1、 290 農業工程學報 2013 年 的生產試驗。 由表 4 發現，采用不加超聲的傳統提取方法提 取 88 min 時紫菜產品得率為（17.950.12）%，其 中 多 糖 得 率 為 （ 10.760.06 ） % ， 蛋 白 得 率 為 （7.190.07）%。周存山等9也采用傳統提取方法對 條斑紫菜中多糖進行了提取研究，獲得的多糖得率 為 22.22%， 明顯高于本文得到的數據， 這主要是因 為不同的提取條件和脫蛋白方法造成的。研究結果 還顯示，傳統提取方法獲得的產品得率僅達到了交 替雙頻逆流超聲輔助提取方法獲得的產品得率的 37.26%；當達到傳統提取得率時，交替雙頻逆流超 聲輔助提取

2、方法僅花費了 32 min， 比傳統提取時間 縮短了 56 min。由此得出，交替雙頻逆流超聲輔助 提取方法與傳統提取方法相比，產品得率增加了 168%， 生產時間縮短了 64%， 不論從產品得率還是 生產周期而言，交替雙頻逆流超聲輔助提取方法均 顯著優于傳統提取方法。這是因為交替雙頻超聲可 以看作為一種變相的脈沖作用，可以實現超聲功率 短時間內的突變，隨后又迅速返回其初始值，這種 脈沖沖擊力可強化超聲波對細胞壁的破壞作用和 傳質效果11。 2.5 交替雙頻逆流超聲輔助提取與單頻逆流超聲 輔助提取效果的比較 在單頻逆流超聲輔助提取的最優條件下，采用 單頻逆流超聲輔助提取方法進行紫菜蛋白和多糖

3、混合產品的生產試驗。 由表 4 發現，單頻逆流超聲輔助提取方法獲得 的紫菜產品得率為（32.140.16）%，其中蛋白得率 為（13.420.14）%，多糖得率為（18.720.03）%。 同樣的，何榮海等8也采用單頻超聲輔助提取技術 提取條斑紫菜中的多糖，獲得的多糖得率為 26.29%，顯著高于本文得到的數據，這是因為他們 采用較高的超聲功率和較長的超聲時間造成的。馬 海樂等 11 還利用單頻超聲輔助提取技術提取條斑 紫菜中的藻紅蛋白， 得率為 3.249%。 研究結果還顯 示，單頻逆流超聲輔助提取方法獲得的產品得率僅 達到交替雙頻逆流超聲輔助提取方法獲得的產品 得率的 66.71%； 實現

4、傳統提取得率， 單頻逆流超聲 輔助提取方法僅花費了 39 min， 比傳統提取時間縮短 了 49 min。相關的研究報道也發現，與傳統提取方法 相比， 超聲輔助提取方法可以大大縮短提取時間2,11。 此外，研究還顯示單頻逆流超聲輔助提取時間比交 替雙頻逆流超聲輔助提取時間多了 7 min。由此得 出，交替雙頻逆流超聲輔助提取方法與單頻逆流超 聲輔助提取方法相比，產品得率增加了 50%，提取 時間縮短了 18%，不論從產品得率還是生產周期而 言，交替雙頻逆流超聲輔助提取方法顯著優于單頻 逆流超聲輔助提取方法。這主要是因為紫菜蛋白和 多糖的結構及存在形式不同，單一頻率的超聲很難 取得理想的溶解效果

5、，而交替雙頻超聲產生比單頻 超聲更強烈的空化效應22，更有利于改變物質的組 織結構和狀態，促進紫菜中蛋白和多糖的溶解，提 高產品得率。 3 結 論 1）交替雙頻超聲模式顯著優于復合雙頻超聲 模式，15 和 20 kHz 交替雙頻超聲是最佳的雙頻超 聲模式。料液比、提取時間和溫度關鍵因素均對交 替雙頻逆流超聲輔助提取效果產生顯著影響。通過 中心組合設計響應面優化試驗確定了交替雙頻逆 流超聲輔助提取的最優工藝條件為：料液比 14 mg/mL、時間 88 min、溫度 41、pH 值 9.0、 15 和 20 kHz 交替雙頻超聲、 超聲交替工作時間各 3 s、功率 200 W/L，在此最優條件下，

6、條斑紫菜蛋白 和多糖混合產品的生產能力最高，單位質量原料生 產出的產品得率為（48.180.08）%，其中蛋白得率 為（24.350.07）%，純度為（45.610.33）%，多 糖得率為（23.830.02）%，純度為（44.640.37） %。 2）從生產成本分析，交替雙頻逆流超聲輔助 提取方法與傳統提取方法相比，產品得率增加了 168%， 生產時間縮短了 64%； 與單頻逆流超聲輔助 提取方法相比，產品得率增加了 50%，提取時間縮 短了 18%。交替雙頻逆流超聲輔助提取技術不僅可 以解決傳統提取技術和現有單頻超聲輔助提取技 術的發展瓶頸，而且可以大大縮短生產周期，提高 產品得率，因此與

7、現有提取技術相比，該技術更適 合工業化生產，工業化推廣價值更高，可應用于農 產品和食品中有效成分提取。 參 考 文 獻 童冠文 . 條斑紫菜經濟性狀研究進展 J. 現代農業科 技，2010，(11：343345. 2 王婷，馬海樂，曲文娟，等. 條斑紫菜蛋白和多糖的 逆流脈沖超聲輔助提取技術研究J. 食品工業科技， 2012，(10：16. Wang Ting, Ma Haile, Qu Wenjuan，et al. Study on counter-current pulsed ultrasound-assisted extraction of protein and polysacchar

8、ide from Porphyra yezoensisJ. Science and Technology of Food Industry, 2012, (10：1 3 6. (in Chinese with English abstract Zhou Cunshan, Yu Xiaojie, Zhang Youzuo. Ultrasonic degradation, purification and analysis of structure and antioxidant activity of polysaccharide from Porphyra yezoensis UdeaJ. C

9、arbohydrate Polymers, 2012, 87(1: 20462051. 1 第1期 4 曲文娟等：交替雙頻逆流超聲輔助提取條斑紫菜蛋白和多糖 291 楊旭東，張杰，林峰. 條斑紫菜多糖對糖尿病大鼠腦 組織的保護作用J.中國康復理論與實踐， 2011， 17(8： 739741. Yang Xudong, Zhang Jie, Lin Feng. Protecting effect of porphyra yezoensis polysaccharide on cerebrum tissue of diabetic ratsJ. Chinese Journal of Rehabi

10、litation Theory and Practice, 2011, 17(8: 739741. (in Chinese with English abstract 時旭，蔡春爾，李春霞，等. 條斑紫菜藻膽蛋白對小 鼠免疫功能及抗氧化活性的影響J. 中國海洋藥物雜 志，2011，30(2：4448. Shi Xu, Cai Chuner, Li Chunxia, et al. Effects of phycobiliprotein from porphyra yezoensis on the mice immunity and antioxidation activityJ. Chinese

11、 Journal of Marine Drugs, 2011, 30(2: 4448. (in Chinese with English abstract Qu Wenjuan, Ma Haile, Pan Zhongli. Preparation and antihypertensive activity of peptides from porphyra yezoensisJ. Food Chemistry, 2010, 123(1: 1420. 呂鐘鐘， 羅建光， 管華詩. 紫菜的生物活性研究進展J. 中國海洋大學學報，2009，39(增刊：4751. Lu Zhongzhong，Luo

12、 Jianguang，Guan Huashi. Progress on chemical constituents and biological activities of PorphyraJ. Periodical of Ocean University of China, 2009, 39(Supp.: 47 51. (in Chinese with English abstract 何榮海，馬海樂，駱新崢. 條斑紫菜多糖超聲提取及 其對 U937 細胞生長抑制影響的研究J. 農業工程學 報，2005，21(8：165168. He Ronghai, Ma Haile, Luo Xinzh

13、eng. Ultrasonic extraction of polysaccharides from porphyra yezoensis and their inhibition effects on U937 cell growthJ. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE, 2005, 21(8: 165168. (in Chinese with English abstract 14 13 6 15 7 8 16 17 9 聲輔助提取J. 農業工

14、程學報， 2007， 23(1： 207210. Ma Haile, Xiao Haifang, Luo Lin. Optimization of the technology for pulse ultrasonic assistant extraction of phycoerythrin from porphyra yazoensisJ. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE, 2007, 23(1: 207210. (in Chinese wit

15、h English abstract Pingret D, Fabiano-Tixier A S, Le Bourvellec C, et al. Lab and pilot-scale ultrasound-assisted water extraction of polyphenols from apple pomaceJ. Journal of Food Engineering, 2012, 111(1: 7381. Xia Enqin, Yu Yingying, Xu Xiangrong, et al. Ultrasound-assisted extraction of oleanol

16、ic acid and ursolic acid from ligustrum lucidum AitJ. Ultrasonics Sonochemistry, 2012, 19(4: 772776. Xie Jianhua, Shen Mingyue, Xie Mingyong, et al. Ultrasonic-assisted extraction antimicrobial and antioxidant activities of cyclocarya paliurus (Batal. iljinskaja polysaccharidesJ. Carbohydrate Polyme

17、rs, 2012, 89(1: 177184. GalvandAlessandro L, Kriaa K, Nikov I, et al. Ultrasound assisted extraction of polyphenols from black chokeberryJ. Separation and Purification Technology, 2012, 93(1: 4247. Pan Zhongli, Qu Wenjuan, Ma Haile, et al. Continuous and pulsed ultrasound-assisted extractions of ant

18、ioxidants from pomegranate peelJ. Ultrasonics Sonochemistry, 2011, 18(5: 12491257. 18 曹雁平，程偉. 多頻超聲連續逆流浸取黃芩中的黃芩 苷J. 食品科學，2008，29(11：219222. Cao Yanping, Chen Wei. Multi-frequency ultrasonic continuous countercurrent extraction of baicalin from scutellaria rootJ. Food Science, 2008, 29(11: 219222. (in

19、 Chinese with English abstract 19 陳俊輝. 生物化學實驗M. 北京：科學出版社，2008： 65. 20 丁鋼強，于村. 食品多糖含量不同測定方法的研究J. 實用預防醫學，2000，7(5：325327. Ding Gangqiang, Yu Cun. The Research of determination on food polysaccharide methodsJ. Practical Preventire Medicine, 2000, 7(5: 325 327. (in Chinese with English abstract 21 曹雁平，矯

20、慶澤. 超聲功率、頻率對姜黃素浸取的影 響與動力學模型J. 高校化學工程學報，2011，25(2： 212218. Cao Yanping, Jiao Qingze. Dynamic model of the ultrasound-assisted extraction of the curcumin from curcuma longa L and the effects of ultrasound intensity and frequency on the extractionJ. Journal of Chemical Engineering of Chinese Universiti

21、es, 2011, 25(2: 212 218. (in Chinese with English abstract 22 崔莉，張培正，孫愛東 . 超聲波輔助對紅曲色素提 取的影響 J. 農業工程學報， 2009 ， 25(1 ： 159 163. 10 周存山，馬海樂，胡文彬. 條斑紫菜多糖提取工藝的 優化J. 農業工程學報，2006，22(9：194197. Zhou Cunshan, Ma Haile, Hu Wenbin. Optimization of the technology for extracting polysaccharides from porphyra yezoe

22、nsisJ. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE, 2006, 22(9: 194197. (in Chinese with English abstract 11 劉青梅，楊性民，鄧紅霞，等. 采用微波技術提取紫 菜多糖的工藝研究J. 農業工程學報，2005，21(2： 153156. Liu Qingmei, Yang Xingmin, Den Hongxia. Technology for extraction of polysaccharide

23、 by using microwave technology from porphyra yezoensisJ. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE, 2005, 21(2: 153156. (in Chinese with English abstract 12 馬海樂，肖海芳，駱琳. 條斑紫菜藻紅蛋白的脈沖超 292 農業工程學報 2013 年 Agricultural Engineering (Transactions of the CSAE,

24、2009, 25(1: 159 163. (in Chinese with English abstract Cui Li, Zhang Peizheng, Sun Aidong. Dynamic model of the effects of ultrasonic assist on extraction of monascus pigmentsJ. Transactions of the Chinese Society of Alternating two-frequency countercurrent ultrasonic-assisted extraction of protein

25、and polysaccharide from Porphyra yezoensis Qu Wenjuan1, Ma Haile1,2, Wang Ting1, Zheng Huihua3 (1. Department of Food & Biological Engineering, Jiangsu University, Zhenjiang, 212013, China; 2. Jiangsu Key Lab of Physical Processing of Agricultural Product, Zhenjiang, 212013, China; 3. China Jiangsu

26、Alphay Biological Technology Co., Ltd, Nantong, 226009, China Abstract: Porphyra yezoensis is rich in high contents of protein and polysaccharide, while that have not been fully used and developed. In order to improve resource utilization rate of Porphyra yezoensis, a new technology of alternating t

27、wo-frequency countercurrent ultrasonic-assisted extraction is developed to produce high-yield protein and polysaccharide mixed products. The effects of two ultrasonic ways were investigated in this research: the composite working and the alternating working of two-frequency (two-frequency of A: 15 a

28、nd B: 20 kHz, A: 15 and C: 28 kHz, and B: 20 and C: 28 kHz, respectively referred to as the AB composition, AB alternation, AC composition, AC alternation, BC composition, and BC alternation, on the product yield (expressed as the total yields of protein and polysaccharide. The response surface opti

29、mization tests for three key factors of material to water ratio, extraction time, and temperature were conducted to optimize the extraction process, based on the central composite design (CCD. Moreover, in order to analyze the feasibility of alternating two-frequency countercurrent ultrasonic-assist

30、ed extraction technique, the extraction performance of the alternating two-frequency countercurrent ultrasonic-assisted extraction method was compared with those of the mono-frequency countercurrent ultrasonic-assisted extraction and the traditional extraction methods. The results showed that the al

31、ternating ultrasonic way was significantly better than the composite one and the alternating two-frequency of 15 and 20 kHz was the most effective. The factors: material to water ratio, extraction time, and temperature had significant effects on the extraction process. Therefore, the established opt

32、imum conditions of alternating two-frequency countercurrent ultrasonic-assisted extraction were as follows: the material to water ratio was 14 mg/mL, the extraction time was 88 min, the temperature was 41oC, the pH value was 9.0, the ultrasonic power was 200 W/L, the alternating two-frequency was 15 and 20 kHz, and an alternating working time of 3 s. Under these