5 11 Vol. 5 No. 11 2014 11 Journal of Food Safety and Quality Nov., 2014 - * 祝洪艳, 林海成, 杨鹤, 王国丽, 赵岩, 郜玉钢, 张连学 (, 130118) 摘要 : 目的 方法, -, SAS, 结果 14 75% 2, 2 h 34.26 mg/g 结论 -,,,, 关键词 : ; A; ; - Optimization of reflux extraction of jujuboside from semen ziziphi spinosae by central composite design-response surface methodology ZHU Hong-Yan, LIN Hai-Cheng, YANG He, WANG Guo-Li, ZHAO Yan, GAO Yu-Gang, ZHANG Lian-Xue * (College of Chinese Medicinal Materials, Jilin Agricultural University, Changchun 130118, China) ABSTRACT: Objective The extraction of jujuboside from semen ziziphi spinosae was optimized. Methods The content of jujuboside was detected by ultraviolet and visible spectrophotometry, and central composite design (CCD)-response surface methodology was used to study the effects of ethanol concentration, ethanol dosage and extraction time as well as their interactions on the content of jujuboside. A predictive model was developed by SAS software, and the optimum extraction conditions were determined by response surface analysis. Results The optimum reflux extraction conditions were 14 times of 75% ethanol extracted 2 h for 2 times. The average content of jujuboside was 34.26 mg/g. Conclusion CCD-response surface methodology for optimizing the reflux extraction press of jujuboside was reliable, and the detected content of jujuboside closed to the predictive value. The predicted extraction process was stabled, and it could be used to direct the industria- 基金项目 : (2011BAI03B010602) (2012ZX09304006) (20130102075JC) (201303111) (2014 60 ) Fund: Supported by National Key Technology Support Program (2011BAI03B010602), National Science and Technology Major Project (2012ZX09304006), the Project Development Plan of Science and Technology of Jilin Province (20130102075JC), National Public Scientific Research (201303111) and the Project of Education Fund of Jilin Province (2014060) * 通讯作者 :,,, E-mail: zlxbooksea@163.com *Corresponding author: ZHANG Lian-Xue, Professor, College of Chinese Medicine Material in Jilin Agricultural University, Changchun 130118, China. E-mail: zlxbooksea@163.com
11, : - 3719 lized extraction of jujuboside. KEY WORDS: jujuboside; jujuboside A; extraction; central composite design -response surface methodology 1 引言 (Semen ziziphi spinosae) [Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H.F. Chou], [1],, [2,3] [4], [5,6], [7-10],,,,, 2 材料与方法 2.1 材料与仪器,, [Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H.F. Chou] ; A (, : 20120423); (, 1306181) UV-6100 ( ); BS210S ( SARTORIUS ); RE-52AA ( ); SHB-Ⅲ ( ),, ( 99.5%, ); ( 99.7%, ); ( 99.0%, ); ( 60~90, ); 2.2 实验方法 2.2.1 酸枣仁总皂苷含量测定 (1) 1 A 2.20 mg, 10 ml, 0.22 mg/ml 2 ( 40 ), 6 h,, 5.0 g,,,,,,,,, 10 ml, (2) A 0.10 0.15 0.20 0.30 0.40 0.50 ml, 10 ml,, 5% 0.20 ml 0.80 ml,, 70 20 min, 5 min, 5 ml,,, UV-6100 546 nm [11] (mg/ml),, Y=26.034X+0.0116, R 2 =0.9995 0.0036~0.0179 mg/ml (3), [(2) ],, (mg/g) 2.2.2 酸枣仁总皂苷提取单因素实验 (1) 6, 70%, 30 min; ( 3:1) ph 4.5, 45 5 h [12] ; 70%, 1 h; 70%, 6 70% ;, 1 h
3720 5,, (2), 30% 50% 70% 80% 95%, 10:1, 2 h, (3), 70%, 8:1 10:1 12:1 15:1 20:1, 2 h, (4), 70%, 10:1, 1 1.5 2 2.5 3 h,, (5), 70%, 10:1, 1 h, 1 2 3 4, 2.2.3 酸枣仁总皂苷提取工艺的中心组合 - 响应面分析实验设计, (X 1 ) (X 2 ) (X 3 ) ( 1),, SAS,,, Origin, 表 1 酸枣仁总皂苷提取工艺的中心组合实验设计因素水平表 Table 1 Factors and levels of CCD design for jujuboside from semen ziziphi spinosae X 1 (%) X 2 (ml/g) X 3 (h) 1.682 50.0 10.0 1.0 1 56.1 11.0 1.2 0 65.0 12.5 1.5 1 73.9 14.0 1.8 1.682 80.0 15.0 2.0 2.2.4 酸枣仁皂苷最优提取工艺的验证实验 3,,, 3 结果与分析 3.1 酸枣仁总皂苷提取单因素实验 3.1.1 不同提取方法对酸枣仁总皂苷含量的影响, 1 1, 70%, [12],,,,,,, Fig. 1 1 (n=3) Effect of different extraction method on the content of jujuboside (n=3)
11, : - 3721 3.1.2 乙醇浓度对酸枣仁总皂苷含量的影响, 2 2, 70%, ; 70%, ; 70%,,,, 50%~80% 3.1.3 液固比对酸枣仁总皂苷含量的影响 70%, 3 3,, 8:1 10:1, ; 10:1,,, 10:1~15:1 3.1.4 提取时间对酸枣仁总皂苷含量的影响 70%,, 4 Fig. 2 2 (n=3) Effect of ethanol concentration on the content of jujuboside (n=3) Fig. 3 3 (n=3) Effect of solvent-to-solid ratio on the content of jujuboside (n=3) Fig. 4 4 (n=3) Effect of extraction time on the content of jujuboside (n=3)
3722 5 4,, 2 h,, 1~2 h 3.1.5 提取次数对酸枣仁总皂苷含量的影响 70%, 5 5, 2 1, 3 4,,, 2 3.2 酸枣仁皂苷提取的中心组合设计实验结果 SAS ( 2),, : Y= 77.44+1.59X 1 +6.18X 2 13.32X 3 0.02X 2 1 +0.10X 2 X 1 0.44X 2 2 +0.06X 3 X 1 +0.35X 3 X 2 +3.02X 2 3 ; 3, 4, 5 Fig. 5 5 (n=3) Effect of extraction times on the content of jujuboside (n=3) 表 2 酸枣仁总皂苷的提取工艺中心组合设计实验设计及结果 Table 2 CCD matrix and results of jujuboside from semen ziziphi spinosae X 1 (%) X 2 (ml/g) X 3 (h) (mg/g) 1 2 3 (mg/g) S1 0(65.0) 0(12.5) 0(1.5) 27.47 29.86 29.31 28.88 30.38 S2 0 0 0 29.25 29.21 27.34 28.60 30.38 S3 0 0 0 27.13 30.23 27.60 28.32 30.38 S4 1(73.9) 1(14.0) 1(1.8) 35.98 37.04 35.43 36.15 37.19 S5 1 1 1(1.2) 33.93 33.97 33.17 33.69 34.14 S6 1 1(11.0) 1 27.86 27.88 27.54 27.76 27.87 S7 1 1 1 27.45 26.34 26.73 26.84 27.17 S8 1(56.1) 1 1 24.03 21.78 24.87 23.56 28.32 S9 1 1 1 25.14 21.67 20.87 22.56 25.92 S10 1 1 1 21.56 21.34 20.67 21.19 24.06 S11 1 1 1 19.88 20.12 19.76 19.92 22.28 S12 1.682(50.0) 0 0 17.55 19.32 18.21 18.36 20.93 S13 1.682(80.0) 0 0 30.63 30.93 31.20 30.92 30.84 S14 0 1.682(10.0) 0 22.07 20.36 22.13 21.52 21.83 S15 0 1.682(15.0) 0 31.24 29.87 29.13 30.08 37.35 S16 0 0 1.682(1.0) 27.16 25.34 27.78 26.76 35.21 S17 0 0 1.682(2.0) 29.56 30.43 35.89 31.96 41.00
11, : - 3723 Table 3 表 3 回归系数显著性检验表 Significance test of each coefficient in the developed regression model t P X 1 1 77.442594 39.675928 1.95 0.0919 1 1.596818 0.562161 2.66 0.0323 * X 2 1 6.178237 3.578923 1.72 0.1282 X 3 1 13.329585 14.944914 0.89 0.4021 X 1 2 1 0.017618 0.003384 5.21 0.0012 ** X 2 X 1 1 0.095572 0.023939 3.99 0.0052 ** X 2 2 1 0.438665 0.121462 3.69 0.0077 ** X 3 X 1 1 0.051850 0.119694 0.43 0.6779 X 3 X 2 1 0.352778 0.711782 0.50 0.6353 X 3 2 1 3.023373 3.036551 1.00 0.3526 : P<0.05, * ; P<0.01, ** Table 4 表 4 回归模型方差分析 Analysis of variance for the developed regression model F P 9 400.575961 0.9859 54.23 <0.0001 ** 3 352.175954 0.8667 143.03 <0.0001 ** 3 34.962769 0.0860 14.20 0.0023 ** 3 13.437238 0.0331 5.46 0.0300 * 7 5.745228 0.820747 5 5.588428 1.117686 14.26 0.0668 2 0.156800 0.078400 Table 5 表 5 模型岭脊回归分析 Ridge analysis for the developed regression model X 1 X 2 X 3 0.0 28.600499 0.521567 65.000000 12.500000 1.500000 0.1 29.435154 0.518235 66.234611 12.630223 1.511316 0.2 30.235925 0.508784 67.413369 12.769914 1.524786 0.3 31.005808 0.494951 68.539117 12.916185 1.540676 0.4 31.747585 0.479948 69.613710 13.066496 1.559261 0.5 32.463956 0.468813 70.637793 13.218561 1.580811 0.6 33.157631 0.468458 71.610853 13.370287 1.605568 0.7 33.831397 0.486704 72.531497 13.519743 1.633712 0.8 34.488141 0.530054 73.397920 13.665181 1.665327 0.9 35.130837 0.601604 74.208489 13.805098 1.700376 1.0 35.762494 0.700998 74.962327 13.938309 1.738695
3724 5 6 Fig. 6 Response surface and contour lines of the content of Jujuboside
11, : - 3725 3, (X 1 ) (P<0.05);, (X 2 ) (X 1 ) (P<0.01);, (X 1 ) (X 2 ) (P<0.01), (X 3 ), (X 1 ) (X 2 ), P>0.3, : Y= 77.44+1.59X 1 +6.18X 2 0.02X 2 1 +0.10X 2 X 1 0.44X 2 2,,, P<0.3, : Y= 77.44+1.59X 1 + 6.18X 2 13.32X 3 0.02X 2 1 +0.10X 2 X 1 0.44X 2 2 +0.05X 3 X 1 + 0.35X 3 X 2 +3.02X 2 3 4, (P<0.01), (P<0.05) R 2 =0.9859, P>0.05, 5, 75.0%, 13.9 ml/g, 2, 1.7 h, 35.76 mg/g 3.3 酸枣仁皂苷提取的响应面优化与预测分析, Origin ( 6), ( 0), 6, (X 1 ) (X 2 ) (X 3 ),,, : (X 1 )75%~80%; (X 2 )14~15 ml/g; (X 3 )1.9~2 h, 75%, 14:1, 2, 2 h 3.4 优选酸枣仁皂苷提取工艺的验证实验, 75%, 14:1, 2, 2 h, 3, 34.12 33.71 34.96 mg/g, 34.26 mg/g,, 4 讨论 [13],, 82.3%, 1:22.3, 78.5 3 h,,,,, 75%, 14:1, 2, 2 h, 3.426%, [14] [15],,, 参考文献 [1]. (2010 Ⅰ )[M]. :, 2010: 343 344. Pharmacopoeia Commission of People s Republic of China. Pharmacopoeia of people s republic of China: 2010 Part Ⅰ[M]. Beijing: Chinese Medical Science and Technology Press, 2010: 343 344. [2]. 60 [J]., 2014, 28(6): 53. Kang J. 60 cases of insomnia treatment efficacy suanzaoren decoction [J]. J Pract Trad Chin Int Med, 2014, 28(6): 53. [3]. [J]., 2014, 29(1): 152 154. Ding DZ. Examples of suanzaoren decoction on mental illness [J]. China J Trad Chin Med Pharm, 2014, 29(1): 152 154. [4],,. [J]., 2013, 28(4): 550 552. Zhang XW, Zhao Q, Wei YH. Research progress of biology characteristics and chemical constituents of semen ziziphi spinosae [J]. China J Chin Med, 2013, 28(4): 550 552.
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