北五味子活性成分高效分离纯化技术及资源多级利用工艺研究
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摘要
本研究以北五味子果实为原料,建立了多级高效利用北五味子药用资源的生产工艺路线,并对各段工艺进行优化。
采用无溶剂微波辅助提取北五味子精油,提取速率快,30min精油提取率为1.10mL/kg;对精油的抗氧化性能进行测试,并与天然抗氧化剂Vc和VE进行比较,无溶剂微波辅助提取精油的抗氧化性优于水中蒸馏和水蒸汽蒸馏提取的精油。无溶剂微波提取得到的精油折射率和密度比水中蒸馏和水蒸汽蒸馏所得精油的折射率和密度稍大,且颜色较深；通过SEM对提取后的五味子果实进行观察发现微波辅助加热过程中,细胞内部由于温度上升以及局部压力升高,使得细胞壁发生破裂从而使精油的溶出更为完全。通过对提取后的五味子果实进行热重分析得出：无溶剂微波辅助提取后的五味子原料,加热过程中失重最少,说明原料中含挥发性成分的量较少,故此法在较短的时间内提取精油更彻底。GC-MS分析五味子精油的主要成分为烯烃类,还有小部分带有羟基的醇类和酯类。无溶剂微波辅助提取法是一种“环境友好”提取精油的方法,其提取率较高,同时避免了使用大量溶剂。
通过响应面法确定了室温搅拌酸性条件提取五味子花色苷的最佳条件,花色苷的提取率可达到29.06mg/g; HPD-300大孔树脂纯化五味子花色苷粗提物,纯度由5.01%提高到29.97%,其抗氧化能力明显提高。通过对五味子花色苷pH值稳定性研究得出pH值为1.0时花色苷稳定性最佳；对五味子花色苷热稳定性研究表明五味子花色苷在低于40℃时降解缓慢,温度越高,加热时问越长,降解越迅速；热降解符合一级反应动力学模型；通过三种波长的紫外光辐射五味子花色苷溶液表明,紫外光的波长越短,对花色苷的降解越严重。超声与微波处理对五味子花色苷含量和颜色的稳定性有一定影响。LC-MS分析表明五味子花色苷的主要成分为矢车菊素-3-O-木糖芸香糖苷,约占花色苷含量的95%。
通过对五味子多糖提取方法的比较,对回流提取,超声辅助提取,微波辅助提取中的影响因素进行单因素分析,在最优条件下,粗多糖的提取率分别可达到85.6g/kg、74.4g/kg.80.2g/kg。依据单因素的实验结果,进行超声与微波交替辅助实验,最佳交替实验的五味子粗多糖提取率为88.7g/kgo用sevag法对五味子粗多糖脱蛋白四次,精多糖得率为51.1g/kg。将精多糖用不同体积分数的乙醇进行沉降分级,得到四种多糖。在低浓度条件下(<0.2mg/mL),多糖对自由基的清除能力与Vc相似；当浓度大于0.2mg/mL时,五味子多糖对羟自由基的清除能力优于Vc近一倍。多糖体外活性试验表明：200μg/mL是五味子多糖促进细胞增长的最佳浓度,且与刀豆蛋白和脂多糖具有协同作用,五味子多糖与脂多糖细胞的协同作用约为和刀豆蛋白细胞协同作用的1.5倍。
建立了UPLC-MS/MS和HPLC-UV方法定性定量检测北五味子木脂素化合物。对不同木脂素提取方法比较得出：索氏提取所得总木脂素的得率最高,但由于长时间加热,杂质的溶出率也很高；50℃热浸和常温冷浸由于太费时,不适用于工业生产；超声与微波辅助提取,尽管总木脂素的得率相对较低,但所得浸膏纯度较高,可作为实验室制备木脂素的方法；80%乙醇回流提取2.5h适合应用于规模化生产制备木脂素粗提物,总木脂素得率为0.344mg/g,浸膏中木脂素纯度为1.60%。微波辅助离子液体提取法可同时获得五味子精油与木脂素。采用水解原位萃取法,将酯化的木脂素在碱性条件下加热水解为游离态的木脂素,增加游离态木脂素的含量,同时富集于有机溶剂中,浓缩有机相得到木脂素粗提物。这种方法水解与萃取同时完成,操作简便,节省时间。其中总木脂素得率为0.506mg/g,浸膏中木脂素纯度为5.46%。采用201*7碱性阴离子交换树脂水解,HPD-5000大孔吸附树脂富集后,总木脂素得率为0.454mg/g,浸膏中木脂素纯度为27.5%。采用硅胶柱层析纯化木脂素,总木脂素的纯度达到96.5%,回收率超过95%。
石油醚超声辅助萃取所得种子油的得率最高为11.39%,颜色浅且透明,乙酸乙酯萃取的种子油表观颜色较浑浊。通过对不同方法不同溶剂提取的种子油进行化学指标检测,微波辅助萃取所得种子油的酸价、皂化值及过氧化值较超声辅助萃取所得种子油略低,石油醚萃取的种子油的酸价(11.2mg KOH/g油)和皂化值(109.0mg KOH/g油)最低；同时碘值(107.6mg KI/g oil)也最低,即不饱和度最低。所以,选取超声波辅助石油醚萃取的五味子种子油为制备生物柴油的原料。分别采用浓硫酸、离子交换树脂和离子液体为催化剂,以加热,微波或超声辅助加热的催化方法对制备生物柴油的工艺条件进行优化。利用热重分析法和红外特征谱图对五味子种子油以及不同方法不同催化剂制备的生物柴油进行分析；利用气相色谱对种子油中的不饱和脂肪酸,以及生物柴油中脂肪酸甲酯的含量进行测定。
在五味子剩余物热化学转化过程中,优化致密成型操作的工艺参数,得到成型棒坯的密度高、表面无裂痕,无明显炭化,无喷料现象。五味子剩余物成型棒坯热解反应,依据醋液的得率和抗氧化性能考虑：热解约200g样品,热解功率200W,终温约为350℃,热解时间为120min,在上述条件下,天然气得率为13-17%,生物质碳得率为28%-32%,醋液得率为29%-35%,焦油得率为18%-23%。采用快速低温沉降法,分离热解后馏出液中的醋液和焦油,品质高于常温下自然沉降法分离得到产品的品质。采用有机溶剂萃取干燥后的醋液,不但可以去除水分,还可以富集多酚类抗氧化物质,其自由基的清除能力和铁还原能力均与合成抗氧化剂BHA相当。通过GC-MS检测醋液和焦油的主要成分。对活化后的活性炭进行比表面积测试,比表面积达到690.47m2/g,孔容积为0.31cm3/g,平均孔径为1.88nm。
本研究建立了北五味子果实综合利用的工艺流程图,将投料量放大进行验证实验,与小试结果基本一致。真正实现了对北五味子果实中天然活性成分的充分利用,环境友好,为北五味子药用资源生态化综合利用提供了理论依据和数据基础。
In this study, Schisandra chinensis as raw materials, production process route of comprehensive utilization of S. chinensis medicinal resources, and optimize each segment of the process.
Four kinds of different extraction methods were used for extracting S. chinensis essential oil, including hydro-distillation (HD), steam-distillation (SD), microwave assisted hydro-distillation (MHD) and solvent-free microwave assisted distillation (SMD). The extraction kinetic curves showed that the efficiency of MHD and MSD was higher, and the extraction yields were1.20and1.10mL/kg after30min, respectively. DPPH radical scavenging experiments, iron reduction ability test and β-carotene and linoleic acid bleaching experiments were investigated to study the antioxidant activity of essential oil extracted from different extraction methods, and compared with the natural antioxidants Vc and VE. The antioxidant activity of essential oil extracted from SMD was better than that from HD and SD. Through the analysis of the physical characteristics of essential oil, it was found that the refractive index and the density of essential oil obtained by SMD were a little higher than that of essential oil obtained by HD and SD, and the color was darker. S. chinensis fruits after extracting by different methods were observed by scanning electron microscope (SEM), and found that the heat emitted by microwave transfer from the interior of the cell after being heated, and then the partial pressure increased, causing the cell walls rupture, and the essential oil was extracted more completely. S. chinensis fruits after extracting by different methods were analyzed by thermal gravimetric analysis (TGA), the weightlessness of the fruits extracted by SMD was least, showed that the essential oil was extracted more completely. The main component of essential oil was analyzed by GC-MS, more than90%of volatile components were olefins, and there is also a small portion of the alcohols and esters with hydroxyl group, so the antioxidant effect of S. chinensis essential oil is perfect good. SMD as an "environmentally friendly" method for extracting essential oil, showed the higher extraction yield and saved the amount of organic solvents, can also instead of HD and SD.
Optimized the conditions of extracting S. chinensis anthocyanins with acidic water stirred at room temperature by response surface method (RSM). Under the optimal conditions, the extraction yield of anthocyanins was up to29.06mg/g. The crude anthocyanins extract was purificated by HPD-300macroporous resin, the purity of anthocyanins increased from5.01%to29.97%, and the antioxidant capacity significantly improved. The iron reduction capacity and the ABTS radical scavenging capacity were respectively1.2times and1.7times more than that before purification. Study on the content and color stability of S. chinensis anthocyanins on the conditions of different pH value, and in acid solution, which pH is1.0, the anthocyanins compound is most stability. The content and color stability of S. chinensis anthocyanins on the conditions of being heated were also investigated, the degradation of anthocyanin was much slow at less than40℃, and the higher temperature, the longer time being heated, the faster degradation rate. Thermal degradation was according to first order kinetics model, and the correlation coefficients were greater than0.99at different temperature. The content and color stability of S. chinensis anthocyanins on the conditions of ultraviolet radiation were also studied. S. chinensis anthocyanins solution was radiated by three wavelength ultraviolet light (UVA, UVB and UVC), the degradation of anthocyanins was more serious at the shorter UV wavelength. The content and color stability of S. chinensis anthocyanins in the process of ultrasound or microwave were investigated, too. The major component of S. chinensis anthocyanins was cyanidin-3-O-xylosylrutinoside, about95%of the total content by LC-MS determination.
Compared with different method for extracting S. chinensis polysaccharides, and optimized the single factor of reflux extraction (RE), ultrasound assisted extraction (UAE), and microwave assisted extraction (MAE). Under the optimal conditions, the extraction yields of crude polysaccharides were85.6g/kg,74.4g/kg, and80.2g/kg, respectively. According to the experimental results of the single factor, alternating UAE and MAE of polysaccharides were operated. Under the optimal experiments conditions, the extraction yield of crude polysaccharides was88.7g/kg. The crude polysaccharides was deproteinized by sevag method4times, and the refined polysaccharides yield was51.1g/kg. And then the refined polysaccharides were graded into four kinds of polysaccharides with different volume fractions of ethanol. Study on hydroxyl radical scavenging ability of polysaccharides, and the results showed that the scavenging ability of polysaccharides was similar with Vc, when the concentration of polysaccharides was less than0.2mg/mL. When the concentration of polysaccharides was higher than0.2mg/mL, the scavenging ability of polysaccharides was nearly2times better than Vc. The in vitro activity test of polysaccharides showed that200μg/mL was the optimal concentration of S. chinensis polysaccharides to promote cell growth and in combination with ConA and Lps, the synergism of Lps was better than that of ConA about1.5times.
UPLC-MS/MS and HPLC-UV detection method of S. chinensis biphenyl cyclooctene lignans were set up. Comparisons of different methods for extracting lignans:the extraction yield of lignans with Soxhlet extraction was the highest of all, but the dissolution yield of impurity was also higher due to the prolonged being heated. Because of too time-consuming,50℃hot-dip and cold soaking at room temperature were not suitable apply to industrial production. Though the yield of lignans was not high with ultrasound assisted extraction (UAE) and microwave assisted extraction (MAE), the purity of extract was relatively higher, can be used as a laboratory preparation method of lignans.80%ethanol reflux extracted2.5h was suitable for large-scale production to prepare lignans crude extract, the lignans yield was0.344mg/g and the purity of extract was1.60%. Ionic liquid MAE method can be obtained essential oil and lignans simultaneously. The ester-bond lignans was hydrolyzed under being heated alkaline conditions by hydrolysis in situ extraction method. The increased content of free-state lignans was enriched in the organic solvent at the same time with hydrolysis, then the organic phase was concentrated and the lignans crude extract was obtained. The lignans yield was0.506mg/g, and the purity of lignans extract was5.46%. After201*7basic anion exchange resin hydrolysis in combination with HPD-5000macroporous resin enrichment, the lignans yield was0.454mg/g, and the purity of lignans extract was27.5%. Purified the Lignans extract using silica gel column chromatography, the purity of lignans was96.5%, and the recovery was more than95%.
Extracted S. chinensis seed oil with different solvent and different methods. Petroleum ether as the extraction solvent, ultrasound assisted extraction (UAE) of seed oil, the yield of seed oil was the highest11.39%, light in color and transparent. The apparent color of seed oil extracted with the ethyl acetate was dark and muddy. Chemical indexes of seed oil extracted by the different solvent and methods were detected, the acid value, saponification value and peroxide value of seed oil extracted by microwave-assisted extraction (MAE) was slightly lower, compared with that by UAE. The acid value (11.2mg KOH/g oil) and saponification value (109.0mg KOH/g oil) of seed oil extracted with petroleum ether was minimum. The iodine value (107.6mg KI/g oil) was also lowest, that to say the unsaturated degree was the lowest. Therefore, the seed oil extracted with petroleum ether by UAE was selected as the raw material for preparation of biodiesel. The seed oil was heated for preparation of biodiesel, with concentrated sulfuric acid, an ion exchange resin, or an ionic liquid as a catalyst, respectively. FT-IR and TG were used to analysis the seed oil and biodiesel. The content of saturated and unsaturated fatty acids in seed oil, and fatty acid methyl esters in biodiesel were measured by GC-MS, high sensitivity, good repeatability, also quantitative accuracy.
In the thermochemical conversion process of S. chinensis residues, under the optimal conditions, the discharging speed was1.9-2.1kg/min, the residues rod bar was high density, no cracks on the surface, no significant carbonization, and no spray phenomenon. In the pyrolysis reaction of S. chinensis residues, the forming rod bar was200g, pyrolysis power was200W, the final temperature was about350℃, and the pyrolysis time was120min. Under the above conditions, the yields of gas, vinegar, tar, and biomass carbon were13-17%,28-32%,18-23%, and13-17%, respectively. The distillate liquid after pyrolysis was separated into vinegar and tar with the rapid low-temperature deposition method. The vinegar was dried by anhydrous sodium sulfate, and then extracted by organic solvent, not only can remove water, also can be enriched polyphenol antioxidants. The radical scavenging ability and iron reducing power of vinegar ethyl formate extract were similar with that of synthetic antioxidants BHA. And then the composition of vinegar and bio-oil was analyzed by GC-MS. BET informed that the surface area, the pore volume, and the average pore size of the activated biomass carbon were690.47m2/g,0.31cm3/g,1.88nm, respectively.
Process diagram of the comprehensive utilization of S. chinensis medicinal resources was set up, enlarged the raw materials amount, and the experimental results were basically consistent with the small test results. The process realized the full advantage of the natural active products in S. chinensis fruits, and zero emissions to the environment. And strive to get the maximum output and economic benefits, with the minimum raw materials input and energy consumption.
采用无溶剂微波辅助提取北五味子精油,提取速率快,30min精油提取率为1.10mL/kg;对精油的抗氧化性能进行测试,并与天然抗氧化剂Vc和VE进行比较,无溶剂微波辅助提取精油的抗氧化性优于水中蒸馏和水蒸汽蒸馏提取的精油。无溶剂微波提取得到的精油折射率和密度比水中蒸馏和水蒸汽蒸馏所得精油的折射率和密度稍大,且颜色较深；通过SEM对提取后的五味子果实进行观察发现微波辅助加热过程中,细胞内部由于温度上升以及局部压力升高,使得细胞壁发生破裂从而使精油的溶出更为完全。通过对提取后的五味子果实进行热重分析得出：无溶剂微波辅助提取后的五味子原料,加热过程中失重最少,说明原料中含挥发性成分的量较少,故此法在较短的时间内提取精油更彻底。GC-MS分析五味子精油的主要成分为烯烃类,还有小部分带有羟基的醇类和酯类。无溶剂微波辅助提取法是一种“环境友好”提取精油的方法,其提取率较高,同时避免了使用大量溶剂。
通过响应面法确定了室温搅拌酸性条件提取五味子花色苷的最佳条件,花色苷的提取率可达到29.06mg/g; HPD-300大孔树脂纯化五味子花色苷粗提物,纯度由5.01%提高到29.97%,其抗氧化能力明显提高。通过对五味子花色苷pH值稳定性研究得出pH值为1.0时花色苷稳定性最佳；对五味子花色苷热稳定性研究表明五味子花色苷在低于40℃时降解缓慢,温度越高,加热时问越长,降解越迅速；热降解符合一级反应动力学模型；通过三种波长的紫外光辐射五味子花色苷溶液表明,紫外光的波长越短,对花色苷的降解越严重。超声与微波处理对五味子花色苷含量和颜色的稳定性有一定影响。LC-MS分析表明五味子花色苷的主要成分为矢车菊素-3-O-木糖芸香糖苷,约占花色苷含量的95%。
通过对五味子多糖提取方法的比较,对回流提取,超声辅助提取,微波辅助提取中的影响因素进行单因素分析,在最优条件下,粗多糖的提取率分别可达到85.6g/kg、74.4g/kg.80.2g/kg。依据单因素的实验结果,进行超声与微波交替辅助实验,最佳交替实验的五味子粗多糖提取率为88.7g/kgo用sevag法对五味子粗多糖脱蛋白四次,精多糖得率为51.1g/kg。将精多糖用不同体积分数的乙醇进行沉降分级,得到四种多糖。在低浓度条件下(<0.2mg/mL),多糖对自由基的清除能力与Vc相似；当浓度大于0.2mg/mL时,五味子多糖对羟自由基的清除能力优于Vc近一倍。多糖体外活性试验表明：200μg/mL是五味子多糖促进细胞增长的最佳浓度,且与刀豆蛋白和脂多糖具有协同作用,五味子多糖与脂多糖细胞的协同作用约为和刀豆蛋白细胞协同作用的1.5倍。
建立了UPLC-MS/MS和HPLC-UV方法定性定量检测北五味子木脂素化合物。对不同木脂素提取方法比较得出：索氏提取所得总木脂素的得率最高,但由于长时间加热,杂质的溶出率也很高；50℃热浸和常温冷浸由于太费时,不适用于工业生产；超声与微波辅助提取,尽管总木脂素的得率相对较低,但所得浸膏纯度较高,可作为实验室制备木脂素的方法；80%乙醇回流提取2.5h适合应用于规模化生产制备木脂素粗提物,总木脂素得率为0.344mg/g,浸膏中木脂素纯度为1.60%。微波辅助离子液体提取法可同时获得五味子精油与木脂素。采用水解原位萃取法,将酯化的木脂素在碱性条件下加热水解为游离态的木脂素,增加游离态木脂素的含量,同时富集于有机溶剂中,浓缩有机相得到木脂素粗提物。这种方法水解与萃取同时完成,操作简便,节省时间。其中总木脂素得率为0.506mg/g,浸膏中木脂素纯度为5.46%。采用201*7碱性阴离子交换树脂水解,HPD-5000大孔吸附树脂富集后,总木脂素得率为0.454mg/g,浸膏中木脂素纯度为27.5%。采用硅胶柱层析纯化木脂素,总木脂素的纯度达到96.5%,回收率超过95%。
石油醚超声辅助萃取所得种子油的得率最高为11.39%,颜色浅且透明,乙酸乙酯萃取的种子油表观颜色较浑浊。通过对不同方法不同溶剂提取的种子油进行化学指标检测,微波辅助萃取所得种子油的酸价、皂化值及过氧化值较超声辅助萃取所得种子油略低,石油醚萃取的种子油的酸价(11.2mg KOH/g油)和皂化值(109.0mg KOH/g油)最低；同时碘值(107.6mg KI/g oil)也最低,即不饱和度最低。所以,选取超声波辅助石油醚萃取的五味子种子油为制备生物柴油的原料。分别采用浓硫酸、离子交换树脂和离子液体为催化剂,以加热,微波或超声辅助加热的催化方法对制备生物柴油的工艺条件进行优化。利用热重分析法和红外特征谱图对五味子种子油以及不同方法不同催化剂制备的生物柴油进行分析；利用气相色谱对种子油中的不饱和脂肪酸,以及生物柴油中脂肪酸甲酯的含量进行测定。
在五味子剩余物热化学转化过程中,优化致密成型操作的工艺参数,得到成型棒坯的密度高、表面无裂痕,无明显炭化,无喷料现象。五味子剩余物成型棒坯热解反应,依据醋液的得率和抗氧化性能考虑：热解约200g样品,热解功率200W,终温约为350℃,热解时间为120min,在上述条件下,天然气得率为13-17%,生物质碳得率为28%-32%,醋液得率为29%-35%,焦油得率为18%-23%。采用快速低温沉降法,分离热解后馏出液中的醋液和焦油,品质高于常温下自然沉降法分离得到产品的品质。采用有机溶剂萃取干燥后的醋液,不但可以去除水分,还可以富集多酚类抗氧化物质,其自由基的清除能力和铁还原能力均与合成抗氧化剂BHA相当。通过GC-MS检测醋液和焦油的主要成分。对活化后的活性炭进行比表面积测试,比表面积达到690.47m2/g,孔容积为0.31cm3/g,平均孔径为1.88nm。
本研究建立了北五味子果实综合利用的工艺流程图,将投料量放大进行验证实验,与小试结果基本一致。真正实现了对北五味子果实中天然活性成分的充分利用,环境友好,为北五味子药用资源生态化综合利用提供了理论依据和数据基础。
In this study, Schisandra chinensis as raw materials, production process route of comprehensive utilization of S. chinensis medicinal resources, and optimize each segment of the process.
Four kinds of different extraction methods were used for extracting S. chinensis essential oil, including hydro-distillation (HD), steam-distillation (SD), microwave assisted hydro-distillation (MHD) and solvent-free microwave assisted distillation (SMD). The extraction kinetic curves showed that the efficiency of MHD and MSD was higher, and the extraction yields were1.20and1.10mL/kg after30min, respectively. DPPH radical scavenging experiments, iron reduction ability test and β-carotene and linoleic acid bleaching experiments were investigated to study the antioxidant activity of essential oil extracted from different extraction methods, and compared with the natural antioxidants Vc and VE. The antioxidant activity of essential oil extracted from SMD was better than that from HD and SD. Through the analysis of the physical characteristics of essential oil, it was found that the refractive index and the density of essential oil obtained by SMD were a little higher than that of essential oil obtained by HD and SD, and the color was darker. S. chinensis fruits after extracting by different methods were observed by scanning electron microscope (SEM), and found that the heat emitted by microwave transfer from the interior of the cell after being heated, and then the partial pressure increased, causing the cell walls rupture, and the essential oil was extracted more completely. S. chinensis fruits after extracting by different methods were analyzed by thermal gravimetric analysis (TGA), the weightlessness of the fruits extracted by SMD was least, showed that the essential oil was extracted more completely. The main component of essential oil was analyzed by GC-MS, more than90%of volatile components were olefins, and there is also a small portion of the alcohols and esters with hydroxyl group, so the antioxidant effect of S. chinensis essential oil is perfect good. SMD as an "environmentally friendly" method for extracting essential oil, showed the higher extraction yield and saved the amount of organic solvents, can also instead of HD and SD.
Optimized the conditions of extracting S. chinensis anthocyanins with acidic water stirred at room temperature by response surface method (RSM). Under the optimal conditions, the extraction yield of anthocyanins was up to29.06mg/g. The crude anthocyanins extract was purificated by HPD-300macroporous resin, the purity of anthocyanins increased from5.01%to29.97%, and the antioxidant capacity significantly improved. The iron reduction capacity and the ABTS radical scavenging capacity were respectively1.2times and1.7times more than that before purification. Study on the content and color stability of S. chinensis anthocyanins on the conditions of different pH value, and in acid solution, which pH is1.0, the anthocyanins compound is most stability. The content and color stability of S. chinensis anthocyanins on the conditions of being heated were also investigated, the degradation of anthocyanin was much slow at less than40℃, and the higher temperature, the longer time being heated, the faster degradation rate. Thermal degradation was according to first order kinetics model, and the correlation coefficients were greater than0.99at different temperature. The content and color stability of S. chinensis anthocyanins on the conditions of ultraviolet radiation were also studied. S. chinensis anthocyanins solution was radiated by three wavelength ultraviolet light (UVA, UVB and UVC), the degradation of anthocyanins was more serious at the shorter UV wavelength. The content and color stability of S. chinensis anthocyanins in the process of ultrasound or microwave were investigated, too. The major component of S. chinensis anthocyanins was cyanidin-3-O-xylosylrutinoside, about95%of the total content by LC-MS determination.
Compared with different method for extracting S. chinensis polysaccharides, and optimized the single factor of reflux extraction (RE), ultrasound assisted extraction (UAE), and microwave assisted extraction (MAE). Under the optimal conditions, the extraction yields of crude polysaccharides were85.6g/kg,74.4g/kg, and80.2g/kg, respectively. According to the experimental results of the single factor, alternating UAE and MAE of polysaccharides were operated. Under the optimal experiments conditions, the extraction yield of crude polysaccharides was88.7g/kg. The crude polysaccharides was deproteinized by sevag method4times, and the refined polysaccharides yield was51.1g/kg. And then the refined polysaccharides were graded into four kinds of polysaccharides with different volume fractions of ethanol. Study on hydroxyl radical scavenging ability of polysaccharides, and the results showed that the scavenging ability of polysaccharides was similar with Vc, when the concentration of polysaccharides was less than0.2mg/mL. When the concentration of polysaccharides was higher than0.2mg/mL, the scavenging ability of polysaccharides was nearly2times better than Vc. The in vitro activity test of polysaccharides showed that200μg/mL was the optimal concentration of S. chinensis polysaccharides to promote cell growth and in combination with ConA and Lps, the synergism of Lps was better than that of ConA about1.5times.
UPLC-MS/MS and HPLC-UV detection method of S. chinensis biphenyl cyclooctene lignans were set up. Comparisons of different methods for extracting lignans:the extraction yield of lignans with Soxhlet extraction was the highest of all, but the dissolution yield of impurity was also higher due to the prolonged being heated. Because of too time-consuming,50℃hot-dip and cold soaking at room temperature were not suitable apply to industrial production. Though the yield of lignans was not high with ultrasound assisted extraction (UAE) and microwave assisted extraction (MAE), the purity of extract was relatively higher, can be used as a laboratory preparation method of lignans.80%ethanol reflux extracted2.5h was suitable for large-scale production to prepare lignans crude extract, the lignans yield was0.344mg/g and the purity of extract was1.60%. Ionic liquid MAE method can be obtained essential oil and lignans simultaneously. The ester-bond lignans was hydrolyzed under being heated alkaline conditions by hydrolysis in situ extraction method. The increased content of free-state lignans was enriched in the organic solvent at the same time with hydrolysis, then the organic phase was concentrated and the lignans crude extract was obtained. The lignans yield was0.506mg/g, and the purity of lignans extract was5.46%. After201*7basic anion exchange resin hydrolysis in combination with HPD-5000macroporous resin enrichment, the lignans yield was0.454mg/g, and the purity of lignans extract was27.5%. Purified the Lignans extract using silica gel column chromatography, the purity of lignans was96.5%, and the recovery was more than95%.
Extracted S. chinensis seed oil with different solvent and different methods. Petroleum ether as the extraction solvent, ultrasound assisted extraction (UAE) of seed oil, the yield of seed oil was the highest11.39%, light in color and transparent. The apparent color of seed oil extracted with the ethyl acetate was dark and muddy. Chemical indexes of seed oil extracted by the different solvent and methods were detected, the acid value, saponification value and peroxide value of seed oil extracted by microwave-assisted extraction (MAE) was slightly lower, compared with that by UAE. The acid value (11.2mg KOH/g oil) and saponification value (109.0mg KOH/g oil) of seed oil extracted with petroleum ether was minimum. The iodine value (107.6mg KI/g oil) was also lowest, that to say the unsaturated degree was the lowest. Therefore, the seed oil extracted with petroleum ether by UAE was selected as the raw material for preparation of biodiesel. The seed oil was heated for preparation of biodiesel, with concentrated sulfuric acid, an ion exchange resin, or an ionic liquid as a catalyst, respectively. FT-IR and TG were used to analysis the seed oil and biodiesel. The content of saturated and unsaturated fatty acids in seed oil, and fatty acid methyl esters in biodiesel were measured by GC-MS, high sensitivity, good repeatability, also quantitative accuracy.
In the thermochemical conversion process of S. chinensis residues, under the optimal conditions, the discharging speed was1.9-2.1kg/min, the residues rod bar was high density, no cracks on the surface, no significant carbonization, and no spray phenomenon. In the pyrolysis reaction of S. chinensis residues, the forming rod bar was200g, pyrolysis power was200W, the final temperature was about350℃, and the pyrolysis time was120min. Under the above conditions, the yields of gas, vinegar, tar, and biomass carbon were13-17%,28-32%,18-23%, and13-17%, respectively. The distillate liquid after pyrolysis was separated into vinegar and tar with the rapid low-temperature deposition method. The vinegar was dried by anhydrous sodium sulfate, and then extracted by organic solvent, not only can remove water, also can be enriched polyphenol antioxidants. The radical scavenging ability and iron reducing power of vinegar ethyl formate extract were similar with that of synthetic antioxidants BHA. And then the composition of vinegar and bio-oil was analyzed by GC-MS. BET informed that the surface area, the pore volume, and the average pore size of the activated biomass carbon were690.47m2/g,0.31cm3/g,1.88nm, respectively.
Process diagram of the comprehensive utilization of S. chinensis medicinal resources was set up, enlarged the raw materials amount, and the experimental results were basically consistent with the small test results. The process realized the full advantage of the natural active products in S. chinensis fruits, and zero emissions to the environment. And strive to get the maximum output and economic benefits, with the minimum raw materials input and energy consumption.
引文
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