Optimization of Ultrasound-Microwave Assisted Extraction of Melanin from Auricularia auricula-judae
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摘要: 以黑木耳（Auricularia auricula-judae）粉末为材料，研究了超声微波联用辅助提取黑木耳黑色素的方法。通过单因素实验考察黑木耳干粉粉碎粒度、超声功率、微波功率和超声微波联用时间对提取效果的影响，并采用响应面法优化工艺条件。通过与传统溶剂提取法及超声辅助提取法进行比较，结合扫描电镜（Scanning electron microscope）观察不同提取方法对于黑木耳干粉组织结构的破坏程度，以此验证超声微波联用辅助提取黑木耳黑色素的优势。结果表明：木耳黑色素的最佳提取工艺条件是：超声功率398 W、微波功率392 W、联合作用时间31 min。此条件下黑木耳黑色素吸光度为0.984，粗黑色素得率达到9.10%，相较传统溶剂法提取法（7.20%）、超声辅助提取法（5.20%）分别提高26.38%、75.00%。经比较不同提取方法后的黑木耳组织的SEM图像，表明黑木耳组织经超声微波联用辅助提取方法后，组织结构破损严重。结果证实：超声微波联用辅助提取黑木耳黑色素的方法相较于传统溶剂法可节约溶剂的使用量，相较于超声辅助提取法所耗费的时间有所减少，并且可提升黑木耳黑色素得率。Abstract: The ultrasound-microwave assisted extraction(UMAE) of melanin from Auricularia auricula-judae powder was studied in this study. The effects of particle size, ultrasonic power, microwave power and ultrasound-microwave combined action time on the extraction of melanin from A. auricula-judae
powder were investigated by single factor experiment, and the process conditions were optimized by response surface methodology.Compared with the traditional solvent extraction method and ultrasound assisted extraction method, combined with scanning electron microscope (SEM), the damage degree of different extraction methods on the structure of A. auricula-judae dry powder was observed, so as to verify the advantages of ultrasound-microwave assisted extraction of A. auricula-judae melanin.The results showed that the optimal extraction conditions were as follows: Ultrasonic power 398 W, microwave power 392 W, combined action time 31 min. Under these conditions, the absorbance of A. auricula-judae melanin was 0.984, and the yield reached 9.10%. Compared with the traditional solvent extraction method (7.20%) and ultrasound-assisted extraction method (5.2%), the extraction yield of Auricularia auricula melanin increased by 26.38% and 75.00%, respectively. Compared the SEM images of the tissue of A. auricula-judae with different extraction methods, it was shown that the tissue structure of A. auricula-judae was seriously damaged after ultrasound-microwave assisted extraction. The results confirmed that the method of ultrasound-microwave assisted extraction of A. auricula-judae melanin could save the amount of solvent compared with the traditional solvent method, reduce the time spent compared with the ultrasound-microwave assisted extraction method, and improve the yield of A. auricula-judae melanin. -
图 1 黑木耳黑色素的紫外扫描图谱
Figure 1. Ultraviolet scanning spectrum of A. auricula-judae melanin
图 2 干粉微粉碎粒度对黑色素吸光度的影响
Figure 2. Effect of dry powder particle size on melanin absorbance
图 5 超声-微波联用时间对黑色素吸光度的影响
Figure 5. Effect of ultrasound-microwave combined action time on the absorbance of melanin
图 6 超声功率和微波功率对黑色素吸光度影响的响应面和等高线图
Figure 6. Response surface and contour diagram of the effect of ultrasonic power and microwave power on the absorbance of melanin
图 8 微波功率和联合作用时间对黑色素吸光度影响的响应面和等高线图
Figure 8. Response surface and contour diagram of the effect of microwave power and combined action time on the absorbance of melanin
图 7 超声功率和联合作用时间对黑色素吸光度影响的响应面和等高线图
Figure 7. Response surface and contour diagram of the effect of ultrasonic power and combined action time on the absorbance of melanin
图 9 不同处理方法的黑木耳组织样品扫描电子显微镜图片
注：A：黑木耳干粉粉碎样品； B：超声辅助提取法处理后黑木耳样品； C：传统溶剂提取法处理后黑木耳样品；D：超声-微波联用辅助提取法处理后黑木耳样品。
Figure 9. Scanning electron microscope images of samples with different treatment methods
表 1 黑木耳黑色素提取 Box-Behnken 试验因素和水平
Table 1. Factors and levels of Box-Behnken test for A. auricula-judae melanin extraction
水平 X1超声功率（W） X2微波功率 (W） X3联合作用时间（ min） 1 350 300 20 2 400 400 30 3 450 500 40 
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表 2 黑木耳黑色素提取 Box-Behnken 试验方案与结果
Table 2. Scheme and results of Box-Behnken experiment for extraction of A. auricula-judae melanin
试验号 X1 X2 X3 OD207 试验号 X1 X2 X3 OD207 1 −1 −1 0 0.841 10 0 1 −1 0.847 2 1 −1 0 0.853 11 0 −1 1 0.797 3 −1 1 0 0.819 12 0 1 1 0.831 4 1 1 0 0.774 13 0 0 0 0.963 5 −1 0 −1 0.768 14 0 0 0 0.977 6 1 0 −1 0.684 15 0 0 0 1.025 7 −1 0 1 0.826 16 0 0 0 0.986 8 1 0 1 0.817 17 0 0 0 1.007 9 0 −1 −1 0.823
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表 3 方差分析
Table 3. Variance analysis
方差来源 自由度 平方和 均方 F值 P值 X1 1 1.985E-003 1.985E-003 1.08 0.3324 X2 1 2.311E-004 2.311E-004 0.13 0.7328 X3 1 2.775E-003 2.775E-003 1.52 0.2579 X1X2 1 8.123E-004 8.123E-004 0.44 0.5266 X1X3 1 1.406E-003 1.406E-003 0.77 0.4098 X2X3 1 2.500E-005 2.500E-005 0.014 0.9102 ${\rm{X}}_1^2 $ 1 0.051 0.051 27.99 0.0011** ${\rm{X}}_2^2 $ 1 0.015 0.015 8.16 0.0245* ${\rm{X}}_3^2 $ 1 0.049 0.049 26.61 0.0013** 模型 9 0.13 0.015 8.16 0.0057** 残差 7 0.013 1.830E-003 失拟项 3 0.010 3.465E-003 5.74 0.0623 纯误差 4 2.415E-003 6.038E-004 总和 16 0.15 注：*表示差异显著（P < 0.05），**表示差异极显著（P < 0.01）。
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