玉米幼芽提取物凝露的研制
摘要
本实验以玉米幼芽提取物(Extracts of maize plumule, EMP)为主要成分,制备了EMP凝露,并对其进行了稳定性评价、卫生评价、安全性评价、体外透皮和抗皮肤衰老功效研究。
1.EMP凝露的制备和稳定性评价:通过正交试验得到EMP凝露的主要成分最佳配比为1,3-丁二醇、丙二醇和NMF-50各占4%、4%和2%,角鲨烷占2%,卡波姆2020占0.5%,均质速度为1500 r/min;用紫外分光光度计建立了EMP含量的测定方法,并对回收率、精密度等进行了研究,结果显示,激动素可以作为EMP的标准品进行试验,275 nm是其最大吸收波长,质量浓度在9～54μg/mL时标准曲线线性关系良好,平均回收率为98.29%,相对标准偏差为1.95%,进样重复性相对标准偏差在0.1%以内,日内精密度试验的相对标准偏差在3%以内,日间精密度试验的相对标准偏差在5%以内,表明此方法可行性强;通过离心试验、耐寒耐热试验和长期储存试验,结果显示,EMP凝露的峰挺值、涂展性、色泽和香气均未发生明显变化,功效成分EMP的含量均保持在90%以上,表明产品具有良好的耐离心性能、耐寒耐热性能和储存性能。
2.EMP凝露的卫生评价:通过对EMP凝露进行微生物检测,结果显示,三个批次样品中含有的菌落总数、霉菌和酵母菌总数分别为210、195、90 CFU/g和15、0、0 CFU/g,均远少于《化妆品卫生规范》中规定的菌落总数不得大于1000 CFU/g、霉菌和酵母菌总数不得大于100 CFU/g的标准,粪大肠菌群、铜绿假单胞菌和金黄色葡萄球菌等三种致病菌均未检出,证明实验室制备的EMP凝露符合卫生标准规范。
3.EMP凝露的安全性评价:通过动物经口和经皮试验,证明EMP凝露无任何刺激性,属实际无毒产品;通过皮肤光毒性和皮肤变态反应试验,证明EMP凝露对皮肤无光毒性,无致敏性;通过细胞毒性试验,结果显示,EMP凝露IC50为3.40 mg/mL,符合欧盟标准,表明EMP凝露细胞毒性极小。
4.EMP凝露的体外透皮和抗皮肤衰老功效研究:通过体外透皮试验比较三种EMP分散体系的透皮吸收效果,结果显示,12 h内EMP凝露总渗透量最高,分别是EMP凝胶和EMP盐酸溶液总渗透量的1.99倍和1.53倍;通过对细胞H2O2诱导后考察细胞相对增殖率,结果显示,EMP凝露保护组对细胞活力的保护作用最为明显,比模型组细胞相对增殖率提高了44.9%,差异极显著(P<0.01);通过对小鼠皮肤组织光刺激后考察抗氧化酶活性和MDA含量,结果显示,EMP凝露保护组能显著(P<0.05)提高光损伤组织中SOD、CAT的活力,较模型组分别提高了21.99%和23.72%,EMP凝露保护组MDA含量较模型组下降了45.35%,差异极显著(P<0.01);体外透皮试验和抗皮肤衰老功效试验结果表明产品具有良好的透皮性能,能有效的抗皮肤自然衰老和光衰老。
综上所述,实验室制备的EMP凝露具有良好的稳定性,符合卫生标准规范,安全无毒副作用,具有良好的透皮性能和抗皮肤衰老功效,能够满足炎热天气人们抗皮肤衰老的需求。
The research prepared fluid gels with extracts of maize plumule(EMP), and then, its stability, microorganism, security, transdermal assimilation out-body and skin anti-aged effect were studied.
1. Preparation and stability study of EMP fluid gels: The best formula of EMP fluid gels was gotted by orthogonal tests, namely 1,3-butanediol was 4%, propylene glycol was 4%, NMF-50 was 2%, squalane was 2%, Carbopol 2020 was 0.5% and the mix rate is 1500 r/min; Spectrophotometer mensuration of EMP was established to study the precision and recovery rate of EMP fluid gels, the results show that, KT could be used as the standard product, 275 nm was its hightest absorption value, calibration curve had fine linear relation between 9μg/mL to 54μg/mL, the mean recovery was 98.29%, RSD of recovery was 1.95%, RSD of repeatability was less than 0.1%, RSD of precision inter-day was less than 3% and intra-day was less than 5%, the analytical method can satisfy the research; The kurtosis, extensibility, colour and fragrance had no changed by centrifugal research, high and low temperature research and long-term storage research, also, the contents of EMP were all more than 90%, its show that, EMP fluid gels had best stability.
2. Microorganism research of EMP fluid gels: Through the microorganism research of EMP fluid gels, the counts of germs, mycete and yeast of three batches of EMP fluid gels were 210, 195, 90 CFU/g and 15, 0, 0 CFU/g, all less than 1000 CFU/g and 100 CFU/g, which were ordered by the standard. The fecal coliforms, pseudomonas and staphylococcus were all un-detected, its show that, EMP fluid gels had conformed with the microorganism standard.
3. Security experiments of EMP fluid gels: The via-skin and via-mouth irritation tests show that, EMP fluid gels had no irritation; The phototoxicity test and allergy test show that, EMP fluid gels had no phototoxicity and no allergy; The cell toxicity test show that, the IC50 of EMP fluid gels was 3.40 mg/mL, which had conformed with the standard of European Union, namely EMP fluid gels had little cell toxicity.
4. Transdermal assimilation out-body test and skin anti-aged effect experiments of EMP fluid gels: The transdermal assimilation out-body tests of three kinds of EMP mixture show that, the transdermal accumulative amounts of EMP fluid gels in 12 h was 0.99 times more than EMP gels and 0.53 times more than EMP Hcl solution; The cells induced by H2O2 tset show that, the RGR of EMP fluid gels protection group was 44.9% more than the model group(P<0.01); The rat skin tissue UVA irritation test show that, compared with UVA model group, the activities of SOD and CAT of EMP fluid gels protection group had increased by 21.99% and 23.72% separately(P<0.05), the amounts of MDA of EMP fluid gels protection group had decreased by 45.35%(P<0.01); Transdermal assimilation out-body test and skin anti-aged effect experiments show EMP fluid gels had best transdermal function and skin anti-aged capability.
Above all, EMP fluid gels was stable, safe and effective, according to the microorganism standard, its can satisfy the people who want to skin anti-age in the hot days.
1.EMP凝露的制备和稳定性评价:通过正交试验得到EMP凝露的主要成分最佳配比为1,3-丁二醇、丙二醇和NMF-50各占4%、4%和2%,角鲨烷占2%,卡波姆2020占0.5%,均质速度为1500 r/min;用紫外分光光度计建立了EMP含量的测定方法,并对回收率、精密度等进行了研究,结果显示,激动素可以作为EMP的标准品进行试验,275 nm是其最大吸收波长,质量浓度在9～54μg/mL时标准曲线线性关系良好,平均回收率为98.29%,相对标准偏差为1.95%,进样重复性相对标准偏差在0.1%以内,日内精密度试验的相对标准偏差在3%以内,日间精密度试验的相对标准偏差在5%以内,表明此方法可行性强;通过离心试验、耐寒耐热试验和长期储存试验,结果显示,EMP凝露的峰挺值、涂展性、色泽和香气均未发生明显变化,功效成分EMP的含量均保持在90%以上,表明产品具有良好的耐离心性能、耐寒耐热性能和储存性能。
2.EMP凝露的卫生评价:通过对EMP凝露进行微生物检测,结果显示,三个批次样品中含有的菌落总数、霉菌和酵母菌总数分别为210、195、90 CFU/g和15、0、0 CFU/g,均远少于《化妆品卫生规范》中规定的菌落总数不得大于1000 CFU/g、霉菌和酵母菌总数不得大于100 CFU/g的标准,粪大肠菌群、铜绿假单胞菌和金黄色葡萄球菌等三种致病菌均未检出,证明实验室制备的EMP凝露符合卫生标准规范。
3.EMP凝露的安全性评价:通过动物经口和经皮试验,证明EMP凝露无任何刺激性,属实际无毒产品;通过皮肤光毒性和皮肤变态反应试验,证明EMP凝露对皮肤无光毒性,无致敏性;通过细胞毒性试验,结果显示,EMP凝露IC50为3.40 mg/mL,符合欧盟标准,表明EMP凝露细胞毒性极小。
4.EMP凝露的体外透皮和抗皮肤衰老功效研究:通过体外透皮试验比较三种EMP分散体系的透皮吸收效果,结果显示,12 h内EMP凝露总渗透量最高,分别是EMP凝胶和EMP盐酸溶液总渗透量的1.99倍和1.53倍;通过对细胞H2O2诱导后考察细胞相对增殖率,结果显示,EMP凝露保护组对细胞活力的保护作用最为明显,比模型组细胞相对增殖率提高了44.9%,差异极显著(P<0.01);通过对小鼠皮肤组织光刺激后考察抗氧化酶活性和MDA含量,结果显示,EMP凝露保护组能显著(P<0.05)提高光损伤组织中SOD、CAT的活力,较模型组分别提高了21.99%和23.72%,EMP凝露保护组MDA含量较模型组下降了45.35%,差异极显著(P<0.01);体外透皮试验和抗皮肤衰老功效试验结果表明产品具有良好的透皮性能,能有效的抗皮肤自然衰老和光衰老。
综上所述,实验室制备的EMP凝露具有良好的稳定性,符合卫生标准规范,安全无毒副作用,具有良好的透皮性能和抗皮肤衰老功效,能够满足炎热天气人们抗皮肤衰老的需求。
The research prepared fluid gels with extracts of maize plumule(EMP), and then, its stability, microorganism, security, transdermal assimilation out-body and skin anti-aged effect were studied.
1. Preparation and stability study of EMP fluid gels: The best formula of EMP fluid gels was gotted by orthogonal tests, namely 1,3-butanediol was 4%, propylene glycol was 4%, NMF-50 was 2%, squalane was 2%, Carbopol 2020 was 0.5% and the mix rate is 1500 r/min; Spectrophotometer mensuration of EMP was established to study the precision and recovery rate of EMP fluid gels, the results show that, KT could be used as the standard product, 275 nm was its hightest absorption value, calibration curve had fine linear relation between 9μg/mL to 54μg/mL, the mean recovery was 98.29%, RSD of recovery was 1.95%, RSD of repeatability was less than 0.1%, RSD of precision inter-day was less than 3% and intra-day was less than 5%, the analytical method can satisfy the research; The kurtosis, extensibility, colour and fragrance had no changed by centrifugal research, high and low temperature research and long-term storage research, also, the contents of EMP were all more than 90%, its show that, EMP fluid gels had best stability.
2. Microorganism research of EMP fluid gels: Through the microorganism research of EMP fluid gels, the counts of germs, mycete and yeast of three batches of EMP fluid gels were 210, 195, 90 CFU/g and 15, 0, 0 CFU/g, all less than 1000 CFU/g and 100 CFU/g, which were ordered by the standard. The fecal coliforms, pseudomonas and staphylococcus were all un-detected, its show that, EMP fluid gels had conformed with the microorganism standard.
3. Security experiments of EMP fluid gels: The via-skin and via-mouth irritation tests show that, EMP fluid gels had no irritation; The phototoxicity test and allergy test show that, EMP fluid gels had no phototoxicity and no allergy; The cell toxicity test show that, the IC50 of EMP fluid gels was 3.40 mg/mL, which had conformed with the standard of European Union, namely EMP fluid gels had little cell toxicity.
4. Transdermal assimilation out-body test and skin anti-aged effect experiments of EMP fluid gels: The transdermal assimilation out-body tests of three kinds of EMP mixture show that, the transdermal accumulative amounts of EMP fluid gels in 12 h was 0.99 times more than EMP gels and 0.53 times more than EMP Hcl solution; The cells induced by H2O2 tset show that, the RGR of EMP fluid gels protection group was 44.9% more than the model group(P<0.01); The rat skin tissue UVA irritation test show that, compared with UVA model group, the activities of SOD and CAT of EMP fluid gels protection group had increased by 21.99% and 23.72% separately(P<0.05), the amounts of MDA of EMP fluid gels protection group had decreased by 45.35%(P<0.01); Transdermal assimilation out-body test and skin anti-aged effect experiments show EMP fluid gels had best transdermal function and skin anti-aged capability.
Above all, EMP fluid gels was stable, safe and effective, according to the microorganism standard, its can satisfy the people who want to skin anti-age in the hot days.
引文
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