Xác định thành phần và hoạt tính sinh học của một số nhóm hợp chất chính trong quả tai chua

Biotechnology and Seedling JOURNAL OF FORESTRY SCIENCE AND TECHNOLOGY NO. 2 - 2018 10 DETERMINATION OF SOME KINDS OF IMPORTANT INGREDIENTS OF COMPONANTS AND BIOLOGICAL ACTIVITY OF GARCINIA COWA FRUIT Nguyen Thi Thu Hang1, Nguyen Van Thanh2, Nguyen Thi Hong Nhung3 1,2,3Vietnam National University of Forestry SUMMARY Tai Chua (Garcinia cowa Roxb.) in the plant family of Clusiaceae was a wealthy medical resource. For the industrial use in fruit processing, we need to understand the components extracted in Tai Chua and their bio- activity as well. We found that Tai Chua fruits had 90.28 - 93.43% humidity, 3.05 - 4.36% total acids, 3.03 - 4.34% total orgnanic acids, 0.7 - 0.74% total carbonhydrates, and 57.75 - 64.75 mg/100 g vitamin C. The extract in ethanol showed the high anti-bactrial effects. Three fruit stages were tested, including the young, ripen and over-ripen. The ripen fruits had high nutrients, vitamin C, and strong antibactrial activities. So it had a poten for the production of the foods originated from Tai Chua. The foods would be able to improve detoxicity, anti-imfamatory, antioxidants, slow aging and immunity. Keywords: Acorn, antimicrobial activity, biochemical composition, organic acid, vitamin C. I. INTRODUCTION Garcinia cowa Roxb., Family Clusiaceae distributed in India, South Thailand, Vietnam, Myanmar. In the world, there have been many published studies on the pharmacological effects of secondary compounds extracted from the tree. Garcinia cowa fruit contain high levels of flavonoids, phloroglucinol and xanthone, have anticancer activity, antioxidant, anti- inflammatory, antimicrobial (Hamidi et al., 2017; Shen et al., 2006). In Vietnam, tilapia can be as high as 30 m, distributed more in northern provinces (Lao Cai, Ha Giang, Vinh Phu, Hoa Binh, Bac Thai, Lang Son), in tropical forests, 200 - 800 m above sea level, average temperature 22 - 260C, rainfall 1,500 - 2,000 mm. In terms of ecological characteristics, acacia is preferred for high humidity but water and weathered soil from sandstone, schist, granite or limestone. Acorn is usually grown for fruit picking, timber, greening, anti-erosion. Among the parts of the acorn, the fruit is rich in acidity with a sour taste, so it is often used as an additive in the food additive, besides, the fruit is also used as Traditional medicine to detoxify, reduce fever (Sriyatep et al., 2015; Wahyuni et al., 2017). Garcinia cowa, despite its many uses, is a valuable source of medicinal herbs that need to be planted and harvested in a directed manner. However, in our country the harvesting and processing of plant products is only by experience. It has not been standardized both quantitatively and qualitatively, so it is difficult to allow the production of high quality sourced, high quality yogurt products. Contribute to the creation of scientific basis for the processing of tangerine fruits into foods that are nutritious and detoxifying, increasing immunity and energy for the body, publishing scientific information. This study demonstrates some of the biochemical and biological effects of acorn fruit. Biotechnology and Seedling JOURNAL OF FORESTRY SCIENCE AND TECHNOLOGY NO. 2 - 2018 11 II. RESEARCH METHODOLOGY 2.1. Research materials The fruits of the mature, unprotected trees, use three types of fruit: (i) fruit buns (large size, blue shells, firm fruit and blue-white); (ii) ripened fruits (large size, bright yellow shell and pulp, firm fruit); (iii) ripe berries (large size, yellow pods and flesh, soft flesh). Test organisms include E. coli, Bacillus subtilis (Gram), Aspergillus niger, Pythium (root rot), yeast Saccharomyces cerevisiae. 2.2. Research Methods Determination of amount of water by drying method: Using heat to evaporate water in the sample. Weigh the sample before and after drying, and then calculate the percentage of water in the sample. - Analysis of total acid content, total organic acid content, total sugar content, vitamin C content by common biochemical methods: total acidity determined by quantitative method with an alkaline solution standard; the organic acid content determined by the method of heating the sample to be combined with the quantity by a standard alkaline solution; Determine total sugar content by DNS method, use glucose to build up the standard graph; Determination of vitamin C content by titration with Iodine 0.001 N solution (Pham Van Cao, Bui Thi Nhu Thuan, 1991). - Determination of antimicrobial activity of the test: Trim the sample of the acorn, soaking the sample in 85% ethanol solvent for 30 days. The extract is then filtered and the aqueous solution is obtained until it is viscous. Highly diluted with water in a high proportion of 9 parts water is high crude solution. Determination of activity against five types of microorganisms tested (Escherichia coli, Bacillus subtillis, Aspergillus niger, Pythium, Saccharomyces cerevisiae) by punching and measuring the diameter of microbial resistance ring formed around the hole after 72 h Inoculate microorganisms on the environment and appropriate temperatures for each type of microorganism. Data collected were processed by Microsoft Excel software to calculate mean, standard deviation. Comparison of data differences between experimental formulas by means of statistical analysis of mean quantities. The mean values were statistically significant different when p < 0.05 (Chu Van Man, 2009). III. RESULTS AND DISCUSSION 3.1. Results of the determination of amount of water of Garcinia cowa fruit The amount of water present in the tangerine fruit at three stages (ripening, ripening, ripe), as shown in table 1. The results show that the average moisture content of the fruit at the fruit stage 92.56% fresh fruit, 90.28% ripening fruit, 93.43% ripe fruit. The moisture content valuescorresponding to the different development periods of Garcinia cowa fruit were statistically analyzed, and the results showed that the difference between the values was statistically significant. Usually, of the same fruit, the moisture content reflects the dry matter content in the fruit: the high moisture content corresponds to the high water content and the dry matter content (nutrient content) is low, and vice versa. Applying this rule in comparison with the dry matter content of the three fruits, the ripening fruit has the highest dry matter content, and the ripe fruit has the lowest dry matter content. Biotechnology and Seedling JOURNAL OF FORESTRY SCIENCE AND TECHNOLOGY NO. 2 - 2018 12 Table 1. Amount water at different stages of fruit development Type of fruit Sample symbol Weight of cup (g) Sample weight (g) Weight of cup and sample (g) Moisture (% w/w) Average moisture (% w/w) Before drying After drying Immatrure Bt1 30.3587 10.0065 40.3652 31.0969 92.6228 92.56b ± 0.10 Bt2 32.4365 10.0098 42.4463 33.1703 92.6692 Bt3 29.7139 10.0645 39.7784 30.4514 92.6723 Bt4 31.1056 10.0005 41.1061 31.8603 92.4534 Bt5 31.2195 9.9958 41.2153 31.9759 92.4328 Bt6 30.9160 10.0033 40.9193 31.6661 92.5015 Matrure Cth1 32.2430 9.9987 42.2417 33.2163 90.2657 90.28c ± 0.17 Cth2 29.8122 10.0261 39.8383 30.8085 90.0629 Cth3 30.3010 10.0258 40.3268 31.3026 90.0098 Cth4 30.5056 10.0343 40.5399 31.4638 90.4508 Cth5 30.3492 10.0383 40.3875 31.3124 90.4047 Cth6 29.9409 10.0160 39.9569 30.8924 90.5002 Over matrure Cm1 31.3740 10.0074 41.3814 32.0250 93.4948 93.43a ± 0.17 Cm2 30.5192 10.0065 40.5257 31.1848 93.3483 Cm3 30.9442 10.0002 40.9444 31.5692 93.7501 Cm4 31.3459 9.9991 41.3450 32.0188 93.2704 Cm5 29.3668 10.0256 39.3924 30.0271 93.4139 Cm6 31.0775 10.0053 41.0828 31.7457 93.3215 Different letters in the same column show statistically significant differences with p < 0.05. 3.2. Analysis of some biochemical components In biochemical norms related to the quality of food (vegetables, fruits), the total acid content of which contains mainly organic acids and vitamins plays an important role. The results showed that the acidity and total organic acids, total sugar, and vitamin C content of Garcinia cowa fruit at different stages of development (Table 2): Total acidity and total organic acids: Total acids include all organic and inorganic acids present in the fruit. Normally, in the plant body, acids usually exist in the form of organic acids. According to Ritthiwigrom et al. (2013), acetic acid in the acorn consists mainly of organic acids such as malic acid, tartaric acid, acetic acid. Analysis of total acidity and total organic acids in sour cherries also showed that, with total acidity as a percentage, when the fruit reached maturity, fruit and fruit ripening was high (3.05% to 4.36%), which accounts for the majority of organic acids (3.03% to 4.34%). In particular, the total acidity and total organic acids at the ripening stage reached the highest and the ripening stage was the lowest. Comparing the organic acidity of the fruit with other nutrient- rich fruits, the organic acidity of the high- yielding of Garcinia cowa fruit is comparable to that of apricots, plums, oranges. The total sugar content in the fruit is low, only 0.7% (fruit cake) to 0.74% (ripe fruit). Thus, the sugar/acid ratio in the taurine is low, about 1/6, and that makes the fruit sour. Vitamin C is also known as ascorbic acid - an organic acid, and in total organic acids with vitamin C. However, given the particularly important role of vitamin C in human and animal health (increasing immune system, anti-aging, antioxidant), vitamin C content is determined Biotechnology and Seedling JOURNAL OF FORESTRY SCIENCE AND TECHNOLOGY NO. 2 - 2018 13 separately. The results showed that vitamin C content in the taurine fruit corresponding to different stages of development is different. The vitamin C content in ripe fruits is higher than that of 64.75 mg/100 g, the fruit is 62.41 mg/100 g, and the fruit is ripe with 57.75 mg/100 g. In general, the vitamin C content of the fruit is high, equivalent to high vitamin C fruits such as grapefruit, orange, lemon, strawberry. The results of the study are also consistent with the publication by Sarma et al. (2014) of the vitamin C content of tilapia. Table 2. Some biochemical norms of the fruit in the development stages Fruit type Full fruit acid (% w/w) Organic Acid (% w/w) Gross Road (% w/w) Vitamin C (mg/100 g) Nine 3.99b ± 0.17 3.97b ± 0.08 0.70b ± 0.03 62.41ab ± 1.82 Nine harvested 4.36a ± 0.19 4.34a ± 0.06 0.73a ± 0.03 64.75a ± 1.34 Nine muzzle 3.05c ± 0.14 3.03c ± 0.06 0.74a ± 0.02 57.75b ± 1.37 Different letters in the same column show statistically significant differences with p < 0.05. 3.3. Results of antimicrobial activity determination Screening of antagonistic activity against microorganisms including bacteria (Bacillus subtilis, E. coli), mold (Aspergillus niger, Pythium), yeast (Saccharomyces cerevisiae) for the three phases of development, there was a strong activity against all five test organisms, with the inverse diameter of the extracted juice when not diluted with each microorganism, as shown in table 3. The antimicrobial activity of the extracted sour cherubic extract was apparent not only with the untreated fruit extract, but also with 10 times dilution (Figure 1). The results were consistent with the findings of Jabit et al. (2009), which confirmed that the fruit was rich in secondary compounds and resistant to microorganisms. Table 3. Vitamin C content of Garcinia cowa fruit according to periods of development Type of fruit Diameter of resistance ring (mm) B. subtilis E. coli A. niger Pythium S. cerevisiae Nine 3.4 ± 0.05 3.0 ± 0.02 4.0 ± 0.01 2.6 ± 0.02 2.4 ± 0.01 Nine harvested 3.4 ± 0.07 3.1 ± 0.03 4.2 ± 0.06 2.6 ± 0.02 2.3 ± 0.01 Nine muzzle 3.2 ± 0.02 3.0 ± 0.05 4.2 ± 0.04 2.5 ± 0.04 2.3 ± 0.03 (A) (B) Fig. 1. Bacillus subtilis (A) and Pythium (B) antimicrobial activity of fruit ripening fruit extracted at the non-diluted fruit (right) and diluted 10 times (left) Biotechnology and Seedling JOURNAL OF FORESTRY SCIENCE AND TECHNOLOGY NO. 2 - 2018 14 IV. CONCLUSION Garcinia cowa (Tai chua) fruit has an organic acid content, high vitamin C content and strong anti-microbial activity. Harvesting of fruit in matrure for nutrient quality and highest vitamin C content. It is suitable as a source of natural ingredients to extract biologically active substances used in medicine and pharmacy, and processed into foods rich in nutritional value, increasing the ability to detoxify and resist inflammation, antioxidant, anti-aging and increased immunity for humans and animals. REFERENCES 1. Chu Van Man (2009). Informatics in Biotechnology. Education publisher. 2. Hamidi D., Aulia H., Susanti M. (2017). High performance thin layer chromatography: densitometry method for determination of rubraxanthone in the stem bark extract of Garcinia cowa Roxb. Pharmacognosy Res, 9(3): 230-233. 3. Jabit M. L., Wahyuni F. S., Khalid R., Israf D. A., Shaari K., Lajis N. H., Stanslas J. (2009). Cytotoxic and nitric oxide inhibitory activities of methanol extracts of Garcinia species. Pharmaceutical Biology, 47(11): 1019-1026. 4. Pham Van Cao, Bui Thi Nhu Thuan (1991). Food Testing. Hanoi Science and Technology Publishing House. 5. Ritthiwigrom T., Laphookhieo S., Pyne S. G. (2013). Chemical constituents and biological activities of Garcinia cowa Roxb. Maejo Int. J. Sci. Technol, 7(2): 212-231. 6. Sarma A., Sarmah P., Kashyap D., Kalita A. (2014). Evaluation of nutraceutical properties and antioxidant activity of Garcinia cowa Roxb ex choisy fruits in Assam (India). World Journal of Pharmacy and Pharmaceutical Sciences, 3(12): 853-859. 7. Shen J., Tian Z., Yang J. (2006). The constituents from the stems of Garcinia cowa Roxb, and their cytotoxic activities. Pharmazie, 62: 549-551. 8. Sriyatep T., Siridechakorn I., Maneerat W., Pansanit A., Ritthiwigrom T., Andersen R. J., Laphookhieo S. (2015). Bioactive prenylated xanthones from the young fruits and flowers of Garcinia cowa. J. Nat. Prod, 78(2): 265-271. 9. Wahyuni F. S., Arisanty D., Hayaty N. F., Juwita D. A., Almahdy (2017). Sub-acute toxicity study of the ethyl acetate fraction of Asam Kandis Rinds (Garcinia cowa Roxb.) on the liver and renal function in mice. Pharmacogn J., 9(3): 345-349. 10. Wahyuni F. S., Hui L. S., Stanslas J., Lajis N. H., Dachriyanus (2017). In vivo study of tetraprenyltoluquinone, an anticancer compounds from Garcinia cowa Roxb. J Young Pharm, 9(2): 296-298. XÁC ĐỊNH THÀNH PHẦN VÀ HOẠT TÍNH SINH HỌC CỦA MỘT SỐ NHÓM HỢP CHẤT CHÍNH TRONG QUẢ TAI CHUA Nguyễn Thị Thu Hằng1, Nguyễn Văn Thanh2, Nguyễn Thị Hồng Nhung3 1,2,3Trường Đại học Lâm nghiệp TÓM TẮT Cho đến nay chưa có nhiều nghiên cứu cơ bản về phân tích, đánh giá các giá trị về thành phần hoạt tính sinh học của quả cây Tai chua (Garcinia cowa fruit). Vì vậy, việc phân tích các thành phần hóa sinh và các hoạt động sinh học trong quả của G. cowa là tiền đề khoa học cho nghiên cứu tiếp theo cho sản xuất và chế biến sử dụng các sản phẩm từ G. cowa. Kết quả nghiên cứu cho thấy: hàm lượng nước 90,28 - 93,43%, tổng hàm lượng axit 3,05 - 4,36%, tổng hàm lượng axit hữu cơ 3,03 - 4,34%, tổng hàm lượng đường 0,70 - 0,74%, nồng độ vitamin C 57,75 - 64,75 mg/100 g. Chiết xuất ethanol của củ quả G. cowa có hoạt tính kháng khuẩn đáng kể. Quả cây Tai chua (Garcinia cowa fruit) là một loại trái cây có tuổi thọ cao và có tính kháng khuẩn cao, thích hợp sử dụng làm nguyên liệu chế biến các thực phẩm chất lượng cao, tăng khả năng giải độc, chống viêm, chống oxy hóa và gia tăng miễn dịch. Từ khoá: Axit hữu cơ, hoạt tính kháng khuẩn, quả Tai chua (Garcinia cowa fruit), thành phần sinh hóa, vitamin C. Received : 02/3/2018 Revised : 23/3/2018 Accepted : 02/4/2018