CN105770525A - 一种益生菌发酵药食两用中药组合物及其制备方法和应用 - Google Patents
一种益生菌发酵药食两用中药组合物及其制备方法和应用 Download PDFInfo
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明公开了一种益生菌发酵药食两用中药组合物及其制备方法,将药食两用中药进行水浸,提取,浓缩,添加碳源、氮源、无机盐,添加或不添加γ‑氨基丁酸,调pH,灭菌,接种多株益生菌,发酵,制得益生菌发酵药食两用中药组合物;本发明所述的药食两用中药为甘草、小麦、大枣。实验结果显示:本发明的益生菌发酵药食两用中药组合物清除自由基效果明显,具有抗氧化和改善睡眠的作用。
Description
技术领域
本发明涉及一种益生菌发酵药食两用中药组合物及其制备方法,属于医药技术领域。
背景技术
近年来,随着社会的巨大变化,生活节奏的日益加快,失眠发病率有逐步上升的趋势,严重影响人们的生活、工作及身心健康。受客观条件所限,当前西医所倡导的对失眠症采取药物、心理、行为综合治疗方法,在实际工作中常难以开展,大多数失眠患者仍需要依靠镇静催眠类药物终特治疗,此类药物有较明显的毒副作用,长期服用容易产生药物依赖,也很难达到彻底治愈的目的。中医治疗失眠的历史源远流长,著述众多,疗效明显, 且副作用小,具有明显优势。
中医认为,失眠是由于阴阳失调引起的,而阴阳失调具体表现为气血失和,气血失调贯穿于失眠症的整个过程,气血失和,使脑海血流不充,脑健失滋,神无所养,亦无所寄。是其重要的病理矛盾,也是其发病和加重的机制。
运用现代科学技术,开发中医经典方剂是对中医精华的传承和发展.。甘麦大枣汤由甘草、小麦、大枣三味药食两用中药组成,出自《金匮要略·妇人杂病脉证并治》,主治“妇人脏躁,喜悲伤欲哭,象如神灵所作,数欠伸,甘麦大枣汤主之。”此方是医圣仲景为脏躁而作,条文主要是论述脏躁,情志郁结,血虚阴亏,心神失养,内火扰神而现喜悲伤欲哭。
甘草,药性甘、平。归心、肺、脾、胃经。其效补脾益气,祛痰止咳,缓急止痛, 清热解毒,调和诸药。现代药理研究:主要活性成分是三萜皂苷和黄酮类化合物。甘草总黄酮与中枢 5-HT 神经功能有关,具有抗应激抑郁的作用。
小麦,药性甘、微寒。归心经。其效养心安神。治心神不宁,烦躁失眠及妇人脏躁证。现代药理研究:从小麦中提取的脂多糖(LPSw)静脉注射或灌胃,均可抑制醋酸引起的小鼠扭体反应,有显著的镇痛作用。
大枣,药性甘、温。归脾、胃心经。其效补中益气,养血安神。能养心安神,为治疗心失充养,心神无主而脏躁的要药,亦能补脾益气。现代药理研究:大枣含有生物碱、三萜酸类化合物、皂甙类化合物、糖类化合物、脂肪酸等成分,有抗肿瘤、抗I型变态反应、中枢神经的抑制作用、增加体重、增强肌力等作用;并确认柚配质(糖甙类)能降低自发运动,并对中枢神经有抑制作用。
目前有少量将甘麦大枣汤制成饮料或制剂的报道。例如,授权公告号为CN102178287 B 的专利公开了及一种甘麦大枣汤饮料的配方及制备方法。其配方在传统甘麦大枣汤的基础上,添加柏子仁、灵芝,制法中含有磁化步骤。公开号为CN 102960815 A 的专利公开了一种即冲型甘麦大枣饮料,是在传统“甘麦大枣汤”经典配方的基础上,增加了一味药食同源的中药枸杞。公开号为CN 104800633 A的专利公开了一种甘麦大枣汤制剂、其制备方法及用途,通过将甘麦大枣汤处方中的部分浮小麦先粉碎为细粉,再与其它药味提取的甘麦大枣汤干浸膏混合,制成固体的甘麦大枣汤制剂,同时,公开了甘麦大枣汤制剂治疗便秘,可以用于制备治疗便秘药物。这些专利虽然有一定的创新,但都仍沿用传统提取制备方法,人体吸收利用率低,存在一定的局限性。
发明内容
本发明在吸取微生态学研究成果的基础上,结合现代微生物工程理论,利用中药发酵技术,制备益生菌发酵药食两用中药组合物。
本发明的一个目的是提供一种益生菌发酵药食两用中药组合物,其技术方案为:将药食两用中药进行水浸,提取,浓缩,添加碳源、氮源、无机盐,添加或不添加γ-氨基丁酸,调pH,灭菌,接种多株益生菌,发酵,制得益生菌发酵药食两用中药组合物。该发酵组合物具有抗氧化、改善睡眠作用。
上述所述的药食两用中药为甘草、小麦、大枣,在益生菌发酵药食两用中药组合物中的重量g/体积ml百分比分别为:甘草4-15%;小麦10-40%;大枣4-35%;所述的益生菌为双歧杆菌和乳杆菌。
优选的,所述的药食两用中药在益生菌发酵药食两用中药组合物中的重量g/体积ml百分比为:甘草8%;小麦24%;大枣28%。
所述的碳源为体积百分比0.5~1.5%的葡萄糖;所述的氮源为体积百分比0.1~0.2%的酵母粉、体积百分比0.1~0.4%的酵母膏和体积百分比0.1~0.5%的蛋白胨。
所述的γ-氨基丁酸(净含量)添加量为体积百分比0~0.1%。γ-氨基丁酸(简称GABA)是一种天然存在的非蛋白质氨基酸,是哺乳动物中枢神经系统中重要的抑制性神经传达物质,约30%的中枢神经突触部位以GABA为递质。GABA在人体大脑皮质、海马、丘脑、基底神经节和小脑中起重要作用,并对机体的多种功能具有调节作用。当人体内GABA缺乏时,会产生焦虑、不安、疲倦、忧虑等情绪。
所述的无机盐为FeSO4•7H2O 1.0mg/100ml、ZnSO4•7H2O 4.4mg/100ml、MgSO4•7H2O70mg/100ml、KH2PO4 100mg/100ml、K2HPO4•3H2O 100mg/100ml。
所述的双歧杆菌选自两歧双歧杆菌、短双歧杆菌、长双歧杆菌、动物双歧杆菌、青春双歧杆菌和婴儿双歧杆菌;所述的乳杆菌选自嗜酸乳杆菌、德氏乳杆菌乳亚种、植物乳杆菌、卷曲乳杆菌、瑞士乳杆菌、唾液乳杆菌、格式乳杆菌、约氏乳杆菌、副干酪乳杆菌、发酵乳杆菌、干酪乳杆菌和鼠李糖乳杆菌。所述菌种来源于中国普通微生物菌种保藏管理中心(CGMCC 中国,北京)或者从正常人的粪便或肠粘膜组织中分离得到这些细菌菌株。
本发明的另一个目的是提供一种益生菌发酵药食两用中药组合物的制备方法,包括如下步骤:
a. 取药食两用中药复方中的3味药,加水浸泡1.0~2.0 小时,煎煮二次,第一次1.0~2.0 小时,第二次1.0~2.0 小时,滤过,合并煎液,滤液减压浓缩至50℃下测定相对密度为1.01 ~1.10,得到浓缩液;
b.上述浓缩液加入葡萄糖、酵母粉、酵母膏、蛋白胨、无机盐,添加或不添加γ-氨基丁酸,调整pH7.0±0.5,并于110~121℃下灭菌30~60分钟,温度降至39℃时,同时接种预培养的体积百分比各为0.2~1.0%的一种或者两种双歧杆菌的菌液和预培养的体积百分比各为0.2~1.0%的两种或者三种乳杆菌的菌液,常规发酵16~48小时,当pH降至4.0以下时结束发酵,稳定化处理,即得到益生菌发酵药食两用中药组合物。
本发明的再一个目的是提供本发明的益生菌发酵药食两用中药组合物在制备抗氧化和改善睡眠的药品、保健品、食品或饮品中的应用。
可将本发明的发酵组合物直接制成保健食品、食品或饮品,用于抗氧化或改善睡眠,适于抗衰老和由于情志郁结或更年期造成的失眠的人使用。
根据本发明的一个优选实施方案,按照本发明的方法制备的发酵组合物,可应用于制备抗氧化或改善睡眠的保健品或药品。例如,可按照已知的常规方法,将本发明的益生菌发酵药食两用中药组合物或其上清液,按照已知的常规方法,添加一种或多种具有相同、相似或不同生物学活性,并对基本活性成分表现有辅助或协同作用,但彼此又互不拮抗的天然或合成的其他药物成份或其混合物,配制成适于口服给药的合剂、乳剂等。
有益效果:为检测本发明的益生菌发酵药食两用中药组合物的抗氧化和改善睡眠的效果。分别进行了体外抗自由基实验和巴比妥钠睡眠潜伏实验。抗自由基实验结果显示:本发明的益生菌发酵药食两用中药组合物清除自由基效果明显。且益生菌发酵药食两用中药组合物的活性高于未发酵的中药组合物。巴比妥钠睡眠潜伏实验结果显示,本发明的益生菌发酵药食两用中药组合物具有改善睡眠功能的作用,且益生菌发酵药食两用中药组合物作用优于未发酵的中药组合物。说明本发明的制备方法提高了中药组合物的抗氧化和改善睡眠的作用。
具体实施方式
实施例1:益生菌发酵药食两用中药组合物A的制备
取甘草80g、小麦240g、大枣280g , 加10倍水浸泡60 分钟,煎煮1.0小时,滤过,滤渣再加8倍水煎煮1.0小时,滤过,合并煎液,滤液减压浓缩至约1000ml,50℃下测定相对密度为1.03;向浓缩液中加入葡萄糖0.5%(重量/ 体积)、酵母粉0.1%(重量/ 体积)、酵母膏0.2%(重量/ 体积)和蛋白胨0.2%(重量/ 体积)。无机盐( 硫酸亚铁•7H2O5.0mg/100ml、硫酸锌•7H2O 4.4mg/100ml、硫酸镁•7H2O 50mg/100ml、磷酸二氢钾 100mg/100ml、磷酸氢二钾•3H2O100mg/100ml)、添加γ-氨基丁酸0.05%(重量/ 体积,由浙江益万生物技术有限公司提供。)。调整pH 7.0并115℃灭菌40 分钟,得发酵底物。当发酵底物温度降至39℃左右时,接入预培养好的短双歧杆菌(AS 1.2213)菌液,接种量为1.0%(体积/体积),再接入预培养的干酪乳杆菌(AS 1.3206)和植物乳杆菌(AS 1.19)菌液,接种量均为0.5%(体积/ 体积)。37℃发酵20~40小时,当pH 值降至4.0以下时,结束发酵。稳定化处理,即得到益生菌发酵药食两用中药组合物A。
实施例2:益生菌发酵药食两用中药组合物B的制备
取甘草40g、小麦400g、大枣160g , 加10倍水浸泡1.0小时,煎煮2.0小时,滤过,滤渣再加8倍水煎煮2.0小时,滤过,合并煎液,滤液减压浓缩至约1000ml,50℃下测定相对密度为1.04;向浓缩液中加入葡萄糖0.7%(重量/ 体积)、酵母粉0.2%(重量/ 体积)、酵母膏0.4%(重量/ 体积)和蛋白胨0.1%(重量/ 体积)。无机盐( 硫酸亚铁•7H2O5.0mg/100ml、硫酸锌•7H2O 4.4mg/100ml、硫酸镁•7H2O 50mg/100ml、磷酸二氢钾 100mg/100ml、磷酸氢二钾•3H2O100mg/100ml)。添加γ-氨基丁酸0.1%(重量/ 体积,由浙江益万生物技术有限公司提供)。调整pH 7.5并121℃灭菌30 分钟,得发酵底物。当发酵底物温度降至39℃左右时,接入预培养好的体积百分比各为1.0%的动物双歧杆菌(CGMCC 1.1852)、青春双歧杆菌(CGMCC1.2190)的菌液,再接种预培养好的体积百分比各约0.5%的嗜酸乳杆菌( CGMCC 1.1854)和鼠李糖乳杆菌( CGMCC 1.104)的菌液。37℃发酵20~40小时,当pH 值降至4.0以下时,结束发酵。稳定化处理,即得到益生菌发酵药食两用中药组合物B。
实施例3:益生菌发酵药食两用中药组合物C的制备
取甘草150g、小麦100g、大枣50g , 加10倍水浸泡2.0小时,煎煮2.0小时,滤过,滤渣再加8倍水煎煮1.0小时,滤过,合并煎液,滤液减压浓缩至约1000ml,50℃下测定相对密度为1.02;向浓缩液中加入葡萄糖1.0%(重量/体积)、酵母粉0.15%(重量/体积)、酵母膏0.3%(重量/体积)和蛋白胨0.5%(重量/体积)。无机盐( 硫酸亚铁•7H2O5.0mg/100ml、硫酸锌•7H2O4.4mg/100ml、硫酸镁•7H2O 50mg/100ml、磷酸二氢钾 100mg/100ml、磷酸氢二钾•3H2O100mg/100ml)。调整pH 6.5并110℃灭菌60 分钟,得发酵底物。当发酵底物温度降至39℃左右时,接入预培养好的体积百分比各为1.0%的两歧双歧杆菌(CGMCC 1.5090)、短双歧杆菌(CGMCC 1.3001)的菌液,再接种预培养好的体积百分比各约0.5%的副干酪乳杆菌( CGMCC1.2468 )、发酵乳杆菌( CGMCC 1.3270) 和德式乳杆菌乳亚种( CGMCC 1.2132)的菌液。37℃发酵16~48小时,当pH 值降至4.0以下时,结束发酵。稳定化处理,即得到益生菌发酵药食两用中药组合物C。
实施例中的重量/体积均为g /ml。
实验例1:益生菌发酵药食两用中药组合物清除自由基试验
选用实施例1、2、3制备的益生菌发酵药食两用中药组合物A、B、C,并以未经益生菌发酵的相同配比的中药组合物a、b、c为对照。分别稀释10倍,离心,取上清液作为供试品。
依次按下表1向试管中加入 FeSO4溶液、样品、 H2O2溶液各1ml、摇匀,静置10min,加水杨酸溶液1ml,混合后,静置30min,加水稀释至10ml,在510nm处测吸收度。
表1实验所需试剂
注:*以纯水代替
结果:清除率=[1-(Ai-Aj)/A0]*100%
表2清除自由基活性比较
由表2可见,在清除自由基实验中,益生菌发酵药食两用中药组合物清除率明显高于未发酵的中药组合物,表明益生菌发酵药食两用中药组合物在清除自由基方面优于未发酵的中药组合物。
实验例2:巴比妥钠催眠小鼠睡眠潜伏期的测定
选用实施例1、2、3发酵组合物A、B、C和未经发酵的相同配比的中药组合物a、b、c作为实验药物,分别根据剂量适当浓缩。
分组及给药:取84只KM小鼠,全雌,体重18-22g,按体重随机分为7个组,每组12只,分别为空白对照组、未发酵中药组合物a组、b组、c组、实施例1、2、3的发酵组合物A、B、C组,(剂量为原生药20g/kg)。 每天给药1次,连续30 d,灌胃容积为O.1ml/lOg。空白对照组给予相应体积的蒸馏水。
实验小鼠末次给药30 min后,采用巴比妥钠390mg/kg剂量对小鼠进行腹腔注射,注射量为0.1ml/10g,以小鼠翻正反射消失为入睡指标,记录小鼠从腹腔注射巴比妥钠溶液开始到进入睡眠的时间,观察中药组合物发酵前后对巴比妥钠催眠下小鼠的睡眠潜伏期的影响。
表3对巴比妥钠催眠小鼠睡眠潜伏期的影响
注:与空白组比较,*p<0.05,**p<0.01
发酵组与未发酵组比较,#p<0.05。
由表3可见,在巴比妥钠睡眠潜伏期实验中,未发酵中药汤组、发酵组合物组睡眠潜伏期均低于空白对照组,与空白对照组比较有显著性差异 (p<0.05或p<O.01),发酵组合物组与未发酵中药组合物组比较也有显著性差异(p<0.05)。 表明发酵组合物组与未发酵中药汤组均能缩短巴比妥钠催眠小鼠的睡眠潜伏期,均具有调节睡眠的作用;且经发酵后调节睡眠作用明显增强。
Claims (8)
1.一种益生菌发酵药食两用中药组合物,其特征在于,将药食两用中药进行水浸,提取,浓缩,添加碳源、氮源、无机盐,添加或不添加γ-氨基丁酸,调pH,灭菌,接种多株益生菌,发酵,制得益生菌发酵药食两用中药组合物;
所述的药食两用中药为甘草、小麦、大枣,在益生菌发酵药食两用中药组合物中的重量g/体积ml 百分比分别为:甘草4-15%;小麦10-40%;大枣4-35%;
所述的益生菌为双歧杆菌和乳杆菌。
2.根据权利要求1所述的益生菌发酵药食两用中药组合物,其特征在于,所述的药食两用中药在益生菌发酵药食两用中药组合物中的重量g/体积ml 百分比为:甘草8%;小麦24%;大枣28%。
3.根据权利要求1所述的益生菌发酵药食两用中药组合物,其特征在于,所述的碳源为体积百分比0.5~1.5%的葡萄糖;所述的氮源为体积百分比0.1~0.2%的酵母粉、体积百分比0.1~0.4%的酵母膏和体积百分比0.1~0.5%的蛋白胨;所述的γ-氨基丁酸0~0.1%。
4.根据权利要求1所述的益生菌发酵药食两用中药组合物,其特征在于,所述的无机盐为FeSO4•7H2O 1.0mg/100ml、ZnSO4•7H2O 4.4mg/100ml、MgSO4•7H2O 70mg/100ml、KH2PO4100mg/100ml、K2HPO4•3H2O 100mg/100ml。
5.根据权利要求1所述的益生菌发酵药食两用中药组合物,其特征在于,所述的双歧杆菌选自两歧双歧杆菌、短双歧杆菌、长双歧杆菌、动物双歧杆菌、青春双歧杆菌和婴儿双歧杆菌;所述的乳杆菌选自嗜酸乳杆菌、德氏乳杆菌、植物乳杆菌、卷曲乳杆菌、瑞士乳杆菌、唾液乳杆菌、格式乳杆菌、约氏乳杆菌、副干酪乳杆菌、发酵乳杆菌、干酪乳杆菌和鼠李糖乳杆菌。
6.权利要求1-5所述的益生菌发酵药食两用中药组合物的制备方法,其特征在于,步骤如下:
a. 取药食两用中药复方中的3味药,加水浸泡1.0~2.0 小时,煎煮二次,第一次1.0~2.0 小时,第二次1.0~2.0 小时,滤过,合并煎液,滤液减压浓缩至50℃下测定相对密度为1.01 ~1.10,得到浓缩液;
b.上述浓缩液加入葡萄糖、酵母粉、酵母膏、蛋白胨、无机盐,添加或不添加γ-氨基丁酸,调整pH7.0±0.5,并于110~121℃下灭菌30~60分钟,温度降至39℃时,同时接种预培养的体积百分比各为0.2~1.0%的一种或者两种双歧杆菌的菌液和预培养的体积百分比各为0.2~1.0%的两种或者三种乳杆菌的菌液,常规发酵16~48小时,当pH降至4.0以下时结束发酵,稳定化处理,即得到益生菌发酵药食两用中药组合物。
7.权利要求1-5所述的益生菌发酵药食两用中药组合物在制备抗氧化和改善睡眠的药品、保健品、食品或饮品中的应用。
8.根据权利要求7所述的应用,其特征在于,按照已知的常规方法,将所述的益生菌发酵药食两用中药组合物,添加一种或多种具有相同、相似或不同生物学活性,并对基本活性成分表现有辅助或协同作用,但彼此又互不拮抗的天然或合成的其他药物成份或其混合物,制成适于口服给药的合剂、乳剂。
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CN114176217A (zh) * | 2021-11-01 | 2022-03-15 | 齐鲁工业大学 | 以药食同源为核心的益生菌发酵功能制品及制备方法 |
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CN108186688B (zh) * | 2018-03-09 | 2021-03-16 | 华大精准营养(深圳)科技有限公司 | 一种促进睡眠的组合物 |
CN108653574A (zh) * | 2018-08-13 | 2018-10-16 | 三株福尔制药有限公司 | 益生菌发酵的抗病毒组合物及其制备方法和应用 |
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CN114176217A (zh) * | 2021-11-01 | 2022-03-15 | 齐鲁工业大学 | 以药食同源为核心的益生菌发酵功能制品及制备方法 |
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