CN110478470A - 一种含葵花盘小分子肽的抗痛风组合物及其制备方法 - Google Patents
一种含葵花盘小分子肽的抗痛风组合物及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种含葵花盘小分子肽的抗痛风组合物及其制备方法,制备所述药物的原料及其重量份数如下:葵花盘10~30份、薏苡仁20~60份、淡竹叶20~60份、白茅根20~60份、杜仲雄花20~60份、山楂20~60份,本发明含葵花盘小分子肽的抗痛风组合物选用的原料均为药食同源的中药,葵花盘小分子肽与这些中药提取物合用,协同作用良好,且无毒副作用,利水渗湿、活血化癖、清热泻火、固本脾肾,可以通过降尿酸治疗痛风性关节炎,并具有抗炎、镇痛的作用,解决了目前西药治疗痛风带来的副作用大的困扰,可以长期服用,降低尿酸,从而治疗痛风。
Description
技术领域
本发明涉及一种抗痛风组合物及其制备方法,具体涉及一种含葵花盘小分子肽的抗痛风组合物及其制备方法,属于中医制药技术领域。
背景技术
痛风(Gout)是一种由于嘌呤代谢紊乱,导致嘌呤代谢的终产物尿酸含量异常升高引起的代谢性疾病。痛风的特征是明显的高尿酸血症,尿酸盐一水合物晶体在关节和肾脏中沉积,导致痛风性关节炎和尿酸肾结石。高尿酸血症(Hyperui~cemia)可被视为各种心血管疾病的临床重要的危险因素。并且高尿酸血症与癌症、肥胖,胰岛素抵抗,高血压和血脂异常(代谢综合征)也有联系。严格控制尿酸水平是预防痛风和其他相关疾病的关键因素。
葵花盘具有降尿酸、抗肿瘤、抗氧化、抗心绞痛、抑菌、镇痛、降血压等等多种疗效,此外,葵花盘经提取得到的小分子肽更具有作用快,疗效显著,用量小等特点;薏苡仁,性味甘淡微寒,有利水消肿、健脾去湿、舒筋除痹、清热排脓等功效;淡竹叶,有清凉,解热、利尿及催产之效;白茅根,气微,味微甜,凉血,止血,清热,利尿;杜仲雄花,对于"强肝、补肾、通便、安眠、降三高"的效果尤为显著;山楂,味微酸涩,具有降血脂、血压、强心、抗心律不齐等作用。
目前抗痛风药物多以西药为主,例如秋水仙碱、吲哚美辛、泼尼松龙、别嘌呤醇、非布索坦、丙磺舒、苯溴马隆等,普遍存在毒副作用及不良反应,因此从植物中寻找疗效好、毒性小的天然活性物质,是目前科研工作者亟待解决的问题之一。
发明内容
有鉴于此,本发明提供了一种含葵花盘小分子肽的抗痛风组合物及其制备方法,该组合物选用的原料均为药食两用,且取毒副作用的中药,几味协同合用利水渗湿、活血化癖、清热泻火、固本脾肾,具有显著的降尿酸作用,从而到到治疗痛风的疗效。
为了实现上述目的,本发明采用如下技术方案:
一种含葵花盘小分子肽的抗痛风组合物,其原料及其重量份数如下:葵花盘10~30份、薏苡仁20~60份、淡竹叶20~60份、白茅根20~60份、杜仲雄花20~60份、山楂20~60份;
优选的,重量份数如下:葵花盘20份、薏苡仁40份、淡竹叶40份、白茅根40份、杜仲雄花40份、山楂40份。
本发明含葵花盘小分子肽的抗痛风组合物原料组合依据如下:
葵花盘药性甘、寒,归肝经,能够平肝止血,清热止痛,可用于治疗高血压、头晕头痛、耳鸣、子宫出血和疮疹等疾病。现代药理研究表明,葵花盘具有降尿酸、抗肿瘤、抗氧化、抗心绞痛、抑菌、镇痛、降血压等多种药理活性。
薏苡仁性味甘淡微寒,有利水消肿、健脾去湿、舒筋除痹、清热排脓等功效,为常用的利水渗湿药。薏仁又是一种美容食品,常食可以保持人体皮肤光泽细腻,消除粉刺、雀斑、老年斑、妊娠斑、蝴蝶斑,对脱屑、痤疮、皲裂、皮肤粗糙等都有良好疗效。
淡竹叶根药用,有清凉,解热、利尿及催产之效;主治:胸中疾热、咳逆上气、吐血、热毒风、止消渴、压丹石毒、消痰、治热狂烦闷、中风失音不语、痛头风、止惊悸、瘟疫迷闷、杀小虫、除热缓脾。
白茅根气微,味微甜。主治:凉血止血,清热解毒。用于吐血,尿血,热淋,水肿,黄疸,小便不利,热病烦渴,胃热呕哕,咳嗽。
杜仲雄花,杜仲是雌雄双株的树,雄花就是杜仲雄树开的花,科学研究发现:"杜仲雄花"特含60多种有效植物成分,如木质素类,环烯醚萜类、苯丙素类活性物质(如京尼平甙酸、绿原酸、桃叶珊瑚甙)、松脂醇双糖甙、黄酮、生物碱、氨基酸、多糖以及矿质元素Zn、Mn、Cu、Fe、Ca、P、B、Mg、K等,杜仲雄花,汇集了所有杜仲的精华,可以说杜仲雄花具备植物杜仲所有的保健功效,而且对于"强肝、补肾、通便、安眠、降三高"的效果尤为显著。
本发明含葵花盘小分子肽的抗痛风组合物选用的原料均为药食同源的中药,葵花盘小分子肽与这些中药提取物合用,协同作用良好,且无毒副作用,利水渗湿、活血化癖、清热泻火、固本脾肾,可以通过降尿酸治疗痛风性关节炎,并具有抗炎、镇痛的作用,解决了目前西药治疗痛风带来的副作用大的困扰,可以长期服用,降低尿酸,从而治疗痛风。
本发明还提供了上述含葵花盘小分子肽的抗痛风组合物的制备方法,步骤如下:
(1)按上述的重量份数称取各原料;
(2)取葵花盘浸泡于葵花盘质量10~20倍的水中,闪式提取器提取5~15次,提取液离心10~30min,取上清液,复合酶酶解,过滤得到分子量小于10kd的提取液,温度-40~50℃,压力0.05~0.08kpa冷冻干燥得到葵花盘小分子肽冻干粉末;
(3)取薏苡仁加薏苡仁质量10~20倍的水浸泡0.5~1h后,提取罐回流提取1~3h得到薏苡仁提取液;
(4)取淡竹叶加淡竹叶质量10~20倍的水浸泡0.5~1h后,提取罐回流提取1~3h得到淡竹叶提取液;
(5)取白茅根加白茅根质量10~20倍的水浸泡0.5~1h后,提取罐回流提取1~3h得到白茅根提取液;
(6)取杜仲雄花加杜仲雄花质量10~20倍的水浸泡0.5~1h后,提取罐回流提取1~3h得到杜仲雄花提取液;
(7)取山楂加山楂质量10~20倍的水浸泡后,提取罐回流提取1~3h得到山楂提取液;
(8)将薏苡仁提取液、淡竹叶提取液、白茅根提取液、杜仲雄花提取液和山楂提取液混合,喷雾干燥后得到干燥提取物粉末;
(9)将葵花盘小分子肽冻干粉末和干燥提取物粉末混合均匀后即得含葵花盘小分子肽的抗痛风组合物。
进一步,步骤(2)中以闪式提取器提取5~15次的操作,每次提取的时间为2~10s。
进一步,复合酶为为胃蛋白酶和胰蛋白酶的混合酶,酶解的温度为35~37℃,酶解时间为4~8h。
具体实施方式
下面将对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1
含葵花盘小分子肽的抗痛风组合物的制备方法,步骤如下:
步骤一:葵花盘30份,15倍量水,闪式提取器提取5次,每次提取10秒钟。提取液离心10分钟,取上清液,复合酶酶解,过超滤装置,得到分子量分别为大于10kd、10kd,小于10kd的提取液。将分子量小于10kd的提取液,经药用真空冷冻干燥机进行干燥,得到葵花盘小分子肽冻干粉末;
步骤二:薏苡仁60份,加入15倍量水,浸泡0.5h,提取罐中回流提取3h,得薏苡仁提取液;
步骤三:淡竹叶60份,加入15倍量水,浸泡0.5h,提取罐中回流提取3h,得淡竹叶提取液;
步骤四:白茅根60份,加入15倍量水,浸泡0.5h,提取罐中回流提取3h,得白茅根提取液;
步骤五:杜仲雄花60份,加入15倍量水,浸泡0.5h,提取罐中回流提取3h,得杜仲雄花提取液;
步骤六:山楂60份,加入15倍量水,浸泡0.5h,提取罐中回流提取3h,得山楂提取液;
步骤七:将步骤二至六提取液合并,经喷雾干燥仪进行喷雾干燥,得干燥提取物粉末;
步骤八:将步骤一与步骤七所得粉末混合均匀,得含葵花盘小分子肽的抗痛风组合物。
实施例2
含葵花盘小分子肽的抗痛风组合物的制备方法,步骤如下:
步骤一:葵花盘20份,10倍量水,闪式提取器提取10次,每次提取5秒钟。提取液离心30分钟,取上清液,复合酶酶解,过超滤装置,得到分子量分别为大于10kd、10kd,小于10kd的提取液。将分子量小于10kd的提取液,经药用真空冷冻干燥机进行干燥,得到葵花盘小分子肽冻干粉末;
步骤二:薏苡仁40份,加入10倍量水,浸泡1h,提取罐中回流提取2h,得薏苡仁提取液;
步骤三:淡竹叶40份,加入10倍量水,浸泡1h,提取罐中回流提取2h,得淡竹叶提取液;
步骤四:白茅根40份,加入10倍量水,浸泡1h,提取罐中回流提取2h,得白茅根提取液;
步骤五:杜仲雄花40份,加入10倍量水,浸泡1h,提取罐中回流提取2h,得杜仲雄花提取液;
步骤六:山楂40份,加入10倍量水,浸泡1h,提取罐中回流提取2h,得山楂提取液;
步骤七:将步骤二至六提取液合并,经喷雾干燥仪进行喷雾干燥,得干燥提取物粉末;
步骤八:将步骤一与步骤七所得粉末混合均匀,得含葵花盘小分子肽的抗痛风组合物。
实施例3
含葵花盘小分子肽的抗痛风组合物的制备方法,步骤如下:
步骤一:葵花盘10份,20倍量水,闪式提取器提取15次,每次提取8秒钟。提取液离心30分钟,取上清液,复合酶酶解,过超滤装置,得到分子量分别为大于10kd、10kd,小于10kd的提取液。将分子量小于10kd的提取液,经药用真空冷冻干燥机进行干燥,得到葵花盘小分子肽冻干粉末;
步骤二:薏苡仁20份,加入20倍量水,浸泡1h,提取罐中回流提取1h,得薏苡仁提取液;
步骤三:淡竹叶20份,加入20倍量水,浸泡1h,提取罐中回流提取1h,,得淡竹叶提取液;
步骤四:白茅根20份,加入20倍量水,浸泡1h,提取罐中回流提取1h,,得白茅根提取液;
步骤五:杜仲雄花20份,加入20倍量水,浸泡1h,提取罐中回流提取1h,,得杜仲雄花提取液;
步骤六:山楂20份,加入20倍量水,浸泡1h,提取罐中回流提取1h,,得山楂提取液;
步骤七:将步骤二至六提取液合并,经喷雾干燥仪进行喷雾干燥,得干燥提取物粉末;
步骤八:将步骤一与步骤七所得粉末混合均匀,得含葵花盘小分子肽的抗痛风组合物。
实施例4
一、本发明实施例1中含葵花盘小分子肽的抗痛风组合物对人脐静脉内皮细胞的增值作用的影响
1材料与方法
1.1实验仪器与材料
C02培养箱、低温低速离心机(关国Hemus公司);超净工作台(苏州净化设备有限公司);电热恒温水浴箱(上海精宏实验设备有限公司);倒置显微镜(日本Olympus公司);微量高速离心机(LahNet公司);全自动酶标仪(Bio~Red,USA);细胞培养箱(NAPCO,USA);流式细胞仪(BD,美国)。
DM EM细胞培养液(GBICO公司);0.25%胰蛋白酶(南京生兴技术有限公司);小牛血清FBS(杭州四季青生物工程材料有限公司);二甲基亚飒DMSO(分析纯,上海凌峰化工);MTT(AERESCO,美国)。
HUVEC(人脐静脉内皮细胞)由南京中医药大学基础医学院提供。
1.2溶液的配置
1.2.1尿酸盐制备
5g尿酸加100mL蒸馏水煮沸,加5%NaOH溶液调pH7.4搅拌,冷却析晶制成尿酸钠结结晶。
1.2.2DMEM培养液尿酸盐溶液制备
将制好的MSU 10mg高压火菌,加不含血清的DMEM培养液10mL,研磨配成1mg/mL的DMEM溶液。实验时,此溶液再加DMEM培养液配成不同浓度DMEM的MSU溶液。
1.3实验方法
1.3.1HUVEC的体外培养
HUVEC株经支原体检测,无支原体污染,细胞经0.25%胰蛋白酶消化,含10%小牛血清的DMEM培养液中和,离心(1000r/min x 6min),去上清液,加含10%小牛血清的DMEM培养液,移入细胞培养瓶中,放37℃、5%C02培养箱中传代培养。
1.3.2HUVEC活力检测
HUVEC在培养瓶中培养,待生长至70%~80%融合时,以0.25%胰蛋白酶消化、用10%小牛血清DMEM培养液调成4x104/mL细胞悬液,植入96孔板(每孔200μL),培养24h,进行以下实验,每组各8孔,分别用终浓度为0、25、50、75、100(μg/mL)的MSU处理HUVEC,加液后继续放37℃、5%C02培养箱中培养24h收集上清液,剩余的HUVEC用于测定细胞活性,每孔再加1mg/mL MTT液100μL,继续放37℃、5%C02培养箱中培养4h后,弃MTT液,加入二甲基亚飒200μL溶解,震荡,于酶标仪读取吸光度值,波长490nm。
2.实验结果
实验结果见表1,以正常对照组细胞存活率(100%)为参照,本发明含葵花盘小分子肽的抗痛风组合物作用根据浓度(50μg/mL和100μg/mL、200μg/mL)时,表现出明显的剂量依赖性,随着剂量的增加,细胞存活率逐渐增强。经计算,作用24h后细胞存活率在浓度50μg/mL和100μg/mL、200μg/mL时分别达到41.21%±0.05%、47.84%±0.02%、54.34%±0.01%。结果表明,本发明含葵花盘小分子肽的抗痛风组合物对HUVEC细胞具有一定的保护作用。
表1含葵花盘小分子肽的抗痛风组合物对HUVEC细胞增值率的实验结果
药物浓度μg/mL | 增值率 |
200浓度 | 54.34% |
100浓度 | 47.84% |
50浓度 | 41.21% |
二、本发明实施例1中含葵花盘小分子肽的抗痛风组合物对PC12细胞(来源于肾上腺嗜铬细胞瘤)的保护作用
1.细胞培养方法
1.1 PC12细胞的原代培养
用已配制好的完全培养液在CO2培养箱中进行常规培养PC12细胞株。CO2培养箱中温度、相对湿度、CO2的相对浓度分别为37℃、90%、5%。每2天更换一次上述培养液,并使用倒置显微镜观察各组细胞的状态。
1.2 PC12细胞传代
待PC12细胞长满培养瓶底3/4的状态时,说明细胞生长是正常的且状态比较好。在生物安全柜中,倒出原来的培养液,用一次性吸管吸取约2mL无菌PBS并来回晃动进行洗涤细胞,重复洗涤两次。加入0.25%胰蛋白酶1~2mL进行消化(胰蛋白酶能覆盖瓶底),可以轻轻拍打培养瓶侧面,并在倒置显微镜下时刻查看细胞变化,当细胞间隙增大,细胞变圆且发生扩散时,迅速加入相同体积的完全培养液来终止此次消化,并使用一次性吸管将瓶壁上的细胞完全吹打下来后,全部移入离心管中。调节离心机的转速为1000r/min,将离心管中的细胞悬液充分离心5分钟后,吸去上清液。并吸取2mL完全培养液加入离心管中,轻轻吹打并混匀成单个细胞悬液,均匀的分装至新的培养瓶中,并在各个培养瓶中分别加入2mL完全培养液,放入CO2培养箱进行培养生长。
1.3 MTT实验检测叶酸对铅暴露下PC12细胞增殖的影响
1.3.1 MTT原理
MTT是一种能够被活细胞线粒体中的琥珀酸脱氢酶还原成难溶性的结晶状紫色产物formazan的黄色染料,而无法在死细胞中发挥此功能。Formazan在特定的溶剂(如DMSO)存在情况下可以被完全溶解,在特定的波长下使用酶标仪等仪器能够测定其吸光度,细胞增殖的越多越快则吸光度值(OD值)越大,可间接反映活细胞的数量。
1.3.2对PC12细胞增值率的影响
挑取分化后细胞长势较好的培养瓶,在其长满瓶底3/4时,加入1~2mL胰蛋白酶来消化瓶底的PC12细胞,制成细胞浓度为5×105/mL的单个细胞悬液,于96孔板每孔加100μl细胞悬液,将处于培养板边缘的各孔加入200μl PBS防止板中各孔培养液挥发。每组各8孔,分别用终浓度为0、25、50、75、100(μg/mL)的PC12细胞,加液后继续放37℃、5%C02培养箱中培养24h收集上清液,于24h时后,分别从CO2培养箱中取出一块96孔板,每孔加入20μl之前配制好的MTT溶液,放入CO2培养箱中反应4h后,拿出96孔板,置于生物安全柜下,将孔内的所有液体弃去,注意枪头不要碰到96孔板各孔底部。最后每孔再加入150μl的DMSO,将96孔培养板放置水平摇床中避光震荡30分钟后,在酶联免疫检测仪下,使用调零孔来调零,同时设置波长490nm来读取并记录各组OD值。
2.实验结果见表2,以正常对照组细胞存活率(100%)为参照,本发明含葵花盘小分子肽的抗痛风组合物作用根据浓度(50μg/mL和100μg/mL、200μg/mL)时,表现出明显的剂量依赖性,随着剂量的减小,细胞存活率逐渐增强。经计算,本发明含葵花盘小分子肽的抗痛风组合物作用24h后细胞存活率在浓度50μg/mL和100μg/mL、200μg/mL时分别达到58.25%±0.05%、47.35%±0.02%、25.22%±0.01%。结果表明,本发明含葵花盘小分子肽的抗痛风组合物对PC12细胞具有一定的保护作用。
表2含葵花盘小分子肽的抗痛风组合物对PC12细胞增值率的实验结果
药物浓度μg/mL | 增值率 |
200浓度 | 25.22% |
100浓度 | 47.35% |
50浓度 | 58.25% |
三、本发明实施例1中含葵花盘小分子肽的抗痛风组合物对RAW264.7巨噬细胞的影响
1.试验方法
1.1试剂的配置
完全培养液:10%灭活胎牛血清+90%DMEM高糖培养基,除菌,4℃保存。
持续培养基:5%灭活胎牛血清+95%DMEM高糖培养基,除菌,4℃保存。
MTT溶液:50mg MTT粉末+10mL PBS,除菌过滤,~20℃保存备用。
细胞冻存液:灭活胎牛血清和DMSO以9:1均匀混合。
1.2 RAW264.7巨噬细胞的复苏、培养、传代及冻存
复苏:冻存细胞取出后37℃速溶,取15mL离心管,加入10mL含血清培养基,将冻存管中液体(通常1~1.5mL)加入上述含培养基离心管中,轻吹混匀,1200r/min常温离心5min,去掉上清,加入3ml含血清培养基,轻吹混匀得细胞悬液,加入培养瓶中,左右摇晃铺平即可。
培养:细胞置于含有完全培养基的培养瓶中贴壁生长,培养条件为恒温37℃和5%CO2。待细胞长满培养瓶底部的80%以上,细胞可进行传代。
传代:弃培养瓶内培养液,加入适量胰蛋白酶液消化使细胞单个分散,约1mL覆盖培养瓶底即可。37℃孵育2~5min,若胞质回缩、细胞间隙增大,立即加入适量含血清的培养液,终止消化。顺序反复地轻轻吹打瓶壁细胞,使贴壁的细胞脱离瓶壁悬浮形成细胞悬液。过程轻柔,防止用力过猛造成细胞损伤。血球计数板计数后,接于新的培养瓶(一传三),放入培养箱培养。
冻存:当细胞处于对数生长期细胞,按上述传代方法制成细胞悬液,使细胞密度在5×107/mL左右,离心,去上清。细胞重悬于细胞冻存液,动作尽量轻,随后分装于无菌冻存管,1mL/管。密封冻存管,做上标记。冻存管以4℃10分钟,~20℃30分钟,~80℃过夜梯度冻存,最后投于液氮槽长期储存。
1.3本发明含葵花盘小分子肽的抗痛风组合物对RAW264.7巨噬细胞增值率的影响
细胞接种于96孔板中,密度为2×105个/mL,加入100μL/孔,培养箱培养2h,细胞完全贴壁后,更换DMEM维持培养液培养。正常组为不加LPS和样品液;LPS组,培养液中加入LPS使其终浓度为1μg/mL;样品组,各样品分别设置20~360μg/m L的浓度梯度。培养24h,每组做5个复孔。之后倒掉培养基,加入20μL/孔MTT溶液,37℃孵育4h。吸去孔中液体后加入DMSO溶剂150μL/孔,酶标板孵育仪轻轻振荡10min,之后540nm处检测吸光度。
2.实验结果见表3,以正常对照组细胞存活率(100%)为参照,本发明含葵花盘小分子肽的抗痛风组合物作用根据浓度(50μg/mL和100μg/mL、200μg/mL)时,表现出明显的剂量依赖性,随着剂量的增加,细胞存活率逐渐增强。经计算,敖东复方作用24h后细胞存活率在浓度50μg/mL和100μg/mL、200μg/mL时分别达到15.2%±0.05%、16.2%±0.02%、23.7%±0.01%。表明,本发明含葵花盘小分子肽的抗痛风组合物对RAW264.7巨噬细胞具有一定的保护作用。
表3含葵花盘小分子肽的抗痛风组合物对RAW264.7巨噬细胞增值率的实验结果
药物浓度μg/mL | 增值率 |
200浓度 | 23.7% |
100浓度 | 16.2% |
50浓度 | 15.2% |
综上所述,本发明中,葵花盘小分子肽与中药提取物组方作用,具有协同效果,对人脐静脉内皮细胞,PC12细胞,RAW264.7巨噬细胞具有增殖作用,表明有保护作用,说明可明显降低尿酸值,并具有抗炎、镇痛的作用。此外,葵花盘、其他几味中药均有药食两用的,无副作用的特点,协同作用明显,对治疗痛风性关节炎效果显著,可长期服用。本发明为中药治疗痛风提供了有效的方法及理论依据。
表4含葵花盘小分子肽的抗痛风组合物体外细胞实验的综合结果
典型病例
通过以下几个典型病例进一步表明本发明的疗效
病例1
李某,男30岁,常规体检检测尿酸值532,喝本发明实施例1含葵花盘小分子肽的抗痛风组合物一个多月后,检测尿酸值431.1,尿酸下降101,说明本发明含葵花盘小分子肽的抗痛风组合物具有较低尿酸值的作用,且效果显著。
病例2
吴某,男64岁,常规体检检测尿酸值494,喝了本发明实施例2含葵花盘小分子肽的抗痛风组合物两个月,复查检测尿酸值287,已经降到正常范围,并且大脚趾头部位的痛风石已经完全溶解。
病例3
林某,男29岁,痛风5年,检测尿酸值535,痛风每年都发作几次,吃了本发明实施例1含葵花盘小分子肽的抗痛风组合物之后,检测尿酸值降低到263,痛风石开始变小变软。
Claims (10)
1.一种含葵花盘小分子肽的抗痛风组合物,其特征在于,制备所述药物的原料及其重量份数如下:葵花盘10~30份、薏苡仁20~60份、淡竹叶20~60份、白茅根20~60份、杜仲雄花20~60份、山楂20~60份。
2.根据权利要求1所述的一种含葵花盘小分子肽的抗痛风组合物,其特征在于,制备所述药物的原料及其重量份数如下:葵花盘20份、薏苡仁40份、淡竹叶40份、白茅根40份、杜仲雄花40份、山楂40份。
3.一种含葵花盘小分子肽的抗痛风组合物的制备方法,其特征在于,步骤如下:
(1)按权利要求1或2所述的重量份数称取各原料;
(2)取葵花盘浸泡于水中,闪式提取器提取5~15次,提取液离心10~30min,取上清液,复合酶酶解,过滤得到分子量小于10kd的提取液,冷冻干燥得到葵花盘小分子肽冻干粉末;
(3)取薏苡仁加水浸泡后,回流提取得到薏苡仁提取液;
(4)取淡竹叶加水浸泡后,回流提取得到淡竹叶提取液;
(5)取白茅根加水浸泡后,回流提取得到白茅根提取液;
(6)取杜仲雄花加水浸泡后,回流提取得到杜仲雄花提取液;
(7)取山楂加水浸泡后,回流提取得到山楂提取液;
(8)将薏苡仁提取液、淡竹叶提取液、白茅根提取液、杜仲雄花提取液和山楂提取液混合,喷雾干燥后得到干燥提取物粉末;
(9)将葵花盘小分子肽冻干粉末和干燥提取物粉末混合均匀后即得所述含葵花盘小分子肽的抗痛风组合物。
4.根据权利要求3所述的一种含葵花盘小分子肽的抗痛风组合物的制备方法,其特征在于,步骤(2)中所述水的用量为葵花盘质量的10~20倍。
5.据权利要求3所述的一种含葵花盘小分子肽的抗痛风组合物的制备方法,其特征在于,步骤(2)中所述以闪式提取器提取5~15次的操作,每次提取的时间为2~10s。
6.据权利要求3所述的一种含葵花盘小分子肽的抗痛风组合物的制备方法,其特征在于,步骤(2)中所述复合酶为胃蛋白酶和胰蛋白酶的混合酶;
所述酶解的温度为35~37℃,酶解时间为4~8h。
7.据权利要求3所述的一种含葵花盘小分子肽的抗痛风组合物的制备方法,其特征在于,步骤(2)中所述冷冻干燥的温度为-40~50℃,压力为0.05~0.08kpa。
8.据权利要求3所述的一种含葵花盘小分子肽的抗痛风组合物的制备方法,其特征在于,步骤(3)中所述水的加入量为薏苡仁质量的10~20倍;
步骤(4)中所述水的加入量为淡竹叶质量的10~20倍;
步骤(5)中所述水的加入量为白茅根质量的10~20倍;
步骤(6)中所述水的加入量为杜仲雄花质量的10~20倍;
步骤(7)中所述水的加入量为山楂质量的10~20倍。
9.据权利要求3所述的一种含葵花盘小分子肽的抗痛风组合物的制备方法,其特征在于,步骤(3)至步骤(7)中任一步骤所述浸泡的时间为浸泡0.5~1h。
10.据权利要求3所述的一种含葵花盘小分子肽的抗痛风组合物的制备方法,其特征在于,步骤(3)至步骤(7)中所述回流提取的操作为以提取罐回流提取1~3h。
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