CN101554234A - 美国蔓越橘提取物及其用途 - Google Patents
美国蔓越橘提取物及其用途 Download PDFInfo
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- CN101554234A CN101554234A CNA2009101178890A CN200910117889A CN101554234A CN 101554234 A CN101554234 A CN 101554234A CN A2009101178890 A CNA2009101178890 A CN A2009101178890A CN 200910117889 A CN200910117889 A CN 200910117889A CN 101554234 A CN101554234 A CN 101554234A
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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Abstract
本发明涉及美国蔓越橘提取物及其用途,还涉及得到该提取物的方法。该提取物含有相对于提取物干重计以原花青素C1当量表示的至少15重量%的原花青素。这种富含原花青素的提取物可以用作饮食制剂或营养品制剂。
Description
技术领域
本发明涉及一种源自越橘(Vaccinium)的提取物和得到该提取物的方法。这种富含原花青素的提取物可以用作饮食制剂或营养品制剂。
背景技术
大果越橘(Vaccinium macrocarpon Aiton)(美国蔓越橘,Americancranberry)是不超过30cm的较小的爬地灌木,只自然生长在从美国卡洛莱纳州到加拿大的北美东部。这种植物旺盛生长在贫瘠而潮湿的泥炭沼、高沼的酸性土壤上或生长在针叶树林中。其果实是直径为10~20mm的小浆果。成熟时,该小浆果颜色为亮红,味道有点酸和涩。
美国蔓越橘在北美通常作为食物被消费已有几百年。它不含有已知毒性的可疑含量的任何组分,并且消费后没有事故被报道。在常用剂量下,很少有不希望的作用。消化系统疾病构成了临床研究中提到的有害作用的主要原因。当浆汁的摄入极为过量(3~4升/天)时会出现消化系统疾病,并且特别是导致痢疾和胃肠痛(Blumenthal M.等人,The ABC clinicalguide to herbs,American Botanical Council,Austin,Texas,2003,pp.73-83)。
美国蔓越橘的特点是富含类黄酮,并且最为特别的是富含A型原花青素低聚物,其是可食用植物中很少遇到的一族化合物。
缩合的单宁或原花青素是诸如儿茶酚、表儿茶酚和它们的衍生物等黄烷醇单元的聚合的酚化合物。
所报道的关于这种浆果治疗用途的第一则信息被认为可追溯到17世纪。美国印第安人似乎以膏状药的形式将其用于伤口。随后,航海家和殖民者将其用于坏血病的预防和用于治疗各种病症:循环系统疾病和肝病、胃痛、发烧等(Blumenthal等人,2003)。在一百多年前出现在美国的文章中已提到美国蔓越橘用于治疗尿路感染的用途。
1984年人们发现将美国蔓越橘汁口服给予小鼠14天产生了能够强烈抑制大肠杆菌(Escherichia coli)的尿路致病菌株粘附到泌尿上皮细胞上去的尿(Sobota A.E.,Inhibition of bacterial adherence by cranberry juice:potential use for the treatment of urinary tract infections.,J.Urol.,1984,131,5,1013-6)。
具有越橘属其他种(尤其是V.myrtillus(蓝莓))的高浓度原花青素的部分被认为具有相似活性(Howell A.B.等人,Inhibition of the adherence ofP-fimbriated Escherichia coli to uroepithelial-cell surfaces by proanthocyanidinextracts from cranberries,New Eng.J.Med.,1998,339,15,1085;Ofek L.等人,Anti-Escherichia coli adhesion activity of cranberry and blueberry juices.,NewEng.J.Med.,1991,324,22,1599)。
在浓度为10~50μg/ml下体外观察到,富含原花青素的大果越橘浆果提取物具有抑制尿路致病大肠杆菌(E.coli)菌株粘附到泌尿上皮细胞的能力(Howell等人,1998)。
在加拿大进行的研究中,含有美国蔓越橘脱水汁的片剂型被认为比蔓越橘汁优选,因为治疗后结果更好,并且浆汁有一种令有些受试者不喜欢的味道(Stothers L.,A.randomised trial to evaluate effectiveness and costeffectiveness of naturopathic cranberry products as prophylaxis against urinarytract infection in women.,Can.J.Urol.,2002,9,3,1558-62)。
然而,这些片剂的低原花青素剂量又迫使病人每天摄取几片片剂,从而使得这种治疗冗长乏味。
在2004年4月6日的2003-SA-0352意见中,Agence Francaise deSécuritéSanitaire des Aliments(AFSSA)(法国食品安全局)认可了以下主张:“Le jus concentréou la poudre de jus de fruits de Vaccinium macrocarponcontribue àdiminuer la fixation de certaines bactéries Escherichia coli sur lesparois des voies urinaires.”(大果越橘果实浓缩的浆汁或浆汁粉末有助于降低某些大肠杆菌对尿路壁的粘附)。AFSSA因此认为主张“contribue àdiminuer la fixation de certaines bactéries E.coli sur les parois des voiesurinaires”(有助于降低某些大肠杆菌对尿路壁的粘附)对于新鲜的/冷冻的蔓越橘和蔓越橘酱来说是可以接受的,条件是该产品的日摄入量可提供至少36mg原花青素(AFSSA,2007)。
因此,仍然需求富含原花青素并基于蔓越橘的组合物,这种组合物易于人类按日剂量摄入,在治疗时不受对病人来说很重要的待摄取的产品味道或待给予的片剂数量等方面的限制,从而每日能很容易地进行所述治疗。
发明内容
本申请人已经研究了一种构成本发明主题的新的越橘提取物。
本发明的主题还在于得到该部分的方法。
另一个主题在于该提取物作为饮食或营养品补品的用途。
经阅读后面的说明和实施例会发现本发明的其他主题。
附图说明
图1显示了作为测试的蔓越橘提取物(0=没有提取物,批号9030=来自Diana Naturals的AFCANN 9030提取物,批号A4124A=商购的提取物)和所用的细菌(DH5α菌株=参考菌株,菌株5和6=尿路致病菌株)的函数并且表示为光密度的粘附于孔板上的细菌数量。
具体实施方式
根据本发明的提取物源自越橘属的植物,含有相对于该提取物干重计以原花青素C1当量表示的至少15重量%的原花青素。
优选地,所述越橘为大果越橘或美国蔓越橘。该提取物优选从该植物的浆果得到。
优选地,该提取物含有相对于该提取物干重计从果实得到的高于5%的糖。
优选地,该提取物的原花青素的平均聚合度(即,单宁链的平均长度)大于5。更优选地,该提取物的原花青素的平均聚合度大于6。再更优选地,本发明提取物中原花青素的平均聚合度大于8。
优选地,该蔓越橘提取物保留有与蔓越橘汁的花青素量相等的花青素量。
由于根据本发明的用于制备富含原花青素的提取物的方法没有改变蔓越橘的花青素比例,因此蔓越橘提取物的性质与处理前浆汁的性质相近。
根据本发明的用于制备提取物的方法的特征在于,将美国蔓越橘果实放到水溶液中,进行固-液分离,加入有机溶剂,然后将所得的产物脱水,从而得到根据本发明的提取物。
优选地,水溶液的pH为2~6,温度低于95℃和压力为1~3atm。
任选地,在水相提取过程中,加入按水溶液体积计浓度为0.01~1%的亚硫酸盐或焦亚硫酸盐,优选浓度为0.3%。可以使用亚硫酸钠、亚硫酸钙、亚硫酸钾、焦亚硫酸钠或焦亚硫酸钾。
由于亚硫酸盐或焦亚硫酸盐的抗氧化和防腐作用,亚硫酸盐或焦亚硫酸盐有助于在制备过程中保留根据本发明的提取物的活性。
优选地,在pH为2~7、温度为15~70℃和压力为1~15atm下进行固-液分离。
优选地,将有机溶剂以相对于水溶液中的蔓越橘体积计0~15的体积比例加入。
优选地,使用的有机溶剂为醇、醛或酯。
更优选地,加入的有机溶剂为乙醇、乙醛或乙酸乙酯。
优选在温度低于60℃、减压(即,低于1atm)下将所得的产物脱水。
任选地,在温度为80~190℃和压力为0.5~2atm下进行最终的脱水步骤。
通过上述方法得到本发明的提取物。
可根据上述方法得到的、具有所述特征的美国蔓越橘提取物可以用作饮食或营养品补品。
根据本发明的富含原花青素的提取物具有防止细菌,具体而言是大肠杆菌细菌粘附到尿路壁表面的特点。
本发明的主题还在于一种组合物,尤其含有上述的富含原花青素的提取物。
根据本发明的组合物可以被摄取。根据给予方式,根据本发明的组合物可以以营养品领域的食品常用的所有形式提供。
根据本发明的组合物尤其可以以明胶胶囊、片剂、粉剂或饮料的形式提供。
本发明的营养品或饮食组合物可根据预期的应用进行常规配制。
以下实施例解释了本发明,而不以任何方式限制本发明。
实施例1:根据本发明的制备AFCANN9030提取物的方法
将美国蔓越橘以每5体积水1体积浆果的量放入水相溶液中。在温度为25℃下磨碎并通过倾析进行固-液分离后,在温度为20℃、pH为4.5、压力为1atm下,以每1体积蔓越橘5体积乙酸乙酯的量将乙酸乙酯加到水相中,然后将所得的两相分离。接下来在减压为0.1~0.5atm、温度为30~40℃下进行脱水,然后在压力为0.9~1.5atm和温度为145℃下进行第二次脱水。
所得的AFCANN9030产物为紫色粉末。
实施例2:分析所得的AFCANN9030提取物中的原花青素
使用经Porter改进的用于分析原花青素的Bate-Smith法(Bate-Smith E.C.Phytochemistry,1973,12,pp907-912和Porter.L.J.等人,Phytochemistry,1986,25,223-230)。
分析反应分两阶段进行。首先,将黄烷-3-醇聚合物在丁醇-HCl介质中水解。水解完全,释放出儿茶酚和表儿茶酚型的单体。其次的分析步骤在于在FeIII作用下使这些单体氧化。氧化产物为花青素,并且使用分光光度计在541nm下进行定量。
通过混合500mL浓度为35%的HCl、500mL正丁醇和150mgFe2(SO4)3来制备Bate-Smith试剂。
将100mg美国蔓越橘提取物溶解于按重量计1/1的软化水和乙醇的100mL混合物中。将2mL这种溶液与6mL Bate-Smith试剂在第一只试管中混合。将该溶液的一半转移至第二只试管。将第一只试管置于暗处,而将第二只试管在温度为100℃的水浴中加热30分钟,然后置于暗处10分钟。用去离子水作空白在541nm下测量两只试管的吸光度值(OD)。
根据下式将结果表示为原花青素C1当量。
测量的AFCANN9030提取物的原花青素以原花青素C1当量表示的浓度为30%±2。
应该指出的是,原花青素的定量考虑了介质中最初存在的花青素,并且在Porter反应后从吸收读数中减去了这些花青素产生的吸收。
实施例3:实施例1的提取物(AFCANN9030)的表征
实施例1的提取物 | |
原花青素%,原花青素C1当量 | 30.5% |
TPP%,儿茶酚当量(UV 280nm) | 17.3% |
黄烷醇%,芦丁当量(UV 354nm) | 2.4% |
黄烷醇%,槲皮素当量(UV 354nm) | 1.4% |
果糖(批号3808001) | 24.4mg/g |
葡萄糖(批号3808001) | 110.8mg/g |
蔗糖(批号3808001) | ND(未检测) |
总的干燥残余物 | 97.5% |
蛋白质(N*6.25)Dumas | 2.7% |
水解的脂肪物质 | <1% |
无机物 | 2.4% |
总饮食纤维 | 13.5% |
TPP:总多酚
实施例4:实施例1所得的提取物(AFCANN9030)的组成
硫代酸解法(thioacidolysis)是可以用于测定原花青素含量和原花青素的平均聚合度的另一种技术(Guyot等人,J.Agric.Food Chem.,1998,1698-1705)。
该技术也基于原花青素聚合物在酸介质(HCl)中的水解。反应介质中存在的α-甲苯硫醇与水解中释放的单体反应,并且通过液相色谱(HPLC)对形成的化合物进行定量。
硫代酸解后的分析结果
实施例1的提取物 | |
mDP | 8.5 |
单宁含量mg/g粉末 | 191.5 |
epi% | 71.5 |
cat% | 0.9 |
dimA% | 27.6 |
mDP指平均聚合度(单宁链的平均长度):
epi%=表儿茶酚%
cat%=儿茶酚%
dimA%=A型二聚体%
实施例5:根据本发明的提取物对生物膜的粘附试验
在该试验中使用没有菌毛的DH5α菌株作为粘附的阴性对照。菌株5和6为从临床分离得到的尿路致病大肠杆菌菌株。
在37℃下、CFA培养基中培养菌株。该培养基的组成如下:将10g酸水解酪蛋白(Difco)、1.5g酵母提取物、0.05g MgSO4和0.005g MnCl2加到1升去离子水中。
对每株菌株,进行只有菌株的粘附测试,或者,在含有30%原花青素的来自根据本发明的批号AFCANN9030的蔓越橘(Diana Naturals)或含有3%原花青素的来自批号A4124A的蔓越橘(商购样品)存在下,进行菌株的粘附测试。制备蔓越橘样品,从而得到浓度为500μg/mL的原花青素,即,16.7mg批号9030或167mg批号A4124A在10mL水中的稀释液。然后在水中将这些溶液稀释10倍,以进行试验。
使用待研究的细菌的过夜培养物,将0.2mL该培养物与待测试的蔓越橘样品一起或不与待测试的蔓越橘样品一起存放在96孔微板(Greiner)上,在水中再稀释5倍。每个试验进行三次。作为培养物的阴性对照,存放没有接种任一种待测试的蔓越橘样品的CFA培养基。
将该微板放到35℃的培养箱中并保持24h。
第二天,除去培养基并用蒸馏水洗涤板孔后,将200μL甲醇加到各孔中并保持15min。除去甲醇后,将板在培养箱中干燥30min。接下来,加入200μL结晶紫(Sigma)并保持10min。除去染色剂并用蒸馏水洗涤板孔后,加入200μL乙酸/乙醇溶液(vol/vol),以使产生的生物膜脱色。用分光光度计在570nm波长下进行读数。光密度值越高,粘附于板上的细菌数量越大。
图1的结果显示出,所测试的根据本发明的提取物其光密度比提取物A4124A的低,因此细菌对板孔的粘附均比批号A4124A的小,而不管测试的细菌如何。此时,在使用根据本发明的提取物的试验和使用A4124A提取物的试验中,培养基中的原花青素浓度相同。因此,对于由培养基中存在的相同浓度的原花青素所产生的抑制细菌对微板粘附的效果而言,根据本发明的提取物比A4124A提取物的高,即使原花青素的浓度相同。因而,在按质量计相同的浓度下,根据本发明的提取物中存在的原花青素对细菌粘附的抑制效果比A4124A提取物的高。
已知该商购提取物(批号A4124A)其原花青素的平均聚合度(即,单宁链的平均长度)为3~4。此外,根据本发明的提取物其原花青素的平均聚合度为8.1。
因此从这些结果可以推断出,根据本发明的蔓越橘提取物中的原花青素的平均聚合度在尿路致病细菌对板孔表面的粘附过程中发挥作用。因而,与商购产品A4124A相比,根据本发明的提取物具有增大的抗粘附功效。
实施例6:明胶胶囊的配制例子
为制造一种总量为250mg的明胶胶囊,将以下物质进行混合:
-240mg根据本发明的提取物,具有相对于提取物干重计以原花青素Cl当量表示的滴度为15%的原花青素,
-10mg微晶纤维素。
每个明胶胶囊含有36mg原花青素。
实施例7:明胶胶囊的配制例子
为制造一种总量为300mg的明胶胶囊,将以下物质进行混合:
-200mg AFCANN9030提取物(相对于提取物干重计以原花青素C1当量表示的滴度为30.5%的原花青素),
-100mg微晶纤维素。
每个明胶胶囊含有61mg原花青素。
实施例8:根据本发明的饮料的例子
准备100g的以下组合物:
量(g) | |
AFCANN9030提取物 | 0.68 |
透明的木莓汁的粉末(由Diana Naturals以名称AFFRAM0013出售) | 56.3 |
透明的草莓汁的粉末(由Diana Naturals以名称AFFRAI0013出售) | 29.3 |
黑加仑汁的粉末(由Diana Naturals以名称AFCASS9003出售) | 11.3 |
三氯蔗糖 | 0.23 |
柠檬酸 | 2.25 |
为制备该饮料,在最终体积为500ml的水中混合22g该粉末状组合物。搅拌后,得到具有明显木莓味的饮料,易于被摄取。
日剂量为500ml的该饮料提供了45mg原花青素。
Claims (18)
1.一种源自越橘属植物的提取物,所述提取物含有相对于提取物干重计以原花青素C1当量表示的至少15重量%的原花青素。
2.如权利要求1所述的提取物,其中所述越橘属植物为大果越橘或美国蔓越橘。
3.如权利要求1和2中任一项所述的提取物,其中使用美国蔓越橘的浆果。
4.如权利要求1~3中任一项所述的提取物,其中原花青素的平均聚合度大于5。
5.如权利要求4所述的提取物,其中原花青素的平均聚合度大于6。
6.如权利要求5所述的提取物,其中原花青素的平均聚合度大于8。
7.一种用于制备如权利要求1~6中任一项所述的提取物的方法,其中将美国蔓越橘放到水溶液中,进行固-液分离,加入有机溶剂,然后将所得的产物脱水,从而得到所述提取物。
8.如权利要求7所述的用于制备提取物的方法,其中进行第二次脱水。
9.如权利要求7和8中任一项所述的用于制备提取物的方法,其中所述有机溶剂为醇。
10.如权利要求9所述的用于制备提取物的方法,其中所述醇为乙醇。
11.如权利要求7和8中任一项所述的用于制备提取物的方法,其中所述有机溶剂为醛。
12.如权利要求11所述的用于制备提取物的方法,其中所述醛为乙醛。
13.如权利要求7和8中任一项所述的用于制备提取物的方法,其中所述有机溶剂为酯。
14.如权利要求13所述的用于制备提取物的方法,其中所述酯为乙酸乙酯。
15.如权利要求7~14中任一项所述的方法,其中将所述溶剂以相对于水溶液中的蔓越橘体积计0~15的体积比例加入。
16.如权利要求1~6中任一项所述的提取物,所述提取物可通过权利要求7~14中任一项所述的方法得到。
17.一种饮食或营养品组合物,其含有如权利要求1~6和16中任一项所述的提取物。
18.如权利要求1~6和16中任一项所述的提取物的用途,用于降低某些大肠杆菌细菌对尿路壁的粘附。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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FR0852386 | 2008-04-09 | ||
FR0852386A FR2929853B1 (fr) | 2008-04-09 | 2008-04-09 | Extrait de canneberge d'amerique et son utilisation |
US12/215,533 | 2008-06-27 |
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CN101554234A true CN101554234A (zh) | 2009-10-14 |
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CNA2009101178890A Pending CN101554234A (zh) | 2008-04-09 | 2009-03-13 | 美国蔓越橘提取物及其用途 |
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US (1) | US20090258940A1 (zh) |
JP (1) | JP2009249384A (zh) |
CN (1) | CN101554234A (zh) |
FR (1) | FR2929853B1 (zh) |
TW (1) | TW200948290A (zh) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101999648A (zh) * | 2010-10-11 | 2011-04-06 | 宝鸡市虹源生物科技有限公司 | 蔓越橘果实中花色苷及原花青素类成分的提取方法 |
CN102807545A (zh) * | 2012-08-08 | 2012-12-05 | 宁波杰顺生物科技有限公司 | 一种蔓越莓中原花青素提取物的制备方法 |
CN112638396A (zh) * | 2018-09-07 | 2021-04-09 | 伊诺弗斯公司 | 抗炎植物提取物 |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20110280851A1 (en) * | 2010-05-12 | 2011-11-17 | Herzlinger Alexander S | Compositions Comprising Cranberry Extract and Methods of Use Thereof |
US9969707B2 (en) | 2011-12-16 | 2018-05-15 | CENTRE DE RECHERCHE INDUSTRIELLE DU QUéBEC | Method for extracting anthocyanin derivatives from a plant source |
WO2017024183A1 (en) * | 2015-08-04 | 2017-02-09 | Ocean Spray Cranberries, Inc. | Subcritical water extraction of fruit material |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
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AU744527B2 (en) * | 1997-09-09 | 2002-02-28 | Rutgers, The State University Of New Jersey | Plant proanthocyanidin extract effective at inhibiting adherence of bacteria with P-type fimbriae to surfaces |
ES2269104T3 (es) * | 1999-04-23 | 2007-04-01 | Kyowa Hakko Kogyo Co., Ltd. | Metodos para purificar oligomeros de proantocianidina. |
US6210681B1 (en) * | 1999-09-07 | 2001-04-03 | Jlb, Inc. | Plant proanthocyanidin extracts |
JP3689413B2 (ja) * | 2003-05-20 | 2005-08-31 | 株式会社東洋新薬 | フラバン化合物含有組成物 |
JP2006282612A (ja) * | 2005-04-01 | 2006-10-19 | Univ Of Tokyo | 脳梗塞の予防又は治療剤、及び機能性食品 |
US8715949B2 (en) * | 2006-09-07 | 2014-05-06 | The United States Of America, As Represented By The Secretary Of The Navy | Applications of the binding interaction of proanthocyanidins with bacteria and bacterial components |
-
2008
- 2008-04-09 FR FR0852386A patent/FR2929853B1/fr not_active Expired - Fee Related
- 2008-06-27 US US12/215,533 patent/US20090258940A1/en not_active Abandoned
-
2009
- 2009-03-13 CN CNA2009101178890A patent/CN101554234A/zh active Pending
- 2009-04-07 TW TW098111451A patent/TW200948290A/zh unknown
- 2009-04-08 JP JP2009093587A patent/JP2009249384A/ja active Pending
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101999648A (zh) * | 2010-10-11 | 2011-04-06 | 宝鸡市虹源生物科技有限公司 | 蔓越橘果实中花色苷及原花青素类成分的提取方法 |
CN101999648B (zh) * | 2010-10-11 | 2012-10-31 | 宝鸡市虹源生物科技有限公司 | 蔓越橘果实中花色苷及原花青素类成分的提取方法 |
CN102807545A (zh) * | 2012-08-08 | 2012-12-05 | 宁波杰顺生物科技有限公司 | 一种蔓越莓中原花青素提取物的制备方法 |
CN102807545B (zh) * | 2012-08-08 | 2014-08-13 | 宁波杰顺生物科技有限公司 | 一种蔓越莓中原花青素提取物的制备方法 |
CN112638396A (zh) * | 2018-09-07 | 2021-04-09 | 伊诺弗斯公司 | 抗炎植物提取物 |
Also Published As
Publication number | Publication date |
---|---|
FR2929853A1 (fr) | 2009-10-16 |
TW200948290A (en) | 2009-12-01 |
FR2929853B1 (fr) | 2010-10-08 |
US20090258940A1 (en) | 2009-10-15 |
JP2009249384A (ja) | 2009-10-29 |
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