLC performance compared with that of conventional particulate stationary phases, with a shorter analysis time and a remarkable reduction of solvent usage. The analytical method optimized in this study was fully validated to show compliance with international requirements. Furthermore, it was applied to the characterization of nine hemp samples and six hemp-based pharmaceutical products. As such, it was demonstrated to be a very useful tool for the analysis of cannabinoids in both the plant material and its derivatives for pharmaceutical and nutraceutical applications.
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                Author and article information

                Journal
                Molecules
                Molecules
                molecules
                Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry
                MDPI
                1420-3049
                03 November 2017
                November 2017
                : 22
                : 11
                : 1894
                Affiliations
                [1 ]School of Pharmacy, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; wxes9050304@ 123456gmail.com (C.-W.C.); date0315@ 123456hotmail.com (C.-C.Y.)
                [2 ]Chinese Medicine Department, Ditmanson Medical Foundation, Chiayi Christian Hospital, Chiayi City 60002, Taiwan; cych07093@ 123456gmail.com
                [3 ]Department of Sports Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; meichich@ 123456kmu.edu.tw
                [4 ]Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung 80708, Taiwan
                Author notes
                [* ]Correspondence: ytwu@ 123456kmu.edu.tw ; Tel.: +886-7-3121101 (ext. 2254); Fax: +886-7-3120683
                Article
                molecules-22-01894
                10.3390/molecules22111894
                6150297
                29099795
                43468b5d-65c5-4d20-bace-ca3b861f7a88
                © 2017 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 07 October 2017
                : 02 November 2017
                Categories
                Article

                δ9-tetrahydrocannabinol,cannabidiol,cannabinol,marijuana,cannabis

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