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2.5 g extract granules 3 times daily (inacceptable for European patients), we were able to develop a novel dry extract for pelletizing, thus concentrating the common Japanese daily dose of secondary metabolites from Shikunshito in just 3 tablets per day (Convenience!). Due to the fact that Shikunshito is used in the EU in
single prescription by Kampo doctors since more than 15 years and since centuries in Japan without negative effects, the presented work opens the possibility to apply for its official registration as a phytopharmacon in Europe.
DEVELOPMENT OF A NOVEL PROANTHOCYANIDIN ENRICHED GINKGO BILOBA L. LEAVE EXTRACT WITH IN VITRO AND IN VIVO NEUROPROTECTIVE EFFECTS
© KuchtaK.1, Qiao H.X.2, Huang H.B.2, Fang L.2, Chen Y.3, Wang R.W.24
1 National Institute of Health Sciences, Division of Pharmacognosy, Phytochemistry & Narcotics, Tokyo, Japan;
2 Zhejiang CONBA Pharmaceutical, Hangzhou, China;
3 Zhejiang University of Technology, Hangzhou, China;
4 Zhejiang Provincial Key Laboratory of TCM Pharmaceutical Technology, Hangzhou, China
A commercial refined extract of Ginkgo biloba L. leaves has been reported to protect brain tissue against cerebral ischemia/reperfusion (I/R) injury [1]. This activity is generally attributed to the antioxidant activity of its flavonoids and proanthocyanidins. However, the latter have not been adequately studied up to now. Therefore, the present study aims to assess the neuroprotective properties of a newly developed proanthocyanidin enriched G. biloba leave extract (GPE) - with >90% proanthocyanidins after resin adsorption - on cerebral I/R injury compared to commercial proanthocyanidins from grape seeds (Vitis vinifera L.) (GSP) and nimodipine as a positive control. In vitro, the tissue protective effect of GPE was measured using PC12 cells, cultured in 96-well plates. After the addition of GPE or GSP (0.625, 1.25, 2.50 ^g/ml) and 12 h of cultivation, cells were incubated with H2O2 (40 mM) for another 24 h. Cell damage was measured using a commercial LDH assay kit. GPE noticeably ameliorated the increase in LDH release and thus the respective decrease in cell viability. For the in vivo assay, male Sprague-Dawley rats were divided into 6 groups
(sham: 8, placebo: 25, GPE 80 mg/kg: 13, GPE 40 mg/kg: 13, GPE 20 mg/kg: 16, GSE 40 mg/kg: 18, nimodipine: 8). All non-sham animals were subjected to cerebral I/R injury by occluding the middle cerebral artery with a nylon suture that was removed after 2 h of ischemia to establish reperfusion. After recovery from anaesthesia, the rats were returned to their cages with free access to water and food. All animals were sacrificed 24 h after reperfusion. Coronal brain sections were stained in order to calculate infarct ratio. Malondialdehyde (MDA) and superoxide dismutase (SOD) levels in the brain tissue were measured using commercial assay kits. Under treatment of cerebral I/R injury in test animals with GPE, the death rate decreased, neurological dysfunctions were reduced, and both average infarct size and concentrations of MDA and SOD were significantly ameliorated as compared to the placebo group.
References:
1. Hu B, Sun S, Mei G, Chen L, Tong E, 2002. Chin Med J (Engl), 115(9):1316-1320.
CURRENT PROGRESS AND FUTURE PROSPECTS OF ANTRODIA CINNAMOMEA RESEARCH IN TAIWAN
© K.J. Senthil Kumar1, Sheng-Yang Wang12
1 Department of Forestry, National Chung Hsing University, Taiwan;
2 Agricultural Biotechnology Research Center, Academia Sinica, Taiwan
Antrodia cinnamomea (Syn. Antrodia camphorate or Taiwanofungus camphorates) is a unique medicinal mushroom endemic to Taiwan. In traditional Chinese medicine (TCM), A. cinnamomea is used for treating various human illness including, food poisoning, drug intoxication, diarrhea, abdominal pain, hypertention, skin irritation, inflammation and cancer. In natural habitat, this mushroom grows the inner sap of age old camphor tree
Cinnamomum kanehira Hay (Lauraceae). Accumulating scientific evidences (nearly 400 research articles) revealed that A. cinnamomea possess various therapeutic effects including, immunomodulation, hepatoprotectiion, neuroprotection, anti-oxidant, anti-inflammation, antihypertensive, anti-hyperlipedemic, anti-diabetic, anti-metastatic and anti-cancer activities. In recent years, this mushroom is starting to attract by pharmaceutical
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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
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