Extraction of Blackcurrant Seed Oil by Ultrasound-Assisted Aqueous Enzymatic Method and Its Quality Analysis
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摘要: 研究了超声波辅助水酶法提取黑加仑籽油的工艺条件，并对产品的品质进行分析。利用超声波对黑加仑籽进行前处理，以油脂提取率为考察指标，通过单因素实验和响应面法优化酶解条件，确定最佳工艺条件为：超声功率300 W，超声温度50 ℃，超声时间25 min，复合酶添加量为2%（蛋白酶:纤维素酶:果胶酶=2:2:1），酶解初始pH6.5，酶解温度55 ℃，液料比5:1（g/g），酶解时间3 h，在最优条件下黑加仑籽油提取率为78.34%。通过理化指标对超声波辅助水酶法提取黑加仑籽油的品质进行分析。结果表明，黑加仑籽油的感官指标、过氧化值和酸价均符合国家食用植物油卫生标准要求；富含不饱和脂肪酸，其中多不饱和脂肪酸比例为67.92%，油中多酚、α-生育酚、γ-生育酚含量分别达到300.49、19.55、41.67 mg/kg，是一种具有高营养价值的功能性植物油脂。本研究为黑加仑籽油工业化生产提供理论参考。Abstract: The process condition of blackcurrant (Ribes nigrum L.) seed oil by microwave-assisted aqueous enzymatic method was studied, and the quality of the product was analyzed. According to extraction rate, the optimum conditions of blackcurrant seed oil by microwave-assisted aqueous enzymatic method were explored by single-factor tests and response surface methodology. The optimum conditions were determined as follows: Ultrasonic power 300 W, ultrasonic temperature 50 ℃, ultrasonic time 25 min, composite enzyme dosage 2% (protease:compound cellulase:pectinase=2:2:1), hydrolysis initial pH6.5, hydrolysis temperature 55 ℃, liquid-solid ratio 5:1 (g/g), hydrolysis time 3 h. Under the optimal conditions, the extraction rate of blackcurrant seed oil was 78.34%. The quality of blackcurrant seed oil were analyzed by physicochemical indexes. The results showed that the sensory index, peroxide value and acid value of blackcurrant seed oil reached the national sanitary standard of edible vegetable oil. Blackcurrant seed oil was a kind of functional vegetable oil with high nutritional value, which contained unsaturated fatty acids (poly unsaturated fatty acids accounted for 67.92%). The contents of polyphenols, α-tocopherols, γ-tocopherols reached 300.49, 19.55 and 41.67 mg/kg. This study would provide a theoretical reference for the industrial production of blackcurrant seed oil.
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Key words:
- blackcurrant seed /
- oil /
- quality analysis /
- ultrasound /
- aqueous enzymatic method
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图 1 酶解时间、酶解初始pH、酶解温度和液料比对黑加仑籽油提取率的影响
Figure 1. Effects of hydrolysis time, hydrolysis initial pH, hydrolysis temperature and liquid-solid ratio on the extraction rate of blackcurrant seed oil
注：不同字母表示样品间存在显著性差异（P<0.05）；图3同。
图 2 酶解初始pH、酶解温度和液料比交互作用的响应曲面图
Figure 2. Response surface diagram for the interaction effect between hydrolysis initial pH, hydrolysis temperature and liquid-solid ratio
图 3 超声波辅助水酶法和水酶法油脂提取率的比较
Figure 3. Comparison of oil extraction rate by ultrasound-assisted aqueous enzymatic method and aqueous enzymatic method
表 1 响应面因素与水平设计
Table 1. Factors and levels of response surface experiment
因素 水平 ?1 0 1 A酶解初始pH 4.0 6.0 8.0 B酶解温度（℃） 45 55 65 C液料比（g/g） 4:1 5:1 6:1 
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表 2 响应面设计方案及结果
Table 2. Design and result of response surface analysis
实验号 因素 提取率（%） A B C 1 0 0 0 79.34±0.87 2 ?1 ?1 0 56.77±1.57 3 0 ?1 1 66.31±0.53 4 1 1 0 68.91±0.45 5 ?1 1 0 61.88±1.10 6 0 0 0 80.03±0.38 7 1 0 1 67.64±0.16 8 1 0 ?1 71.96±0.33 9 0 1 ?1 68.93±0.68 10 ?1 0 1 67.21±0.78 11 1 ?1 0 60.82±0.52 12 0 1 1 71.24±0.59 13 0 0 0 78.86±0.37 14 0 0 0 80.69±0.46 15 ?1 0 ?1 62.91±0.12 16 0 ?1 ?1 66.29±0.27 17 0 0 0 78.61±0.39 
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表 3 回归模型方差分析
Table 3. Variance analysis of the regression equation
来源 自由度df 平方和 均方 F值 P 显著性 模型 9 870.48 96.72 85.14 <0.0001 ** A 1 52.82 52.82 46.49 0.0002 ** B 1 53.75 53.75 47.31 0.0002 ** C 1 0.68 0.68 0.60 0.463 AB 1 2.21 2.21 1.94 0.2062 AC 1 18.58 18.58 16.36 0.0049 ** BC 1 1.35 1.35 1.19 0.3121 A2 1 347.39 347.39 305.79 <0.0001 ** B2 1 291.97 291.97 257.00 <0.0001 ** C2 1 37.65 37.65 33.14 0.0007 ** 残差 7 7.95 1.14 失拟项 3 5.01 1.67 2.27 0.2221 纯误差 4 2.94 0.73 总和 16 878.43 注：*表示差异显著（P<0.05）；**表示差异极显著（P<0.01）。 
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表 4 黑加仑籽油的主要理化指标
Table 4. Main physicochemical index of blackcurrant seed oil
理化指标 商品油 黑加仑籽油 国标 色泽 浅黄绿色 浅黄绿色 具有产品应有的色泽 状态 澄清、透明 澄清、透明 具有产品应有的
状态，无正常视力
可见的外来异物滋味和气味 具有清甜的
香味，无异味具有清甜的
香味，无异味具有产品应有的
滋味和气温，无焦臭、
酸败及其他异味酸值（mg KOH/g） 0.76±0.07b 0.87±0.08a ≤4.0 过氧化值（mmol/kg） 0.19±0.05a 0.15±0.03a ≤0.25 碘值（g I2/100 g） 198.97±2.58a 196.51±3.31a ? 皂化值（mg KOH/g） 192.48±4.15a 188.67±5.02a ? 相对密度 0.95±0.08a 0.94±0.05a ? 折光指数（n40） 1.48±0.25ab 1.61±0.33a ? 不溶性杂质（%） ≤0.05 ≤0.05 ? 注：同行不同小写字母表示样品间存在显著性差异（P<0.05）；表5~表7同。 
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表 5 黑加仑籽油脂肪酸组成及相对含量（%）
Table 5. Fatty acid composition and relative content of blackcurrant seed oil (%)
脂肪酸 商品油 黑加仑籽油 豆蔻酸C14:0 0.17±0.02b 0.22±0.01a 棕榈酸C16:0 4.24±0.25a 4.57±0.53a 棕榈一烯酸C16:1 0.25±0.01b 0.29±0.03a 硬脂酸C18:0 0.86±0.18b 1.85±0.38a 油酸C18:1 16.89±1.08a 17.51±1.21a 亚油酸C18:2 39.89±1.14a 34.54±1.89b 亚麻酸C18:3 31.55±1.01ab 33.38±2.03a 花生酸C20:0 0.08±0.01a 0.05±0.01b 花生一烯酸C20:1 2.12±0.03b 3.09±0.07a 不饱和脂肪酸比例（%） 96.05±1.78a 95.51±2.13a 
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表 6 黑加仑籽油反式脂肪酸含量（%）
Table 6. Trans-fatty acid content in blackcurrant seed oil (%)
反式脂肪酸 商品油 黑加仑籽油 反式油酸 0.48±0.03b 1.56±0.01a 反式亚油酸 0.33±0.02b 0.47±0.01a 反式亚麻酸 0.64±0.06a 0.53±0.04b 总量 1.45±0.11b 2.56±0.18a 
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表 7 黑加仑籽油中活性成分含量（mg/kg）
Table 7. Active ingredient content in blackcurrant seed oil (mg/kg)
活性成分（mg/kg） 商品油 黑加仑籽油 α-生育酚 18.68±0.17ab 19.55±0.35a γ-生育酚 43.89±0.89a 41.67±1.03ab 角鲨烯 26.65±1.48a 25.27±2.07a 多酚 255.56±3.57b 300.49±2.86a β-胡萝卜素 5.08±0.02a 1.79±0.08b 
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