超声波提取技术是指以超声波辐射产生的机械振动效应、空化效应和热效应，引起机械搅拌、加速扩散溶解的一种利用外场介入的强化溶剂提取方法。“空化现象”产生巨大压力的会破坏原料细胞壁，加速提取物的溶出。“机械振动”和“热效应”使溶出成分进一步扩散，最终实现实现固--液萃取分离，具有时间短，效率高，成本低的特点。目前利用超声波提取猴头菇多糖的研究很多，也取得了较为理想的结果。柴军红等人采用超声波法提取猴头菇下脚料多糖，得率为9.56%；张素斌等人对比7种提取方法对猴头菇多糖得率的影响，结果表明：超声波与复合酶法的协同作用可使多糖提取率大为提高，达到15.59％。

1 料液比对猴头菇多糖提取效果的影响

猴头菇子实体粉碎成过20目筛，按重量的20倍、30倍、40倍加入蒸馏水，分别在超声波中提取15分钟，用3号玻璃坩埚过滤并洗涤残渣得清液，合并清液并定容至250mL，清液用超纯水再稀释10倍后，取1mL滤液测定多糖浓度,每个处理3个重复。

2 超声时间对猴头菇多糖提取效果的影响

称取等量的子实体粉末（2.00克），按重量的20倍加入蒸馏水，于超声波中分别提取10、15、20分钟，用3号玻璃坩埚过滤并洗涤残渣得清液，合并清液并定容至250mL，清液用超纯水再稀释10倍后，取1mL滤液测定多糖浓度,每个处理3个重复。

3 超声水浴温度对猴头菇多糖提取效果的影响

称取同等量的子实体粉末（2.00克）三份，按重量的20倍加入蒸馏水，于，超声波的水浴温度分别为40℃、50℃、60℃中提取二次，用3号玻璃坩埚过滤并洗涤残渣得清液，合并清液并定容至250mL，清液用超纯水再稀释10倍后，取1mL滤液测定多糖浓度,每个处理3个重复。

4 正交试验

为进一步探讨浸提参数中的料液比、时间和浸提次数对子实体多糖的影响，进行3因素3水平的正交试验。

结果与讨论

1、料液比对猴头菇多糖提取效果的影响

以猴头菇粉末为实验材料，在水浴温度为50℃、超声时间为20min的条件下，研究料液比对猴头菇粗多糖浸提效果的影响，实验结果如图1。由图1可知，当料液比为1：20时，猴头菇多糖浸出量较大；随着料液比加大，粗多糖提取率提高不显著，为了节约成本及提高效率，故选择料液比为1：20作为水浴浸提的最佳条件。

表1 料液比对猴头菇粗多糖浸提的影响

Table1 The effect of the mixture ratio on theextraction yield of H. erinaceus polysaccharide

料液比	1：10	1：20	1：30	1：40	1：50	1：60
第一次提取率（%）	14.78	15.86	15.41	15.41	15.67	15.28
第一次提取率（%）	14.31	15.50	15.61	15.41	15.28	15.34
第一次提取率（%）	14.29	15.41	15.54	15.41	15.19	15.19
平均提取率（%）	14.46	15.59	15.52	15.41	15.38	15.27
差异性显著分析	Aa	Bb	Bb	Bb	Bb	Bb

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图1 料液比对猴头菇粗多糖浸提的影响

Fig1 The effect of solid-liquid ratio on theextraction yield of H. erinaceus polysaccharide

2 水浴温度对猴头菇多糖提取效果的影响

以剪碎后的猴头菇子实体为实验材料，在料液比为1：20、超声时间为20 min的条件下，研究温度对猴头菇粗多糖浸提效果的影响，实验结果如图2。由图2可知，当水浴温度为50℃时，猴头菇多糖浸出量较大；随着温度的升高，粗多糖提取率反而下降，这是因为提取温度过高时，粗多糖类化合物容易降解失活，故选择水浴温度为50℃作为水浴浸提的最佳条件。

表2 温度对猴头菇粗多糖浸提的影响

Table2 Theeffect of temperature on the extraction yield of hericium polysaccharide

温度（℃）	30	40	50	60	70
第一次提取率（%）	9.89	12.09	15.56	14.04	11.7
第一次提取率（%）	9.69	11.87	15.62	13.92	11.54
第一次提取率（%）	9.79	11.98	15.59	13.98	11.62
平均提取率（%）	9.79	11.98	15.59	13.98	11.62
差异性显著分析	Aa	Bb	Bb	Bb	Bb

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图2 温度对猴头菇粗多糖浸提的影响

Fig2 The effect of temperature on the extractionyield of hericium polysaccharide

3 浸提时间对猴头菇多糖提取效果的影响

以剪碎后的猴头菇子实体为实验材料，在料液比为1：20、温度为50℃的条件下，研究浸提时间对猴头菇粗多糖浸提效果的影响，实验结果如图3。由图3可知，当浸提时间为20min时，猴头菇多糖浸出量较大；随着浸提时间的延长，粗多糖提取率反而下降，这是因为浸提时间过长时，粗多糖类化合物易降解失活，故选择浸提时间为20min作为水浴浸提的最佳条件。

表3 超声时间对猴头菇粗多糖浸提的影响

Table3 Theeffect of ultrasonic time on the extraction yield of H. erinaceus polysaccharide

时间（min）	10	15	20	25	30
第一次提取率（%）	13.94	14.87	15.68	15.61	15.58
第二次提取率（%）	14.21	15.16	15.63	15.42	15.41
第三次提取率（%）	14.18	15.09	15.46	15.68	15.39
平均提取率（%）	14.11	15.04	15.59	15.57	15.46
差异性显著分析	Aa	Bb	Bb	Bb	Bb

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图3 超声时间对猴头菇粗多糖浸提的影响

Fig3 Theeffect of ultrasonic time on the extraction yield of hericium polysaccharide

4 最佳条件下水浴提取法的多糖提取率

在最佳的因素下，进行了浸提取，结果见表4。

表4 超声提取最优条件下多糖提取率

Table4 Theextraction rate of polysaccharide under the optimum condition by ultrasonic

实验次数	多糖提取率（%）		平均提取率（%）		RSD（%）
1		15.01		15.13		0.76
2		15.24
3		15.14

在最佳的超声波条件下：料液比为1：20、水浴温度：50℃、浸提时间：20分钟的条件下，进行三次平行实验，得到平均粗多糖提取率为15.13%，RSD符合要求。

表5 L9（3³）正交实验结果及其分析

Table5 Results and analysis of orthogonal experiment

实验号		因素				实验指标
实验号		料液比A	时间B	温度C	误差	提取率(%)
1		（1）1:20	（1）10	（1）30	1	9.61
2		（1）1:20	（2）15	（2）40	2	12.24
3		（1）1:20	（3）20	（3）50	3	15.59
4		（2）1:30	（1）10	（2）40	3	13.21
5		（2）1:30	（2）15	（3）50	1	15.52
6		（2）1:30	（3）20	（1）30	2	9.79
7		（3）1:40	（1）10	（3）50	2	14.11
8		（3）1:40	（2）15	（1）30	3	9.97
9		（3）1:40	（3）20	（2）40	1	11.98
总糖	K₁₁	37.44	36.93	29.37	37.11
	K₁₂	38.52	37.73	37.43	36.14
	K₁₃	36.06	37.36	45.22	38.77
	k1	12.48	12.3	9.8	12.4
	k2	12.8	12.6	12.5	12.0
	k3	12.0	12.5	15.1	12.9
	R	2.46	0.8	15.85	2.63
	K₂₁²	1401.754	1363.825	862.5969	1377.152
	K₂₂²	1483.79	1423.553	1401.0049	1306.1
	K₂₃²	1300.324	1395.77	2044.8484	1503.113
	Q	1395.289	1394.382	1436.150067	1395.455
	S	1.01	0.11	41.87	1.18

表6 正交试验方差分析表

Table6 Analysis of variance table for orthogonalexperiment

变异来源	平方和	自由度	均方	F值	显著水平
料液比A	1.01	2	0.51	0.86	F0.05（2，2）=19.0
时间B	0.1	2	0.1	0.09
温度C	41.9	2	20.9	35.51
误差	1.2	2	0.6
总和	44.2	8