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3-carene. The antioxidant activities of total EOs were lower than those of the corresponding fractions, with F2 the strongest in all cases. EOs and fractions showed different degrees of antibacterial efficacy against different microbial pathogens (moderately strong antimicrobial activity against C. albicans and C. krusei, while low or no activity against E. faecalis and E. coli strains). The detected inhibition zones and MICs for
the EOs and fractions were in the range of 14-35 mm and 0.125 - 1% (v/v), respectively. The components responsible for the antioxidant and antimicrobial activity were oxygenated monoterpenes and sesquiterpenes recovered in the polar EO fractions. These activities seem to be regulated by reciprocal interactions among the different subclasses of phytochemical species present in the EOs.
THE COMPOSITION OF HEMP-BASED FOOD SUPPLEMENTS IN THE SLOVENIAN MARKET
© Kusar A.1. Pravst I.1, Blaznik U.2, Ucakar V.2, Ivanos J.3, Jeretin B.3
1 Nutrition Institute, Slovenia,
2 National Institute of Public Health, Slovenia;
3 National Laboratory of Health, Environment and Food, Slovenia
The use and consumption of hemp-containing food supplements (HFS) in the EU is rapidly increasing and concerns have been raised about the quality and safety of products on the market, especially regarding their tetrahydrocannabinol (THC) and cannabidiol (CBD) content. Although the lowest observed effect level (LOEL) for THC in food products was given in the EFSA's scientific opinion (2.5 mg/kg) [1], Slovenian authorities [2] suggest a maximum daily dose of 2 mg of CBD (MDD) since there is a lack of appropriate safety studies. A study was performed to detect the availability of hemp food supplements in the Slovenian market and to determine the content of cannabinoids.
In the study, all HFS available in the Slovenian market were collected and purchased, totalling 13 different products. Products were in the form of hard capsules, tablets or drops. Samples for analyses were prepared using the QuEChERS extraction method. Determination of cannabinoids was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS), using previously reported method [3, 4].
The source of hemp in HFS was either sprout extract, seeds extract, oil, juice, seeds or sprouts. Higher amounts of THC were detected in four HFS (n = 4), ranging from 220 mg/kg to 1,000 mg/kg, while in the remaining products
(n = 9) the THC content was below LOEL (2.5 mg/kg) (1). When observing the CBD content, in eight HFS (n = 8) the content was negligible, while in the remaining products (n = 5) the amounts of CBD were higher, varying from 14,000 mg/kg to 62,000 mg/kg. With respect to data available from declarations and analytical data (the recommended daily dose was not provided for 2 HFSs), the LOEL and acceptable MDD for THC or CBD is exceeded in at least five HFS products (n = 5). The use of hemp-based food supplements in the EU is rapidly increasing and concerns have been raised about the quality and safety of products on the market. To ensure the safe use of hemp food supplements, it is recommended that the legislative limitations be enforced.
References:
1. EFSA Journal. 2015. 3(6):4141, 10.2903/j.efsa.2015.4141.
2. NIJZ pdf document 2016. Available online: https://goo.gl/ wLfqQu.
3. UCT: Determination of 35 Pesticides and 3 Cannabinoids in Marijuana Edibles.
4. Phenomenex: THC and Metabolites from Whole Blood on Strata-x-Drug B & Kinetex C18 by LC-MS-MS.
The work was supported by the Slov. Ministry of Health and Slov. Research Agency (V3-1639; P3-0395).
RAPID IDENTIFICATION OF BIOACTIVE CONSTITUENTS FROM VITIS AMURENSIS ROOTS USING LC-MS COUPLED WITH TYROSINASE INHIBITORY ASSAY
© Kyung-Eon Oh, Hyeji Shin and Ki Yong Lee
College of Pharmacy, Korea University, Republic of Korea
In the course of searching for tyrosinase inhibitory constituents from natural products, the methanolic extract of Vitis amurensis showed significant tyrosinase inhibitory activity. V. amurensis is family Vitaceae and
widely distributed in Asian countries. LC-MS coupled with tyrosinase inhibitory assay for simultaneous identification of active compounds from the extracts was used. Two peaks of total extracts of V. amurensis showed high
Obzory po kliniceskoj farmacologii i lekarstvennoj terapii [Reviews of clinical pharmacology and drug therapy]
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tyrosinase inhibitory activity. Two oligostilbenes (1-2) were isolated from the ethyl acetate fraction of V. amurensis by silica gel column chromatography. The structures of two compounds were determined on E-e-viniferin (1) and E-vitisin B (2) based on the analysis of the 1H-NMR, 13C-NMR and LC-MS data. All compounds exhibited
significant tyrosinase inhibitory activity (IC50 3.5 and 10.5 |M, respectively) and active compounds were well correlated with predicted them.
This research was supported by the National Research Foundation of Korea (NRF) funded by the Korea Government (No. 2017R1A2B4003403)
ANTI-PROLIFERATIVE ACTIVITY OF TELECTADIUM DONGNAIENSE AND ITS CONSTITUENTS VIA INHIBITION OF THE WNT/B-CATENIN SIGNALING PATHWAY IN HUMAN COLON CANCER CELLS
© Won Kyung Kim1, Jedo Oh1, Duc-Hiep Bach1, Hyung Won Ryu2, Hyen Joo Park1, Ji-Young Hong1, Hyuk-Hwan Song2, Sangmi Eum3, Tran The Bach4, Sang Kook Lee 1
1 College of Pharmacy, Seoul National University, Republic of Korea;
2 Natural Medicine Research Center, Korea Research Institute of Bioscience & Biotechnology, Republic of Korea;
3 International Biological Material Research Center, Korea Research Institute of Bioscience & Biotechnology, Republic of Korea;
4 Institute of Ecology and Biological Resources, Vietnam Academy of Sciences and Technology, Hanoi, Vietnam
Wnt/p-catenin signaling pathway is a potential target for the treatment of human colon cancer [1]. Thus, the inhibitory effects of various plant extracts on cell proliferation and Wnt signal transduction were evaluated to discover a Wnt signaling inhibitor. The present study aimed to investigate the cytotoxicity involved in Wnt pathway of the MeOH extract from Telectadium dongnaiense bark (TDB) and to identify its bioactive constituents by bioassay-guided fractionation.
The sulforhodamine B-based proliferation assay and the p-catenin/TCF-responsive reporter gene assay were employed as screening systems. The isolation and identification of compounds were elucidated on the basis of spectroscopic methods. Inhibitory effects on the expression levels of Wnt target genes were determined by real-time PCR and western blotting.
The extract of TDB most strongly inhibited cell proliferation and TOPflash activity (IC50 = 1.5 and 2.0 |ig/mL), which was correlated with its inhibitory effects on the expression of Wnt target genes. Three major compounds were isolated from bioactive fractions and were identified as 1,4-dicaffeoylquinic acid (1), quercetin 3-rutinoside (2), and periplocin (3). Only compound 3 showed anti-proliferative activity (IC50 = 0.06 |M) and exhibited Wnt signaling inhibitory effects in HCT116 colon cancer cells.
This study contributes to understanding the cytotoxic properties of TDB extract and its constituents and provides a potent strategy for its further application.
References:
1. Morin PJ, Sparks AB, Korinek V, Barker N, Clevers H, Vogelstein B, Kinzler KW. 1997. Science, 275:1787-1790.
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