CN115005442B - 一种天山雪莲多糖提取物制备方法与应用 - Google Patents
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Abstract
本发明公开了一种天山雪莲多糖提取物的制备方法与应用。该方法以天山雪莲全草为原料,粉碎、提取、浓缩后,经乙醇沉淀、后获得雪莲多糖提取物。本发明制备的天山雪莲多糖提取物对青春双歧杆菌、粪肠球菌和德氏保加利亚乳杆菌具有显著的促进作用。本发明得到的天山雪莲多糖提取物可以起到促进益生菌生长的作用,能够作为原料或添加剂广泛应用于各类营养与保健用品及日化产品中。
Description
技术领域
本发明属于营养与保健品技术领域,具体涉及一种天山雪莲多糖提取物制备方法与应用。
背景技术
益生菌被称作为人体健康微生态调节剂,具有广泛的应用价值,从人体皮肤表面到与外界相通的呼吸道、口腔、肠道、生殖泌尿道等都存在着微生物菌群,为人体健康发挥着重要作用。益生菌是活的微生物,摄入适量有助于保持健康。微生物在人类健康中的重要性可以追溯到路易·巴斯德第一次发现发酵的重要性的时代,也让人们注意到食用发酵食品可能有益于健康和长寿。近年来,养生保健品的制作过程中直接添加益生菌或添加促进益生菌生长的原料,制成口服液、软/硬胶囊、颗粒等益生菌制剂保健品。研究表明,有益微生物的任何失衡也会导致疾病的发生,如菌群紊乱与炎症性皮肤病之间存在一定的关联性,从而增加了补充益生菌作为炎性皮肤病治疗选择的可能性。然而,对局部应用益生菌产品的功效研究少有报道。
多糖是由各种单糖通过糖苷键结合而组成的天然高分子碳水化合物,具有广泛的生物学效应。研究表明,多糖在抗肿瘤、抗炎、抗病毒、降血糖、抗衰老、促进免疫等方面都具有生物活性。另外,天然植物多糖对人体较为安全,吸收、利用率较高。因此,随着分子生物学的发展,多糖所具有的各种生理功能逐渐为人们所重视,尤其是增强免疫力的功能被广泛认同。
天山雪莲是新疆特有的一种名贵的生草本植物,为新疆稀有名贵中草药,生长于新疆天山海拔3000米以上终年积雪的悬崖峭壁上。目前,以天山雪莲提取物及雪莲果为主成分的营养与保健产品很多,其功效主要集中在抗疲劳,活血化瘀,免疫调节,抗肥胖等。然而关于天山雪莲多糖及其促进益生菌生长,调节微生态的研究尚未见报道。
发明内容
本发明的目的在于,克服现有技术中存在的不足,提供一种天山雪莲多糖提取物制备方法与应用,该方法取天山雪莲全草,粉碎、提取、浓缩后,经乙醇沉淀后获得天山雪莲粗多糖,通过本发明所述方法获得的天山雪莲多糖提取物对青春双歧杆菌、粪肠球菌和德氏保加利亚乳杆菌具有显著的促进作用。可以起到促进益生菌生长的作用,能够作为原料或添加剂广泛应用于各类营养与保健用品及日化产品中。
本发明所述的一种天山雪莲多糖提取物的制备方法,按下列步骤进行:
a、热水提取:将选取的天山雪莲全草粉碎得到雪莲全草粉,按料液比1:10-1:50加入水,在温度为60-90℃的中提取2次,提取时间为1.5-3.5h,过滤,合并两次的上清液,得到总提取液;
b、提取液浓缩:将步骤a得到的提取液进行浓缩,浓缩至原提取液的1/40-1/10,浓缩温度为40-60℃,得雪莲多糖浓缩液;
c、醇沉处理:将步骤b得到的浓缩液按体积比1:3-5与食用酒精沉淀,得到雪莲多糖沉淀物;
d、将步骤c得到的沉淀物用食用酒精洗涤,温度20-55℃干燥,粉碎,得到天山雪莲多糖提取物。
所述方法获得的天山雪莲多糖提取物在制备促进益生菌生长中的用途。
所述的益生菌为青春双歧杆菌、粪肠球菌和德氏保加利亚乳杆菌。
具体实施方式
为了进一步理解本发明,下面结合实施列对本发明做进一步说细描述,但不仅限于所描述的施例。
实施例1
a、将选取的天山雪莲全草药材50克进行粉碎得到雪莲全草粉,加入原料重量的20倍纯净水1000mL,在温度75℃水中提取2次,提取时间为2小时,过滤,合并两次的上清液,得到总提取液;
b、将步骤a得到的总提取液进行浓缩,采用旋转蒸发仪在温度50℃下进行减压浓缩至1/10体积,得到浓缩液;
c、将步骤b得到得浓缩液用4倍体积的食用酒精沉淀并过夜,再将沉淀物离心,得到沉淀物;
d、将步骤c得到的沉淀物用食用酒精洗涤,温度为50℃干燥,粉碎得到天山雪莲提取物;即为雪莲多糖粗品9.8g,多糖提取率为19.6%;分析结果表明,提取物的多糖含量为22.5%,蛋白质含量为23.1%。
实施例2
a、将选取的天山雪莲全草药材50克进行粉碎得到雪莲全草粉,加入原料重量的25倍纯净水1250ml,在温度为80℃水中提取2次,提取时间为2.5小时,过滤,合并两次的上清液,得到总提取液;
b、将步骤a得到的总提取液进行浓缩,采用旋转蒸发仪在温度45℃下对提取液进行减压浓缩至1/30体积,得到浓缩液;
c、将步骤b得到得浓缩液用4倍体积的食用酒精中沉淀并过夜,将沉淀物抽滤,得到沉淀物;
d、将步骤c得到的沉淀物用食用酒精洗涤,温度为55℃干燥,粉碎得到天山雪莲提取物;即为雪莲多糖粗品10.5g,多糖提取率为21%;分析结果表明,提取物的多糖含量为23.15%、蛋白质含量为24.5%。
实施例3
a、将选取的天山雪莲全草药材50克进行粉碎得到雪莲全草粉,加入原料重量的30倍纯净水1500ml,于温度85℃提取2次,每次提取3.0h;过滤,合并2次上清液,得到总提取液;
b、将步骤a得到的总提取液进行浓缩,采用旋转蒸发仪在温度40℃下对提取液进行减压浓缩至1/35体积,得到浓缩液;
c、将步骤b得到得浓缩液在4.5倍体积的食用酒精中沉淀并过夜,将沉淀物离心,得到沉淀物;
d、将步骤c得到的沉淀物用食用酒精洗涤,温度为20℃干燥,粉碎得天山雪莲提取物,即为雪莲多糖粗品9.45g,多糖提取率为18.9%;分析结果表明,提取物的多糖含量为24.15%、蛋白质含量为22.7%。
实施例4
a、将选取的天山雪莲全草药材50克进行粉碎得到雪莲全草粉,加入原料重量的50倍纯净水2500ml,于温度90℃提取2次,每次提取1.5h,过滤、合并2次上清液,得到总提取液;
b、将步骤a得到的总提取液进行浓缩,采用旋转蒸发仪在60℃下对提取液进行减压浓缩至1/30体积,得到浓缩液;
c、将步骤b得到得浓缩液在4倍体积的食用酒精中沉淀并过夜,将沉淀物抽滤,得到沉淀物;
d、将步骤c得到的沉淀物用食用酒精洗涤,温度为50℃干燥、粉碎得到天山雪莲提取物;即为雪莲多糖粗品7.60g,多糖提取率为15.2%;分析结果表明,提取物的多糖含量为20.10%、蛋白质含量为19.7%。
实施例5
a、将选取的天山雪莲全草药材50克进行粉碎得到雪莲全草粉,加入原料重量的10倍纯净水500ml,于温度60℃提取2次,每次提取2.0h,过滤,合并2次上清液,得到总提取液;
b、将步骤a得到的总提取液进行浓缩,采用旋转蒸发仪在温度50℃下对提取液进行减压浓缩至1/20体积,得到浓缩液;
c、将步骤b得到的浓缩液用4倍体积的食用酒精中沉淀并过夜,将沉淀物离心,得到沉淀物;
d、将步骤c得到的沉淀物用食用酒精洗涤,温度为20℃干燥、粉碎得到天山雪莲提取物;即为雪莲多糖粗品6.50g,多糖提取率为13.0%;分析结果表明,提取物的多糖含量为17.10%、蛋白质含量为18.7%。
实施例6
选用皮肤益生菌青春双歧杆菌、保加利亚乳杆菌、粪链球菌,观察天山雪莲多糖对益生菌的作用,具体方案如下:
(1)菌种活化:从保存的保加利亚乳杆菌、青春双歧杆菌、粪链球菌菌种制剂中分别取一环接种于基础培养基、TPY培养基和THY培养基,于温度37℃培养箱培养48h后,取0.1mL接种于相应液体培养基培养48h,传代两次后得到相应活化的菌种,其中青春双歧杆菌为厌氧型微生物,故将其置于装有厌氧产气袋的不透气乐扣箱培养;
(2)多糖对三种益生菌体外生长的影响试验:分别取0.5mL的保加利亚乳杆菌、青春双歧杆菌、粪链球菌菌液分别接种至添加雪莲多糖提取物的溶液中,使雪莲多糖提取物最终浓度为1、2、3mg/mL,分别于相应的培养条件培养16h,各取10μL菌液使用计数仪计算菌落数,以80mg/mL低聚果糖(FOS)作为阳性对照,每组试验重复3次,结果见表1、2、3;
表1.天山雪莲多糖对青春双岐杆菌生长的影响
注:所有天山雪莲多糖提取物溶液样品在接种青春双歧杆菌前已用0.22μM滤过膜完成过滤除菌;
结果表明:天山雪莲多糖提取物在1、2、3mg/mL具有显著提高青春双岐杆菌活菌数量,说明雪莲多糖提取物具有显著促进青春双岐杆菌生长作用;
表2.天山雪莲多糖对粪肠球菌生长的影响
注:所有天山雪莲多糖提取物溶液样品在接种粪肠球菌前已用0.22μM滤过膜完成过滤除菌;
结果表明:天山雪莲多糖提取物在1、2、3mg/mL具有显著提高粪肠球菌数量,说明天山雪莲多糖提取物具有显著促进粪肠球菌生长作用;
表3.天山雪莲多糖对德氏乳杆菌生长的影响
注:所有天山雪莲多糖提取物溶液样品在接种德氏乳杆菌前已用0.22μM滤过膜完成过滤除菌;
天山雪莲多糖提取物在2、3mg/mL具有显著提高德氏乳杆菌数量,说明天山雪莲多糖提取物具有显著促进德氏乳杆菌生长作用;
结论:天山雪莲多糖提取物对益生菌的生长具有促进作用。
Claims (1)
1.一种天山雪莲多糖提取物在制备促进益生菌为青春双歧杆菌、粪肠球菌和德氏保加利亚乳杆菌生长产品中的用途,所述天山雪莲多糖提取物的制备方法,按下列步骤进行:
a、热水提取:将选取的天山雪莲全草粉碎得到雪莲全草粉,按料液比1:10-1:50加入水,在温度为60-90℃的中提取2次,提取时间为1.5-3.5h,过滤,合并两次的上清液,得到总提取液;
b、提取液浓缩:将步骤a得到的提取液进行浓缩,浓缩至原提取液的1/40-1/10,浓缩温度为40-60℃,得雪莲多糖浓缩液;
c、醇沉处理:将步骤b得到的浓缩液按体积比1:3-5与食用酒精沉淀,得到雪莲多糖沉淀物;
d、将步骤c得到的沉淀物用食用酒精洗涤,温度20-55℃干燥,粉碎,得到天山雪莲多糖提取物。
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