Nghiên cứu ảnh hưởng của quá trình thu hoạch và chế biến đến hàm lượng các chất có hoạt tính sinh học trong cây thuốc dòi (pouzolzia zeylanica l benn) tóm tắt tiếng anh

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MINISTRY OF EDUCATION AND TRAINING CAN THO UNIVERSITY DOCTORAL THESIS SUMMARY Specialized: Food Technology Code: 9540101 NGUYEN DUY TAN EFFECT OF HARVESTING AND PROCESSING ON BIOACTIVE COMPOUNDS CONTENT IN MEDICINAL PLANT, CASE STUDY OF Pouzolzia zeylanica L Benn Can Tho, 2018 THE PhD THESIS WAS COMPLETED AT CAN THO UNIVERSITY Advisor: Assoc Prof Dr Nguyen Minh Thuy The doctoral thesis was evaluated by The Board of Examiners at basic level Meeting at: B007 Hall, School of Agriculture and Applied Biology, Can Tho University At 15.30 p.m., date 09/11/2017 Reviewer 1: Prof Dr Le Van Viet Man Reviewer 2: Assoc Prof Dr Truong Thi Minh Hanh You can find thesis at the library: Learning Resource Center, Can Tho University National Library of Vietnam LIST OF PUBLISHED WORKS Nguyen Duy Tan and Nguyen Minh Thuy, 2016 Optimization of concentration process on Pouzolzia zeylanica extract by response surface methodology Proceeding of The 23rd Regional Symposium on Chemical Engineering, 27-28 October 2016 at Vung Tau, Vietnam, page 196-205 Vietnam National UniversityHo Chi Minh Publishing House (ISBN 978-604-73-4690-5) Nguyen Duy Tan and Nguyen Minh Thuy, 2016 Optimal operating conditions of spray drying Pouzolzia zeylanica extract using maltodextrin and arabic gum as carrier Proceedings in the 18th Food Innovation Asia Conference, 16-18 June 2016 at BITEC Bangkok Thailan, page 499-509 Nguyen Duy Tanand Nguyen Minh Thuy, 2016 Optimization of carrier content (Maltodextrin, xanthan gum) for spray drying of Pouzolzia zeylanica extract using response surface methodology Science and Technology Journal of Agriculture and Rural Development, Ministry of Agriculture and Rural Development, Vietnam,11: 167-174 Nguyen Duy Tan and Nguyen Minh Thuy, 2017 Effect of extraction parameters to bioactive compounds content and antioxidant ability of Pouzolzia zeylanica extract Science and Technology Journal of Agriculture and Rural Development, Ministry of Agriculture and Rural Development, Vietnam, 8: 53-61 Nguyen Duy Tan, Le Quoc Viet, Vo Tan Thanh and Nguyen Minh Thuy, 2017 Optimization of polyphenol, flavonoid and tannin extraction conditions from Pouzolzia zeylanica L Benn using response surface methodology Can Tho University Journal of Science, 5: 122-131 Nguyễn Duy Tan and Nguyen Minh Thuy, 2017 Effect of drying temperature/sun drying and material size on dehydration and quality properties of dried Pouzolzia zeylanica L Benn i Science and Technology Journal of Agriculture and Rural Development, Ministry of Agriculture and Rural Development, Vietnam, 13: 90-98 Nguyễn Duy Tan and Nguyen Minh Thuy, 2017 Effect of maltodextrin concentration and spray drying conditions on physical properties and bioactive compounds of Pouzolzia zeylanica powder product Journal of Food and Nutrition Sciences, Vietnam Nutrition Association, 13 (5): 66-74 Nguyễn Duy Tan, Vo Thi Xuan Tuyen and Nguyen Minh Thuy, 2017 Comparative analysis of bioactive compounds content in red-purple and wild green Pouzolzia zeylanica collected from An Giang province Journalof Vietnam Agricultural Science and Technology, Vietnam Academy of Agricultural Sciences, (79): 80-84 Nguyen Duy Tan and Nguyen Minh Thuy, 2017 Optimization of carriers (maltodextrin, arabic gum) for spray-drying of Pouzolzia zeylanica extracts using response surface methodology Can Tho University Journal of Science, 6: 102-110 10 Nguyễn Duy Tan and Nguyen Minh Thuy, 2017 Optimization of carboxymethyl cellulose, sucrose and citric acid content added in concentration process of Pouzolzia zeylanica extract using response surface methodology Vietnam Journal of Chemistry, Vietnam Academy Science and Technology, 55 (4E23): 229-234 11 Nguyen Duy Tan, Vo Thi Xuan Tuyen and Nguyen Minh Thuy, 2017 Effect of cultivating season and harvesting time on antioxidant constituents in Pouzolzia zeylanica (L.) Benn Journal of Vietnam Agricultural Science and Technology, Vietnam Academy of Agricultural Sciences, (81): 61-67 12 Nguyen Duy Tan and Nguyen Minh Thuy, 2017 Effect of different gum types on stability of antioxidant components and physical properties of spray dried Pouzolzia zeylanica powder Vietnam Journal of Science and Technology, 55 (5A): 10-17 ii INTRODUCTION THE NECESSARY OF THESIS Pouzolzia zeylanica is one of the medicinal plants which people from many Asian countries have used to treat various kinds of diseases by traditional method In Vietnam, this plant was popularly cultivated in the Mekong Delta region, it can be used as fresh or dried plant, decoction drink to treat cough with phlegm, pulmonary tuberculosis, sore throat, etc A great number of studies showed that Pouzolzia zeylanica extract contains many bioactive compounds with antioxidant, antimicrobial and antifungal properties However, this species has not been fully researched on the possibility of growing and harvesting; preliminarily processing and preserving materials recently; as well as processing into convenient food products that can assist in the prevention and treatment of diseases Therefore, the essential content of this research is to meet the requirements the above issues OBJECTIVE OF THESIS Determining the harvesting time and postharvest treatment methods suitable for Pouzolzia zeylanica plant to obtain maximum the content of bioactive compounds and to develop the production process of two types of products (concentrated and spray dried powder) high quality and able to support health THE NEW CONTRIBUTIONS OF THESIS Providing useful information on the therapeutic effect of Pouzolzia zeylanica that is a medicinal plant commonly used in folk medicine; It has not been studied and found much Determining the harvesting time and appropriate drying methods to obtain the dried material with high levels of bioactive compounds, as well as the loss of these compounds during storage Using vacuum concentrating technology and spray drying to produce two value added products (concentrated and powder), convenient from Pouzolzia zeylanica plants Testing antioxidant activity in vitro and respiratory antibacterial ability, effect on reducing sputa and cough cure in the experimental mouse model The results of the research can guide people to plant in accordance with planning and promoting the production of products for commercialization, and increasing the economic value of Pouzolzia zeylanica plants LAYOUT OF THESIS The thesis consists of 160 pages with five chapters: Chapter 1: Introduction (pages 1÷4); Chapter 2: Literature review (page 5÷44); Chapter 3: Materials and method of research (page 45÷68); Chapter 4: Results and discussion (page 69÷158); Chapter 5: Conclusions and recommendations (page 159÷160) and 36 pages of references (page161÷196) Appendix consists of 68 pages with five parts: Appendix A: Some pictures in research process (page 1÷8); Appendix B: Some results of exploring studies (page 9÷36), Appendix C: Analytical methods (page 37÷42); Appendix D: Results statistical analysis of experimental data (page 43÷68) THE CONTENT OF THESIS Chapter 1: Overview 1.1 Introduction about Pouzolzia zeylanica plants Pouzolzia zeylanica plants belong to the Urticaceae family, the English name is Graceful Pouzolzsbush, originating in the tropics of southern Asia Nowadays, Pouzolzia zeylanica plants are present in many Asian countries and in other places of the world, are perennial herbaceous species that can be harvested year-around, but are best harvested at the end of the dry season Harvesting the trunk on the ground, cleaned, picking shoot plants to eat as raw vegetable or cut into small pieces, dry use gradually The whole plant is used as medicine, often used to treat diseases such as cough or tuberculosis, sore throat, lung disease, dysentery, enteritis, urinary tract infection, urinary retention, toothache, fungal infections Extract fom Pouzolzia zeylanica plants contains anthocyanin, flavonoid, polyphenol, tannin, alkaloid, saponin, isoflavone and other compounds These are compounds with many biological activities such as anti-cancer, antioxidant, strengthening the immune system, reducing the risk of cardiovascular disease, diabetes, antibacterial, anti-fungal The content of bioactive compounds in plants depends on the cultivating season, the growth time These compounds are also very sensitive to preparing process and processing conditions (drying, extraction, concentrating and spray drying) 1.2 Techological processes used in research Drying methods by sun drying/oven drying can damage some of the phytochemical compounds if the drying conditions are not appropriate The instability of phenolic compounds from plants can show the sensitivity of these compounds to drying methods However, in some cases, drying may be more favorable in the protection of bioactive compounds Extracting is method of obtaining bioactive compounds from different parts of plants by selective solvents The obtained extract is very complex and unstable, the content of active ingredients changes because it depends on a lot of factors: climate and geographic factors, cultivation mode, parts used, growth stage, level of processing, moisture content and material size, storage time In food technology the solvent is commonly used as water, followed by ethanol because it is environmentally friendly and less toxic Vacuum concentration is the process of evaporating water in food under the influence of heat in order to increase the content of the dry matter of the food In the concentration process, the input material is always in liquid form and the initial dry matter content is usually around 1035% After the concentration, the obtained product is also liquid and the dry matter content can be as high as 80% The vacuum concentrating spends a lot of energy cost, but the remarkable advantage of the method is that the dry matter content in the end can be very high, maintaining better than the flavor and the bioactive substance in the product Spray drying is a common process for transferring materials from liquid to powder for preservation, storage, transportation and distribution The spray drying process is suitable for drying sensitive heat-sensitive materials because of the product's thermal exposure time in the dryer is shortened One of the most popular advantages of spray drying is the ability to process a variety of materials and obtain good quality, high-quality compounds such as phenolic compounds, flavonoid, carotenoid, anthocyanin and tannin Some factors can affect the quality of spray dried powder such as dry matter concentration in extract before spray drying, type of drying agent, drying temperature, input flow rate, drying air flow rate Chapter 2: Materials and methods 2.1 Materials Materials: Pouzolzia zeylanica L Benn planted in An Giang University; sucrose RE (Bien Hoa Sugar Company); some food additives used as carboxymethyl cellulose (CMC), maltodextrin DE15, xanthan gum, k-carrageenan, arabic gum (LD Carlson, France); citric acid and ascorbic acid (Analar Normapur, Belgium) The chemicals used in the study include gallic acid, quercetin, tannic acid, folin-cioalteau, folin-denis, DPPH, TPTZ, and some other analytical chemicals were purchased from Sigma/Aldrich, USA and Merck, Germany or AR, China and Himedia, India 2.2 Research conetents Part 2.1: Study on effect of harvesting and post-harvesting treatment on the content of bioactive compounds in Pouzolzia zeylanica 2.1.1 Investigation ofthe effect of cultivating season and harvesting time on bioactive compounds The experiment was conducted on an area of 200 m2 at An Giang University Each plot was m2 and arranged in lots for one treatment Cutting (using red-purple trunk) was collected from households in Hoa Binh Commune, Cho Moi District, An Giang Province Planting of crop with a period of months from 1÷4 in 2015 (in the sunny season) and crop with a period of months from 7÷10 in 2015 (in the rainy season) The process of planting, tending and fertilizing is carried out according to a reference procedure, when the medicinal plants have the growth time (30, 45, 60, 75 and 90 days after planting) The physico-chemical characteristics of the tree were measured, such as: height, yield, moisture content, content of anthocyanin, flavonoid, tannin, polyphenol and antioxidant activity of extract (DPPH, FRAP and AAI) 2.1.2 Investigation of the effect of drying temperature/sun drying and material size on bioactive compounds With a sample weight of 350 grams, the medicinal plants harvested with optimum growth time in study 2.1.1 were washed, then cut to size of 3÷5 cm or the whole plant,spreading over a stainless steel tray (40 x 60 cm), drying at a temperature of 60°C to 100°C in a Hot-air Sterilizer (Model YCO-010, Taiwan) until the moisture content reached balance and control sample was sun drying Then, the samples were ground into fine flour through a mm diameter sieve, the powder was packed in PP and stored frozen, ready for determination of moisture content, carotenoids content, chlorophyll, anthocyanin, flavonoid, polyphenol, tannin and antioxidant extracts (DPPH, FRAP and AAI) 2.1.3 Investigation of the effect moisture content and material size on change bioactive compounds during storage time The samples were dried at the optimum drying temperature of study 2.1.2 until the moisture content reached 10.5% and a sample was 13.5% Then cut into pieces (3÷5 cm) or grinded into fine flour through a mm diameter sieve, the samples were stored in PP bags and in a black plastic bag at room temperature After each month, the samples were analyzed and evaluated for obtaining the content of bioactive compounds Part 2.2: Study on processing two products (concentrate and spray dried powder) from Pouzolzia zeylanica 2.2.1 Investigation of the effect of extracting conditions on bioactive compounds The extraction process of dried medicinal plants with water was carried out with the investigated factors including temperature (63, 70, 80, 90, 97oC), time (13, 20, 30, 40, 47 minutes) and the water/material ratio (17/1, 20/1, 25/1, 30/1, 33/1 v/w) Using the surface response method from the STATGRAPHIC PLUS XVI software for experimental layout and factors optimization Each factor was examined with levels (-α, -1, 0, +1, + α); with α = ± 1.682 The experiments were designed in a Central Composite Design 23+star model Total sample: 20 samples including replicates at the centeral point Extractor with 10 litters capacity, stirrer, temperature controller Each sample was taken liters Then filter through cotton and filter paper (Whatman's No.1); The analysis color absorption (Abs), soluble dry matter and the content of bioactive compounds 2.2.2 Investigation of the effect of blending citric acid, carboxymethyl cellulose (CMC) and oBrix into extraxt on sensory value and the content of bioactive compounds The blending process had strong effect on sensory values (color, taste and appearance) of concentrated product as well as stability of bioactive compounds The experimental design was conducted by using citric acid content ranging from 0.1 to 0.5%, Brix degree from 12 to 28oBrix and CMC content of 0.1 to 0.5% The experimental layout was presented similar in section of 2.2.1 Each sampleof 15 liters of extract was applied with optimal extraction conditions in the study 2.2.1 The concentration was conducted at a vacuum level of 600 mmHg for 40 minutes The sensory score and physico-chemical characteristics of the product were evaluated 2.2.3 Investigation of the effect of vacuum concentration process on sensory value and the content of bioactive compounds During vacuum concentration process, the pressure and time are two parameters which had a great influence on the quality of the product Thus, the experiment was performed with two factors: vacuum level (550, 600, 650 mmHg) and concentration time (30, 35, 40 minutes) Response surface methodology was applied to (a) 50,67a 60 Anthocyanin content (mgCE/100g FW) Anthocyanin content (mgCE/100g FW) 60 70 48,05b 44,66c 50 35,53d 30,55e 40 30 20 40 33,87b 30 20 10 10 45 60 Growth time (day) Flavonoid (mgQE/g FW) 75 Dry season 90 Polyphenol (mgGAE/g FW) Tannin (mgTAE/g FW) (c) 4,99a 4,51b 5,18a 4,34b 3,98c 3,97a 3,48b 3,55b 3,52b 2,93c 2,2ab 2,29a 2,11b 1,97c 1,59d 45 60 Growth time (day) 75 Dry season Wet season (d) 5,19a 4,01b 3,70a 3,29b 2,20a 1,86b 0 30 Wet season Cultivating season Bioactive compounds content 30 Bioactive compounds content (b) 49,91a 50 Flavonoid Polyphenol Tannin (mgQE/g FW) (mgGAE/g FW) (mgTAE/g FW) 90 5,67a 5,50a Antioxidant index (AAI) Antioxidant index (AAI) Figure 3.2: The average content of anthocyanin (a, b), flavonoid, polyphenol and tannin (c, d) with different growth time and cultivating season 4,95b 4,96b (a) 3,89c 5,48a 4,50b (b) 2 30 45 60 75 Dry season 90 Wet season Cultivating season Growth time (day) (c) (d) Figure 3.3: Antioxidant index AAI (a, b) and antioxidant activity (DDPH, FRAP) (c,d) with different growth time and cultivating season 3.1.2 Effect of drying temperature/sun drying and material size on dehydrated and quality properties of Pouzolzia zeylanica Results in Figure 3.4, Table 3.1 and 3.2 showed that the dehydrate ability during initial drying stage of cut samples was more slowly than whole plants, however the dehydrate was more quickly at the end of drying stage and the moisture content was low As the drying temperature increased from 10 60÷100oC, the drying time was reduced from 720 to 230 minutes The content of bioactive compounds and anitoxidant activity decreased the most at drying temperature of 70÷80oC The bioactive compounds contents and antioxidant activity of dried whole Pouzolzia zeylanica plants were maintained much more than the cut samples In addition, the study also analyzed the dynamics of thermal decomposition of bioactive compounds The results showed that the constant of degradation rate (k) of bioactive compounds increased with the drying temperature, but a difference between dried material size was found The activation energy (Ea) for degradation of anthocyanin, flavonoid, polyphenol and tannin contents ranged from 28.07÷46.37 kJ/mol and followed the first order model The drying temperature of 60oC or sun drying maintained well the quality of Pouzolzia zeylanica material and were selected as a parameter for subsequent studies (a) (b) Figure 3.4: Plots show moisture reducing of Pouzolzia zeylanica plants during drying process in whole plants (a) and cut short forms (b) Table 3.1: Effect of temperature and material size on the content of bioactive compounds of Pouzolzia zeylanica Material forms Whole plant Cut short sample Sun drying/drying temperature (oC) Sun drying 60 70 80 90 100 Anthocyanin (mgCE/100g) 22.68a 17.69b Anthocyanin (mgCE/100g) 37.54a 35.87b 22.39c 9.35d 8.31e 7.64e Flavonoid (mgQE/g) 5.30a 4.91b Flavonoid (mgQE/g) 5.68a 5.83a 4.76bc 4.52c 4.85bc 4.98b Polyphenol (mgGAE/g) 10.35a 9.43b Polyphenol (mgGAE/g) 12.00a 11.67a 8.17d 8.41d 9.14c 9.97b Tannin (mgTAE/g) 6.78a 6.22b Tannin (mgTAE/g) 7.10a 7.25a 5.86c 5.92c 6.15bc 6.70ab Notes: theexperimental meanings with different characters in the same column showed statistically significant differences (P0.01) 11 Table 3.2: Effect of temperature and material size on the antioxidant activity FRAP DPPH Chlorophyl Carotenoid Material forms AAI (%) a (mg/g) (mg/g) (MFeSO4/g) Whole plant 7.95a 64.89a 85.30a 0.455a 1.714a b b b b Cut short sample 7.31 62.50 82.91 0.429 1.627b FRAP Sun drying/drying DPPH Chlorophyl Carotenoid AAI temperature (oC) (%) (mg/g) (mg/g) (MFeSO4/g) Sun drying 8.62a 66.37ab 87.31a 0.468a 1.663c a a a a 60 8.52 66.80 88.35 0.467 1,644c c c c b 70 6.48 59.46 80,97 0.447 1,492d c c c b 80 6.57 60.42 79.73 0.440 1.609c b ab b c 90 7.67 64.82 84.41 0.428 1.757b ab b b d 100 7.93 64.31 83.87 0.402 1.859a Notes: the experimental meanings with different characters in the same column showed statistically significant differences (P0.01) 3.1.3 Effect of storage time on changes of bioactive compounds in dried Pouzolazia zeylanica material The results in Figure 3.5 showed that the content of bioactive compounds decreased after 12 months of storage at room temperature In particular, the anthocyanin content in the samples had the highest loss of 68.21÷83.48%; next is tannin 33.88÷64.81%; flavonoid from 38.04 to 56.68% and polyphenol from 30.42÷55.68% CK1 CK2 B1 B2 Flavonoid content (mgQE/g) Anthocyanin content (mgCE/100g) 60 50 40 (a) 30 20 10 CK1 B1 B2 (b) 0 10 12 14 10 12 14 Storage time (month) Storage time (month) 10 18 CK1 15 CK2 B1 Tannin content (mgTAE/g) Polyphenol content (mgGAE/g) CK2 B2 12 (c) CK1 CK2 B1 B2 (d) 0 10 12 14 10 12 14 Storage time (month) Storage time (month) Figure 3.5: Plots shows loss of anthocyanin (a), flavonoid (b), polyphenol (c) and tannin content (d) of samples during 12 months storage 12 Part 3.2: Research processing concentrated and spray dried powder products from Pouzolzia zeylanica plant 3.2.1 Effect of extract conditions on the content of bioactive compounds, color absorbance level Abs and total soluble solids The results showed that the extraction parameters (temperature, time and water/material ratio) were influenced by the second-order model to the levels of bioactive compounds (anthocyanin, flavonoid, polyphenol and tannin) in the obtained extracts As the temperature and time of extraction were increased, the content of bioactive compounds had a tendency to increase, but increasing to the optimum point and then tended to decrease At the same time, the ratio of water /material increased, the content of bioactive compounds had trend to increase and increasing to the optimal point then balance or mitigate As the temperature, the time and the water/material ratio increased, the color absorption of the extract and the dry matter content increased Through the ANOVA analysis, it was found that the predictive models of target functions had a coefficient of correlation R2 and P-value of Lack of fit greater than 0.05 The correlation between the predicted and experimental data was also very high with R20.0981 The simultaneous optimization of multiple surfaces was found to be 81.17oC for 30.10 minutes and 26.86 v/w (Figure 3.6) Figure 3.6: Plots show multiple response optimization with temperature and time (a), temperature and water/material ratio (b) (one factor was kept at central point) The optimal parameters obtained from the prediction model were re-tested and the results showed that at an extracting temperature of 81oC, for 30 minutes with water/material of 27/1 (v/w) The values of important target functions such as anthocyanin, flavonoid, polyphenol, tannin were 22.46/23.39 mgCE/100g; 566.6/563 mgQE/100g; 904/921 mgGAE/100g; 654/643 mgTAE/100g DM 13 3.2.2 Effect of blending process of taste and viscosity agents on the content of bioactive compounds and sensory value of product The study was similar to the one in section 3.2.1 This result showed that the bioactive compounds would be well maintained in the product when there was a balance between CMC gel matrix and supportive substances such as sucrose and citric acid In addition, anthocyanin, flavonoids, polyphenol and tannin compounds often contain multiple OH-phenol groups in their molecular structure, thus being able to bind to each other or sugars to form stable glycosides Results of ANOVA statistical analysis showed that the predictive models for the physicochemical indexes of the product had a correlation coefficient R20.976 and P-value for Lack of fit greater than 0.0717 Simultaneous optimization results were found to be 0.307% citric acid, 19.437% Brix and 0.281% CMC In addition, the results of sensory analysis by logistic regression model showed that the correlation between the Odd ratio and the citric acid content, oBrix and CMC were added into the extract with the acceptable level of the product (Odd ratio) were highest at 0.284%, 21.5oBrix and 0.29% respectively (Figure 3.7) Selected results for subsequent studies were 0.29% citric acid, 20oBrix and 0.29% CMC (a) (b) Figure 3.7: Relation of Odd ratio with oBrix and CMC (a), acid citric and oBrix (b) of concentrated product, while one factor was kept at central point 3.2.3 Effect of vacuum concentration process on the content of bioactive compounds and sensory of product The results showed that most of the compounds were reduced by decreasing the vacuum from 650 to 550 mmHg, corresponding to an increase in concentrating temperature from 60÷70oC However, the duration of the concentration was influenced by the unclear quadratic model on the content of bioactive compounds When the extract was 14 concentrated at 550 mmHg (70±2°C), the concentration time was more significant effect when concentrating at a vacuum of 650 mmHg (60±2°C) Simultaneous optimization of multiple surfaces result found concentration process parameters of 618.446 mmHg and 32.713 min, soluble solids at 45oBrix, maintaining the highest bioactive compounds content (Figure 3.8) However, the sensory results of 13 samples with different concentration conditions had three samples achieved the requirements of the high soluble solids (oBrix 50÷60) as samples concentrated at 550 mmHg for 30 minutes; 600 mmHg for 35 minutes and 600 mmHg for 40 minutes The sensory evaluation results (Figure 3.9) showed that the sample was concentrated at 600 mmHg for 40 minutes and the final dry matter was obtained 60oBrix The anthocyanin, flavonoid, polyphenol and tannin levels were 2.85 mgCE/100g; 3.81 mgQE/g; 8.05 mgGAE/g and 6.12 mg/g DM 7.22a 6.33b 6.56ab Preferred level Color 4.06a 3.46b Flavor 3.38b 4.08a State Sample 3.79a 3.93a 3.71b Figure 3.8: Contour plot shows simultaneous optimizationof multiple surface response with different concentrating pressure and time 3.82a Sample 3.91a Sample Figure 3.9: Plot shows sensory evaluation score of samples selected in 13 experimental samples 3.2.4 Effect of blending process carrier on the content of bioactive compounds in product Results in Table 3.3 and Figure 3.10 and 3.11showed that when maltodextrin was combined with three types of gum (arabic, carrageenan and xanthan) during the spray drying process of Pouzolzia zeylanica extract, the obtained powder had physicochemical properties such as particle size, noenzyme browning index (NEB), moisture content, water activity, color values (L, a, b and E) and the content of different bioactive compounds (anthocyanin, flavonoid, polyphenol and tannin) were different The combination of maltodextrin and arabic was optimal 15 Arabic Carrageenan Xanthan Figure 3.10: SEM micrograph of spray dried powder samples with different gum types Table 3.3: The content of bioactive compounds, antioxidant activity and color values (L, a, b and E) of spray dried powder samples with different gum types Anthocyanin Flavonoid Polyphenol Tannin Gum types (mgCE/100g) (mgQE/g) (mgGAE/g) (mgTAE/g) Arabic 7.56±0.106a 31.15±1.035a 27.96±0.380a 25.79±0.530b b b a Carrageenan 5.62±0.578 28.36±0.366 27.59±0.430 27.98±1.005a b b a Xanthan 6.37±0.793 27.42±0.855 28.01±0.800 26.73±0.275ab L a b Gum types E Arabic 78.43±0.189b 7.55±0.065a 6.96±0.180b 18.86±0.580a Carrageenan 79.79±0.085a 6.98±0.040b 7.30±0.080a 17.82±0.157b b a c Xanthan 78.52±0.103 7.57±0.030 6.64±0.025 18.62±0.245a DPPH (%) AAI Gum types FRAP (M FeSO4) a a Arabic 75.55±0.65 102.87±3.85 38.17±1.24a b b Carrageenan 72.14±1.06 95.75±2.39 37.55±1.79a b ab Xanthan 72.83±1.28 97.39±1.12 36.57±0.96a Notes: reported means (n=3) and standard deviation (±SD) Means within a same column followed by the same letter were not significantly different (P0.05) 0.5 NEB ( = 440 nm) Particle size (µm) 7.0 6.8 6.54a 6.6 6.4 6.26b 6.32b 6.2 (a) Carrageenan 0.298b 0.3 0.2 0.1 (b) Arabic Xanthan Xanthan 0.65 6.67a 0.591a 6.54a Water activity Moisture content (%) 7.0 6.6 Carrageenan Different gum types Different gum types 6.8 0.311b 0.0 6.0 Arabic 0.363a 0.4 6.42a 6.4 (c) 6.2 6.0 0.565b 0.60 0.55 0.514c 0.50 (d) 0.45 Arabic Carrageenan Xanthan Arabic Different gum types Carrageenan Xanthan Different gum types Figure 3.11: Plots show particle size (a), noenzyme browning index NEB (b), moisture content (c) and aw (d) of spray dried powder with different types 16 3.2.5 Effect of spray drying condition on the content of bioactive compounds in product Feed flow rate and spray drying temperature were important factors of the spray drying process If the spray drying process was carried out in high feed flow rate and low temperature, the moisture content of powder product could be high Otherwise, if this process was performed in low feed flow rate and high temperature, the quality of powder product would be affected such as the brownness of the product, no more decrease of moisture content of the product and the bioactive compounds could be destroyed The bioactive compounds were the main objective in spray drying process Besides, moisture content and particle size distribution of the products might also be interested Research results showed that spray drying conditions had effect on bioactive compounds content and physical characteristics of product The content of anthocyanin range from 4.64 to 8.12 mgCE/100g, flavonoid content ranged from 28.02 to 31.82 mgQE/g, polyphenol content varied from 26.06 to 27.95 mg GAE/g and tannin content changed from 23.85 to 25.51 mgTAE/g (Figure 3.12) The moisture content, particle size, nonenzyme browning index NEB, a value of product modified from 6.42 to 7.32%, 6.05 to 7.56 àm, 0.281ữ0.305, 7.21ữ8.46, respectively Result of the simultaneous optimization of multiple responses found spray drying temperatureof 179.832oC and feed flow rateof 18.264 rpm Figure 3.12: Response surface and contour plot of anthocyanin (a), flavonoid (b), polyphenol (c) and tannin (d) with spray drying temperature and feed flow rate 17 Results of re-testing the predict models showed that the anthocyanin, flavonoid, polyphenol and tannin contents of the samples were not significantly different from 1.1 to 2.8%, these compounds contents were 8.06 mgCE/100g; 30.06 mgQE/g; 28.12 mgGAE/g and 24.85 mgTAE/g product, respectively 3.2.6 Effect of blending process sucrose and ascorbic acid on sensory value and acceptable levels of consumer The study found that if the blending with high sugar content and low ascorbic acid for tea was too sweet In contrast, high acid and low sugar content, the tea is too sour, resulting in less acceptance If sugar and acid content added at the low level, tea was light taste Similarly, the taste of the tea would be too strong when both are added at high levels The results shown in Figures 3.13 and 3.14 showed that most of the consumers agreed (the highest Odd ratio) with the blended spray dried powder with sugar and ascorbic acid content were 15 and 0.1% Tea had a pH value of 4.3; oBrix 15.6 and color difference E of 56.35 Figure 3.13: Plot shows color difference E, o Brix and pH of tea samples blended with different rate of sucrose and ascorbic acid Figure3.14: Response surface plot shows opyimization of rate of sucrose and ascorbic acid with Odd ratio 3.2.7 Changing the content of bioactive compounds in product (concentrate and powder) during storage The content of anthocyanin and flavonoid had higher loss than polyphenol and tannin (Figure 3.15) At room temperature storage, the loss of bioactive compoundsin the products was higher than samplesat low temperature storage, approximatelyabout 1.38÷1.55 times The polyphenol and tannin contents were slightly reduced 18 MC-BQ-T Anthocyanin content (mgCE/100g) Anthocyanin content (mgCE/100g) during storage for the first 1st to 4th months and slightly increased in 5th to 6th months and then decreased gradually The anthocyanin and flavonoid contents decreased with time of storage For spray dried powder, the loss of bioactive compounds was found about 33.86÷51.85% after 12 months of storage MC-BQ-L y = -0,007x2 - 0,016x + 2,863 R² = 0,988 2 y = 0,005x - 0,217x + 2,911 R² = 0,991 0 10 12 10 MB-BQ-T y = -0,014x - 0,202x + 8,163 R² = 0,995 0 14 4 y = 0,011x2 - 0,253x + 3,755 R² = 0,954 35 y = 0,018x2 - 0,397x + 3,665 R² = 0,984 14 MB-BQ-T 25 20 15 10 y = 0,130x2 - 2,872x + 30,65 R² = 0,990 10 12 14 Storage time (month) Polyphenol content(mgGAE/g) MC-BQ-L y = -0,038x2 + 0,281x + 7,775 R² = 0,954 y = -0,043x2 + 0,274x + 7,588 R² = 0,915 0 10 12 35 12 14 MB-BQ-T 25 20 y = -0,114x2 + 0,741x + 27,23 R² = 0,945 15 10 14 10 Storage time (month) (e) 12 14 (f) 30 MC-BQ-L Tannin content (mgTAE/g) MC-BQ-T y = -0,027x + 0,194x + 5,949 R² = 0,968 10 30 Storage time (month) 8 (d) MC-BQ-T 10 Storage time (month (c) Polyphenol content (mgGAE/g) 12 0 Tannin content (mgTAE/g) 10 30 (b) MC-BQ-L Flavonoid content (mgQE/g) Flavonoid content (mgQE/g) (a) MC-BQ-T Storage time (month Storage time (month) y = -0,034x + 0,239x + 5,828 R² = 0,903 MB-BQ-T 25 20 y = -0,098x2 + 0,670x + 23,72 R² = 0,913 15 10 10 12 14 10 12 14 Storage time (month) Storage time (month) (i) (j) Figure 3.15: Plots showed changing content of anthocyanin (a, b), flavonoid (c, d), polyphenol (e, f) and tannin (i, j) of product (concentrate and powder) during storage 19 Part 3.3: Analysis functional characteristics and commercialitic ability of product 3.3.1 Determining antioxidant ability of product The results showed thatfree radical scavenging ability DPPH, reducing ferrous FRAP and antioxidant index AAI of product (concentrate and powder) were higher than some standard substances as well as some extracts from herbs has reported (Figure 3.16) Sp ray dried powder 75 59,73 69,9 68,05 64,55 Reducing ability DPPH (%) Reducing ability DPPH (%) Concentrateed product 90 56,68 60 63,78 60,2 45 49,39 37,24 30 (a) 26,02 15 Antioxidant index ( AAI) FRAP (M FeSO4/100mg) 4.165,52 3.962,37 3.000 4.171,24 2.000 1.000 6,687 116,198 70 65 (b) 60 55 0,1 4.144,95 4.000 Ascorbic acid Tannic acid 75 50 Concentration (mg/ml) 5.000 Quercetin Gallic acid 80 (c) 0,2 0,3 0,4 Concentration (mg/ml) 0,5 50 41,792 40 30 20 14,333 (d) 10 0,02 0,05 Concentration (mg/ml) Figure 3.16: Free radical scavenging ability DPPH (a, b), reducing ferrous FRAP (M FeSO4/100g) (c) and AAI (d) of concentrate and spray dried powder product, and some standard substances 3.3.2 Initial survey the level of consumer acceptance Both products are highly appreciated by 150 consumers for color, flavor and taste of the product; the quality of the products was assessed from good to very good; and most consumers were willing to buy products when they were present in the market (Figure 3.17) Figure 3.17: Plots showed quality level of concentrate product (a) and willing to buy product when they were present in market by 150 consumers 20 3.3.3 Testing the ability against respiratory bacteria and effect on cough treatment of product The obtained results showed that the spray driedpowder product had antibacterial activity on strains of Streptococcus pyogenes with MIC of 50 mg/ml; at human dose of g/day, it had effect on reducing sputa and cough symptom due to stimulation of cough model with capsaicin In contrast, the concentrated product did not show antimicrobial activity in bacterial strains related with respiratory disease; at a dose of g/day converted in humans, it had effect on reducing sputa in experimental mice affected with capsaicin, but the effect of it on cough symptoms caused by the capsaicin model was not typical in comparison with spray dried powder (Table 3.3, 3.4, 3.5 3.6) Table 3.3: MIC of test samples in survey condition MIC (mg/ml) Baterial strains Concentrate product Spray dried powder Staphyloccocus aureus n/a n/a Streptoccocus pyogenes n/a 50 Streptoccocus pneumoniae n/a n/a Klebsiella pneumoniae n/a n/a Pseudomonas aeruginosa n/a n/a Note: (n/a) not determine activity Table 3.4: Result evaluation percent increased phenol redness of normal mice group and mice group were caused cough using a capsaicin model Experimental group Oral dose % increased Optical density P-value n = 8-10 (g/kg) phenol redness Distilled water control 0.083 ± 0.005 Ambroxol 0.24 0.166 ± 0.027 0.004 99.70 Concentrate product 1.0 0.150 ± 0.021 0.026 80.96 2.0 0.102 ± 0.013 0.409 23.01 Spray dried powder 1.5 0.164 ± 0.022 0.008 97.47 3.0 0.102 ± 0.014 0.682 22.13 Oral dose Experimental group % increased Optical density P-value (g/kg) n=9 phenol redness Physiological control 0.086 ± 0.004 0.060 ± 0.007 0.008* Capsaicin control  30.27 tablet/kg 0.093 ± 0.010 0.016# Neo-codion®  54.55 1.0 0.093 ± 0.013 0.043# Concentrate product  55.29 1.5 0.099 ± 0.010 0.006# Spray dried powder  65.68 Notes: *: compare with physiological control; #: compare with pathological control 21 Table 3.5: Result evaluation time that appear cough symptoms of mice group were caused cough using a capsaicin model Experimental group Oral dose Time P-value n=8 (g/kg) (seconds) Capsaicin control 1.63 ± 0.26 Neo-codion® tablet/kg 1.75 ± 0.25 0.736 Concentrate product 1.0 2.38 ± 0.26 0.063 Spray dried powder 1.5 2.88 ± 0.52 0.049 Table 3.6: Result evaluation number stimulus jumps of mice group were caused cough using a capsaicin model Experimental group Oral dose Number of P-value n=8 (g/kg) stimulus jumps Capsaicin control 21.38 ± 4.02 Neo-codion® tablet/kg 10.13 ± 2.89 0.039 Concentrate product 1.0 12.0 ± 3.53 0.101 Spray dried powder 1.5 9.25 ± 3.10 0.032 Chapter 4: Conclusion and request 4.1 Conclusion The Pouzolzia zeylanica plants were planted and growing very well with optimum harvesting time of 4÷8 weeks after planting, medicinal plants were planted from January to April 2015 (for dry season) contained more bioactive compounds than plants from July to October (in rainy season); the whole plants were dehydrated at 60oC or sun drying until the moisture of material was less 12%, then cut into 3÷5 cm and kept in PE package (2 layers) They could be stored for 12 months at ambient temperature The optimal extraction conditions of bioactive compounds in dried Pouzolzia zeylanica plants were performed at temperature of 81oC for 30 minutes and water/material ratio of 27/1 v/w The processing of concentrated product as followed: extract was blended with citric acid of 0.29%, 20°Brix and 0.29% carboxymethyl cellulose; vacuum cooking at 600 mmHg for 40 minutes The Brix degree in the final product was 60 with high viscosity (538 mPas), the water activity (aw) was 0.9085 and pH value of 3.9 The contents of bioactive compounds (anthocyanin, flavonoid, polyphenol and tannin) 22 were 2.85 mgCE/100g; 3.81 mgQE/g; 8.05 mgGAE/g and 6.12 mgTAE/g, respectively The production process of spray dried powder: The extract was blended with 9% maltodextrin and 0.08% arabic gum, spray dried process was conducted at inlet drying temperature of 179oC, feed flow rate of 18 rpm, then mixed with the ratio of spray dried powder/sucrose/ascorbic acid as 3/15/0.1 (w/w/w) in 100 ml hot water to make tea with harmonious taste The bioactive compounds content (anthocyanin, flavonoid, polyphenol and tannin) and moisture of the product were 8.06 mgCE/100g; 30.86 mgQE/g; 28.12 mgGAE/g and 24.85 mgTAE/g; and 6.5%, respectively The concentrated product was stored at room temperature had higher loss of anthocyanin, flavonoid, polyphenol and tannin of 64.21; 63.52; 45.59 and 40.52% than the samples stored at low temperatures; this sample had loss of 42.11; 40.94; 32.30 and 29.58% respectively The bioactive compounds had loss of spray dried powder were 51.85; 50.97; 36.64 and 33.86%; respectively The soluble powder had higher antioxidant activity than the concentrate product with an IC50 value of DPPH free radical scavenging activity that was 0.98 and 3.18 mg/ml; the ability of reducing ferrous following FRAP method was 116.198 and 6.687 μFeSO4/100mg; the AAI index determined by total reducing power method was 41.792 and 14.333 respectively for two products In addition, the spray dried powder had antibacterial activity on strains of Streptococcus pyogenes with MIC of 50 mg/ml; at human dose of 6g/day, it had effect on reducing sputa and cough symptom due to stimulation of cough model with capsaicin In contrast, the concentrated product did not show antimicrobial activity in bacterial strains related with respiratory disease; at a dose of g/day converted in humans, it had effect on reducing sputa in experimental mice affected with capsaicin, but the effect of it on cough symptoms caused by the capsaicin model was not typical in comparison with spray dried powder All both products was observed that 75÷85% consumers evaluated the color, flavor and taste of products from “like” to “extremely like”; approximately 93÷94% consumers evaluated the 23 product quality from “good” to “very good”; and around 89÷91% of consumers agreed with the cost price of 30,000 VND (concentrated product in 250 ml glass bottle and 100g of soluble powder in plastic box were produced from Pouzolzia zeylanica extract) and around 80÷84% of consumers were willing to buy in case of the product is available in the market 4.2 Recommendations Further studies on the effects of human cough experiments, anti-inflammatory, anti-edema effects and the model of allergic asthma with albumin Calculating economic efficiency in production and large-scale production testing in order to transfer technology of production to local enterprise 24 ... Pouzolzia zeylanica plants Pouzolzia zeylanica plants belong to the Urticaceae family, the English name is Graceful Pouzolzsbush, originating in the tropics of southern Asia Nowadays, Pouzolzia zeylanica. .. ability of Pouzolzia zeylanica extract Science and Technology Journal of Agriculture and Rural Development, Ministry of Agriculture and Rural Development, Vietnam, 8: 53-61 Nguyen Duy Tan, Le... 23+star model Total sample: 20 samples including replicates at the centeral point Extractor with 10 litters capacity, stirrer, temperature controller Each sample was taken liters Then filter through
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Xem thêm: Nghiên cứu ảnh hưởng của quá trình thu hoạch và chế biến đến hàm lượng các chất có hoạt tính sinh học trong cây thuốc dòi (pouzolzia zeylanica l benn) tóm tắt tiếng anh, Nghiên cứu ảnh hưởng của quá trình thu hoạch và chế biến đến hàm lượng các chất có hoạt tính sinh học trong cây thuốc dòi (pouzolzia zeylanica l benn) tóm tắt tiếng anh, Part 2.2: Study on processing two products (concentrate and spray dried powder) from Pouzolzia zeylanica, Part 3.2: Research processing concentrated and spray dried powder products from Pouzolzia zeylanica plant, Part 3.3: Analysis functional characteristics and commercialitic ability of product

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