摘要
新几内亚凤仙(Impatiens hawkeri)属凤仙花科凤仙花属多年生常绿草本植物,是目前风靡世界的草花新秀。作为新型的花卉品种,目前国内外的研究范围还只限于栽培学方面,而对其花色素的研究甚少。作为一种天然的色素资源,提取纯化后可广泛应用于食品、制药和化妆品等行业,因此对于它的研究有着重要的意义。本文对新几内亚凤仙花色素的提取、精制、稳定性以及结构组成等方面进行了研究,结果如下:
1 通过经典的显色反应试验、溶解性试验以及可见光谱数据分析,得出该色素属于花色素苷类色素。
2 从加工利用的角度对新几内亚凤仙花色素苷提取和精制过程中的几个重要问题进行了研究。正交试验和极差分析表明:浸提温度和浸提次数是影响色素提取的主要因素,而浸提时间和料液比为次要因素。对色素精制方法的比较研究证明:用XAD-4阳离子交换树脂法要优于HPD300、HPD600大孔吸附树脂法,应优先予以考虑。
3 新几内亚凤仙花色素苷的稳定性研究表明,该花色素苷对室内自然光、温度(低于80℃)、蔗糖、葡萄糖、食盐、还原剂、防腐剂、K~+、Na~+、Ca~(2+)、Zn~(2+)、Mg~(2+)等金属离子在通常情况下是十分稳定的;而对紫外光、室外自然光、氧化剂、柠檬酸、维生素C、pH、Cu~(2+)、Mn~(2+)、Al~(3+)、Fe~(2+)、Fe~(3+)的耐受性较差,在生产和加工过程中应予以注意。
4 利用高压液相色谱-质谱联用(LC-MS)技术对该花色素苷的化学成分和结构进行了鉴定分析。其中色谱条件:ODS C_(18)柱(5μm,3.9×150mm);流动相:85%的0.1%三氟乙酸水溶液(V/V)+15%乙腈;流速:0.3mL/min;进样量:20μL;PDA检测器,检测波长:510nm。质谱采用电喷雾电离(ESI)方式,正离子检测;电离电压:35.00psi;离子源温度:350℃;电喷雾接口干燥气(N_2)流速:9.00L/min。利用以上条件得出的数据、谱图结合有关资料得出:本品种新几内亚凤仙花色素苷中含有矢车菊素-3-葡糖苷(cyanidin-3-glucoside),可能含有矢车菊素-3-芸香糖苷(cyanidin-3-rutinoside)、天竺葵素-3,5-二葡糖苷(pelargonidin-3, 5-diglucoside)、天竺葵素-3-芸香糖苷(pelargonidin-3-rutinoside)、天竺葵素-3-(2~G-葡糖基芸香糖苷)(pelargonidin-3-(2~G-glucosylrutinoside))以及其它一些以矢车菊素为母体色素的花色素苷。
Impatiens hawkeri, as one of the popular flowers in the world, not only has high horticultural value, but also contains rich anthocyanins, which can be used in many important fields, such as in the food, pharmaceutical and cosmetic industries. The development of natural anthocyanins can meet with the consumer demand instead of synthetic colorants. Up to now, most of the research work in flowers focused on flower cultivation. How to use the natural pigment resourch is a big problem. The extraction, purification, stability and components of the anthocyanins in Impatiens hawker were studied in this paper. Main results showed as follows.
1 Through the color respond solvability experiments and the analysis of spectrum data, the results indicated that the pigment of Impatiens hawker flower belongs to the anthocyanins.
2 An Orthogonal experiment was designed to explore the extraction process of Impatiens hawkeri anthocyanins. The results indicated that the effect of temperature and immersing times on pigment extraction was significant, while immersing time and the value of solute/solvent was not significant. For purification method, the contrast tests were performed, and the results showed that the ion exchange resin XAD-4 method was better than HPD300, HPD600 method.
3 The research has been made on stability of the anthocyanins in Impatiens hawkeri. Conclusion was drawn that the indoors nature light, temperature( lower than 80 C ), cane sugar, glucose, salt, reductant, antiseptic, K+, Na+, Ca2+, Zn2+, Mg2+ etc. have no significant effects on the stability of the anthocyanins; the ultraviolet, outdoors nature light, hydroperoxide, lemon acid, vitamin C, pH, Cu2+, Mn2+, A13+, Fe2+, Fe3+, etc. have badly destroying power to the anthocyanins.
4 The high pressure liquid chromatography-mass spectrometry (LC-MS) was used to isolate anthocyanins and to determinate the molecular weight. Of those the column: Hypersil ODS C18(5 m , 3.9 150mm); solvent: (A) 0.1%TFA(concentrated TFA/water, 0.1/99.9, V/V), (B)acetonitrile. Conditions: isocratic elution at 0.3mL/min with 85%A and 15%B at room temperature; injection volume 20 L; PDA spectrophotometry, detection at 510 nm: the MS conditions: the ion source type : ESI:
mass range mode : Std/Normal; ion polarity: Positive; dry temperature: 350 C; nebulizer: 35 psi; dry gas: 9.00L/min. The results showed that it conclude cyanidin-3-glucoside, may be conclude pelargonidin-3,5-diglucoside, cyanidin-3-rutinoside, Pelargonidin-3-(2G-glucosylrutinoside), Pelargonidin-3-rutinoside, and other cyanidins.
引文
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