CN105395919B - 一种含有黑木耳提取物、具有降血脂作用的组合物及其制备方法 - Google Patents
一种含有黑木耳提取物、具有降血脂作用的组合物及其制备方法 Download PDFInfo
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- CN105395919B CN105395919B CN201510816925.8A CN201510816925A CN105395919B CN 105395919 B CN105395919 B CN 105395919B CN 201510816925 A CN201510816925 A CN 201510816925A CN 105395919 B CN105395919 B CN 105395919B
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Abstract
本发明公开了一种含有黑木耳提取物、具有降血脂作用的组合物及其制备方法,所述组合物由以下成分组成:黑木耳多糖400~1100mg、黑木耳黑色素35~100mg、红曲提取物95~350mg、银杏叶提取物75~180mg、山楂提取物55~170mg、蒲黄提取物55~170mg、甘草提取物70~170mg。本发明中组合物采用黑木耳提取物、红曲提取物、银杏叶提取物、山楂提取物、蒲黄提取物等多种具有降血脂功能的生物活性物质,增强了降血脂效果,而且增加了具有解毒排毒功能的甘草提取物作为组合物,具有协同增效、减毒排毒的效果。在降低血清总胆固醇、血清甘油三酯和低密度脂蛋白胆固醇方面存在着明显的协同性作用,在降低副作用方面具有明显优势。
Description
技术领域
本发明涉及一种具有降血脂作用的组合物及其制备方法。
背景技术
高血脂症严重威胁人体健康。高血脂症能直接导致脑梗塞、血栓性脉管炎、肥胖症、动脉粥样硬化症、脂肪肝、胆石病及动脉粥样硬化冠状动脉性心脏病等心脑血管疾病,而心脑血管疾病是人类最大死因之一,全球每年约有1200万人死于心脑血管病,因此高脂血症的预防和治疗至关重要。目前对高血脂的治疗以西药为主,副作用大,因此研制疗效显著、副作用低的以预防为主的食品或药品具有十分重要的意义。
降血脂复方制剂中成分复杂,或多或少有一定的副作用。因此研制真正具有降血脂功效且无毒副作用的保健品或者药品具有十分重要的意义和较好的应用前景。
黑木耳是真菌门担子菌纲木耳目木耳科木耳属的药食兼用真菌,含有多糖、黑色素等多种活性成分。传统医学记载,黑木耳具有“性平、味甘”、“益气不饥,轻身强志”、“清肺润肺”、“利五脏、排毒气”之功效。现代医学研究证明,黑木耳具有提高免疫力、抗衰老、抗癌、抗辐射、抗血栓形成、降血脂、降血糖、提升肠胃消化功能等功效。黑木耳无论在疾病的预防、治疗还是日常保健方面,都凸显出其独到的、难以替代的诸多功效。黑木耳具有降血脂活性的可能机制是黑木耳可增强机体免疫力,调节免疫功能,维持人体内各器官的正常运作,提高参与酶的活性,促进肝胆的正常循环作用。这样使得血脂中各种成分达到正常状态,从而降低血脂在血液中的堆积量,保证人体健康。
目前黑木耳的降血脂活性成分研究主要集中于多糖,而针对黑木耳多糖提取的常规水提醇沉工艺的缺点如下:①得率较低,而成本较高,黑木耳多糖提取收率一般为3~5%,且多为实验室阶段,成本较高,难以放大生产。②由于提取时高温导致有效成分的降血脂活性降低,限制其广泛应用。
发明内容
本发明的目的是提供一种含有黑木耳提取物、具有降血脂作用的组合物及其制备方法,该组合物含有黑木耳提取物、红曲米提取物、银杏叶提取物、山楂提取物、蒲黄提取物、甘草提取物,具有降血脂作用,且其降血脂效果显著优于单独使用黑木耳提取物、红曲提取物、银杏叶提取物、山楂提取物或甘草提取物,具有协同增效、协同减毒的效果。
本发明的目的是通过以下技术方案实现的:
一种含有黑木耳提取物、具有降血脂作用的组合物,由以下成分组成:黑木耳多糖400~1100mg、黑木耳黑色素35~100mg、红曲提取物95~350mg、银杏叶提取物75~180mg、山楂提取物55~170mg、蒲黄提取物55~170mg、甘草提取物70~170mg。
上述含有黑木耳提取物、具有降血脂作用的组合物的制备方法,包括以下步骤:
(1)将黑木耳粉碎,用30~60倍体积的去离子水搅拌均匀,放置1~3h,常温采用高压差法(溶液加压至1.0~1.5Mpa,在0.1~0.3s内迅速释压至常压状态)提取1~3次,过滤,滤液真空减压浓缩,干燥,粉碎,即得黑木耳多糖提取物;滤渣加入2~5倍体积的氢氧化钠溶液调整溶液pH值为10.0~11.0,静置1~2h,过滤,上清液用盐酸调整pH值为3.0~3.5,静置1~2h,过滤,沉淀冷冻干燥,粉碎,得到黑木耳黑色素。
(2)银杏叶、山楂和蒲黄均按照以下提取工艺制备提取物:原料粉碎,用10~20倍体积分数为50~95%的乙醇溶液超声波200~500W下提取2~3次,每次0.5~1.0h,合并提取液,过滤,滤液回收乙醇,浓缩,干燥,粉碎,即得醇提取物。
(3)甘草中加入6~20倍体积的氨乙醇溶液(0.1%氨水,25%乙醇)于70~90℃温度下进行回流提取1~2h,过滤,烘干,粉碎,即得甘草提取物。
(4)红曲米粉碎,采用15~25倍体积的60~85%的乙醇溶液,超声波200~500W下提取2~3次,每次0.5~1.0h,合并提取液,过滤,滤液回收乙醇,浓缩,干燥,粉碎,即得醇提取物。
(5)将黑木耳多糖、黑木耳黑色素、山楂提取物、银杏提取物、甘草提取物、蒲黄提取物和红曲提取物混合,即得含有黑木耳提取物、具有降血脂作用的组合物。
本发明中,所述黑木耳原料采用黑29品种在黑龙江省尚志市亚布力地区进行秋季栽培生产的黑木耳子实体,经过比较分析发现该原料的功效成分如黑木耳多糖和黑色素含量均比其他地区生产的黑木耳高。
本发明中,所述黑木耳提取物中黑木耳多糖含量30~40%(多糖溶液超滤后,采用苯酚硫酸法测定),黑色素含量3%。
本发明中,所述黑木耳多糖和黑色素的纯度为30~40%。
本发明中,所述红曲提取物中有效成分Monacolin-K(洛伐他汀)含量达到1.8%以上(HPLC法),其中开环(酸式)Monacolin-K含量占85%以上。
本发明中,所述银杏叶提取物中银杏黄酮≥30%(HPLC法),银杏总内酯≥8%(HPLC法),银杏酸≤3ppm。总黄酮醇苷的含量测定参照《中国药典》银杏叶提取物法,组合物中总黄酮含量采用高压液相色谱法测定。
本发明中,所述甘草提取物中甘草酸含量15%以上,三萜类和黄酮类化合物总含量20~30%,多羟基酚类化合物含量5~35%。
本发明中,所述蒲黄提取物中总黄酮含量20~25%(HPLC法)。
本发明中,所述山楂提取物中总黄酮含量3~25%。
本发明中,所述黑木耳的粉碎粒度为150~240目,最佳粒度200目。
本发明中,所述银杏叶、山楂、蒲黄和红曲米的粉碎粒度为100~150目。
红曲是由红曲霉发酵产生,红曲霉发酵可产生多种代谢产物,主要有红曲色素、酶类活性物质、降血压物质等。红曲提取物的应用在我国历史悠久,自古被用于酿酒、酿醋、烤鸭等食物着色。现代医学发现红曲具有降低胆固醇的作用。由于红曲中起到降血脂作用的有效成分不仅是Monacolin类(洛伐他汀),还包括羟基酸洛伐他汀、麦角甾醇、色素类、γ-氨基丁酸、Dimerumicacid等及其他成分,由于红曲提取物的其他成分也在发挥其独特的作用,因此红曲提取物在降血脂、抗癌及抗氧化方面的作用效果可能会大于单用洛伐他汀的效果。
银杏叶为银杏科植物银杏的干燥叶,具有活血化疲、通络止痛等功效,在临床上多用于心脑血管疾病。到目前为止,在银杏叶中分离得到黄酮、萜内酯、有机酸、氨基酸、多糖等成分。总黄酮和萜类内酯作为银杏叶中的特征性成分,有明确生物活性。
山楂是蔷薇科植物,含有多种生物活性物质,如三萜酸、黄酮等,其中黄酮类成分有降压、降血脂、增加冠脉流量、强心、抗心律不齐等药理作用。根据山楂的药理作用,现在已有很多山楂及山楂复方制剂应用于临床来治疗高血脂症、冠心病、心绞痛、心律失常等心血管疾病,我国山楂资源丰富,成本低廉,有很好的开发利用价值。
蒲黄为香蒲科植物或同属植物的干燥花粉,具有止血、化瘀、通淋的功能,用于心脑血管疾病的防治。
甘草提取物有防治高脂血症、抗病毒、防治艾滋病、抗癌、增强细胞免疫调节及减毒排毒功效,甘草的主要有效成份是甘草酸及甙元甘草次酸。
本发明具有如下优点:
1)黑木耳提取物中粗多糖提取收率达12~18%以上,纯度30%以上,且为低温提取,充分保留了生物活性;黑色素提取率3%以上,纯度40%以上。
2)本发明中组合物采用黑木耳提取物、红曲提取物、银杏叶提取物、山楂提取物、蒲黄提取物等多种具有降血脂功能的生物活性物质,增强了降血脂效果,而且增加了具有解毒排毒功能的甘草提取物作为组合物,具有协同增效、减毒排毒的效果。在降低血清总胆固醇、血清甘油三酯和低密度脂蛋白胆固醇方面存在着明显的协同性作用,在降低副作用方面具有明显优势。按照国家保健食品功能试验中降血脂试验操作,结果表明组合物在降血脂效果方面显著优于单独使用黑木耳提取物、红曲提取物或银杏叶提取物。
3)黑木耳提取物采用高效高压差低温连续式提取分离浓缩技术,全程采用物理方式、低温提取,提取率高且实现活性成分的原生态保留。高低压差提取技术使得细胞瞬间爆裂,与溶媒充分接触,使黑木耳中85%以上的多糖均被提取出来,使多糖收率达到12~18%。
4)该组合物经动物试验和人体试食结果证明具有辅助降血脂作用,可应用于中成药、保健食品或食品,降低胆固醇和血液粘度,促进心脑血管的微循环,改善体内脂肪代谢、增强机体免疫力,具有辅助降血脂功能。
具体实施方式
下面结合实施例对本发明的技术方案作进一步的说明,但并不局限于此,凡是对本发明技术方案进行修改或者等同替换,而不脱离本发明技术方案的精神和范围,均应涵盖在本发明的保护范围中。
实施例1:
本实施例提供的含有黑木耳提取物、具有降血脂作用的组合物由以下成分组成:黑木耳提取物(包括纯度40%的黑木耳多糖550mg和纯度为40%的黑色素70mg)、红曲提取物330mg、银杏叶提取物90mg、山楂提取物150mg、蒲黄提取物150mg、甘草提取物150mg。
实施例2:
本实施例提供的含有黑木耳提取物、具有降血脂作用的组合物由以下成分组成:黑木耳提取物(包括纯度30%的黑木耳多糖1000mg和纯度为40%的黑色素90mg)、红曲提取物110mg、银杏叶提取物160mg、山楂提取物80mg、蒲黄提取物80mg、甘草提取物80mg。
实施例3:
本实施例提供的含有黑木耳提取物、具有降血脂作用的组合物由以下成分组成:黑木耳提取物(包括纯度40%的黑木耳多糖800mg和纯度为30%的黑色素50mg)、红曲提取物250mg、银杏叶提取物120mg、山楂提取物100mg、蒲黄提取物100mg、甘草提取物100mg。此为1人次的日用量(1.52g),分2次服用。
制备方法:
(1)将黑木耳粉碎至200目,用40倍体积的去离子水搅拌均匀,放置2h,常温采用高压差法(溶液加压至1.0Mpa,在0.1s内迅速释压至常压状态)提取2次,过滤,滤液真空浓缩,干燥,粉碎,即得黑木耳多糖提取物;滤渣加入4倍体积的氢氧化钠溶液调整溶液pH值为10.5,静置1.5h,过滤,上清液用盐酸调整pH值为3.2,静置1.5h,过滤,沉淀冷冻干燥,粉碎,得到黑木耳黑色素。
(2)银杏叶、山楂和蒲黄均按照以下提取工艺制备提取物:原料粉碎至100目,用15倍体积分数为60%的乙醇溶液超声波300W下提取3次,每次0.5h,合并提取液,过滤,滤液回收乙醇,浓缩,干燥,粉碎,即得醇提取物。
(3)甘草中加入10倍体积的氨乙醇溶液(0.1%氨水,25%乙醇)于80℃温度下进行回流提取1.5h,过滤,烘干,粉碎,即得甘草提取物。
(4)红曲提取物制备工艺为:五常有机大米、浸泡、蒸煮、冷却、接种紫叶红曲菌、天然培养发酵、生产红曲米,粉碎至100目,采用20倍体积的80%的乙醇溶液,超声波500W下提取2次,每次1.0h,合并提取液,过滤,滤液回收乙醇,浓缩,干燥,粉碎,即得醇提取物。本工艺生产的功能红曲有效成分的含量更明确,效果更确切。不含桔霉素、无毒副作用,减少肝脏负担,吸收更容易,效果更卓越。
(5)将黑木耳多糖、黑木耳黑色素、山楂提取物、银杏提取物、甘草提取物、蒲黄提取物和红曲提取物,制备硬胶囊,或与药用辅料一起混合按常规方法制成口服制剂、硬胶囊、软胶囊、片剂、颗粒剂、散剂、丸剂、袋泡茶、液体饮料中的一种或多种,可在制备降血脂药品、保健食品中应用。
动物实验:
实验方法:采用SD大鼠由上海斯莱克实验动物有限责任公司提供,生产许可证号为SCXK(沪)2012-0002;体重200-250g,雄性,随机分组,分别为正常组、模型组、阳性对照药血脂康0.2g·kg-1组,以及黑木耳提取物复方制剂0.135、0.27、0.54g·kg-1低、中、高3个剂量组,每组10只。除正常组给予饮用水外,各组大鼠均以高脂饲料(南通特洛菲饲料)饲喂,每天3次,连续3周。模型组与正常组比较,TC和LDL-C均明显升高,有显著性差异(P<0.01),表明高脂饲料造大鼠实验性高血脂动物模型成功。检验合格的大鼠开始试验:灌胃给药,每日,除正常对照组外,各组大鼠继续以高脂饲料饲喂。各给药组给黑木耳提取物复方制剂低、中、高剂量组(剂量分别为0.135、0.27、0.54g·kg-1),灌胃给药容量20mL·kg-1。阳性药血脂康0.2g·kg-1,灌胃给药容量10mL·kg-1。连续8周。各组大鼠采血,分离血清,采用甘油三酯、总胆固醇、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)测试盒(北京北化泰临床试剂有限公司)测定各含量。
实验结果:与模型组比较,阳性对照血脂康组能有效降低TG、TC和LDL-C水平,差异有显著性意义(P<0.01);各剂量组血清高密度脂蛋白胆固醇(HDL-C)不显著低于模型对照组;复方黑木耳提取物低、中、高剂量组能明显降低TG、TC和LDL-C水平,除低剂量组降低低密度脂蛋白胆固醇(LDL-C)差异较显著(P<0.05)之外,其他各组与模型组比较差异均极显著(P<0.01),而各组HDL-C水平与模型组差异不显著。各实验组血脂水平检测结果如表1所示。
表1 黑木耳复方制剂对高脂血症大鼠血脂水平的影响()mmol·L-1
注:与空白组比较aP<0.01,bP<0.05;与模型组比较,cP<0.01,dP<0.05。
组合物具有降低血清甘油三酯、总胆固醇、低密度脂蛋白的作用,而使高密度脂蛋白胆固醇稍有提高,具有显著降血脂功能,而且降血脂效果优于单用组合物中任意1种提取物的降血脂效果(见表2)。
表2 黑木耳复方制剂与单方制剂对高脂血症大鼠血脂水平的影响()mmol·L-1
注:与空白组比较aP<0.01,bP<0.05;与模型组比较,cP<0.01,dP<0.05。
人体试食试验:
为检验本发明的治疗效果,选取200例高血脂患者,所选患者均符合高脂血症诊断标准。其中男性75例,女性75例,年龄40~63岁,平均年龄48岁。随机分为4组,每组50人。第一组用实施例1制备的组合物治疗;第二组用实施例2制备的组合物治疗;第三组用实施例3制备的组合物治疗;第四组为空白对照组给以安慰剂。疗程90d,分别在治疗前后均空腹12h后于清晨抽取静脉血测定TC、TG、LDL-C、HDL-C、心、肝、肾功能及血、尿、便常规等。在试验开始前进行胸透、心电图、腹部B超检查。结果见表3。
各项指标判定如下:
(1)有效:TC降低>10%;TG降低>15%;HDL-C上升>0.104mmol/L。
(2)无效:未达到有效标准者。
表3 人体试吃试验组试验结果(n=50)
通过对试验后的各组个体进行血脂指标的检查和血、尿、便常规、肝、肾功能检查的结果进行分析发现各实施例总有效率为96.0%以上,副反应发生率和半年复发率均为0。肝、肾功能检查结果表明试验组肝肾功能比服用组合物之前的生理指标还健康,整体表现了组合物具有增效减毒的突出特点。
结论:
本发明对高脂血症的发生和治疗有较好作用。
通过动物实验表明:1)模型对照组和空白对照组比较,血清总胆固醇或低密度脂蛋白胆固醇升高,差异有显著性,判定模型成立;2)各剂量组与模型对照组比较,中、高剂量组血清总胆固醇、血清甘油三酯、低密度脂蛋白胆固醇降低,差异有显著性,并且各剂量组血清高密度脂蛋白胆固醇不显著低于模型对照组,表明该受试样品辅助降血脂功能动物实验结果阳性。
通过人体试食试验检验结果表明:1)组合物各实施例总有效率为96.0%以上,副反应发生率和半年复发率均为0;2)人体生理指标和肝、肾功能等检查结果表明组合物整体表现了具有增效减毒的突出特点。
Claims (5)
1.一种含有黑木耳提取物、具有降血脂作用的组合物的制备方法,含有黑木耳提取物、具有降血脂作用的组合物,由以下成分组成:黑木耳多糖400~1100mg、黑木耳黑色素35~100mg、红曲提取物95~350mg、银杏叶提取物75~180mg、山楂提取物55~170mg、蒲黄提取物55~170mg、甘草提取物70~170mg;
其特征在于所述制备方法步骤如下:
(1)将黑木耳粉碎,用30~60倍体积的去离子水搅拌均匀,放置1~3h,常温采用高压差法提取1~3次,过滤,滤液真空减压浓缩,干燥,粉碎,即得黑木耳多糖提取物;滤渣加入2~5倍体积的氢氧化钠溶液调整溶液pH值为10.0~11.0,静置1~2h,过滤,上清液用盐酸调整pH值为3.0~3.5,静置1~2h,过滤,沉淀冷冻干燥,粉碎,得到黑木耳黑色素;
(2)银杏叶、山楂和蒲黄均按照以下提取工艺制备提取物:原料粉碎,用10~20倍体积分数为50~95%的乙醇溶液超声波200~500W下提取2~3次,每次0.5~1.0h,合并提取液,过滤,滤液回收乙醇,浓缩,干燥,粉碎,即得醇提取物;
(3)甘草中加入6~20倍体积的氨乙醇溶液于70~90℃温度下进行回流提取1~2h,过滤,烘干,粉碎,即得甘草提取物;
(4)红曲米粉碎,采用60~85%的乙醇溶液,超声波200~500W下提取2~3次,每次0.5~1.0h,合并提取液,过滤,滤液回收乙醇,浓缩,干燥,粉碎,即得醇提取物;
(5)将黑木耳多糖、黑木耳黑色素、山楂提取物、银杏提取物、甘草提取物、蒲黄提取物和红曲提取物混合,即得含有黑木耳提取物、具有降血脂作用的组合物。
2.根据权利要求1所述的含有黑木耳提取物、具有降血脂作用的组合物的制备方法,其特征在于所述黑木耳为黑29品种进行秋季栽培的黑木耳子实体。
3.根据权利要求1所述的含有黑木耳提取物、具有降血脂作用的组合物的制备方法,其特征在于所述黑木耳多糖和黑色素的纯度为30~40%。
4.根据权利要求1所述的含有黑木耳提取物、具有降血脂作用的组合物的制备方法,其特征在于所述黑木耳的粉碎粒度为150~240目。
5.根据权利要求1所述的含有黑木耳提取物、具有降血脂作用的组合物的制备方法,其特征在于所述银杏叶、山楂、蒲黄和红曲米的粉碎粒度为100~150目。
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