CN106616177A - 一种具有降尿酸疗效的复合多肽固体饮料及其制备方法 - Google Patents
一种具有降尿酸疗效的复合多肽固体饮料及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种具有降尿酸疗效的复合多肽固体饮料及其制备方法。本发明一种具有降尿酸疗效的复合多肽固体饮料从减少尿酸生成的机理入手,以抑制黄嘌呤氧化酶活性为评价指标,从我国药食同源天然植物中提取功能因子,并与动植物蛋白源活性多肽制备而成。本发明具有降尿酸疗效的复合多肽固体饮料配方结构合理,天然绿色,安全无毒无副作用,口味甘醇,携带服用方便,能有效抑制黄嘌呤氧化酶的活性,具有良好的降尿酸功效。
Description
技术领域
本发明涉及具有降尿酸疗效的固体饮料,具体涉及一种具有降尿酸疗效的复合多肽固体饮料及其制备方法。
背景技术
人类最早关于痛风的描述可追溯至数千年前,因其多见于少数富人阶层,故被冠以“帝王病”、“王者之疾”等称谓。几百年来随着社会经济的发展和生活方式的改变,痛风现已成为最常见的慢性非传染性疾病之一,其疾病负担越来越重。痛风的科学定义为遗传性和/或获得性因素引起长期嘌呤代谢紊乱,导致的一组异质性、代谢性疾病。临床表现为高尿酸血症,反复发作的急、慢性关节炎,即关节疼痛,关节畸形及功能障碍,产生痛风石甚至出现间质性肾炎、尿酸性泌尿系统结石,有些病人还伴发心脑血管疾病而危及生命。
痛风的流行病学表现为:随着我国人民生活水平的不断提高,高尿酸血症(HUA)的患病率呈逐年上升的趋势。目前中国HUA呈现高流行、年轻化、男性高于女性、沿海高于内地的趋势。HUA是糖尿病、代谢综合征、高脂血症等代谢性疾病、慢性肾病、心血管疾病、脑卒中的独立危险因素,与痛风密不可分。HUA与摄入过多含嘌呤高的海产品、动物内脏、肉类以及大量饮用啤酒等因素有关。
医学上痛风急性期治疗方法为:24小时内,服用如COX-2抑制剂的非甾体类药物(NSAIDs)或秋水仙碱或类固醇药物。痛风急性发作的预防:小剂量秋水仙碱或NSAIDs,连续使用6个月;无效或不能耐受或有禁忌症,改用小剂量强的松或强的松龙,连用6个月;同时,持续降尿酸治疗。降尿酸药物可分为促尿酸排泄、抑制尿酸合成及新型的促进尿酸分解3大类。促尿酸排泄药包括苯溴马隆、丙磺舒或苯磺唑酮。抑制尿酸合成药有别嘌醇和非布司他(febuxostat)。促进尿酸分解药包括拉布立酶或聚乙二醇尿酸酶(PEG-uricase)。尽管医学上促尿酸排泄和抑制尿酸生成的药物有很多,但是都存在肾毒性或者皮疹等副反应,而促进尿酸分解的药会产生抗体反应且在我国并未应用。于是寻找新的黄嘌呤氧化酶抑制剂药物至关重要。我国有着非常丰富的中药资源和使用历史,从中寻找黄嘌呤氧化酶抑制剂可以为以黄嘌呤氧化酶为靶点的抗痛风药或保健食品开发提供理论依据。黄嘌呤氧化酶的分子量较大,约27万,并含有两分子黄素腺嘌呤二核苷酸(FAD)、两个钼原子和八个铁原子。黄嘌呤氧化酶中的钼以钼蝶呤辅因子的形式存在(MOS),是酶的活性位点。铁原子则为[2Fe-2S]铁氧还蛋白铁硫簇的一部分(FES),参与电子转移反应。
本发明从食品或者药食同源天然中草药中提取功能因子,原理同非布司他和别嘌呤醇,都是抑制黄嘌呤氧化酶活性,从而减少体内尿酸生成,具有排毒消炎,降低尿酸的功效,同时又不会产生任何副作用。
发明内容
本发明目的在于提供一种具有降尿酸疗效的复合多肽固体饮料及其制备方法。
本发明一种具有降尿酸疗效的复合多肽固体饮料从减少尿酸生成的机理入手,以抑制黄嘌呤氧化酶活性为评价指标,从我国药食同源天然植物中提取功能因子,并与动植物蛋白源活性多肽制备而成。
本发明一种具有降尿酸疗效的复合多肽固体饮料由芹菜籽、蒲公英、葛根和油橄榄叶经提取工艺得到天然产物功能因子,并与从鲣鱼和大豆蛋白中酶解得到的活性多肽复配成固体饮料形式。
芹菜籽:芹菜籽为伞形科植物芹菜的种子,常应用于调味品和食品中,它含有丰富的维生素及挥发性芳香成分,有很好的药用价值。研究发现芹菜籽提取物含有木犀草素、芹菜甲素、芹菜乙素、新蛇床内酯和苯酞类,有很好的抗炎、镇痛和显著降低血清尿酸的作用。黄嘌呤氧化酶抑制活性研究中木樨草素活性较好,50ug/ml时抑制率达到74.23%;芹菜甲素、芹菜乙素和新蛇床内酯对黄嘌呤氧化酶也有抑制活性;苯酞类的体内降尿酸和镇痛活性实验结果表明,苯酞类有明显降尿酸和镇痛作用,说明苯酞类也是抗痛风活性成分。
鲣鱼:俗称炸弹鱼,属鲈形总目、金枪鱼亚目、金枪鱼科、鲣属。鲣鱼肽是深海鱼鲣鱼肉的酶解产物,鲣鱼肽中包括肌肽和鹅肌肽在内的二肽是有效活性成分。除了抗疲劳和消除活性氧外,鲣鱼肽还可以控制罹患痛风个体中的尿酸含量,并改善与高量尿酸相关的症状,包括肌肉痉挛、局部肿胀、发炎、关节疼痛、肌肉疲倦或压力感及心肌梗塞。
葛根:豆科植物野葛的干燥根,有解肌退热,透疹,生津止渴,升阳止泻之功。常用于表证发热,项背强痛,麻疹不透,热病口渴,阴虚消渴,热泻热痢,脾虚泄泻。葛根中葛根素可以通过抑制黄嘌呤氧化酶活性抑制尿酸生成,并增加尿酸在尿液中的溶解度,保持尿酸的排泄量,从而抑制机体的血尿酸水平升高,有益于改善高尿酸血症患者健康状况。
蒲公英:菊科,蒲公英属多年生草本植物,其提取物活性成分主要包括有黄酮类、萜类、酚酸类、蒲公英色素、植物甾醇类、倍半萜内酯类和香豆素类。蒲公英提取物具有良好的广谱抗菌、抗自由基、抗病毒、抗感染、抗肿瘤作用,有养阴凉血、舒筋固齿、通乳益精、利胆保肝、增强免疫力等功效。蒲公英提取物与其它中药配合用于治疗烧伤合并感染、胃痛、急性胆道感染、腮腺炎等多种疾病。蒲公英提取物有广谱抗菌作用,可用于治疗上呼吸道感染、急性扁桃体炎、腮腺炎、急性鼻窦炎、腥红热、尿毒症、阑尾脓肿、剖腹产后高热、宫颈糜烂、脓皮病、肛周炎、外科炎症、外伤性继发性脓毒血症、骨髓炎、龋齿、烫伤、急慢性肝炎、急慢性胃肠炎及溃疡等,口腔炎、淋巴结炎、滴虫性阴道炎、灼伤合并感染,慢性胆囊炎及急性乳腺炎、慢性痢疾、尿路感染、化脓性中耳炎、慢性阻塞性肺疾患合并感染。
油橄榄叶:橄榄叶提取物是从油橄榄叶中提取的组分,主要成分为黄酮类和萜类内酯化合物,近年来药理研究发现黄酮类和萜类内酯化合物具有很好的抗氧化、降血脂,通过阻断COX-2的表达来减轻海洛因造成的肾组织损伤,改善肾功能的作用。
一种具有降尿酸疗效的复合多肽固体饮料,按质量百分比,包括如下组分:
进一步地,所述具有降尿酸疗效的复合多肽固体饮料是天然健康食品,应用于治疗或预防高尿酸血症和痛风。
进一步地,所述芹菜籽提取物、蒲公英提取物、葛根提取物和橄榄叶提取物通过如下方法得到:
(1)选取优质芹菜籽、蒲公英、葛根和油橄榄叶,分别进行除杂、干燥后,粉碎并过20~40目筛;
(2)将步骤(1)过筛后得到的芹菜籽粉末、蒲公英粉末、葛根粉末和橄榄叶粉末分别用乙醇进行热提取,过滤,提取液分别进行喷雾干燥,得到所述芹菜籽提取物、蒲公英提取物、葛根提取物和橄榄叶提取物。
更进一步地,步骤(2)中,所述乙醇的体积分数为60%~80%。
更进一步地,步骤(2)中,所述芹菜籽粉末、蒲公英粉末、葛根粉末和橄榄叶粉末与乙醇的料液比均为1:15~1:25g/mL。
更进一步地,步骤(2)中,所述热提取是:在50℃~70℃下提取5~8小时,重复三次,合并三次的提取液。
进一步地,所述鲣鱼多肽和大豆多肽通过如下方法得到:
将优质鲣鱼和大豆蛋白分别用碱性蛋白酶进行酶解,所得酶解液分别进行喷雾干燥,得到所述鲣鱼多肽和大豆多肽。
更进一步地,所述酶解是在55℃~65℃进行酶解7h~9h。
更进一步地,所述酶解过程中,pH值为7.5~9.0,碱性蛋白酶的添加量为鲣鱼或大豆蛋白质量的0.5%~1.5%。
制备上述任一项所述的一种具有降尿酸疗效的复合多肽固体饮料的方法,包括如下步骤:
按所述质量百分比,将各组分混合均匀,杀菌,得到所述具有降尿酸疗效的复合多肽固体饮料,将得到的固体饮料在内包装无菌车间分装入袋。
与现有技术相比,本发明具有如下有益效果:
本发明具有降尿酸疗效的复合多肽固体饮料配方结构合理,天然绿色,安全无毒无副作用,口味甘醇,携带服用方便,能有效抑制黄嘌呤氧化酶的活性,具有良好的降尿酸功效。
附图说明
图1为实施例4中测得的各提取物及多肽的黄嘌呤氧化酶抑制率比较。
具体实施方式
下面通过实施例的方式进一步说明本发明,但并不因此将本发明限制在所述的实施例范围之中,下列实施例中未注明具体条件的实验方法,均按照常规方法和条件,或按照商品说明书选择,如下实施例中的原料均为市售。
实施例1
具有降尿酸疗效的复合多肽固体饮料的制备。
芹菜籽提取物20%,鲣鱼多肽6%,蒲公英提取物5%,葛根提取物2%,橄榄叶提取物0.2%,大豆多肽35%,麦香红茶粉2%,菊粉29.5%,甜菊糖0.3%;
(1)选取优质芹菜籽、蒲公英、葛根和油橄榄叶,分别进行除杂、干燥后,粉碎并过20目筛;
(2)将步骤(1)过筛后所得的芹菜籽粉末、蒲公英粉末、葛根粉末和橄榄叶粉末分别与体积分数70%的乙醇按照料液比1:15g/mL进行混合,60℃进行提取8小时,提取三次,合并三次提取液,过滤,将不同原料的提取液分别进行喷雾干燥,得到芹菜籽提取物、蒲公英提取物、葛根提取物和橄榄叶提取物;
(3)将优质鲣鱼和大豆蛋白分别用碱性蛋白酶在55℃酶解9h;酶解过程中,pH=8.0,碱性蛋白酶添加量为鲣鱼或大豆蛋白质量的1.0%;所得酶解液分别进行喷雾干燥,得鲣鱼多肽和大豆多肽;
(4)将步骤(2)和步骤(3)制得的提取物与多肽,以及麦香红茶粉、菊粉和甜菊糖按照所述质量百分比进行混合均匀,杀菌,得到所述具有降尿酸疗效的复合多肽固体饮料;
(5)包装复合多肽固体饮料:在内包装无菌车间内,把得到的复合多肽固体饮料在不锈钢包装容器上,用度铝箔膜食品用塑料袋自动分装成5g/袋的小袋固体饮料;将分装好的小袋消毒灭菌并20袋/盒装盒装箱,入库。
实施例2
具有降尿酸疗效的复合多肽固体饮料的制备。
芹菜籽提取物15%,鲣鱼多肽4%,蒲公英提取物3%,葛根提取物2.5%,橄榄叶提取物0.3%,大豆多肽25%,麦香红茶粉5%,菊粉45%,甜菊糖0.2%;
(1)选取优质芹菜籽、蒲公英、葛根和油橄榄叶,分别进行除杂、干燥后,粉碎并过30目筛;
(2)将步骤(1)过筛后所得的芹菜籽粉末、蒲公英粉末、葛根粉末和橄榄叶粉末分别与体积分数60%的乙醇按照料液比1:20g/mL进行混合,50℃进行提取5小时,提取三次,合并三次提取液,过滤,将不同原料的提取液分别进行喷雾干燥,得到芹菜籽提取物、蒲公英提取物、葛根提取物和橄榄叶提取物;
(3)将优质鲣鱼和大豆蛋白分别用碱性蛋白酶在60℃酶解8h;酶解过程中,pH=7.5,碱性蛋白酶添加量为鲣鱼或大豆蛋白质量的0.5%;所得酶解液分别进行喷雾干燥,得鲣鱼多肽和大豆多肽;
(4)将步骤(2)和步骤(3)制得的提取物与多肽,以及麦香红茶粉、菊粉和甜菊糖按照所述质量百分比进行混合均匀,杀菌,得到所述具有降尿酸疗效的复合多肽固体饮料;
(5)包装复合多肽固体饮料:在内包装无菌车间内,把得到的复合多肽固体饮料在不锈钢包装容器上,用度铝箔膜食品用塑料袋自动分装成5g/袋的小袋固体饮料;将分装好的小袋消毒灭菌并20袋/盒装盒装箱,入库。
实施例3
具有降尿酸疗效的复合多肽固体饮料的制备。
芹菜籽提取物10%,鲣鱼多肽4.35%,蒲公英提取物6%,葛根提取物3%,橄榄叶提取物0.4%,大豆多肽15%,麦香红茶粉5%,菊粉56%,甜菊糖0.25%;
(1)选取优质芹菜籽、蒲公英、葛根和油橄榄叶,分别进行除杂、干燥后,粉碎并过40目筛;
(2)将步骤(1)过筛后所得的芹菜籽粉末、蒲公英粉末、葛根粉末和橄榄叶粉末分别与体积分数80%的乙醇按照料液比1:25g/mL进行混合,70℃进行提取7小时,提取三次,合并三次提取液,过滤,将不同原料的提取液分别进行喷雾干燥,得到芹菜籽提取物、蒲公英提取物、葛根提取物和橄榄叶提取物;
(3)将优质鲣鱼和大豆蛋白分别用碱性蛋白酶在65℃酶解7h;酶解过程中,pH=9.0,碱性蛋白酶的添加量为鲣鱼或大豆蛋白质量的1.5%;所得酶解液分别进行喷雾干燥,得鲣鱼多肽和大豆多肽;
(4)将步骤(2)和步骤(3)制得的提取物与多肽,以及麦香红茶粉、菊粉和甜菊糖按照所述质量百分比进行混合均匀,杀菌,得到所述具有降尿酸疗效的复合多肽固体饮料;
(5)包装复合多肽固体饮料:在内包装无菌车间内,把得到的复合多肽固体饮料在不锈钢包装容器上,用度铝箔膜食品用塑料袋自动分装成5g/袋的小袋固体饮料;将分装好的小袋消毒灭菌并20袋/盒装盒装箱,入库。
实施例4
实施例1中得到的芹菜籽提取物、蒲公英提取物、葛根提取物及鲣鱼多肽的体外黄嘌呤氧化酶抑制活性。
(1)试剂配制
配制0.2mol/L的磷酸缓冲液(pH=7.5),0.2mM的黄嘌呤溶液,0.1U/mL的黄嘌呤氧化酶溶液,1M的HCl,1M的NaOH,待用。
阳性对照药:别嘌呤醇。
(2)给药溶液配制
测试样品溶液:将四种样品原料分别配制成2.5mg/mL溶液,再用磷酸缓冲溶液稀释,得到各提取物及鲣鱼多肽200g/mL的样品溶液。
阳性对照品溶液:别嘌呤醇先用NaOH(1M)溶解,再用0.2mol/L磷酸缓冲液(pH=7.5)配成40g/mL的别嘌呤醇溶液,备用。
(3)操作步骤
将50μL浓度为0.1U/mL的黄嘌呤氧化酶溶液、50μL浓度为0.2mol/L(PH=7.5)的磷酸盐缓冲液、200g/mL的样品溶液依次加入到96微孔版中,振摇30s后37℃保温15min(平衡时间对结果有影响,此时间保证操作可行性),采用多道移液器快速96微孔板加入150μL浓度为0.2mM的黄嘌呤溶液,在酶标仪中反应30min;后加入50μL浓度为1M的HCl终止反应,于290nm处测定吸光值。
(4)按照公式计算提取物样品对XOD的抑制活性
抑制率(%)=[(A-B)-(C-D)]/(A-B)×100%
A:50μL黄嘌呤氧化酶溶液+50μL缓冲液+150μL黄嘌呤溶液
B:50μL缓冲液+50μL缓冲液+150μL黄嘌呤溶液
C:50μL黄嘌呤氧化酶溶液+50μL样品溶液+150μL黄嘌呤溶液
D:50μL缓冲液+50μL样品溶液+150μL黄嘌呤溶液
用配制好的40g/mL的别嘌呤醇溶液代替样品溶液,进行阳性对照。
(5)测定结果
得到的各原料提取物及鲣鱼多肽对黄嘌呤氧化酶抑制活性的结果如图1所示,图1结果表明各原料提取物及鲣鱼多肽具有较好的黄嘌呤氧化酶抑制活性,比临床药别嘌呤醇稍低。
实施例5
固体饮料产品动物实验
1.动物:SPF级SD大鼠72只,雄性,体重200±20(g),广州中医药大学实验动物中心提供(许可证号:SCXK(粤)2013-0020)。
2.药品和试剂:别嘌醇片(广东彼迪药业有限公司,批准文号:国药准字H44021368);氧嗪酸钾(山东中科泰斗化学有限公司,批号:Batch No.120901);羧甲基纤维素钠(上海赛璐珞厂,产品标准号:GB2760);尿酸、检测试剂盒(南京建成生物工程研究所,生产批号:20140306);A组受试药:实施例1制备的固体饮料。B组受试药:实施例2制备的固体饮料。C组受试药:实施例3制备的固体饮料。
3.仪器:TGL-16G型高速冷冻离心机(上海安亭科学仪器厂);多功能酶标仪(BioTelc USA,Synergy HT)。
4.动物饲养状况:动物饲养于暨南大学SPF级实验动物中心,饲养饲料、垫料均由暨南大学实验动物中心提供。自由食水,灯照12h,室温为(20±2)℃,相对湿度(40~70)%。
5.实验方法
动物分组和造模:SPF级SD雄性大鼠72只,随机分为正常对照组(12只)和造模组(60只),造模组大鼠每天灌胃氧嗪酸钾(2g·kg-1),正常对照组大鼠灌胃给予等容积的溶媒;连续7天后,3%戊巴比妥钠(30mg·kg-1)腹腔注射麻醉,眼眶后静脉丛取血(0.5ml),以4℃、3000rpm、离心15min,取上层血清测定尿酸含量;将尿酸含量高于110umol·L-1的大鼠确定为造模成功。
将造模成功的大鼠按尿酸含量随机分为模型对照组、别嘌呤组(25mg·kg-1)、A组、B组、C组,每组大鼠12只,灌胃给药(给药容积10ml/kg),模型对照组大鼠给予等容积的蒸馏水。处理7d、14d时,于末次给药50min,3%戊巴比妥钠(30mg·kg-1)腹腔注射麻醉后,眼眶后静脉丛取血(0.5ml),测定血清尿酸含量;处理第21d时,3%戊巴比妥钠麻醉后腹主动脉取血5ml,测定血清尿酸含量。
不同药物处理不同时间时模型大鼠血清尿酸含量的变化如表1所示。
表1 抗痛风产品处理不同时间对高尿酸血症大鼠血清尿酸含量的影响(±s)
由表1可见,造模组给药前大鼠经过氧嗪酸钾处理1周后,血清尿酸含量明显高于正常对照组大鼠(p<0.01)。而模型对照组大鼠在后续处理7d,14d和21d时,血尿酸含量仍明显高于正常对照组大鼠(p<0.01),且尿酸含量有增加的趋势;别嘌醇组具有非常明显的降低模型大鼠血清尿酸的作用(p<0.01),受试药A、B、C组也表现出降低血清尿酸的作用(p<0.05)。
Claims (10)
1.一种具有降尿酸疗效的复合多肽固体饮料,其特征在于,按质量百分比,包括如下组分:
芹菜籽提取物 10%-20%,
鲣鱼多肽 4%-6%,
蒲公英提取物 3%-6%,
葛根提取物 2%-3%,
橄榄叶提取物 0.2%-0.4%,
大豆多肽 15%-35%,
麦香红茶粉 2%-5%,
菊粉 29.5%-56%,
甜菊糖 0.2%-0.3%。
2.根据权利要求1所述的一种具有降尿酸疗效的复合多肽固体饮料,其特征在于,所述具有降尿酸疗效的复合多肽固体饮料是天然健康食品,应用于治疗或预防高尿酸血症和痛风。
3.根据权利要求1所述的一种具有降尿酸疗效的复合多肽固体饮料,其特征在于,所述芹菜籽提取物、蒲公英提取物、葛根提取物和橄榄叶提取物通过如下方法得到:
(1)选取优质芹菜籽、蒲公英、葛根和油橄榄叶,分别进行除杂、干燥后,粉碎并过20~40目筛;
(2)将步骤(1)过筛后得到的芹菜籽粉末、蒲公英粉末、葛根粉末和橄榄叶粉末分别用乙醇进行热提取,过滤,提取液分别进行喷雾干燥,得到所述芹菜籽提取物、蒲公英提取物、葛根提取物和橄榄叶提取物。
4.根据权利要求3所述的一种具有降尿酸疗效的复合多肽固体饮料,其特征在于,步骤(2)中,所述乙醇的体积分数为60%-80%。
5.根据权利要求3所述的一种具有降尿酸疗效的复合多肽固体饮料,其特征在于,步骤(2)中,所述芹菜籽粉末、蒲公英粉末、葛根粉末和橄榄叶粉末与乙醇的料液比均为1:15~1:25 g/mL。
6.根据权利要求3所述的一种具有降尿酸疗效的复合多肽固体饮料,其特征在于,步骤(2)中,所述热提取是:在50℃~70℃下提取5~8小时,重复三次,合并三次的提取液。
7.根据权利要求1所述的一种具有降尿酸疗效的复合多肽固体饮料,其特征在于,所述鲣鱼多肽和大豆多肽通过如下方法得到:
将优质鲣鱼和大豆蛋白分别用碱性蛋白酶进行酶解,所得酶解液分别进行喷雾干燥,得到所述鲣鱼多肽和大豆多肽。
8.根据权利要求7所述的一种具有降尿酸疗效的复合多肽固体饮料,其特征在于,所述酶解是在55℃~65℃进行酶解7h~9h;所述酶解过程中,pH值为7.5~9.0。
9.根据权利要求7所述的一种具有降尿酸疗效的复合多肽固体饮料,其特征在于,所述碱性蛋白酶的添加量为鲣鱼或大豆蛋白质量的0.5%~1.5%。
10.制备权利要求1~9任一项所述的一种具有降尿酸疗效的复合多肽固体饮料的方法,其特征在于,包括如下步骤:
按所述质量百分比,将各组分混合均匀,杀菌,得到所述具有降尿酸疗效的复合多肽固体饮料,将得到的固体饮料在内包装无菌车间分装入袋。
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