【作者】 王化；

【导师】 祖元刚；

【作者基本信息】 东北林业大学 ， 植物学， 2011， 博士

【摘要】 随着对人工合成抗氧化剂的副作用与潜在危害的不断揭示,消费者开始对安全无毒、副作用的植物源天然抗氧化剂亲睐有加。本文分别从迷迭香的三个主要天然活性成分——迷迭香精油、鼠尾草酸和迷迭香酸着手,建立了提高迷迭香天然抗氧化剂综合得率的提取分离工艺；并通过对本工艺获得的三种迷迭香天然活性成分在应用中抗氧化活性的深入研究,评价了其抗氧化效果,为迷迭香抗氧化剂能更广的应用提供了参考。本文主要结论如下：1、建立了一种可同时测定迷迭香中的鼠尾草酸和迷迭香酸的HPLC方法,采用HiQ ODS C18反相色谱柱(4.6mm×250mm,5μm),以甲醇和0.1%磷酸水溶液为流动相进行梯度洗脱,检测波长为230nm和330nm,流速为1ml/min,进样量10μL。测定的迷迭香酸和鼠尾草酸的出峰时间分别为7.9min和31.5min,鼠尾草酸和迷迭香酸能够和其它峰很好的分离,峰形良好,方法学考察结果良好,适用于对原植物、粗提取物等中含有鼠尾草酸和迷迭香酸的样品进行测定,可以节省时间、节约试剂。2、迷迭香精油提取实验结果表明,水蒸汽蒸馏法为最佳提取方法,最佳蒸馏时间为1.5 h,此时精油的得率为1.93ml/100g,提取精油后样品中鼠尾草酸和迷迭香酸的含量分别为13.45%和2.51%。经此法中试提取迷迭香精油的毛油(包括头馏油、成品油和釜底油)中共鉴定出31种化合物,主要为烯萜类化合物,占毛油总量的94.74%。其中三种优势成分为α-蒎烯、1,8-桉叶素和莰烯。从馏出水中通过萃取和复镏两种方法得到化合物共50种,主要为含氧类化合物和芳烃类化合物。对迷迭香挥发油含量和有效成分的分析评价,为进一步开发利用迷迭香资源在创制中成药新品种、天然香料及综合利用等方面提供了依据。3、首次将不同浓度迷迭香精油与角鲨烯调配成功能性复合油,通过对这种复合油的氧化稳定性与添加抗氧化剂(VE和BHT)角鲨烯的氧化稳定性进行比较,在60℃加速氧化条件下深入研究了迷迭香精油成品油的抗氧化效果。结果显示,所有调配的复合油氧化稳定性良好,最佳比例为迷迭香精油：角鲨烯为2mg：1g。60℃氧化15d,2mg/g迷迭香精油对角鲨烯过氧化值的抑制率为46.08%；对丙二醛含量抑制率为42.43%；对于羟基增量的抑制率为53.66%,能够有效抑制角鲨烯氧化的发生;15d时角鲨烯含量为96.81%,仅降低了3.18%,有力的证明了迷迭香精油较强的抗氧化活性,可以考虑作为保健食品的复配原料。4、采用超声辅助法从去油迷迭香原料中提取以鼠尾草酸和迷迭香酸为目标成分的抗氧化剂,同时以鼠尾草酸和迷迭香酸在浸膏中的质量百分含量为响应因子进行了提取,考察了超声提取过程中各因素对提取效果的影响。得到的最佳超声辅助提取工艺条件为：原料粉碎程度200目,乙醇体积分数80%,料液比1：9,超声功率200W,超声时间40min,鼠尾草酸含量12.74±0.50%,迷迭香酸含量1.74±0.07%。以迷迭香原料干重计算,鼠尾草酸得率2.84g/100g,迷迭香酸得率为0.39g/100g。整个实验过程以鼠尾草酸和迷迭香酸总含量为考察目标,优化得到的工艺条件能实现迷迭香原料的最佳利用率。5、验证了鼠尾草酸作为食品添加剂对红松籽油的抗氧化效果,首次将鼠尾草酸应用到红松籽油当中,通过定期取样检测油脂初级氧化产物和次级氧化产物指标(过氧化值、共轭二烯值、硫代巴比妥酸值及游离脂肪酸值)；通过GC-MS对红松籽油中标志性成分——皮诺敛酸含量变化进行了监测；同时,通过FTIR谱图分析了红松籽油顺式双键、反式双键、羰基键含量的变化,对比研究了不同浓度的鼠尾草酸与合成抗氧化剂(TBHQ.BHA和VE)在加速氧化条件下(60℃)对红松籽油氧化稳定性的影响。研究结果表明,随着鼠尾草酸浓度的增加,其对红松籽油的抗氧化能力增强。3个浓度鼠尾草酸的抗氧化效果均优于VE；0.1mg/g CA的抗氧化效果近似于BHA；0.2和0.3mg/g CA弱于或近似于TBHQ。6、我们选定的迷迭香酸泡腾片制备配方为：每100g泡腾片中含20%迷迭香提取物20g,柠檬酸12.41g,碳酸氢钠27.59g,淀粉17.5g,β-环糊精17.5g,木糖醇5g。此配方制备的迷迭香酸泡腾片表面光洁完整无缺损,崩解时限为2min,溶解后pH值稳定。迷迭香酸泡腾片清除DPPH自由基的SC50值为4.338mg/ml,抗氧化性较好。便于保存和携带的迷迭香酸泡腾片应用前景较好。更多还原

【Abstract】 The major objectives of this study were to investigate the high effective extraction on the combined yield of rosemary essential oil (REO), camosic acid (CA) and rosmarinic acid (RA), and via further study of the antioxidant capacity of REO, CA and RA to offer reference for more extensive application of rosemary antioxidants. The main results obtained in this paper were as follows:1. A method for simultaneous determination of the contents of CA and RA in rosemary samples using HPLC was established. HiQ ODS C18 column (4.6mm×250mm,5μm) and gradient elution were used with methanol and 0.1% phosphoric acid solution as the mobile phase. The flow rate was 1.0ml/min. Sample volume was 10μl. Detection wavelengths were set at 230nm and 330nm. Retention time of RA and CA are 7.9min and 31.5min, respectively. Results Under the chosen condition, RA and CA could be separated from other compounds, and the methods precision, repeatability are up to the requirement through methodology inspection. This method is suitable for the determination of the contents of RA and CA in rosemary samples and extracts simultaneously.2. The results of REO extraction show that the steam distillation is the best method on REO extraction. The optimal distillation time is 1.5h with the REO yield is 1.93ml/100g, and the contents of CA and RA s are 13.45% and 2.51%, respectively in the residual rosemary samples. The pilot test by this method was carried out with 20kg rosemary materials, and the REO were divided into five types by refined treatment. They are the first distillate oil, product oil, residue distillate oil from the crude oil, and extraction oil and second distillation oil from the distilled water. The 31 compounds in rosemary crude oil were identified and the main constituents were terpenoids (94.74%). And 50 compounds in the distilled water from REO extraction were identified, most of which are organic compounds containing O and aromatic compounds. Study on volatile oil content and active ingredients of rosemary will helpful for the further development and utilization of rosemary resources.3. Functional composite oil of squalene with different contents of REO was studied for the first time. Under 60℃, REO and the other antioxidants (VE and BHT) on the oxidation of squalene were studied by determination the index of primary oxidation products and secondary oxidation products, such as peroxide value and TBA. Results showed that the optimum proportion of REO to squalene is 2mg:1g. At the 15th day, the inhibition rate of 2mg/g REO on the peroxide value of squalene was 46.08%; on the malondialdehyde was 42.43; and on the hydroxy increase was 53.66%. REO of this concentration could inhibit the oxidation of squalene efficiently. The content of squalene was 96.81% after 15 days (only 3.18% reduced), which indicated that REO has a strong antioxidant activity. And REO could be considered as a raw material of compound health food.4. The ultrasound-assisted extraction of CA and RA from the raw materials after REO extraction was established, and the optimun extraction parameters were as follows:extraction solvent:brokenness of powder 200 eye,80% EtOH, ratio of solid to liquid is 1:9, ultrasonic power:200W, ultrasonic time:40min. Results Under the chosen condition, the contents of CA and RA were 12.74±0.50% and 1.74±0.07%, respectively. And the extraction yields of CA and RA in dry weight of raw materials were 2.84g/100g and 0.39g/100g, respectively. The total content of CA and RA were investigated throughout the experiment, and the optimized process conditions will achieve an optimum utilization of rosemary materials.5. Antioxidant capacities of CA on pine seed oil with different concentration of carnosic acid were studied for the first time. CA and the artificially synthesized antioxidants on the oxidation of pine seed oil were studied by determination the index of primary oxidation products and secondary oxidation products, such as peroxide value, TBA, conjugated diene value and free fatty acid. Gas chroamatography-mass spectrometry (GC-MS) was conducted to analyze the variation of pine seed oil amount during the storage. And the changes of cis fatty acid (CFA), trans fatty acid (TFA) and aldehyde compounds contents were investigated by FTIR at 60℃over a period of 15 days. Results showed that the relationship of the activities and the content of active constituents in CA indicted positive relevance between the content of carnosic acid and the activities of CA. And the antioxidant capacity of CA was lower than TBHQ, but it was greater than BHA and VE. That is, we could substitute CA for VE and BHA to prevent the lipid oxidation in pine seed oil, even with a low content of carnosic acid.6. The selected formulation of rosmarinic acid effervescent tablets were as follows:Per 100g effervescent tablets has rosemary extract 20g, citric acid 12.41g, sodium bicarbonate 27.59g, starch 17.5g,β-cyclodextrin 17.5g, xylitol 5g. Effervescent tablets according to the formulation have an excellent surface finish and the disintegration time was 2min. Its PH value was stability after dissolved. The antioxidant activity effervescent tablets was well with the SC50 value of scavenging of DPPH radical was 4.338mg/ml. Rosmarinic acid effervescent tablets will have a good prospect by its outstanding advantage of easy to preservation and carrying.更多还原