乳状液膜法分离富集稻谷壳和甘蔗渣提取液中阿魏酸
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摘要
稻谷壳和甘蔗渣是农产品的废弃物,我国拥有丰富资源。稻谷壳和甘蔗渣中含有多种具有生理功能的活性物质,其中包括经济价值较高的药用有效成分阿魏酸,如将其深度开发,价值大幅提高。但由于缺乏快速检测稻谷壳和甘蔗渣中阿魏酸的手段和提取方法,使稻谷壳和甘蔗渣不能得到充分的开发。为了更好的利用这两种废弃物,研究从中提取分离阿魏酸的有效方法,具有很大的意义。
本课题研究了以下四个方面内容:
1.根据阿魏酸的性质,建立了测定甘蔗渣中阿魏酸含量的高效毛细管电泳(HPCE)方法。对缓冲溶液的pH和离子强度等因素进行了研究,得到阿魏酸的HPCE分析条件:毛细管柱为未涂层的石英毛细管柱(长50cm,内径75um,距检测窗:40cm);缓冲溶液为50mmol·L-1的硼砂溶液,pH值8.6;分离电压25kV;柱温25℃。在上述条件下,阿魏酸浓度在0.08g·L-1—2.4g·L-1范围内线性关系良好,R=0.9995,该方法对样品的处理简单。其回收率可达98.6%。
2.用正交设计试验优化碱法提取稻谷壳中阿魏酸,通过考察确定NaOH浓度、提取温度和提取时间三因素,三水平的试验,方差分析后得到的提取条件是:温度55℃, NaOH浓度2%,提取时间6h,可以从100g稻谷壳中提取150mg左右的阿魏酸。
建立了检测稻谷壳中阿魏酸含量的高效液相色谱(HPLC)方法,以乙腈—1%乙酸溶液为流动相,采用梯度洗脱的方式,在柱温30℃,流速0.8mL·min-1的条件下,对稻谷壳提取液中阿魏酸进行分离测定,结果如下:阿魏酸浓度在1.760X10-3g·L-1—2.200 X10-2g·L-1的范围内线性关系良好,R=0.9976,回收率93.0%。
3.借阿魏酸分子结构中的酚羟基和羧基与三正辛胺分子形成氢键的原理,建立了含流动载体的乳状液膜体系,分别用于分离富集甘蔗渣和稻谷壳提取液中阿魏酸。研究了Span-80和三正辛胺的质量浓度、外相HCl和内相磷酸钠的浓度等因素对阿魏酸迁移率的影响,得出最优条件:膜相由5%三辛胺、8%Span-80和膜溶剂煤油组成、内水相含0.16mol·L-1磷酸钠溶液,外水相含0.024 mol·L-1HCl溶液,膜相和内相比为1:1,在2500rpm速度下制乳,乳液与外相比为1.2:2,低速搅拌10min。在此条件下,对水中阿魏酸迁移率最大可达99.0%。采用该乳状液膜体系对甘蔗渣中阿魏酸进行富集,富集倍数2.1,分离系数1.6。该乳状液膜体系对稻谷壳中阿魏酸进行富集,富集倍数2.0,分离系数3.6。以上参数表明,相比富集甘蔗渣中阿魏酸,该体系富集稻谷壳中阿魏酸的效果较好。
4.阿魏酸在不同酸度条件下,会形成离子和分子两种型体。根据他们在水和有机相中溶解度的大小不同,建立了无载体的乳状液膜体系,以Span-80为表面活性剂,液体石蜡为膜增强剂,磷酸钠为内相组成。考察了Span-80和液体石蜡质量浓度、外相HCl和内相磷酸钠浓度等因素,得出的富集条件为:膜相含5% Span-80、3%液体石蜡和膜溶剂煤油,内相为0.08mol·L-1的磷酸钠溶液,外相为0.1mol·L-1HCl溶液,膜相和内相比1:1.2,在2000rpm的速度下制乳,乳液和外相比1:2,低速搅拌20min。在此条件下,对水中阿魏酸迁移率可达99.0%。用该乳状液膜体系从甘蔗渣中富集阿魏酸,富集倍数2.5,分离系数6.6。该体系对稻谷壳中阿魏酸进行富集,富集倍数3.2,分离系数10.4。以上参数值表明,该体系不仅富集稻谷壳中阿魏酸的能力强于富集甘蔗渣中的阿魏酸,而且消除干扰的能力也较强。
Rice shell and sugar cane bagasse are wastes of agricultural products,which have rich resources in china. They contain some active substances with various physiological function and higher economic value such as ferulic acid. If these wastes could be deeply researched, the value may be increased tens of times. However, due to the lack of rapid measure methods and extraction methods of ferulic acid in rice shell and sugar cane bagasse, it is difficult to make well application of rice shell and sugar cane bagasse. In order to apply these waste well, providing and developing more effective methods of extraction and separation of ferulic acid from these waste become a great significance.
The mean contents involve four aspects:
1.According to the property of ferulic acid, a method for the measure of ferulic acid content in sugar cane bagasse is provided with high-performance capillary Electrophoresis. By studying the pH value of buffer solution、the concentration of buffer solution and other factors , provided the HPCE analysis conditions of ferulic acid have been developed; capillary column is the uncoated quartz capillary column (length 50cm, diameter of 75μm, from the detection window: 40cm); the concentration of buffer solution is 50mmol·L-1 Borax buffer solution; the pH value of buffer solution is 8.6; separation voltage : 25kV and column temperature is at 25℃. In the above-mentioned conditions, the concentration of ferulic acid have a good linear relationship between 0.08g·L-1 and 2.4g·L-1. R=0.9995, The method of sample processing is simple. The recovery can be up to 98.6%.
2.Alkali extraction conditions of ferulic acid in rice shell is optimized by orthogonal design test. Three factors and levels were tested including the NaOH concentration, extraction temperature and extraction time. Finally, the extraction conditions after analysis is received. The conditions are that extraction temperature is at 55℃, the concentration of NaOH is 2% and extraction time is for 6h. Under this condition , 150mg Ferulic acid can be got from 100g rice shell .
A method for determination of ferulic acid content in rice shell is established with high-performance liquid chromatography. In order to separating and determining the content of ferulic acid in the extraction solution of rice shell,the gradient elution consisting of 1% acetic acid solution and acetonitrile was applied with flow rate at 0.8 mL·min-1、column temperature at 30℃and detection wavelength at 313 nm. The results are as follows: the concentration of ferulic acid in the range from 1.760X10-3g·L-1 to 2.200X10-2g·L-1 have a good linear relationship, R=0.9976, the recovery can be up to 93.0 %.
3.The phenolic hydroxyl and carboxyl in the molecular structure of ferulic acid can form hydrogen bonds with Tri-n-octylamine molecules. By this principle, the emulsion liquid membrane system with a mobile carrier is developed. The separation and enrichment of ferulic acid in the extraction solution of sugar cane bagasse and rice shell were carried out by using the system. The optimal conditions are as follows: the membrane phase contains 5% trioctylamine, 8% span-80 and 87% kerosene;the internal and external aqueous phase is 0.16mol·L-1 Na3PO4 solution and 0.024 mol·L-1HCl solution; the ratio of membrane phase to internal aqueous phase is 1:1; the speed of making emulsion liquid is 2500rpm; the ratio of emulsion liquid membrane to external aqueous phase is 1.2:2, the stirring time is for 10min. Under above-discussing conditions, the migration rate of ferulic acid in the water can be as high as 99.0% of the maximum. Ferulic acid in the extraction solution of sugar cane bagasse is enriched by the emulsion liquid membrane system. Enrichment multiples is 2.1 and the separation factor is 1.6. The emulsion liquid membrane system enrich ferulic acid in the extraction solution of rice shell. Enrichment multiples reach 2.0 and the separation factor is 3.6. The results showed that ferulic acid in the extraction solution of rice shell can be enriched more effectively than that of sugar cane bagasse with this system.
4.In the different acidity, Ferulic acid can form ions and molecules type. According to their different solubility in water and organic phase, a carrier-free emulsion liquid membrane system is made up with Span-80 as surfactant, paraffin as the membrane enhancer and Na3PO4 solution as the internal phase. The optimal extraction conditions are as follows: membrane phase containing 5% Span-80, 3% liquid paraffin and kerosene membrane solvent, 0.08mol·L-1 Na3PO4 solution in the internal phase, 0.1 mol·L-1HCl solution in the external phase, 1:1.2 of the ratio of membrane phase to internal phase, 2000rpm making emulsion speed, 1:2 of emulsion liquid membrane to external phase, stirring time for 20min in low-speed. Under the above-mentioned conditions, the migration rate of ferulic acid in the water can be as high as 99.0%. When this emulsion liquid membrane system is used for enriching ferulic acid in the extraction solution of sugar cane bagasse, enrichment multiples is 2.5 , and separation factor is 6.6. when this system is made use of enriching ferulic acid in the extraction solution of rice shell, enrichment multiples are up to 3.2, separation factor is 10.4. The results tell us that the system can not only enrich ferulic acid in the extraction solution of rice shell stronger than that in sugar cane bagasse, but also do better capablity of reducing interference.
本课题研究了以下四个方面内容:
1.根据阿魏酸的性质,建立了测定甘蔗渣中阿魏酸含量的高效毛细管电泳(HPCE)方法。对缓冲溶液的pH和离子强度等因素进行了研究,得到阿魏酸的HPCE分析条件:毛细管柱为未涂层的石英毛细管柱(长50cm,内径75um,距检测窗:40cm);缓冲溶液为50mmol·L-1的硼砂溶液,pH值8.6;分离电压25kV;柱温25℃。在上述条件下,阿魏酸浓度在0.08g·L-1—2.4g·L-1范围内线性关系良好,R=0.9995,该方法对样品的处理简单。其回收率可达98.6%。
2.用正交设计试验优化碱法提取稻谷壳中阿魏酸,通过考察确定NaOH浓度、提取温度和提取时间三因素,三水平的试验,方差分析后得到的提取条件是:温度55℃, NaOH浓度2%,提取时间6h,可以从100g稻谷壳中提取150mg左右的阿魏酸。
建立了检测稻谷壳中阿魏酸含量的高效液相色谱(HPLC)方法,以乙腈—1%乙酸溶液为流动相,采用梯度洗脱的方式,在柱温30℃,流速0.8mL·min-1的条件下,对稻谷壳提取液中阿魏酸进行分离测定,结果如下:阿魏酸浓度在1.760X10-3g·L-1—2.200 X10-2g·L-1的范围内线性关系良好,R=0.9976,回收率93.0%。
3.借阿魏酸分子结构中的酚羟基和羧基与三正辛胺分子形成氢键的原理,建立了含流动载体的乳状液膜体系,分别用于分离富集甘蔗渣和稻谷壳提取液中阿魏酸。研究了Span-80和三正辛胺的质量浓度、外相HCl和内相磷酸钠的浓度等因素对阿魏酸迁移率的影响,得出最优条件:膜相由5%三辛胺、8%Span-80和膜溶剂煤油组成、内水相含0.16mol·L-1磷酸钠溶液,外水相含0.024 mol·L-1HCl溶液,膜相和内相比为1:1,在2500rpm速度下制乳,乳液与外相比为1.2:2,低速搅拌10min。在此条件下,对水中阿魏酸迁移率最大可达99.0%。采用该乳状液膜体系对甘蔗渣中阿魏酸进行富集,富集倍数2.1,分离系数1.6。该乳状液膜体系对稻谷壳中阿魏酸进行富集,富集倍数2.0,分离系数3.6。以上参数表明,相比富集甘蔗渣中阿魏酸,该体系富集稻谷壳中阿魏酸的效果较好。
4.阿魏酸在不同酸度条件下,会形成离子和分子两种型体。根据他们在水和有机相中溶解度的大小不同,建立了无载体的乳状液膜体系,以Span-80为表面活性剂,液体石蜡为膜增强剂,磷酸钠为内相组成。考察了Span-80和液体石蜡质量浓度、外相HCl和内相磷酸钠浓度等因素,得出的富集条件为:膜相含5% Span-80、3%液体石蜡和膜溶剂煤油,内相为0.08mol·L-1的磷酸钠溶液,外相为0.1mol·L-1HCl溶液,膜相和内相比1:1.2,在2000rpm的速度下制乳,乳液和外相比1:2,低速搅拌20min。在此条件下,对水中阿魏酸迁移率可达99.0%。用该乳状液膜体系从甘蔗渣中富集阿魏酸,富集倍数2.5,分离系数6.6。该体系对稻谷壳中阿魏酸进行富集,富集倍数3.2,分离系数10.4。以上参数值表明,该体系不仅富集稻谷壳中阿魏酸的能力强于富集甘蔗渣中的阿魏酸,而且消除干扰的能力也较强。
Rice shell and sugar cane bagasse are wastes of agricultural products,which have rich resources in china. They contain some active substances with various physiological function and higher economic value such as ferulic acid. If these wastes could be deeply researched, the value may be increased tens of times. However, due to the lack of rapid measure methods and extraction methods of ferulic acid in rice shell and sugar cane bagasse, it is difficult to make well application of rice shell and sugar cane bagasse. In order to apply these waste well, providing and developing more effective methods of extraction and separation of ferulic acid from these waste become a great significance.
The mean contents involve four aspects:
1.According to the property of ferulic acid, a method for the measure of ferulic acid content in sugar cane bagasse is provided with high-performance capillary Electrophoresis. By studying the pH value of buffer solution、the concentration of buffer solution and other factors , provided the HPCE analysis conditions of ferulic acid have been developed; capillary column is the uncoated quartz capillary column (length 50cm, diameter of 75μm, from the detection window: 40cm); the concentration of buffer solution is 50mmol·L-1 Borax buffer solution; the pH value of buffer solution is 8.6; separation voltage : 25kV and column temperature is at 25℃. In the above-mentioned conditions, the concentration of ferulic acid have a good linear relationship between 0.08g·L-1 and 2.4g·L-1. R=0.9995, The method of sample processing is simple. The recovery can be up to 98.6%.
2.Alkali extraction conditions of ferulic acid in rice shell is optimized by orthogonal design test. Three factors and levels were tested including the NaOH concentration, extraction temperature and extraction time. Finally, the extraction conditions after analysis is received. The conditions are that extraction temperature is at 55℃, the concentration of NaOH is 2% and extraction time is for 6h. Under this condition , 150mg Ferulic acid can be got from 100g rice shell .
A method for determination of ferulic acid content in rice shell is established with high-performance liquid chromatography. In order to separating and determining the content of ferulic acid in the extraction solution of rice shell,the gradient elution consisting of 1% acetic acid solution and acetonitrile was applied with flow rate at 0.8 mL·min-1、column temperature at 30℃and detection wavelength at 313 nm. The results are as follows: the concentration of ferulic acid in the range from 1.760X10-3g·L-1 to 2.200X10-2g·L-1 have a good linear relationship, R=0.9976, the recovery can be up to 93.0 %.
3.The phenolic hydroxyl and carboxyl in the molecular structure of ferulic acid can form hydrogen bonds with Tri-n-octylamine molecules. By this principle, the emulsion liquid membrane system with a mobile carrier is developed. The separation and enrichment of ferulic acid in the extraction solution of sugar cane bagasse and rice shell were carried out by using the system. The optimal conditions are as follows: the membrane phase contains 5% trioctylamine, 8% span-80 and 87% kerosene;the internal and external aqueous phase is 0.16mol·L-1 Na3PO4 solution and 0.024 mol·L-1HCl solution; the ratio of membrane phase to internal aqueous phase is 1:1; the speed of making emulsion liquid is 2500rpm; the ratio of emulsion liquid membrane to external aqueous phase is 1.2:2, the stirring time is for 10min. Under above-discussing conditions, the migration rate of ferulic acid in the water can be as high as 99.0% of the maximum. Ferulic acid in the extraction solution of sugar cane bagasse is enriched by the emulsion liquid membrane system. Enrichment multiples is 2.1 and the separation factor is 1.6. The emulsion liquid membrane system enrich ferulic acid in the extraction solution of rice shell. Enrichment multiples reach 2.0 and the separation factor is 3.6. The results showed that ferulic acid in the extraction solution of rice shell can be enriched more effectively than that of sugar cane bagasse with this system.
4.In the different acidity, Ferulic acid can form ions and molecules type. According to their different solubility in water and organic phase, a carrier-free emulsion liquid membrane system is made up with Span-80 as surfactant, paraffin as the membrane enhancer and Na3PO4 solution as the internal phase. The optimal extraction conditions are as follows: membrane phase containing 5% Span-80, 3% liquid paraffin and kerosene membrane solvent, 0.08mol·L-1 Na3PO4 solution in the internal phase, 0.1 mol·L-1HCl solution in the external phase, 1:1.2 of the ratio of membrane phase to internal phase, 2000rpm making emulsion speed, 1:2 of emulsion liquid membrane to external phase, stirring time for 20min in low-speed. Under the above-mentioned conditions, the migration rate of ferulic acid in the water can be as high as 99.0%. When this emulsion liquid membrane system is used for enriching ferulic acid in the extraction solution of sugar cane bagasse, enrichment multiples is 2.5 , and separation factor is 6.6. when this system is made use of enriching ferulic acid in the extraction solution of rice shell, enrichment multiples are up to 3.2, separation factor is 10.4. The results tell us that the system can not only enrich ferulic acid in the extraction solution of rice shell stronger than that in sugar cane bagasse, but also do better capablity of reducing interference.
引文
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