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and nutraceutical industries because it is one of best source of bioactive components including, triterpinoids, polysaccharides, benzenoids, benzoquinone derivatives, and maleic/succinic acid derivatives. Due to the rareness and cost effect, scientist developed several
culture techniques for the mass production. Recent pre-clinical and clinical studies on human strongly suggest that Antrodia cinnamomea could be novel alternative phytotherapeutic agent, or a synergizer in the treatment of cancer and other immune-related diseases.
DETECTION OF COUMARIN BY HPLC ANALYSIS IN CINNAMON SPICES FROM TURKISH MARKETS
© N. Yagmur Kumser Diker1, Gurbet Ünal2, Zehra Büyüktuncer Demirel2, I. Irem Tatli Qankaya
1 Hacettepe University, Faculty of Pharmacy, Department of Pharmaceutical Botany, Ankara, Turkey;
2 Hacettepe University, Faculty of Health Sciences, Department of Nutrition and Dietetics, Ankara, Turkey
Cinnamon is generally used in many desserts to regulate insulin and blood glucose in Turkey. However, it can contain coumarin and the coumarin is a hepatotoxic natural compound found in different Cinnamomum species, specially in Cinnamomum cassia (L.) J. Presl. In this study, 22 spice samples and known cinnamon species [Cinnamomum cassia(L.) J. Presl and Cinnamomum verum J. Presl.] were performed using HPLC-DAD method. Each ground cinnamon sample (0.5 g) from different brands of the markets was extracted with 25 mL methanol (80%) by stirring with a magnetic stirrer at room temperature for 30 min. Samples were filtered and evaporated to dryness. Remain extract was adjusted to 5 mL in volumetric flask after dissolved in distillated water, then, the samples were lyophilized. Stock solutions of coumarin and samples (1 mg/mL) were prepared in methanol (50%). Although, the analysis method developed by Ding, Y. et al. was directly used for fingerprint analysis [1], for the detection of coumarin, a modified HPLC separation [2] was carried out using a reversed phase (RP-18, 5 ^m, 250 mm x 4.6 mm i.d.) column. The mobile phase made up of water: 5 mM ammonium acetate buffer, (0.2% (v/v) acetic acid)
(A) and acetonitrile/methanol 1:2 (v/v) (B) in a gradient elution: 0-2 min: 70% A; 2-30 min: 70-20% A; 30-35 min: 20-0% A; 35-45 min: 0% A; 45-55 min: 70% A. The sample injection volume was 5 and the flow rate was
0.8.mL/min. The analysis was performed at a wavelength of 279 nm. Even though, coumarin was not detected in standards and 4 samples, it was shown that 18 samples contained coumarin in the range between 0.3 and 4.9 mg in 1 g powder cinnamon. It is suggested that spice products should be evaluated for their quality according to tolerable daily intake (TDI) of 0.1 mg coumarin/kg body weight that limited by European Food Safety Authority (EFSA) [3]. Additionally, in fingerprint analysis, 2-methoxy cinnamaldehyde and eugenol peaks that were shown in known spices were observed in only one sample among 22 samples. This was also thought to depend on the quality of the product.
References:
1. Ding Y et al., 2011. Food Chemistry, 127:755-760.
2. Sproll C et al., 2008. Food Chemistry, 109:462-496.
3. EFSA. 2004. EFSA Journal, 104:1-36.
CHEMICAL COMPOSITION, ANTIOXIDANT AND ANTIMICROBIAL ACTIVITIES OF THE ESSENTIAL OILS (EOS) OF DIFFERENT PINUS SPECIES FROM KOSOVO
© Fatbardhe Kurti1,2, Giangiacomo Beretta2, Behxhet Mustafa13, Fabrizio Gelmini2, Avni Hajdari13
1 Department of Biology. Faculty of Mathematical and Natural Science. University of Prishtina, Prishtine, Kosovo;
2 Department of Pharmaceutical Sciences, Universita degli Studi di Milano, Milan, Italy;
3 Institute of Biological and Environmental Research. University of Prishtina, Prishtine. Kosovo
Chemical profile, antioxidant and antimicrobial activity of total and fractionated EOs (F1 - hexane, F2 -hexane/diethyl ether, F3 - diethyl ether) derived from five Pinus species (Pinus heldreichii, P. peuce, P. mugo, Pinus nigra, P. sylvestris), were investigated. The hydrodistilled
EOs and their chromatographic fractions (direct solid phase extraction, SPE) were analysed by GC-MS and 112 compounds separated and identified. The main constituents were a-pinene, p-pinene, D-limonene, p-caryophyllene, germacrene D, bornyl acetate and
Obzory po kliniceskoj farmacologii i lekarstvennoj terapii [Reviews of clinical pharmacology and drug therapy] vol. 15/2017/suppLement 1
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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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