基于超微细处理的猕猴桃籽油保健功能特性及应用研究
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摘要
以猕猴桃果渣中的果籽为原料,采用超临界CO：萃取技术提取猕猴桃籽油,研究萃取分离条件与抗氧化稳定性,分析猕猴桃籽油的理化特性与质量指标,研究使其油滴粒度达到超微细状态的超高压处理条件及粒径分布特征,并通过动物试验模型进一步开展超微细猕猴桃籽油的保健功效特性研究与安全性评价,然后在此基础上对其开展软胶囊与护肤品类产品的开发应用研究。研究结果表明：
①超临界CO2萃取猕猴桃籽油的最佳条件为：果籽粉碎度40-60目,萃取压力30MPa,萃取温度48℃,CO2流量300kg/h,萃取时间210min,出油率34.4%；选择二级降压分离,可把超临界CO2萃取出的猕猴桃籽油中的游离脂肪酸与水分进一步分离出来,最佳分离条件为：分离釜Ⅰ的压力与温度分别为9MPa、42℃,分离釜Ⅱ的压力与温度分别为6MPa、室温,猕猴桃精油出油率可达32.3%；温度与光照条件均可加速氧化猕猴桃籽油,应注意低温避光保存,添加由0.1‰TBHQ、0.1‰BHT、0.05‰V。组成的复合抗氧化剂可提高其抗氧化稳定性。
②对猕猴桃籽油的不合格品进行精炼的最佳脱酸条件为：碱炼初温35℃,碱液浓度9%,下碱时间5min,搅拌转速60r/min,保温沉降8h；最佳脱水条件为：脱水温度105℃,脱水时间55min,搅拌转速45r/min,脱水真空度0.08Mpa；脱色条件为：加活性白土2.5%,脱色温度95℃,真空度0.08MPa,搅拌转速55-60r/min,时间30min,然后采用板框过滤机进行过滤；脱臭条件为：脱臭温度150℃,时间2.5h,真空度0.08Mpa,脱臭完成后充入N2破真空度,采用0～5℃低温贮藏。
③猕猴桃籽油的理化特征指标为：相对密度0.927,折光指数1.484,烟点160℃,皂化值182mgKOH/g,碘值171g I2/100g；脂肪酸组成中,主要含亚麻酸64.3%、亚油酸13.3%、油酸14.5%、棕榈酸5.6%、硬脂酸1.3%,是当今较为理想的天然多烯酸来源。
④采用超高压纳米均质技术制备超微细猕猴桃籽油的最佳条件为：操作压力135MPa,样品流量70L/h,对猕猴桃籽油处理3次,可使其油滴粒径最小；采用光子相关光谱法对普通猕猴桃籽油的油滴粒度进行检测,粒度大于2μm的油滴占97.891%,100%的油滴粒度小于6p m,平均粒径为3.0536μ m,表明普通猕猴桃籽油的油滴粒度处于微米级范围；100%超微细猕猴桃籽油的油滴粒径小于0.1972μ m,其中小于100nm的油滴所占比例高达88.594%,油滴平均粒径0.0634μ m,平均粒径比超微细处理之前的原油缩小48.2倍；经扫描电镜(SEM)分析,普通猕猴桃籽油的油滴杂乱无章,没有规整的几何形状,而超微细猕猴桃油的油滴分布较为分散,外形呈球型,比表面积相对较大。
⑤普通猕猴桃籽油和超微细猕猴桃籽油都能降低高脂饲料模型大鼠的血清TC和TG水平及提高HDL-C水平,但两者都对降低TG水平更为有效；超微细猕猴桃籽油比普通猕猴桃籽油调节血脂的功效更为显著,尤以低剂量的超微细猕猴桃籽油(0.335g/kg·BW)的功效更为突出,对高脂大鼠TC水平的降幅为48.67%,对TG水平的降幅为53.25%,对HDL-C水平的增幅为201.41%,比普通猕猴桃籽油对TC的降幅提高10.5倍,对TG的降幅提高2.3倍,对HDL-C的增幅提高9.2倍；超微细猕猴桃籽油和普通猕猴桃籽油都能降低溴代苯肝损伤模型小鼠组织中MDA的含量,但前者比对MDA的降幅比后者提高4.8倍；两种猕猴桃籽油均能极显著增强小鼠血清SOD的活力,前者增幅达到190.05%,比后者提高3.2倍；果蝇生存试验结果表明,两种猕猴桃籽油均可均显著延长果蝇的平均寿命和平均最高寿命,但前者功效更为明显。
⑥以超微细猕猴桃籽油喂饲大鼠30d,对大鼠的生长发育、血液学指标、血液生化指标没有影响,也未见病理形态学改变,可初步认定对人体安全。
⑦依据超微细猕猴桃籽油保健功能特性与安全性评价结果,以其为原料制备超微细猕猴桃籽油软胶囊,囊壳配比(水：明胶：甘油)为1：1：0.5时,软胶囊质量最佳；超微细猕猴桃籽油软胶囊的囊壳厚度定为0.60mm为宜,可防止崩解迟缓；软胶囊的最佳干燥条件为干燥温度25℃,相对湿度30%,干燥时间30h；软胶囊的崩解时限随储存温度的升高而延长,长期储存时控制温度低于35℃以下；以超微细猕猴桃籽油为基料,制备出乳液与膏霜护肤产品,人体斑贴试验结果表明对人体皮肤未引起不良反应。
通过开展上述研究,进一步明确了超微细猕猴桃籽油更为显著的营养保健价值,扩宽了应用范围,也有利于进一步提高我国猕猴桃产业应用基础研究水平,完善超微细功能性油脂的功效研究理论体系。
The conditions of extraction, separation and anti-oxidative stability of the kiwi fruit seed oil which was extracted by the supercritical CO2were studied with the kiwi fruit seed from the fruit residue as raw material, and its physicochemical characters and quality indexes were determined and analyzed; the ultra-high pressure homogenizing conditions that made the kiwi fruit oil particle size reached the ultra fine state and the distribution characteristics of the particle diameter were researched, then further carried out the research on the health care effects of the kiwi fruit seed oil by ultra-fine processing and its safety evaluation through constructing the animal experimental model; and on this basis, also performed relevant application research, such as soft capsule and skin care product. The research results were as follows:
①Optimal conditions for extracting the kiwi fruit oil by the supercritical CO2method:fruit seed grinding degree,40-60mesh; extraction pressure,30MPa; extraction temperature,48℃; CO2flow rate,300kg/h; extraction time,210min; and oil yield,34.4%; the free fatty acid and water could be further separated from the kiwi fruit seed oil which was extracted by the supercritical CO2if two-step depressurization separation method was used, and optimal separation conditions were as follows:the pressure and temperature of the separation pot Ⅰ, respectively9MPa and42℃; those of the separation pot Ⅱ, respectively6MPa and room temperature; kiwi fruit oil rate,32.3%; both the temperature and illumination condition could accelerate the oxidation of the kiwi fruit seed oil, so it shall be low temperature stored preserved without light and the anti-oxidative stability could be improved by adding the composite antioxidant which was made up of0.1‰TBHQ,0.1‰BHT and0.05‰VC.
②The refining technology of the rejected product of the kiwi fruit seed oil was optimized. And optimal acid removal conditions were as follows:initial temperature of the alkali refining,35℃; concentration of the alkali liquor,9%;alkali refining time,5min;whisk speed,60r/min; thermal subsidence,8h; and optimal dehydration conditions were as follows:temperature,105℃; time,55min; whisk speed,45r/min; vacuum degree,0.08Mpa; and decolorizing conditions were as follows:adding activated clay,2.5%; decolorizing temperature,95℃; vacuum degree,0.08MPa; whisk speed,55-60r/min; time,30min, and then filtering through the frame filter: and deodorization conditions were as follows:deodorization temperature,150℃; time,2.5h; vacuum degree,0.08Mpa; filling in N2to remove the vacuum degree after deodorization had been finished and storing at a low temperature of0-5℃.
③Physicochemical characteristic indexes of kiwi fruit seed oil were as follows: relative density,0.927; refraction Index,1.484; smoke point,160℃; saponification ratio,182mgKOH/g; iodine value,171g I2/100g; and main compositions of the fatty acid included:linolenic acid,64.3%; linoleic acid,13.3%; oleic acid,14.5%; palmitic acid,5.6%; and stearic acid,1.3%; and the kiwi fruit seed oil was the nowadays main source of the most ideal natural polyenoic acid.
④Optimal conditions for preparing the ultra-fine kiwi fruit seed oil through the ultra-high pressure homogenizing technology were as follows:operating pressure,135MPa; flow rate of sample,70L/h; the oil particle size can be made minimum if the kiwi fruit seed oil was treated3times; the photon correlation spectroscopy method was used to test the common kiwi fruit seed oil particle size:the oil droplets with the particle size more than2μm took up97.891%, the particle size of100%oil droplets was less than6μm and the average particle diameter was3.0536μm, all of which showed that the oil particle size of kiwi fruit seed oil was within the micron range; and oil particle size of100%ultra-fine kiwi fruit seed oil was less than0.1972μm, thereinto, oil droplets with the particle size less than100nm reached88.594%and the average particle size was0.0634μm, which was shrunk48.2times than that before the ultra-fine processing; SEM analyzed that the common kiwi fruit seed oil droplets were disorderly without regular geometric shapes, while those of the ultra-fine kiwi fruit seed oil were more dispersed and the shape was ball-type and the specific surface area was relatively large.
⑤The experiment of regulating blood lipid showed that both the common and ultra-fine kiwi fruit seed oils could reduce the TC and TG level, and enhance the HDL-C level of the white rats in the hyperlipemia animal model, however, both of them could more effectively reduce the TG level; the effectiveness for adjusting the blood fat of the ultra-fine kiwi fruit seed oil was more remarkable than that of the common kiwi fruit seed oil, especially the effectiveness of low-dosage ultra-fine kiwi fruit seed oil (0.335g/kg-BW) was outstanding; in details, the decreasing amplitude for TC level of the hyperlipidemic rats was48.67%, that for TG level was53.25%and the increasing amplitude for HDL-C level was201.41%, which were respectively10.5times of the TC and2.3times of the TG decreasing amplitude, and9.2times of the HDL-C increasing amplitude of the common kiwi fruit seed oil; and the oxidation resistance test showed that both the ultra-fine and common kiwi fruit seed oils could reduce the MDA content of the model mice of liver injury by monobromo-benzene, however, the decreasing amplitude of MDA of the former was4.8times of that of the later; both of the kiwi fruit seed oils could remarkably strengthen the SOD vitality in the blood serum of mice, but the increasing amplitude of the former can reach190.05%, which was3.2times of that of the later; the survival test results of the drosophila showed that both of the kiwi fruit seed oils could prolong the average life span and maximum life span, but the effectiveness of the former was more obvious than that of the later.
⑥After rats were fed by ultra-fine kiwi fruit seed oil for30d, their growth&development, hematology index and blood biochemical index were not affected, and pathomorphological changes were also not found, all of which could offer a preliminary determination that the ultra-fine kiwi fruit seed oil was safe for human body.
⑦Prepared the ultra-fine kiwi fruit seed oil soft capsule with the ultra-fine kiwi fruit seed oil as raw material based on its health care functions and safety evaluation results. When the mixture ratio of the capsule shell (water:gelatin:glycerol) was1:1:0.5, the quality of the soft capsule was the best; it's proper to determine the thickness of the ultra-fine kiwi fruit seed oil capsule shell as0.60mm, which could avoid delay of disintegration; drying conditions of the soft capsule were as follows: drying temperature,25℃; relative humidity,30%; and drying time,30h; the disintegration time limited of the soft capsule was prolonged as the storage temperature increased, so the long-time storage temperature shall be controlled below35℃; prepared the skin protector serum and cream with skin care effect by using the ultra-fine kiwi fruit seed oil as the base material, and the cosmetic patch test results showed that adverse reactions for human skin were not caused by this product.
The above study could not only further make clear the remarkable nutrition and health care value of the ultra-fine kiwi fruit seed oil so as to widen its range of application, but also be favorable to further improve the application basic research level of the kiwi fruit industry of China and perfect the theoretical system of effect research on the ultra-fine functional oils.
①超临界CO2萃取猕猴桃籽油的最佳条件为：果籽粉碎度40-60目,萃取压力30MPa,萃取温度48℃,CO2流量300kg/h,萃取时间210min,出油率34.4%；选择二级降压分离,可把超临界CO2萃取出的猕猴桃籽油中的游离脂肪酸与水分进一步分离出来,最佳分离条件为：分离釜Ⅰ的压力与温度分别为9MPa、42℃,分离釜Ⅱ的压力与温度分别为6MPa、室温,猕猴桃精油出油率可达32.3%；温度与光照条件均可加速氧化猕猴桃籽油,应注意低温避光保存,添加由0.1‰TBHQ、0.1‰BHT、0.05‰V。组成的复合抗氧化剂可提高其抗氧化稳定性。
②对猕猴桃籽油的不合格品进行精炼的最佳脱酸条件为：碱炼初温35℃,碱液浓度9%,下碱时间5min,搅拌转速60r/min,保温沉降8h；最佳脱水条件为：脱水温度105℃,脱水时间55min,搅拌转速45r/min,脱水真空度0.08Mpa；脱色条件为：加活性白土2.5%,脱色温度95℃,真空度0.08MPa,搅拌转速55-60r/min,时间30min,然后采用板框过滤机进行过滤；脱臭条件为：脱臭温度150℃,时间2.5h,真空度0.08Mpa,脱臭完成后充入N2破真空度,采用0～5℃低温贮藏。
③猕猴桃籽油的理化特征指标为：相对密度0.927,折光指数1.484,烟点160℃,皂化值182mgKOH/g,碘值171g I2/100g；脂肪酸组成中,主要含亚麻酸64.3%、亚油酸13.3%、油酸14.5%、棕榈酸5.6%、硬脂酸1.3%,是当今较为理想的天然多烯酸来源。
④采用超高压纳米均质技术制备超微细猕猴桃籽油的最佳条件为：操作压力135MPa,样品流量70L/h,对猕猴桃籽油处理3次,可使其油滴粒径最小；采用光子相关光谱法对普通猕猴桃籽油的油滴粒度进行检测,粒度大于2μm的油滴占97.891%,100%的油滴粒度小于6p m,平均粒径为3.0536μ m,表明普通猕猴桃籽油的油滴粒度处于微米级范围；100%超微细猕猴桃籽油的油滴粒径小于0.1972μ m,其中小于100nm的油滴所占比例高达88.594%,油滴平均粒径0.0634μ m,平均粒径比超微细处理之前的原油缩小48.2倍；经扫描电镜(SEM)分析,普通猕猴桃籽油的油滴杂乱无章,没有规整的几何形状,而超微细猕猴桃油的油滴分布较为分散,外形呈球型,比表面积相对较大。
⑤普通猕猴桃籽油和超微细猕猴桃籽油都能降低高脂饲料模型大鼠的血清TC和TG水平及提高HDL-C水平,但两者都对降低TG水平更为有效；超微细猕猴桃籽油比普通猕猴桃籽油调节血脂的功效更为显著,尤以低剂量的超微细猕猴桃籽油(0.335g/kg·BW)的功效更为突出,对高脂大鼠TC水平的降幅为48.67%,对TG水平的降幅为53.25%,对HDL-C水平的增幅为201.41%,比普通猕猴桃籽油对TC的降幅提高10.5倍,对TG的降幅提高2.3倍,对HDL-C的增幅提高9.2倍；超微细猕猴桃籽油和普通猕猴桃籽油都能降低溴代苯肝损伤模型小鼠组织中MDA的含量,但前者比对MDA的降幅比后者提高4.8倍；两种猕猴桃籽油均能极显著增强小鼠血清SOD的活力,前者增幅达到190.05%,比后者提高3.2倍；果蝇生存试验结果表明,两种猕猴桃籽油均可均显著延长果蝇的平均寿命和平均最高寿命,但前者功效更为明显。
⑥以超微细猕猴桃籽油喂饲大鼠30d,对大鼠的生长发育、血液学指标、血液生化指标没有影响,也未见病理形态学改变,可初步认定对人体安全。
⑦依据超微细猕猴桃籽油保健功能特性与安全性评价结果,以其为原料制备超微细猕猴桃籽油软胶囊,囊壳配比(水：明胶：甘油)为1：1：0.5时,软胶囊质量最佳；超微细猕猴桃籽油软胶囊的囊壳厚度定为0.60mm为宜,可防止崩解迟缓；软胶囊的最佳干燥条件为干燥温度25℃,相对湿度30%,干燥时间30h；软胶囊的崩解时限随储存温度的升高而延长,长期储存时控制温度低于35℃以下；以超微细猕猴桃籽油为基料,制备出乳液与膏霜护肤产品,人体斑贴试验结果表明对人体皮肤未引起不良反应。
通过开展上述研究,进一步明确了超微细猕猴桃籽油更为显著的营养保健价值,扩宽了应用范围,也有利于进一步提高我国猕猴桃产业应用基础研究水平,完善超微细功能性油脂的功效研究理论体系。
The conditions of extraction, separation and anti-oxidative stability of the kiwi fruit seed oil which was extracted by the supercritical CO2were studied with the kiwi fruit seed from the fruit residue as raw material, and its physicochemical characters and quality indexes were determined and analyzed; the ultra-high pressure homogenizing conditions that made the kiwi fruit oil particle size reached the ultra fine state and the distribution characteristics of the particle diameter were researched, then further carried out the research on the health care effects of the kiwi fruit seed oil by ultra-fine processing and its safety evaluation through constructing the animal experimental model; and on this basis, also performed relevant application research, such as soft capsule and skin care product. The research results were as follows:
①Optimal conditions for extracting the kiwi fruit oil by the supercritical CO2method:fruit seed grinding degree,40-60mesh; extraction pressure,30MPa; extraction temperature,48℃; CO2flow rate,300kg/h; extraction time,210min; and oil yield,34.4%; the free fatty acid and water could be further separated from the kiwi fruit seed oil which was extracted by the supercritical CO2if two-step depressurization separation method was used, and optimal separation conditions were as follows:the pressure and temperature of the separation pot Ⅰ, respectively9MPa and42℃; those of the separation pot Ⅱ, respectively6MPa and room temperature; kiwi fruit oil rate,32.3%; both the temperature and illumination condition could accelerate the oxidation of the kiwi fruit seed oil, so it shall be low temperature stored preserved without light and the anti-oxidative stability could be improved by adding the composite antioxidant which was made up of0.1‰TBHQ,0.1‰BHT and0.05‰VC.
②The refining technology of the rejected product of the kiwi fruit seed oil was optimized. And optimal acid removal conditions were as follows:initial temperature of the alkali refining,35℃; concentration of the alkali liquor,9%;alkali refining time,5min;whisk speed,60r/min; thermal subsidence,8h; and optimal dehydration conditions were as follows:temperature,105℃; time,55min; whisk speed,45r/min; vacuum degree,0.08Mpa; and decolorizing conditions were as follows:adding activated clay,2.5%; decolorizing temperature,95℃; vacuum degree,0.08MPa; whisk speed,55-60r/min; time,30min, and then filtering through the frame filter: and deodorization conditions were as follows:deodorization temperature,150℃; time,2.5h; vacuum degree,0.08Mpa; filling in N2to remove the vacuum degree after deodorization had been finished and storing at a low temperature of0-5℃.
③Physicochemical characteristic indexes of kiwi fruit seed oil were as follows: relative density,0.927; refraction Index,1.484; smoke point,160℃; saponification ratio,182mgKOH/g; iodine value,171g I2/100g; and main compositions of the fatty acid included:linolenic acid,64.3%; linoleic acid,13.3%; oleic acid,14.5%; palmitic acid,5.6%; and stearic acid,1.3%; and the kiwi fruit seed oil was the nowadays main source of the most ideal natural polyenoic acid.
④Optimal conditions for preparing the ultra-fine kiwi fruit seed oil through the ultra-high pressure homogenizing technology were as follows:operating pressure,135MPa; flow rate of sample,70L/h; the oil particle size can be made minimum if the kiwi fruit seed oil was treated3times; the photon correlation spectroscopy method was used to test the common kiwi fruit seed oil particle size:the oil droplets with the particle size more than2μm took up97.891%, the particle size of100%oil droplets was less than6μm and the average particle diameter was3.0536μm, all of which showed that the oil particle size of kiwi fruit seed oil was within the micron range; and oil particle size of100%ultra-fine kiwi fruit seed oil was less than0.1972μm, thereinto, oil droplets with the particle size less than100nm reached88.594%and the average particle size was0.0634μm, which was shrunk48.2times than that before the ultra-fine processing; SEM analyzed that the common kiwi fruit seed oil droplets were disorderly without regular geometric shapes, while those of the ultra-fine kiwi fruit seed oil were more dispersed and the shape was ball-type and the specific surface area was relatively large.
⑤The experiment of regulating blood lipid showed that both the common and ultra-fine kiwi fruit seed oils could reduce the TC and TG level, and enhance the HDL-C level of the white rats in the hyperlipemia animal model, however, both of them could more effectively reduce the TG level; the effectiveness for adjusting the blood fat of the ultra-fine kiwi fruit seed oil was more remarkable than that of the common kiwi fruit seed oil, especially the effectiveness of low-dosage ultra-fine kiwi fruit seed oil (0.335g/kg-BW) was outstanding; in details, the decreasing amplitude for TC level of the hyperlipidemic rats was48.67%, that for TG level was53.25%and the increasing amplitude for HDL-C level was201.41%, which were respectively10.5times of the TC and2.3times of the TG decreasing amplitude, and9.2times of the HDL-C increasing amplitude of the common kiwi fruit seed oil; and the oxidation resistance test showed that both the ultra-fine and common kiwi fruit seed oils could reduce the MDA content of the model mice of liver injury by monobromo-benzene, however, the decreasing amplitude of MDA of the former was4.8times of that of the later; both of the kiwi fruit seed oils could remarkably strengthen the SOD vitality in the blood serum of mice, but the increasing amplitude of the former can reach190.05%, which was3.2times of that of the later; the survival test results of the drosophila showed that both of the kiwi fruit seed oils could prolong the average life span and maximum life span, but the effectiveness of the former was more obvious than that of the later.
⑥After rats were fed by ultra-fine kiwi fruit seed oil for30d, their growth&development, hematology index and blood biochemical index were not affected, and pathomorphological changes were also not found, all of which could offer a preliminary determination that the ultra-fine kiwi fruit seed oil was safe for human body.
⑦Prepared the ultra-fine kiwi fruit seed oil soft capsule with the ultra-fine kiwi fruit seed oil as raw material based on its health care functions and safety evaluation results. When the mixture ratio of the capsule shell (water:gelatin:glycerol) was1:1:0.5, the quality of the soft capsule was the best; it's proper to determine the thickness of the ultra-fine kiwi fruit seed oil capsule shell as0.60mm, which could avoid delay of disintegration; drying conditions of the soft capsule were as follows: drying temperature,25℃; relative humidity,30%; and drying time,30h; the disintegration time limited of the soft capsule was prolonged as the storage temperature increased, so the long-time storage temperature shall be controlled below35℃; prepared the skin protector serum and cream with skin care effect by using the ultra-fine kiwi fruit seed oil as the base material, and the cosmetic patch test results showed that adverse reactions for human skin were not caused by this product.
The above study could not only further make clear the remarkable nutrition and health care value of the ultra-fine kiwi fruit seed oil so as to widen its range of application, but also be favorable to further improve the application basic research level of the kiwi fruit industry of China and perfect the theoretical system of effect research on the ultra-fine functional oils.
引文
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