摘要
火棘果(Pyracantha fortuneana)是一种药食两用、储量丰富的野生植物资源,除营养成分丰富外,还富含多种类黄酮成分,具有美白、降血脂、增强免疫力、促进消化等多种功效。但目前国内外对火棘果的资源化学研究并不深入,缺乏科学有效的成分分析及活性评价方法等问题,致使火棘果产品开发缺乏资源特色,产业化发展仍未成气候。本文围绕火棘果皮肉资源化学成分分析、动态变化规律及其体外抗氧化活性等方面展开了系统研究,为火棘果资源的产业化利用提供了理论基础和科学依据。现已取得如下研究成果:
1.建立了一套火棘果资源化学成分的分析方法。
包括火棘果多种类黄酮成分的分组分析方法;火棘果HPLC指纹图谱的整体性评价分析方法;基于体外抗氧化活性评价火棘果提取物功效的分析方法。该分析方法可从成分分析和活性评价两个角度对火棘果资源化学成分进行评价,为火棘果原料及产品的品质分析及质量监控提供了有效手段。
2.研究了火棘果中的资源化分成分特征,利用LC-MS技术并参考文献报道,对火棘果HPLC指纹图谱中部分指纹峰进行归属,包括:绿原酸、二聚儿茶素、矢车菊素-3-葡萄糖苷、矢车菊素-3-半乳糖苷、芦丁、金丝桃甙、异金丝桃甙等;
火棘果提取物的体外抗氧化活性明显优于模式抗氧化剂BHT,可广泛作为抗氧化功能性保健品、化妆品、植物药等原料。
3.研究了火棘果发育成熟过程中资源化学成分的动态积累规律,不同时期各成分含量均有所差异,并存在一定的贮失规律,该研究对火棘果资源的综合利用和最佳采收期的确定具有很大的指导意义。
4.优化了火棘果抗氧化成分的提取工艺:乙醇浓度为71%;提取液pH=3.2,提取温度为51℃。火棘果醇提液在弱酸性体系、低温避光条件下稳定性良好,外源添加适量的绿原酸、茶多酚等黄酮类化合物可起到辅助效果,提示火棘果中原有的黄酮成分与花青素、原花青素间也存在辅色作用和协同增效作用。
Pyracantha fortuneana, a kind of abundant wild plant resource, is usd as both food and medicine. It is not only rich in nutrients, but also contains abundant isoflavonoid components with lots of health maintenance efficacy, such as excellent whitening effect, antiatheroscloresis, increasing immunity and facilitating digestion. But some bottlenecks, for example, few study in the resources chemistry of P. fortuneana, lacking in scientifically effectively methods about component analysis and bioassay evaluation, resulted in its products lacking in resource characteristics and its industrialization has not been a noticeable trend. In this paper, we analysed the bioresource chemistry constituent of P. fortuneana, and studied their dynamic change law and vitro antioxidant activity. This would promote the industrialization development and efficient utilization of P. fortuneana. Some results were obtained as follows:
1. We established serial analytical methods about P. fortuneana bioresource chemistry compounds.
Established the fractional analysis method of multiple isoflavonoid components by spectrophotometry; Established the overall evaluation method of group characteristics of phytochemical compounds using HPLC finger-print; Established the activity-evaluation method of P. fortuneana extracts by vitro antioxidant activity.
This method could be used to study the components and activity of bioresource chemistry compounds, which supplied an efficient method for the attributional analysis and quality monitoring of P. fortuneana and its products.
2. We researched the characteristics of bioresource chemical components of P. fortuneana systematically. After analysis of LC-MS results and comparison with published reports, some major common peaks were identified as chlorogenic acid, dicatechin, glucoside, galactose, rutin, hyperoside and isohyperoside;
The antioxidant activity of P. fortuneana extracts were better than BHT obviously. It can be used as the raw material for functional products, cosmetic and botanical drug.
3. We monitored the dynamic changes of bioresource chemical components of P. fortuneana and found that their contents are different during ripening period. This provide theoretical basis for the comprehensive utilization of P. fortuneana and the determination of optimal harvesting time.
4. We optimized the extraction process of antioxidant components of P. fortuneana: ethanol concentration 71%, pH 3.2 and extracting temperature 51℃. The extract was more stable in acid condition and low temperature evades under light, adding some flavonoid compounds such as chlorogenic acid and tea polyphenol could increase the stability, which indicated that there were intermolecular co-pigmentation and synergistic effect between its internal flavonoid and anthocyanin components.
引文
[1]中国科学院植物志编委会.中国植物志[M].北京:科学出版社, 1974: 36.
[2]王三根,邓如福.野生植物火棘的开发利用及生理生态[J].中国野生植物, 1988, 3, l3-l5.
[3]蒋利华,熊远福,李霞,等.野生火棘果有效成分研究进展[J].中国野生植物资源, 2007, 26(2): 8-10.
[4]蔡金腾,丁筑红.贵州省火棘资源调查研究[J].贵州农业科学, 1994, 52-55.
[5]邓如福,王三根,李关荣.野生植物-火棘果营养成分[J].营养学报, 1900, 12(1): 79-84.
[6]黄祖良,韦国锋,何有成,等.桂西火棘果实特性和营养成分的研究[J].食品研究与开发, 2004, 25(2): 75-76.
[7]柴立,郑亚玉,谢宝忠,等.赤阳子开发应用系列研究之一赤阳子营养成分及保健作用研究[J].贵阳中医学院学报, 1988, 1(23): 39-41.
[8]四川中药志编写组.四川中药志[M].四川人民出版社, 1982.
[9]薛慧,李玉山,等.火棘对高脂饮食大鼠血液流变学的影响[J].湖北民族学院学报:医学版, 2001, 18(4): 8-9.
[10]侯建军,刘希林,魏文科,等.火棘消食健脾功效的动物试验[J].湖北农业科学, 2003, 4(34): 84-86.
[11]韦忠恒,胡柳,黄剑娴.火棘果汁干预砷中毒小鼠肝功能损害指标的变化[J].中国临床康复, 2006, 10(23): 99-101.
[12]续洁琨,姚新生,郑洁静,等.火棘果实提取物对拘束应激负荷引起小鼠肝损伤的保护作用[J].中草药, 2007, 38(12): 1849-1853.
[13]蔡金腾,吴翔.刺梨,金樱子,火棘果实特性及营养成分[J].贵州农业科学, 1997, 25(3): 17-21.
[14]侯建军,覃红斌,魏文科.不同产地火棘果实营养成分分析及评价[J].湖北农业科学, 2003, 2(31): 83-85.
[15]王军宪,牛娟芳.秦岭产火棘果实的成分分析[J].西北药学杂志, 1994, 9(5): 204-206.
[16]刘世彪,易浪波,李宽,等.湘西自治州不同产地火棘果实的营养成分[J].中国野生植物资源, 2007, 26(3): 58-60.
[17]王三根,邓如福.火棘果发育与贮藏期间营养成分和色素的变化研究[J].营养学报, 1996, 18(3): 371-374.
[18]牟君富,王树平.红子果实营养成分含量变化及其利用研究[J].西南农业学报, 1992, 5(3): 42-47.
[19]王军宪,陈新民.火棘化学成分的研究[J].天然产物研究与开发, 1990, 2(3): 63-66.
[20]王军宪,牛娟芳.火棘果化学成分研究[J].西北药学杂志, 1994, 9(6): 253-255.
[21]梅兴国,万国晖,等.火棘果化学成分研究[J].中药材, 2002, 25(5): 329-330.
[22]梁淑芳,马绒利,马柏林.火棘黄酮类化合物的提取及微乳薄层色谱分离鉴定[J].西北林学院学报, 2003, 18(3): 60-62.
[23] Dai Y, Zhou G X, Kurihara H, et al. Biphenyl glycosides from the fruit of Pyracantha fortuneana[J]. Journal of Natural Products, 2006, 69(7): 1022-1024.
[24] Dai Y, Zhou G, Kurihara H, et al. Fortuneanosides G-L, dibenzofuran glycosides from the fruit of Pyracantha fortuneana [J]. Chemical and Pharmaceutical Bulletin, 2008, 56(4): 439-442.
[25] Fico G, Bilia A R, Morelli I, et al. Flavonoid distribution in Pyracantha coccinea plants at different growth phases[J]. Biochemical Systematics and Ecology, 2000, 28(7): 673-678.
[26] Lampe J W. Health effects of vegetables and fruits: assessing mechanism of action in human experimental studies. American Journal of Clinical Nutrition, 1999, 70, 475-490.
[27] Huxley R R, Neil H. The relationship between dietary flavonol intake and coronary heart disease mortality: a metaanalysis of prospective cohort studies. European Journal of Clinical Nutrition, 2003, 57, 904–908.
[28]蒋利华,熊远福,李霞,等.野生火棘果中红色素的提取研究[J].中国食品添加剂, 2007, 1(11): 58-61.
[29]蒋新龙,蒋益花.微波法提取火棘红色素的工艺研究[J].食品科技, 2006, 31(7): 182-185.
[30] Roy M, Sen S, Chakraborti A S. Action of pelargonidin on hyperglycemia and oxidative damage in diabetic rats: implication for glycation-induced hemoglobin modification[J]. Life Science, 2008, 82(21-22): 1102-1110.
[31] Butelli E, Titta L, Giorgio M, et al. Enrichment of tomato fruit with health-promoting anthocyanins by expression of select transcription factors[J]. Nature Biotechnology, 2008, 26(11): 1301-1308.
[32] Vattem D A, Ghaedian R, Shetty K. Enhancing health benefits of berries through phenolic antioxidant enrichment: focus on cranberry[J]. Asia Pacific Journal of Clinical Nutrition, 2005, 14(2): 120-130.
[33]卫亚丽,杨江.野生火棘果红色素提取工艺优化[J].中国野生植物资源, 2010, 29(4): 50-53.
[34]何玲,陈翠.超声波提取火棘红色素工艺参数的优化[J].西北林学院学报, 2010, 25(6): 148-151.
[35]蒋利华,黄忠良,熊远福,等.野生火棘果中黄色素的提取研究[J].中国食品添加剂, 2010(2): 138-142.
[36]李伟,程超,张应团,等.不同火棘果实功效成分的聚类分析[J].食品科学, 2008, 29(9): 207-210.
[37]陈瑛,鲁周民,李志西.火棘果汁饮料生产工艺及生产过程中HACCP质量控制研究[J].食品科学, 2007, 28(8): 598-601.
[38]周文斌,王崇均,甘小龙.火棘果醋生产工艺研究[J].食品科学, 2009, 30(6): 282-284.
[39] Correa M, de Melo G O, Costa S S. Natural products from plant origin potentially usefull in the asthma therapy[J]. Revista Brasileira De Farmacognosia-Brazilian Journal of Pharnacognosy, 2008, 18Suppl. S: 785-797.
[40] Hoi C P, Ho Y P, Baum L, et al. Neuroprotective effect of honokiol and magnolol, compounds from magnolia officinalis, on beta-amyloid-induced toxicity in PC12 cells[J]. Phytotherapy Research, 2010, 24(10): 1538-1542.
[41] Menghini L, Epifano F, Genovese S, et al. Antiinflammatory activity of coumarins from Ligusticum lucidum Mill. subsp. cuneifolium (Guss.) Tammaro (Apiaceae)[J]. Phytotherapy Research, 2010, 24(11): 1697-1699.
[42] Srivastava J K, Shankar E, Gupta S. Chamomile: A herbal medicine of the past with a bright future (Review)[J]. Molecular Medicine Reports, 2010, 3(6): 895-901.
[43]何兰,姜志宏,周荣汉.天然产物资源化学[M].北京.科学出版社, 2008, 84-92.
[44] Pan Y M, He C H, Wang H S, et al. Antioxidant activity of microwave-assisted extract of Buddleia officinalis and its major active component[J]. Food Chemistry, 2010, 121(2): 497-502.
[45] Awang D. Standardization of herbal medicinal products[M]. Leuven1: International Society Horticultural Science, 2004, 111-114.
[46] Chen J H, Lee F, Li L, et al. Standardized extracts of Chinese medicinal herbs: Case study of Danshen (Salvia miltiorrhiza Bunge)[J]. Journal of Food and Drug Analysis, 2007, 15(4): 347-364.
[47]吴兴壮,李利峰,李晓东,等.越桔果实资源的开发利用现状及前景预测[J].辽宁农业科学, 2003, 6, 23-24.
[48] Pappas E, Schaich K M. Phytochemicals of cranberries and cranberry products: characterization, potential health effects, and processing stability[J]. Critical Reviews in Food Science and Nutrition, 2009, 49(9): 741-781.
[49]张玉萍.越橘的保健作用及其在我国的开发利用前景[J].山西农业科学, 2006, 34(4): 22-25.
[50] Yan X J, Murphy B T, Hammond G B, et al. Antioxidant activities and antitumor screening of extracts from cranberry fruit (Vaccinium macrocarpon)[J]. Journal of Agricultural and Food Chenistry, 2002, 50(21): 5844-5849.
[51] Neto C C. Cranberry and its phytochemicals: A review of in vitro anticancer studies[J]. Journal of Nutrition 2007, 137Suppl. S(1): 186-193.
[52] Vattem D A, Ghaedian R, Shetty K. Enhancing health benefits of berries through phenolic antioxidant enrichment: focus on cranberry[J]. Asia Pacific Journal of Clinical Nutrition, 2005, 14(2): 120-130.
[53] Chun O K, Kim D O, Lee C Y. Superoxide radical scavenging activity of the major polyphenols in fresh plums[J]. Journal of Agricultural and Food Chenistry, 2003, 51(27): 8067-8072.
[54] Prior R L, Cao G H, Martin A, et al. Antioxidant capacity as influenced by total phenolic and anthocyanin content, maturity, and variety of Vaccinium species[J]. Journal of Agricultural and Food Chenistry, 1998, 46(7): 2686-2693.
[55] Li Y F, Guo C J, Yang J J, et al. Evaluation of antioxidant properties of pomegranate peel extract in comparison with pomegranate pulp extract[J]. Food Chemistry, 2006, 96(2): 254-260.
[56]王少波,杜永峰,姚秉华. pH示差法测定黑豆皮中的花青素[J].化学分析计量, 2008, 17(1): 46-47, 49.
[57] Lee J, Rennaker C, Wrolstad R E. Correlation of two anthocyanin quantification methods: HPLC and spectrophotometric methods[J]. Food Chemistry, 2008, 110(3): 782-786.
[58] Kirakosyan A, Seymour E M, Llanes D E U, et al. Chemical profile and antioxidant capacities of tart cherry products[J]. Food Chemistry, 2009, 115(1): 20-25.
[59]严赞开.紫外分光光度法测定植物黄酮含量的方法[J].食品研究与开发, 2007, 28(9): 164-167.
[60] Khan M K, Abert-Vian M, Fabiano-Tixier A S, et al. Ultrasound-assisted extraction of polyphenols (flavanone glycosides) from orange (Citrus sinensis L.) peel[J]. Food Chemistry, 2010, 119(2): 851-858.
[61]李晓丹,肖娅萍. 5种药用石韦总黄酮的提取及测定[J].中国野生植物资源, 2009, 28(1): 59-61.
[62]李巨秀,往柏玉.福林酚比色法测定桑椹中总多酚[J].食品科学, 2009, 30(18): 292-295.
[63] Silva E M, Rogez H, Larondelle Y. Optimization of extraction of phenolics from Inga edulis leaves using response surface methodology[J]. Separation and Purification technlogy, 2007, 55(3): 381-387.
[64]肖付才,李华,王华.葡萄籽原花青素的提取和检测方法[J].食品研究与开发, 2007, 28(6): 165-169.
[65]周玮婧,孙智达,谢笔钧,等.荔枝皮原花青素提取工艺优化[J].农业工程学报, 2009, 25(增刊1): 175-179.
[66] Souquet J M, Cheynier V, Brossaud F, et al. Polymeric proanthocyanidins from grape skins[J]. Phytochemistry, 1996, 43(2): 509-512.
[67] Castaneda-Ovando A, Pacheco-Hernandez M D, Paez-Hernandez M E, et al. Chemical studies of anthocyanins: A review[J]. Food Chemistry, 2009, 113(4): 859-871.
[68] Wu X L, Prior R L. Systematic identification and characterization of anthocyanins by HPLC-ESI-MS/MS in common foods in the United States: Fruits and berries[J]. Journal of Agricultural and Food Chenistry, 2005, 53(7): 2589-2599.
[69] Downey M, Rochfort S. Simultaneous separation by reversed-phase high- performance liquid chromatography and mass spectral identification of anthocyanins and flavonols in Shiraz grape skin[J]. Journal of Chromatography A. 2008, 1201(1): 43-47.
[70] Clifford M N, Kirkpatrick J, Kuhnert N, Roozendaal H, et al. LC–MSn analysis of the cis isomers of chlorogenic acids. Food Chemistry. 2008, 106, 379–385.
[71] Prasad K N, Yang B, Yang S Y, et al. Identification of phenolic compounds and appraisal of antioxidant and antityrosinase activities from litchi (Litchi sinensis Sonn.) seeds[J]. Food Chemistry. 2009, 116(1): 1-7.
[72] Prior R L, Wu X L, Schaich K. Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements[J]. Journal of Agricultural and Food Chenistry, 2005, 53(10): 4290-4302.
[73] Pulido R, Bravo L, Saura-Calixto F. Antioxidant activity of dietary polyphenols as determined by a modified ferric reducing/antioxidant power assay[J]. Journal of Agricultural and Food Chenistry, 2000, 48(8): 3396-3402.
[74] Markakis, P. Stability of anthocyanins in foods. In Anthocyanins as Food Colors; Markakis, P., Ed.; Academic Press Inc.:London, UK, 1982; pp 163-180.
[75] Kirca A, Ozkan M, Cemeroglu B. Effects of temperature, solid content and pH on the stability of black carrot anthocyanins[J]. Food Chemistry. 2007, 101(1): 212-218.
[76] Pacheco-Palencia L A, Talcott S T. Chemical stability of acai fruit (Euterpe oleracea Mart.) anthocyanins as influenced by naturally occurring and externally added polyphenolic cofactors in model systems[J]. Food Chemistry. 2010, 118(1): 17-25.
[77] Mollov P, Mihalev K, Shikov V, et al. Colour stability improvement of strawberry beverage by fortification with polyphenolic copigments naturally occurring in rose petals[J]. Innovative Food Science and Emerging Technologies. 2007, 8(3): 318-321.
[78] Eiro M J, Heinonen M. Anthocyanin color behavior and stability during storage: effect of intermolecular copigmentation[J]. Journal of Agricultural and Food Chemistry, 2002, 50(25): 7461-7466.