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1、?544?Chin J Mod Appl Pharm,2014 May,Vol.31 No.5?2014?5?31?5?*(?510006)?Box-Behnken?3?1?40?pH 5.5?49.94?1.53 h?2.00%?R284.2?B?1007-7693(2014)05-0544-06 DOI:10.13748/ki.issn1007-7693.2014.05.007 Optimization of Enzymatic Extraction Technology for Polysaccharides from Desmodium Styracifolium by Respons
2、e Surface Method CHENG Xuanxuan,GUO Chuchu,ZOU Fengru,YANG Quan*(School of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006,China)ABSTRACT:OBJECTIVE To study the optimal technological conditions for extracting Desmodium styracifolium polysaccharides.METHODS According
3、 to Box-Behnken design principle and results of single-factor experiments,the quadratic polynomial mathematical model,response surface and contour line map were analyzed.The effects of enzymatic hydrolysis time,temperature and enzyme amount on the polysaccharides yield were explored by using yield a
4、nd content of polysaccharides as evaluating indicators.RESULTS The degree of fitting and confidence level for mathematical model was satisfactory.Optimum extraction technology conditions were as follows:the ratio of liquid to solid 1?40,pH 5.5,enzymatic hydrolysis temperature 49.94?,time of 1.53 h a
5、nd cellulase amount of 2.00%.The effect order of three factors on polysaccharides yield was as follows:enzymatic hydrolysis timeenzymatic hydrolysis temperatureenzyme amount.The interaction effects of different factors were highly significant.CONCLUSION Optimized extraction technology is stable,feas
6、ible,highly efficient.It could provide references for the industrial production.KEY WORDS:Desmodium styracifolium polysaccharides;enzymatic extraction;response surface method;extraction technology?(20101H020)?Tel:(020)39352327 E-mail: *?Tel:(020)39352173 Email:?1?Desmodium styracifolium(Osb.)Merr.?2
7、?3?4?5-6?7?2014?5?31?5?Chin J Mod Appl Pharm,2014 May,Vol.31 No.5?545?1?YJ-10?(?)?UV-2450?(?)?FA2204B?(?)?PHS-25?pH?(?)?PH2T-92VOA?(?)?DK-S26?(?)?KQ-500DE?(?)?EYELA-N-1100D-WD?(?)?Desmodium styracifolium(Osb.)Merr.?(30 000 Ug-1?20130506)?(20 000 Ug-1?20130420)?(?20130509?98%)?(?)?2?2.1?8?2?1:8?2?40
8、min?45?2.2?-?7-8?10 mg?100 mL?0.1 mgmL1?0.0?0.1?0.2?0.3?0.5?0.6?0.7 mL?1.0 mL?5%?1.0 mL?5.0 mL?30 min?1.0 mL?490 nm?X(mgmL1)?Y?Y=48.275X+0.187 6(r=0.999 6)?1.4310.0 gmL1?2.3?10 mg?50 mL?2 mL?5%?2.0 mL?10.0 mL?30 min?490 nm?2.4?2 g?pH?5 min?80%?24 h?7?(%)=100%?(g)/?(g)?(%)=100%?C?D/M?C?(gmL1)?D?M?(mg
9、)?100?X?Y?Z=0.5X+0.5Y?Z?2.5?2.5.1?6?2 g?5?0.5%?1(?=3?2)?2(?=1?1)?3(?=2?3)?1?20?pH?4.5?45?2 h?5 min?2.4?2.3?2?1?546?Chin J Mod Appl Pharm,2014 May,Vol.31 No.5?2014?5?31?5?1?Fig 1 Effect of different enzymes on colligation scores 2.5.2?5?2 g?1?10?1?20?1?30?1?40?1?50?0.5%?pH 4.5?45?2 h?2.5.1?2?1?10?1?4
10、0?1?40?2?Fig 2 Effect of different liquid/solid ratio on colligation scores 2.5.3?5?2 g?1?40?0.5%?pH?4.5?40?45?50?55?60?2 h?2.5.1?9?50?3?3?Fig 3 Effect of different enzyme hydrolysis temperatures on colligation scores 2.5.4?5?2 g?1?40?0.5%?pH 4.5?50?1?1.5?2?2.5?3 h?2.5.1?4?1.5 h?4?Fig 4 Effect of di
11、fferent enzymatic hydrolysis time on colligation scores 2.5.5?5?2 g?1?40?0.5%?1%?1.5%?2%?2.5%?pH 4.5?50?1.5 h?2.5.1?5?2%?2%?5?Fig 5 Effect of different enzyme amounts on colligation scores 2.5.6 pH?5?2 g?1?40?2%?pH?4.0?4.5?5.0?5.5?6.0?50?1.5 h?2.5.1?6?pH?5.5?2.6?Design Expert 8.0.5?Box-Behnken?5-6,8
12、?3?2014?5?31?5?Chin J Mod Appl Pharm,2014 May,Vol.31 No.5?547?3?3?1?2?6 pH?Fig 6 Effect of different pH values on colligation scores?1?Tab 1 Factors and levels of response surface method?A.?/?B.?/h C.?/h 1 45 1.0 1.5 0 50 1.5 2.0 1 55 2.0 2.5?2?Tab 2 Design scheme and results of response surface ana
13、lysis?A B C?/%?/%?1 50 1.5 2.0 2.51 31.09 98.07 2 45 1.0 2.0 1.90 21.57 71.11 3 55 2.0 2.0 2.00 21.92 73.61 4 55 1.5 2.5 2.25 26.60 85.89 5 50 2.0 1.5 2.29 24.19 82.84 6 50 1.0 2.5 2.11 24.91 80.49 7 45 2.0 2.0 2.40 25.67 87.32 8 50 1.5 2.0 2.49 31.27 97.97 9 50 1.0 1.5 2.13 23.34 78.39 10 50 1.5 2.
14、0 2.50 31.27 98.16 11 50 1.5 2.0 2.58 30.19 97.90 12 55 1.5 1.5 2.18 26.13 83.79 13 50 2.0 2.5 2.24 23.77 81.20 14 55 1.0 2.0 2.29 25.99 85.70 15 45 1.5 1.5 2.35 25.09 85.43 16 45 1.5 2.5 2.31 24.22 83.27 17 50 1.5 2.0 2.48 31.45 98.06?Z?Z=97.999+0.230A+1.156B+0.049C7.079AB+1.065AC0.939BC7.352A211.2
15、14B26.052C2?A?B?C?(?)?(h)?(%)?3?3?ProbF?0.05(?)?R2=0.999 6?R2Adj=0.997 7?CV=0.20?10?ProbF?A?B?AB?AC?BC?A2?B2?C2?(P?79?11?79?0?7?2%?-10?89?0?2?(PF P?1 227.885 9 136.431 6 4 502.471 0.00 01 0.01 A?0.423 798 1 0.423 798 13.986 05 0.007 3 0.01 B?10.683 06 1 10.683 06 352.558 7 0.00 01 0.01 C?0.019 286 1
16、 0.019 286 0.636 486 0.451 2 AB 200.433 4 1 200.433 4 6 614.635 0.000 1 0.01 AC 4.534 344 1 4.534 344 149.640 9 0.000 1 0.01 BC 3.523 129 1 3.523 129 116.269 1 0.000 1 0.01 A2 227.604 8 1 227.604 8 7 511.335 0.000 1 0.01 B2 529.441 3 1 529.441 3 1 7472.44 0.000 1 0.01 C2 154.192 9 1 154.192 9 5 088.
17、620 0.000 1 0.01?0.212 111 7 0.030 302?0.172 391 3 0.057 464 5.787 003 0.061 5?0.039 719 4 0.009 930?1 228.097 16?7?Fig 7 Contour line and response curved surface of temperatures and time to colligation scores?8?Fig 8 Contour line and response curved surface of temperatures and enzyme amounts to col
18、ligation scores?9?Fig 9 Contour line and response curved surface of time and enzyme amounts to colligation scores?2014?5?31?5?Chin J Mod Appl Pharm,2014 May,Vol.31 No.5?549?3?(?)?(?)?(?1 h)?1.71%?2.70%?3.04%?REFERENCES 1 FAN W C,MEI Q X,LAI H B.A research review of pharmacological function and clini
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22、012,29(6):498-503.8 HUANG X X,LAI H F,LUO L C.Optimization of extraction technology for polysaccharide from Lobelia chinensis by ultrasonic-composite enzyme synergistic method J.Chin J Exp Tradit Med Form(?),2012,18(5):44-46.9 LIN J Y,JI L H.Optimization of flavonoids from Gingo biloba using respons
23、e surface analysis J.J Chin Inst Food Sci Technol(?),2013,13(2):83-90.10 HUANG X B,LIN L J,ZHOU Y M.Optimization of ultrasonic extraction technology of flavonoids from Longan(Dimocarpus longan)seed by response surface methodology J.Acta Agric Jiangxi(?),2012,24(4):116-119,123.11 SUN P,PEI G L,SUN X
24、F.Optimization of enzymatic extraction of polysaccharides from Sterculia lychnophora by central composite design J.Sci Technol Food Ind(?),2013,34(2):269-272.?2013-09-28?()?(?050035)?(Alpinia oxyphylla Miq.)?Sephadex LH-20?5?(1)?(2)?(3)?()-?(4)?(+)-?(5)?1?3?4?5?R284.1?B?1007-7693(2014)05-0549-03 DOI
25、:10.13748/ki.issn1007-7693.2014.05.008 Chemical Constituents from the Water Extract of Alpinia Oxyphylla Miq.()?CHANG Qingxian,WANG Zongquan,JIA Jiming(Shijiazhuang Yiling Pharmaceutical Co.,Ltd,Shijiazhuang 050035,China)ABSTRACT:OBJECTIVE To study the chemical constituents from the water extract of Alpinia oxyphylla Miq.METHODS The chemical constituents were isolated by solvent and Sephadex LH-20 methods.The structures were identified on the basis of?Tel:15232123326 E-mail: