CN114947119B - 一种具辅助降血糖血脂作用的核桃花发酵食品及其制备方法 - Google Patents
一种具辅助降血糖血脂作用的核桃花发酵食品及其制备方法 Download PDFInfo
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- CN114947119B CN114947119B CN202210604118.XA CN202210604118A CN114947119B CN 114947119 B CN114947119 B CN 114947119B CN 202210604118 A CN202210604118 A CN 202210604118A CN 114947119 B CN114947119 B CN 114947119B
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Abstract
本发明公开了一种具辅助降血糖血脂作用的核桃花发酵食品的制备方法，包括以下步骤：(1)菌种的制备：从乳饼中筛选分离具降脂作用的植物乳杆菌；并与干酪乳杆菌复配，获得复合菌液；(2)护色和脱涩：对核桃花进行护色、脱涩；(3)发酵：将护色脱涩后的核桃花和蒸馏水经高压蒸汽灭菌处理，冷却后，接种复合菌液发酵，再加入小米辣、生姜、大蒜、八角、花椒、盐、白酒浸泡发酵；(4)杀菌：发酵后的核桃花经称量、真空包装后，杀菌，冷却，获得核桃花发酵食品。本发明通过发酵、调配和杀菌等工序，去除了核桃花天然的苦涩味，所得产品口感脆，有嚼劲，味道鲜香开胃，保质期可在室温条件下延长至4个月，且具降血糖血脂功效。
Description
技术领域
本发明属于食品加工技术领域，尤其涉及一种具辅助降血糖血脂作用的核桃花发酵食品及其制备方法。
背景技术
糖脂代谢性疾病是以糖代谢、脂代谢紊乱为特征，以2型糖尿病、肥胖、高血压、高脂血症、高血脂和动脉粥样硬化等为主要表型的流行性慢性疾病，严重威胁人类身体的健康。糖尿病以高血糖为主要特征，采用各种手段降低血液中葡萄糖含量是其主要治疗手段。糖尿病发病机制十分复杂，目前尚无根治的办法，目前采用的各种口服或者注射降糖药物存在价格昂贵、疗效有限、存在不同程度毒副作用等缺陷。因此寻找安全、高效、毒副作用小，成本低廉的药物资源具有重要意义。
核桃花是核桃产业的主要副产物之一，2019年我国核桃产量居世界首位，为362.7万吨，核桃花产量高达60万吨。核桃花具有很高的营养价值，我国西南地区一直将其作为绿色蔬菜食用。如能高值化利用我国的大量核桃花资源，将增加核桃种植农户和企业的销售收入，增加其市场竞争力。
发明人研究发现，核桃花含有较高含量的酚类化合物，以及较强的抗氧化活性，且可有效抑制糖代谢关键酶(α-葡萄糖苷酶)的活性，因此具有开发为降血糖食品的潜质。本发明从云南特产——自然发酵山羊乳饼中筛选得到一种具有降血脂等益生功效的乳酸菌，将该菌与干酪乳杆菌复配后对核桃花进行发酵，发现其α-葡萄糖苷酶抑制活性显著提高。发明人建立了高糖饮食诱导的糖尿病果蝇模型，对其饲喂发酵核桃花，发现核桃花可延长糖尿病果蝇的寿命，降低果蝇体内的葡萄糖、海藻糖含量，并降低其体内甘油三酯水平，调节其氧化应激程度，表明所发明的核桃花发酵食品具有辅助降血糖血脂作用。
发明内容
本发明提供了一种具辅助降血糖血脂作用的核桃花发酵食品的制备方法，包括以下步骤：
(1)菌种的制备：从乳饼中筛选分离具降脂作用的植物乳杆菌，配制为活菌数1×109CFU/mL的菌液；并与相同活菌数的干酪乳杆菌按体积比1：1复配，获得复合菌液；
(2)护色和脱涩：核桃花用清水冲洗2遍，冲洗后，再经乙二胺四乙酸二钠与氯化钙复配护色后，煮沸保持8～15分钟进行脱涩，脱涩完成后将核桃花冲洗干净；
(3)发酵：将护色脱涩后的核桃花和蒸馏水经高压蒸汽灭菌处理，冷却后，接种所述复合菌液并于37℃发酵3天，再加入小米辣、生姜、大蒜、八角、花椒、盐、白酒浸泡发酵10天；
(4)杀菌：发酵后的核桃花经称量、真空包装后，于85℃水浴中加热20min 杀菌，并立即淋冷水冷却，获得所述核桃花发酵食品。
进一步地，所述复合菌液的制备方法为：
1)乳饼中乳酸菌的分离纯化：从乳饼中心部分取10克研碎后，用无菌生理盐水制成悬液，采用梯度稀释法分离其杆状及球状乳酸菌；挑取平板上具溶钙圈的菌落划线以获得纯菌落；采用革兰氏染色、镜检、生理生化试验初步鉴定菌种后，采用-80℃甘油保藏法贮藏备用；
2)降血脂乳酸菌的体外筛选：分别将分离纯化得到的乳酸菌以4％的接种量接种至胆固醇培养基、甘油三酯培养基、MRS及M17液体培养基及含胆盐、抗生素的培养基中，检测乳酸菌的降血脂和耐酸耐胆盐的能力，筛选出降血脂和耐酸耐胆盐性能高的菌株；
3)乳酸菌的鉴定：将筛选得到的菌株进行PCR扩增，得到特异性扩增产物，通过凝胶电泳检测扩增产物后，测序，通过BLAST比对分析，鉴定该菌株为植物乳杆菌；
4)扩培：首先将所述植物乳酸菌活化，按1％接种量将植物乳酸菌接入MRS 液体培养基中，并在37℃下摇床培养至对数生长，获得所述具降脂作用的植物乳杆菌的菌液；
5)复合菌液的制备：将扩培后的植物乳杆菌菌液梯度稀释后均匀涂布到MRS 固体乳酸菌选择培养基上，用接种环挑取菌群到0.85％生理盐水中，并通过10倍梯度稀释法进行计数，得到活菌数1×109CFU/mL的植物乳杆菌菌悬液，将所述植物乳杆菌菌悬液与相同活菌数的干酪乳杆菌菌悬液按体积比1：1复配，制成所述复合菌液。
进一步地，所述核桃花的护色方法为：
1)护色液的配制：所述护色液中的护色剂为乙二胺四乙酸二钠和氯化钙，乙二胺四乙酸二钠与氯化钙的质量比m(乙二胺四乙酸二钠)∶m(氯化钙)＝1∶1，护色剂的质量浓度为5.23mg/mL；
2)核桃花的护色：新鲜核桃花以清水冲洗2遍，在配置好的护色剂溶液中，于25.5℃护色2.3h，取出沥干表面水分，完成护色。
进一步地，所述发酵的技术参数为：
1)第一次发酵：每500g核桃花加入1L蒸馏水，接种5mL所述复合菌液，于37℃发酵3d；
2)第二次发酵及浸泡入味：第一次发酵完成后再加入小米辣180～220g、生姜180～220g、大蒜150～180g、八角4～6个、花椒8～12g、盐100～120g、白酒15～20g，于室温条件下浸泡发酵10d。
进一步地，所述杀菌的技术参数为：
1)每40g发酵核桃花装至蒸煮袋中，经真空封口后，于85℃水浴中加热 20min，立即于冷水中冷却，擦干水分，在每个包装箱中装入20袋发酵核桃花产品。
进一步地，所述复合菌液的活菌数为1×109CFU/mL。
本发明的有益效果在于：本发明所选用的核桃花原料富含酚类、黄酮、膳食纤维等活性成分，采用的发酵菌株——植物乳杆菌为从自然发酵乳饼中自主分离的，与干酪乳杆菌复配后用于发酵核桃花，发酵的核桃花经体外试验和动物试验验证具有降血糖血脂效应；且制备的发酵核桃花产品口感脆，有嚼劲，具有发酵的鲜香风味，满足了人们对发酵蔬菜的口感和功能需要，并可解决目前核桃花资源的深加工问题。本发明的降血糖血脂效应主要体现在：
(1)核桃花具有较强的α-葡萄糖苷酶抑制能力：α-葡萄糖苷酶是一种重要的治疗靶标酶，因为它参与复杂膳食碳水化合物的消化过程阶段，在该阶段代谢更简单的寡糖以供肠道吸收。研究其抑制参与碳水化合物消化的α-葡萄糖苷酶的潜力是一种治疗糖尿病的新方法。如图1所示，核桃花具有较强的α-葡萄糖苷酶抑制活性，IC50值为679.91±8.64μg/mL，强于核桃分心木、藜蒿和高良姜等其他植物。
(2)发酵可以提高核桃花的α-葡萄糖苷酶抑制能力：经本发明所述的益生菌发酵后，核桃花的α-葡萄糖苷酶抑制活性相比于发酵前提高到了321.74％，如图2所示。因此，本发明所述的发酵过程是提高核桃花α-葡萄糖苷酶抑制活性的有效手段。
(3)果蝇体内的海藻糖、葡萄糖含量是评估其血糖水平的一项重要指标，同时，高血糖环境会增加线粒体活性氧(ROS)的产生，导致氧化应激和器官损伤，使丙二醛(MDA)水平上升，而超氧化物歧化酶(SOD)、过氧化氢酶(CAT) 等指标下降。果蝇体内的甘油三酯(TG)则是评估其血脂水平的一项重要指标。发酵核桃花可显著降低果蝇的体内血糖水平，同时显著降低果蝇体内的ROS、 MDA和TG水平(p<0.05)，并改善果蝇体内SOD、CAT等水平，其中高剂量发酵核桃花的修复效果更明显。
附图说明
图1为核桃花对α-葡萄糖苷酶抑制活性的IC50值；
图2为发酵前和各实施例及对比例的核桃花α-葡萄糖苷酶抑制能力对比图；
图3为饲喂实施例1制备的核桃花发酵食品后果蝇体内糖含量和生化指标。
具体实施方式
下面结合实施例进行详细的说明：
实施例1
一种具辅助降血糖血脂作用的核桃花发酵食品的制备方法，包括以下步骤：
(1)菌种的制备：从乳饼中筛选分离具降脂作用的植物乳杆菌，配制为活菌数1×109CFU/mL的菌液；并与相同活菌数的干酪乳杆菌(Lactobacillus casei， ATCC 334)按体积比1：1复配，获得复合菌液(活菌数为1×109CFU/mL)；
(2)护色和脱涩：核桃花用清水冲洗2遍，冲洗后经乙二胺四乙酸二钠与氯化钙复配的护色液进行护色，然后煮沸保持10分钟进行脱涩，脱涩完成后将核桃花冲洗干净；
(3)发酵：将护色脱涩后的核桃花500g和1L蒸馏水经高压蒸汽灭菌处理，冷却后，接种5mL所述复合菌液，于37℃发酵3天，再加入200g小米辣、200g 生姜、170g大蒜、5个八角、10g花椒、100g盐、15g白酒，于室温条件下浸泡发酵10天；
(4)杀菌：每40g发酵核桃花装至蒸煮袋中，经真空封口后，于85℃水浴中加热20min，然后立即于冷水中冷却，擦干水分，获得所述核桃花发酵食品。
其中所述复合菌液的制备方法为：
1)乳饼中乳酸菌的分离纯化：从乳饼中心部分取10克研碎后，用无菌生理盐水制成悬液，采用梯度稀释法分离其杆状及球状乳酸菌；挑取平板上具溶钙圈的菌落划线以获得纯菌落；采用革兰氏染色、镜检、生理生化试验初步鉴定菌种后，采用-80℃甘油保藏法贮藏备用；
2)降血脂乳酸菌的体外筛选：分别将分离纯化得到的乳酸菌以4％的接种量接种至胆固醇培养基、甘油三酯培养基、MRS及M17液体培养基及含胆盐、抗生素的培养基中，检测乳酸菌的降血脂和耐酸耐胆盐的能力，筛选出降血脂和耐酸耐胆盐性能高的菌株；
3)乳酸菌的鉴定：将筛选得到的菌株进行PCR扩增，得到特异性扩增产物，通过凝胶电泳检测扩增产物后，测序，通过BLAST比对分析，鉴定该菌株为植物乳杆菌；
4)扩培：首先将所述植物乳酸菌活化，按1％接种量将植物乳酸菌接入MRS 液体培养基中，并在37℃下摇床培养至对数生长，获得所述具降脂作用的植物乳杆菌的菌液；
5)复合菌液的制备：将扩培后的植物乳杆菌菌液梯度稀释后均匀涂布到 MRS固体乳酸菌选择培养基上，用接种环挑取菌群到0.85％生理盐水中，并通过10倍梯度稀释法进行计数，得到活菌数1×109CFU/mL的植物乳杆菌菌悬液，将所述植物乳杆菌菌悬液与相同活菌数的干酪乳杆菌菌悬液按体积比1：1复配，制成所述复合菌液。
所述核桃花的护色方法为：
1)护色液的配制：所述护色液中的护色剂为乙二胺四乙酸二钠和氯化钙，乙二胺四乙酸二钠与氯化钙的质量比m(乙二胺四乙酸二钠)∶m(氯化钙)＝1∶1，护色剂的质量浓度为5.23mg/mL；
2)核桃花的护色：新鲜核桃花以清水冲洗2遍，在配置好的护色剂溶液中，于25.5℃护色2.3h，取出沥干表面水分，完成护色。
对比例1
一种核桃花发酵食品的制备方法，包括以下步骤：
(1)制备活菌数为1×109CFU/mL的干酪乳杆菌(Lactobacillus casei，ATCC 334)生理盐水；
(2)护色和脱涩：核桃花用清水冲洗2遍，冲洗后经乙二胺四乙酸二钠与氯化钙复配的护色液进行护色，然后煮沸保持10分钟进行脱涩，脱涩完成后将核桃花冲洗干净；
(3)发酵：将护色脱涩后的核桃花500g和蒸馏水1L经高压蒸汽灭菌处理，冷却后，接种5mL所述干酪乳杆菌生理盐水，于37℃发酵3天，再加入200g 小米辣、200g生姜、170g大蒜、5个八角、10g花椒、100g盐、15g白酒，于室温条件下浸泡发酵10天；
(4)杀菌：每40g发酵核桃花装至蒸煮袋中，经真空封口后，于85℃水浴中加热20min，然后立即于冷水中冷却，擦干水分，获得本对比例所述核桃花发酵食品。
其中所述核桃花的护色方法为：
1)护色液的配制：所述护色液中的护色剂为乙二胺四乙酸二钠和氯化钙，乙二胺四乙酸二钠与氯化钙的质量比m(乙二胺四乙酸二钠)∶m(氯化钙)＝1∶1，护色剂的质量浓度为5.23mg/mL；
2)核桃花的护色：新鲜核桃花以清水冲洗2遍，在配置好的护色剂溶液中，于25.5℃护色2.3h，取出沥干表面水分，完成护色。
对比例2
一种核桃花发酵食品的制备方法，包括以下步骤：
(1)菌种的制备：制备活菌数为1×109CFU/mL的鼠李糖乳杆菌 (Lactobacillusrhamnosus,ATCC 53013)生理盐水；
(2)护色和脱涩：核桃花用清水冲洗2遍，冲洗后经乙二胺四乙酸二钠与氯化钙复配的护色液进行护色，然后煮沸保持10分钟进行脱涩，脱涩完成后将核桃花冲洗干净；
(3)发酵：将护色脱涩后的核桃花500g和蒸馏水1L经高压蒸汽灭菌处理，冷却后，接种5mL所述鼠李糖乳杆菌生理盐水，于37℃发酵3天，再加入200g 小米辣、200g生姜、170g大蒜、5个八角、10g花椒、100g盐、15g白酒，于室温条件下浸泡发酵10天；
(4)杀菌：每40g发酵核桃花装至蒸煮袋中，经真空封口后，于85℃水浴中加热20min，然后立即于冷水中冷却，擦干水分，获得本对比例所述核桃花发酵食品。
其中所述核桃花的护色方法为：
1)护色液的配制：所述护色液中的护色剂为乙二胺四乙酸二钠和氯化钙，乙二胺四乙酸二钠与氯化钙的质量比m(乙二胺四乙酸二钠)∶m(氯化钙)＝1∶1，护色剂的质量浓度为5.23mg/mL；
2)核桃花的护色：新鲜核桃花以清水冲洗2遍，在配置好的护色剂溶液中，于25.5℃护色2.3h，取出沥干表面水分，完成护色。
对比例3
一种核桃花发酵食品的制备方法，包括以下步骤：
(1)菌种的制备：将活菌数为1×109CFU/mL的干酪乳杆菌(Lactobacillus casei，ATCC 334)与相同活菌数的鼠李糖乳杆菌(Lactobacillus rhamnosus，ATCC 53013)按体积比1：1复配，获得复合菌液(活菌数为1×109CFU/mL)；
(2)护色和脱涩：核桃花用清水冲洗2遍，冲洗后经乙二胺四乙酸二钠与氯化钙复配的护色液进行护色，然后煮沸保持10分钟进行脱涩，脱涩完成后将核桃花冲洗干净；
(3)发酵：将护色脱涩后的核桃花500g和蒸馏水1L经高压蒸汽灭菌处理，冷却后，接种5mL所述复合菌液，于37℃发酵3天，再加入200g小米辣、200g 生姜、170g大蒜、5个八角、10g花椒、100g盐、15g白酒，于室温条件下浸泡发酵10天；
(4)杀菌：每40g发酵核桃花装至蒸煮袋中，经真空封口后，于85℃水浴中加热20min，然后立即于冷水中冷却，擦干水分，获得所述核桃花发酵食品。
其中所述核桃花的护色方法为：
1)护色液的配制：所述护色液中的护色剂为乙二胺四乙酸二钠和氯化钙，乙二胺四乙酸二钠与氯化钙的质量比m(乙二胺四乙酸二钠)∶m(氯化钙)＝1∶1，护色剂的质量浓度为5.23mg/mL；
2)核桃花的护色：新鲜核桃花以清水冲洗2遍，在配置好的护色剂溶液中，于25.5℃护色2.3h，取出沥干表面水分，完成护色。
对比例4
一种核桃花发酵食品的制备方法，包括以下步骤：
(1)菌种的制备：从乳饼中筛选分离具降脂作用的植物乳杆菌；配置活菌数为1×109CFU/mL的所述植物乳杆菌生理盐水；
(2)护色和脱涩：核桃花用清水冲洗2遍，冲洗后经乙二胺四乙酸二钠与氯化钙复配的护色液进行护色，然后煮沸保持10分钟进行脱涩，脱涩完成后将核桃花冲洗干净；
(3)发酵：将护色脱涩后的核桃花500g和蒸馏水1L经高压蒸汽灭菌处理，冷却后，接种5mL所述植物乳杆菌生理盐水，于37℃发酵3天，再加入200g 小米辣、200g生姜、170g大蒜、5个八角、10g花椒、100g盐、15g白酒，于室温条件下浸泡发酵10天；
(4)杀菌：每40g发酵核桃花装至蒸煮袋中，经真空封口后，于85℃水浴中加热20min，然后立即于冷水中冷却，擦干水分，获得本对比例所述核桃花发酵食品。
其中所述植物乳杆菌生理盐水的制备方法为：
1)乳饼中乳酸菌的分离纯化：从乳饼中心部分取10克研碎后，用无菌生理盐水制成悬液，采用梯度稀释法分离其杆状及球状乳酸菌；挑取平板上具溶钙圈的菌落划线以获得纯菌落；采用革兰氏染色、镜检、生理生化试验初步鉴定菌种后，采用-80℃甘油保藏法贮藏备用；
2)降血脂乳酸菌的体外筛选：分别将分离纯化得到的乳酸菌以4％的接种量接种至胆固醇培养基、甘油三酯培养基、MRS及M17液体培养基及含胆盐、抗生素的培养基中，检测乳酸菌的降血脂和耐酸耐胆盐的能力，筛选出降血脂和耐酸耐胆盐性能高的菌株；
3)乳酸菌的鉴定：将筛选得到的菌株进行PCR扩增，得到特异性扩增产物，通过凝胶电泳检测扩增产物后，测序，通过BLAST比对分析，鉴定该菌株为植物乳杆菌；
4)扩培：首先将所述植物乳酸菌活化，按1％接种量将植物乳酸菌接入MRS 液体培养基中，并在37℃下摇床培养至对数生长，获得所述具降脂作用的植物乳杆菌的菌液；
5)植物乳杆菌菌液的制备：将扩培后的植物乳杆菌菌液梯度稀释后均匀涂布到MRS固体乳酸菌选择培养基上，用接种环挑取菌群到0.85％生理盐水中，并通过10倍梯度稀释法进行计数，得到所述植物乳杆菌生理盐水。
所述核桃花的护色方法为：
1)护色液的配制：所述护色液中的护色剂为乙二胺四乙酸二钠和氯化钙，乙二胺四乙酸二钠与氯化钙的质量比m(乙二胺四乙酸二钠)∶m(氯化钙)＝1∶1，护色剂的质量浓度为5.23mg/mL；
2)核桃花的护色：新鲜核桃花以清水冲洗2遍，在配置好的护色剂溶液中，于25.5℃护色2.3h，取出沥干表面水分，完成护色。
实施例2
分别测试各实施例和对比例所制备的核桃花的相对α-葡萄糖苷酶抑制活性，结果如图2所示。由图2可知，本发明以核桃花和自主筛选的具降血脂作用的植物乳杆菌及干酪乳杆菌作为主要原材料，相比于其他发酵菌种，所得核桃花的相对α-葡萄糖苷酶抑制活性明显更好，且复合菌液的发酵效果显著好于单种菌液的发酵效果。且本发明原料具有来源易得、安全绿色等优点，且通过体外实验和动物试验表明所发明的发酵核桃花产品具降血糖血脂功效。
以上对本发明所提供的技术方案进行了详细介绍，对于本领域的一般技术人员，依据本发明实施例的思想，在具体实施方式及应用范围上均会有改变之处，综上所述，本说明书内容不应理解为对本发明的限制。
Claims (5)
1.一种具有辅助降血糖血脂作用的核桃花发酵食品的制备方法，其特征在于，包括以下步骤：
（1）菌种的制备：从乳饼中筛选分离具有降脂作用的植物乳杆菌，配制为活菌数1×109 CFU/mL的菌液；并与相同活菌数的干酪乳杆菌按体积比1：1复配，获得复合菌液；
（2）护色和脱涩：核桃花用清水冲洗2遍，冲洗后，再经乙二胺四乙酸二钠与氯化钙复配护色后，煮沸保持8～15分钟进行脱涩，脱涩完成后将核桃花冲洗干净；
（3）发酵：将护色脱涩后的核桃花经高压蒸汽灭菌处理，冷却后，接种所述复合菌液并于37℃发酵3天，再加入小米辣、生姜、大蒜、八角、花椒、盐、白酒浸泡发酵10天；
（4）杀菌：发酵后的核桃花经称量、真空包装后，于85 ℃水浴中加热20 min，并立即淋冷水冷却，获得所述核桃花发酵食品；
其中所述复合菌液的制备方法具体为：
1）乳饼中乳酸菌的分离纯化：从乳饼中心部分取10克研碎后，用无菌生理盐水制成悬液，采用梯度稀释法分离其杆状及球状乳酸菌；挑取平板上具有溶钙圈的菌落划线以获得纯菌落；采用革兰氏染色、镜检、生理生化试验初步鉴定菌种后，采用-80℃甘油保藏法贮藏备用；
2）降血脂乳酸菌的体外筛选：分别将分离纯化得到的乳酸菌以4%的接种量接种至胆固醇培养基、甘油三酯培养基、MRS及M17液体培养基及含胆盐、抗生素的培养基中，检测乳酸菌的降血脂和耐酸耐胆盐的能力，筛选出降血脂和耐酸耐胆盐性能高的菌株；
3）乳酸菌的鉴定：将筛选得到的菌株进行PCR扩增，得到特异性扩增产物，通过凝胶电泳检测扩增产物后，测序，通过BLAST比对分析，鉴定该菌株为植物乳杆菌；
4）扩培：首先将所述植物乳酸菌活化，按1%接种量将植物乳酸菌接入MRS液体培养基中，并在 37℃下摇床培养至对数生长，获得所述具有降脂作用的植物乳杆菌的菌液；
5）复合菌液的制备：将扩培后的植物乳杆菌菌液梯度稀释后均匀涂布到MRS固体乳酸菌选择培养基上，用接种环挑取菌群到0.85%生理盐水中，并通过10 倍梯度稀释法进行计数，得到活菌数1×109 CFU/mL的植物乳杆菌菌悬液，将所述植物乳杆菌菌悬液与相同活菌数的干酪乳杆菌菌悬液按体积比1：1复配，制成所述复合菌液。
2.根据权利要求1所述的一种具有辅助降血糖血脂作用的核桃花发酵食品的制备方法，其特征在于，所述核桃花的护色方法为：
1）护色液的配制：所述护色液中的护色剂为乙二胺四乙酸二钠和氯化钙，乙二胺四乙酸二钠与氯化钙的质量比m(乙二胺四乙酸二钠)∶m(氯化钙)=1∶1，护色剂的质量浓度为5.23 mg/mL；
2）核桃花的护色：新鲜核桃花以清水冲洗2遍，在配置好的护色剂溶液中，于25.5 ℃护色2.3 h，取出沥干表面水分，完成护色。
3.根据权利要求1所述的一种具有辅助降血糖血脂作用的核桃花发酵食品的制备方法，其特征在于，所述发酵的技术参数为：
1）第一次发酵：每500 g核桃花加入1 L蒸馏水，接种5 mL所述复合菌液，于37 ℃发酵3d；
2）第二次发酵及浸泡入味：第一次发酵完成后再加入小米辣180～220 g、生姜180～220 g、大蒜150～180 g、八角4～6 个、花椒8～12 g、盐100～120 g、白酒15～20 g，于室温条件下浸泡发酵10 d。
4.根据权利要求1所述的一种具有辅助降血糖血脂作用的核桃花发酵食品的制备方法，其特征在于，所述杀菌的技术参数为：
1） 每40 g发酵核桃花装至蒸煮袋中，经真空封口后，于85 ℃水浴中加热20 min，立即于冷水中冷却，擦干水分，在每个包装箱中装入20袋发酵核桃花产品。
5.根据权利要求1所述的一种具有辅助降血糖血脂作用的核桃花发酵食品的制备方法，其特征在于，所述复合菌液的活菌数为1×109 CFU/mL。
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