• Title/Summary/Keyword: response surface optimization

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Optimization of Conditions for the Microencapsulation of ${\alpha}-Tocopherol$ and Its Storage Stability (${\alpha}-Tocopherol$ 미세캡슐화의 최적화 및 저장안정성 규명)

  • Chang, Pahn-Shick;Ha, Jae-Seok;Roh, Hoe-Jin;Choi, Jin-Hwan
    • Korean Journal of Food Science and Technology
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    • v.32 no.4
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    • pp.843-850
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    • 2000
  • We have produced the microcapsule composed of ${\alpha}-tocopherol$ as a core material (Cm) and the gelatinized polysaccharide as a wall material (Wm). Firstly, we have developed a simple, sensitive, and quantitative analysis method of the microencapsulation product using 5% cupric acetate pyridine solution. We could then optimize all the conditions for the microencapsulation process such as the ratio of [Cm] to [Wm], the temperature of dispersion fluid, and the emulsifier concentration using response surface methodology (RSM). As for the microencapsulation of ${\alpha}-tocopherol$, the regression model equation for the yield of microencapsulation (YM, %) to the change of an independent variable could be predicted as follows : YM=99.77-1.76([Cm]:[Wm])-1.72$([Cm]\;:\;[Wm])^2$. From the ridge of maximum response, the optimum conditions for the microencapsulation of ${\alpha}-tocopherol$ were able to be determined as the ratio of [Cm] to [Wm] of 4.6:5.4(w/w), the emulsifier concentration of 0.49%, and dispersion fluid temperature of $25.5^{\circ}C$, respectively. Finally, the microcapsules produced under the optimal conditions were applied for the analysis of storage stability. The optimal conditions for the storage were found to be the values of pH 9.0 and $25{\sim}35^{\circ}C$. And the storage stability of the microcapsules containing ${\alpha}-tocopherol$ were higher than 99% for a week at pH 9.0 and $25^{\circ}C$.

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Strain Improvement and Bioprocess Optimization for Enhanced Production of Haluronic Acid(HA) in Bioreactor Cultures of Streptococcus zooepidemicus (히알루론산 생산성 향상을 위한 Streptococcus zooepidemicus 균주 개량 및 발효조 배양공정 최적화)

  • Kim, Soo Yeon;Chun, Gie-Taek
    • Microbiology and Biotechnology Letters
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    • v.48 no.3
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    • pp.344-357
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    • 2020
  • Strain improvement and bioprocess development were undertaken to enhance hyaluronic acid(HA) production by Streptococcus zooepidemicus cells. Using a high-yielding mutant strain, statistical medium optimization was carried out in shake flask cultures, resulting in 52% increase in HA production (5.38 g/l) at the optimal medium composition relative to the parallel control cultures. For sufficient supply of dissolved oxygen (DO), which turned out to be crucial for enhanced production of HA, agitation system and speed were intensively investigated in 5 L bioreactor cultures. Increase in oxygen mass transfer coefficient (kLa) through increment of agitation speed (rpm) and 35% expansion of diameter of the newly-designed impellers showed significantly positive effects on HA production. By installing an expanded Rushton-turbine impeller for efficient break-down of sparged air, and an extended marine impeller above the Rushton-turbine impeller for efficient mixing of the air-born viscous fermentation broth, maximum amount of HA (9.79 g/l) was obtained at 450 rpm, 1.8 times higher level than that of the corresponding flask culture. Subsequently, the possibility of bioprocess scale-up to a 50 L bioreactor was investigated. Despite almost identical maximum HA production (9.11 vs 9.25 g/l), the average HA volumetric productivity (rp) of the 50 L culture turned out only 74% compared to the corresponding 5 L culture during the exponential phase, possibly caused by shear damages imposed on the producing cells at the high stirring in the 50 L culture. The scale-up process could be successfully achieved if a scale-up criterion of constant oxygen mass transfer coefficient (kLa) is applied to the 50 L pilot-scale bioreactor system.

A study on the Degradation and By-products Formation of NDMA by the Photolysis with UV: Setup of Reaction Models and Assessment of Decomposition Characteristics by the Statistical Design of Experiment (DOE) based on the Box-Behnken Technique (UV 공정을 이용한 N-Nitrosodimethylamine (NDMA) 광분해 및 부산물 생성에 관한 연구: 박스-벤켄법 실험계획법을 이용한 통계학적 분해특성평가 및 반응모델 수립)

  • Chang, Soon-Woong;Lee, Si-Jin;Cho, Il-Hyoung
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.1
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    • pp.33-46
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    • 2010
  • We investigated and estimated at the characteristics of decomposition and by-products of N-Nitrosodimethylamine (NDMA) using a design of experiment (DOE) based on the Box-Behken design in an UV process, and also the main factors (variables) with UV intensity($X_2$) (range: $1.5{\sim}4.5\;mW/cm^2$), NDMA concentration ($X_2$) (range: 100~300 uM) and pH ($X_2$) (rang: 3~9) which consisted of 3 levels in each factor and 4 responses ($Y_1$ (% of NDMA removal), $Y_2$ (dimethylamine (DMA) reformation (uM)), $Y_3$ (dimethylformamide (DMF) reformation (uM), $Y_4$ ($NO_2$-N reformation (uM)) were set up to estimate the prediction model and the optimization conditions. The results of prediction model and optimization point using the canonical analysis in order to obtain the optimal operation conditions were $Y_1$ [% of NDMA removal] = $117+21X_1-0.3X_2-17.2X_3+{2.43X_1}^2+{0.001X_2}^2+{3.2X_3}^2-0.08X_1X_2-1.6X_1X_3-0.05X_2X_3$ ($R^2$= 96%, Adjusted $R^2$ = 88%) and 99.3% ($X_1:\;4.5\;mW/cm^2$, $X_2:\;190\;uM$, $X_3:\;3.2$), $Y_2$ [DMA conc] = $-101+18.5X_1+0.4X_2+21X_3-{3.3X_1}^2-{0.01X_2}^2-{1.5X_3}^2-0.01X_1X_2+0.07X_1X_3-0.01X_2X_3$ ($R^2$= 99.4%, 수정 $R^2$ = 95.7%) and 35.2 uM ($X_1$: 3 $mW/cm^2$, $X_2$: 220 uM, $X_3$: 6.3), $Y_3$ [DMF conc] = $-6.2+0.2X_1+0.02X_2+2X_3-0.26X_1^2-0.01X_2^2-0.2X_3^2-0.004X_1X_2+0.1X_1X_3-0.02X_2X_3$ ($R^2$= 98%, Adjusted $R^2$ = 94.4%) and 3.7 uM ($X_1:\;4.5\;$mW/cm^2$, $X_2:\;290\;uM$, $X_3:\;6.2$) and $Y_4$ [$NO_2$-N conc] = $-25+12.2X_1+0.15X_2+7.8X_3+{1.1X_1}^2+{0.001X_2}^2-{0.34X_3}^2+0.01X_1X_2+0.08X_1X_3-3.4X_2X_3$ ($R^2$= 98.5%, Adjusted $R^2$ = 95.7%) and 74.5 uM ($X_1:\;4.5\;mW/cm^2$, $X_2:\;220\;uM$, $X_3:\;3.1$). This study has demonstrated that the response surface methodology and the Box-Behnken statistical experiment design can provide statistically reliable results for decomposition and by-products of NDMA by the UV photolysis and also for determination of optimum conditions. Predictions obtained from the response functions were in good agreement with the experimental results indicating the reliability of the methodology used.

Optimization of Betacyanin Production by Red Beet (Beta vulgaris L.) Hairy Root Cultures. (Red Beet의 모상근 배양을 이용한 천연색소인 Betacyanin 생산의 최적화)

  • Kim, Sun-Hee;Kim, Sung-Hoon;Lee, Jo-No;An, Sang-Wook;Kim, Kwang-Soo;Hwnag, Baik;Lee, Hyeong-Yong
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.435-441
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    • 1998
  • Optimal conditions for the production of natural color, betacyanin were investigated by varying light intensity, C/N ratio, concentrations of phosphate and kinds of elicitors. Batch cultivation was employed to characterize cell growth and betacyanin production of 32 days. The maximum specific growth rate, ${\mu}$$\sub$max/, was 0.3 (1/day) for batch cultivation. The maximum specific production rate, q$\^$max/$\sub$p/, was enhanced 0.11 (mg/g-cell/day) at 3 klux. A light intensity of 3 klux was shown to the best for both cell growth and betacyanin production. The maximum specific production rate was 0.125 (mg/g-cell/day) at 0.242 (1/day), the maximum specific growth rate. The dependence of specific growth rate on the light lintensity is fit to the photoinhibition model. The correlation between ${\mu}$ and q$\sub$p/ showed that the product formation parameters, ${\alpha}$ and ${\beta}$$\sub$p/ were 0.3756 (mg/cell) and 0.001 (mg/g-cell/day), respectively. The betacyanin production was partially cell growth related process, which is different from the production of a typical product in plant cell cultures. In C/N ratio experiment, high carbon concentration, 42.1 (w/w) improved cell growth rate while lower concentration, 31.6 (w/w) increased the betacyanin production rate. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.26 (1/day) and 0.075 (mg/g-cell/day), respectively. Beta vulgaris L. cells under 1.25 mM phosphate concentration produced 10.15 mg/L betacyanin with 13.46 (g-dry wt./L) of maximum cell density. The production of betacyanin was elongated by adding 0.1 ${\mu}$M of kinetin. This also increased the cell growth. Optimum culture conditions of light intensity, C/N, phosphate concentration were obtained as 5.5 klux, 27 (w/w), 1.25 mM, respectively by the response surface methodology. The maximum cell density, X$\sub$max/, and maximum production, P$\sub$max/, in optimized conditions were 16 (g-dry wt./L), 12.5 (mg/L) which were higher than 8 (g-dry wt./L), 4.48 (mg/L) in normal conditions. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.376 (1/day) and 0.134 (mg/g-cell/day) at the optimal condition. The overall results may be useful in scaling up hairy root cell culture system for commercial production of betacyanin.

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Mass Screening of Lovastatin High-yielding Mutants through Statistical Optimization of Sporulation Medium and Application of Miniaturized Fungal Cell Cultures (Lovastatin 고생산성 변이주의 신속 선별을 위해 통계적 방법을 적용한 Sporulation 배지 개발 및 Miniature 배양 방법 개발)

  • Ahn, Hyun-Jung;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • KSBB Journal
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    • v.22 no.5
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    • pp.297-304
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    • 2007
  • For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

Optimization of Production Medium by Response Surface Method and Development of Fermentation Condition for Monascus pilosus Culture (Monascus pilosus 배양을 위한 반응표면분석법에 의한 생산배지 최적화 및 발효조건 확립)

  • Yoon, Sang-Jin;Shin, Woo-Shik;Chun, Gie-Taek;Jeong, Yong-Seob
    • KSBB Journal
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    • v.22 no.5
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    • pp.288-296
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    • 2007
  • Monascus pilosus (KCCM 60160) in submerged culture was optimized based on culture medium and fermentation conditions. Monacolin-K (Iovastatin), one of the cholesterol lowing-agent which was produced by Monascus pilosus may maintain a healthy lipid level by inhibiting the biosynthesis of cholesterol. Plackett-Burman design and response surface method were employed to study the culture medium for the desirable monacolin-K production. As a result of experimental designs, optimized production medium components and concentrations (g/L) were determined on soluble starch 96, malt extract 44.5, beef extract 30.23, yeast extract 15, $(NH_4)_2SO_4$ 4.03, $Na_2HPO_4{\cdot}12H_2O$ 0.5, L-Histidine 3.0, $KHSO_4$ 1.0, respectively. Monacolin-K production was improved about 3 times in comparison with shake flask fermentation of the basic production medium. The effect of agitation speed (300, 350, 400 and 450 rpm) on the monacolin-K production were also observed in a batch fermenter. Maximum monacolin-K production with the basic production medium was 68 mg/L when agitation speed was 500 rpm. And it was found that all spherical pellets (average diameter of $1.0{\sim}1.5mm$) were dominant during fermentation. Based on the results, the maximum production of 185 mg/L of monacolin-K with the optimized production medium was obtained at pH (controlled) 6.5, agitation rate 400 rpm, aeration rate 1 vvm, and inoculum size 3%.

Optimization of Compound K Production from Ginseng Extract by Enzymatic Bioconversion of Trichoderma reesei (Trichoderma reesei 유래 산업효소를 이용한 인삼추출물로부터 Compound K 생산 최적화)

  • Han, Gang;Lee, Nam-Keun;Lee, Yu-Ri;Jeong, Eun-Jeong;Jeong, Yong-Seob
    • The Korean Journal of Food And Nutrition
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    • v.25 no.3
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    • pp.570-578
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    • 2012
  • Compound K(ginsenoside M1) is one of saponin metabolites and has many benefits for human health. This study was to investigate Compound K produced from ginseng crude saponin extract with commercial cellulolytic complex enzyme(cellulase, ${\beta}$-glucanase, and hemicellulase) obtained from Trichoderma reesei. The effect factors(temperature, pH, ginseng crude saponin extract and enzyme concentration, and reaction time) on production of Compound K from ginseng crude saponin extract were determined by one factor at a time method. The selected major factor variables were ginseng crude saponin extract of 2%(w/v), enzyme of 7%(v/v), reaction time of 48 hr. Based on the effect factors, response surface method was proceeded to optimize the enzymatic bioconversion conditions for the desirable Compound K production under the fixed condition of pH 5.0 and $50^{\circ}C$. The optimal reaction condition from RSM was ginseng crude saponin extract of 2.38%, enzyme of 6.06%, and reaction time of 64.04 hr. The expected concentration of Compound K produced from that reaction was 840.77 mg/100 g. Production of Compound K was 1,017.93 mg/100 g and 862.31 mg/100 g, by flask and bench-scale bioreactor($2.5{\ell}$) system, respectively.

Exploration of optimum conditions for production of saccharogenic mixed grain beverages and assessment of anti-diabetic activity (잡곡당화음료 제조 최적 조건 탐색 및 항당뇨 활성 평가)

  • Lee, Jae Sung;Kang, Yun Hwan;Kim, Kyoung Kon;Yun, Yeong Kyeong;Lim, Jun Gu;Kim, Tae Woo;Kim, Dae Jung;Won, Sang Yeon;Bae, Moo Hoan;Choi, Han Seok;Choe, Myeon
    • Journal of Nutrition and Health
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    • v.47 no.1
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    • pp.12-22
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    • 2014
  • Purpose: This study was conducted to establish the production conditions through optimization of the production process of beverages using Aspergillus oryzae CF1001, and to analyze volatile compounds and antidiabetic activity. Methods: The optimum condition was selected using the response surface methodology (RSM), through a regression analysis with the following independent variables gelatinization temperature (GT, $X_1$), saccharogenic time (ST, $X_2$), and dependent variable; ${\Delta}E$ value (y). The condition with the lowest ${\Delta}E$ value occurred with combined 45 min ST and $50^{\circ}C$ GT. The volatile compounds were analyzed quantitatively by GC-MS. Results: Assessment of antidiabetic activity of saccharogenic mixed grain beverage (SMGB) was determined by measurement of ${\alpha}$-glucosidase inhibition activity, and glucose uptake activity and glucose metabolic protein expression by reverse transcriptase polymerase chain reaction (RT-PCR) and western blot analysis. Results of volatile compounds analysis, 62 kinds of volatile compounds were detected in SMGB. Palmitic acid (9.534% ratio), benzaldehyde (8.948% ratio), benzyl ethyl ether (8.792% ratio), ethyl alcohol (8.35% ratio), and 2-amyl furan (4.826% ratio) were abundant in SMGB. We confirmed that ${\alpha}$-glucosidase inhibition activity, glucose uptake activity, and glucose-metabolic proteins were upregulated by SMGB treatment with concentration dependent manner. Conclusion: Saccharogenic mixed grain beverage (SMGB) showed potential antidiabetic activity. Further studies will be needed in order to improve the taste and functionality of SMGB.

Optimization of Interesterification Reaction for the Continuous Production of trans-Free Fat in a Packed Bed Enzyme Bioreactor with Immobilized Lipase (고정화 리파제를 이용한 충진형 효소생물반응기 내에서의 무-트랜스 유지 연속 생산을 위한 에스테르 교환 반응의 최적화)

  • Kim, Sang-Woo;Park, Kyung-Min;Ha, Jae-Uk;Lee, Jae-Hwan;Chang, Pahn-Shick
    • Korean Journal of Food Science and Technology
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    • v.41 no.2
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    • pp.173-178
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    • 2009
  • Epidemiological studies showed that high trans-fat consumption is closely associated with getting the risks of cardiovascular disease. The objective of this study was to produce trans-free fat through lipase-catalyzed interesterification, as a substitute for the cream margarine commonly used in industry. Optimum conditions for interesterification in a packed bed enzyme bioreactor (PBEB) were determined using response surface methodology (RSM) based on central composite design. Three kinds of reaction variables were chosen, such as substrate flow rate (0.4-1.2 mL/min), reaction temperature (60-70$^{\circ}C$), and ratio of fully hydrogenated canola oil (FHCO, 35-45%) to evaluate their effects on the degree of interesterification. Optimum conditions from the standpoint of solid fat content (SFC) were found to be as follows: 0.4 mL/min flow rate, 64.7$^{\circ}C$ reaction temperate, and 42.8% (w/w) ratio of FHCO, respectively. The half-life of immobilized lipase in PBEB with two stages at 60$^{\circ}C$ ($1^{st}$ stage) and 55$^{\circ}C$ ($2^{nd}$ stage) was about more than 30 days as estimated by extrapolating the incubation time course of tristearoyl glycerol (TS) conversion, whereas the half-life of the enzyme in PBEB with single stage at 65$^{\circ}C$ was only about 15 days. Finally, the results from SFC analysis suggest that trans-free fat produced in this study seems to be a suitable substitute for the cream margarine commonly used in industry.

Optimization of Hot-Water Extraction Conditions for Preparation of Polyphenol and Gallic Acid from Acorn (도토리의 Polyphenol 및 Gallic Acid 성분의 열수 추출조건 최적화)

  • Kim, Seong-Ho;Lee, Jin-Man
    • Food Science and Preservation
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    • v.15 no.1
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    • pp.58-65
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    • 2008
  • The Acorn (Quercus acutissima CARRUTHERS), which contains a large quantity of tannin, should be developed as a processed food as the acorn is rich in natural antioxidants and other valuable components. Accordingly, acorn extraction conditions for polyphenol and gallic acid (both antioxidants) were investigated by response surface methodology. The content of polyphenols were determined under 16 different extraction conditions based upon a central composite design. The parameters varied over $30-70^{\circ}C$ of extraction temperature, 1-5 h of extraction time, and 5-25 mL/g of solvent ratio, Gallic acid extraction was optimal at $60-100^{\circ}C$ extraction temperature, 1-5 h of extraction time, and 5-25 mL/g of solvent ratio, Epicatechin content was highest at $56.77^{\circ}C$, 4.16 hand 22.38 mL/g. Catechin content was highest at $52.37^{\circ}C$, 2h and 23.59 mL/g. The maximum catechin content was $91.30{\mu}g/mL$. Epigallocatechin content was influenced by extraction temperature and time. The maximum epigallocatechin content was $1,066.56{\mu}g/mL$ at $61.42^{\circ}C$, 4.17h, and 9.25 mL/g. The maximum value of epicatechingallate content was $125.39{\mu}g/mL$ at $47.72^{\circ}C$, 3.04h, and 24.93mL/g. Epigallocatechingallate content was influenced principally by solvent ratio and the maximum content was $61.38{\mu}g/mL$ at $48.11^{\circ}C$, 2.96h, and 24.95mL/g. The total polyphenol content was maximal at $1,332.75{\mu}g/mL$, after extraction at $61.50^{\circ}C$, 4.24h, at 9.71mL/g. The higher the extraction temperature and the longer the extraction time, the greater the polyphenol content. Gallic acid content was highest, the maximal level was $30.51{\mu}g/mL$ after $65.84^{\circ}C$, 1.65h at 17.17 mL/g, and this was influenced principally by extraction time and solvent ratio.