CN117243227B - 一种柠檬草-百里香复合精油微胶囊的制备方法和应用 - Google Patents
一种柠檬草-百里香复合精油微胶囊的制备方法和应用 Download PDFInfo
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Abstract
本发明提供了一种柠檬草‑百里香复合精油微胶囊及其制备方法和应用，属于防腐技术领域。本发明所述的柠檬草‑百里香复合精油中，柠檬草精油和百里香精油的体积比为1～3:1～3。本发明确定了柠檬草‑百里香复合精油对同时抑制块菌贮藏中的枯草芽孢杆菌、粪小短杆、冻土毛霉、粉红粘帚菌和腐皮镰孢菌的最佳体积比，可以为块菌采后贮、运、销各环节的保鲜措施提供新的技术思路。
Description
技术领域
本发明涉及防腐技术领域，尤其涉及一种柠檬草-百里香复合精油微胶囊的制备方法和应用。
背景技术
块菌(truffle)也称松露、猪拱菌、无娘藤、土茯苓、土菇等，属真菌界(Eumycetes)、子囊菌亚门(Ascomycota)、子囊菌纲(ascomycetes)、盘菌亚纲(Pezizomycetidae)、盘菌目(Pezizales)、块菌目(Pezizales)、块菌科(Tuberaceae)、块菌属(Tuber)。因其产量低、营养丰富、香味独特，在欧美售价可达500-1000美元/kg，在国内售价可达500-1000元/kg，享有“黑色钻石”“地下黄金”等美誉，具有极高的药用价值和营养价值。
由于采后的块菌子实体仍具有生理后熟作用，加上表面没有明显的保护结构，容易失水和受到微生物的侵染，出现变软、变黏和发臭等现象，导致块菌的贮、运、销环节受到影响，给产业发展带来较大的经济损失。目前，块菌保鲜主要采用低温保藏，可明显延长货架期，但因能耗高、设备要求高，经济投资较大。
现有技术记载了柠檬草精油和百里香精油各自具有一定的抗菌功能。但是，单一植物精油的抑菌谱较窄，因此，本文旨在探讨适宜块菌保鲜的柠檬草-百里香复合精油及其抑菌最佳体积配比，并利用微胶囊技术将液态精油转变成固态的粉末，在提高精油稳定性的同时，便于运输及后续使用。
发明内容
本发明的目的在于提供一种柠檬草-百里香复合精油微胶囊的制备方法和应用，旨在为块菌采后贮、运、销各环节的保鲜措施提供新的技术思路。
为了实现上述发明目的，本发明提供以下技术方案：
本发明提供了一种柠檬草-百里香复合精油在块菌保鲜中的应用，所述柠檬草-百里香复合精油中柠檬草精油和百里香精油的体积比为1～3:1～3。
优选的，所述复合精油抑制块菌贮藏中的枯草芽孢杆菌、粪小短杆、冻土毛霉、粉红粘帚菌和腐皮镰孢菌。
本发明还提供了柠檬草-百里香复合精油的微胶囊的制备方法，包括以下步骤：
(1)将柠檬草精油和百里香精油混合，得到复合精油；
(2)将所述复合精油与乙醇混合，得到精油乙醇混合溶液；
(3)将β-环糊精与水混合，水浴搅拌，得到β-环糊精过饱和溶液；
(4)所述β-环糊精过饱和溶液冷却后，搅拌，同时滴加所述精油乙醇混合溶液，水浴搅拌进行包埋，得到柠檬草-百里香复合精油微胶囊初品；
(5)用乙醇和水清洗柠檬草-百里香复合精油微胶囊初品，真空抽滤后进行真空冷冻干燥，得到柠檬草-百里香复合精油微胶囊。
优选的，步骤(2)所述复合精油与乙醇混合的体积比例为1：20～25，所述乙醇的体积分数为90％～98％。
优选的，步骤(3)所述β-环糊精与水的质量体积比为1：20～50，水浴的温度为60℃～80℃，所述搅拌的转速为500-1000r/min。
优选的，步骤(4)所述精油乙醇混合溶液与所述β-环糊精过饱和溶液的质量比为1:8～12。
优选的，步骤(4)所述水浴的温度为30℃～50℃，所述搅拌的时长为1.5～2.5h。
优选的，步骤(5)所述乙醇的体积分数为90％～98％，所述乙醇和水清洗的次数分别为2～3次，所述真空冷冻干燥的温度为-30～-50℃之间，时长为10-14h。
本发明还提供了所述制备方法制备的柠檬草-百里香复合精油微胶囊。
本发明还提供了所述的柠檬草-百里香复合精油微胶囊在块菌保鲜中的应用。
本发明确定了柠檬草-百里香复合精油对同时抑制块菌贮藏中的枯草芽孢杆菌、粪小短杆、冻土毛霉、粉红粘帚菌和腐皮镰孢菌的最佳体积比，可以为块菌采后贮、运、销各环节的保鲜措施提供新的技术思路。
附图说明
图1为实施例1中柠檬草和百里香复合精油在不同配比下的抑菌效果图；
图2为实施例2中柠檬草和百里香复合精油的全波长扫描谱图；
图3为实施例3中单因素分析包埋温度、时间、壁芯比对微胶囊包埋效果的影响结果；
图4为实施例4中β-环糊精、85.11％包埋率和24.34％包埋率微胶囊的电镜扫描图；
图5为实施例4中柠檬草-百里香复合精油微胶囊的热重曲线图。
具体实施方式
下面结合实施例对本发明提供的技术方案进行详细的说明，但是不能把它们理解为对本发明保护范围的限定。
实施例1
滤纸片法测定柠檬草精油和百里香精油在不同配比下的抑菌效果
在9cm平皿上(倾倒20mL的PDA培养基后，剩余平皿的空间体积约是0.07L)，分别涂布2株致腐细菌的菌悬液(107CFU/mL)，接种3株致腐真菌的5mm菌饼。在培养皿盖中心贴上边长1cm的方形灭菌滤纸片，在其上滴加0.7μL的复合精油(柠檬草精油和百里香精油按图1所示不同体积比混合得到)，由于精油是挥发性物质，通过微量移液器加入滤纸片之后，依靠挥发性布满平皿的整个空间，皿内浓度约为10μL/L。以无菌水为对照，各做3个重复。培养6d后，测定抑菌圈直径。利用SPSS26.0软件对试验结果进行分析。抑菌率＝(对照菌落直径-处理菌落直径)/(对照菌落直径-菌饼直径)*100％×100％。结果如图1所示。由图1可知，当平皿内精油浓度为10μL/L，V柠檬草精油:V百里香精油＝1:3时的抑菌效果最好，且对图1中5种主要致腐菌的抑菌率分别达到86.7％、76.7％、73.9％、67.3％和87.1％。
预实验显示，单一的柠檬草精油对枯草芽孢杆菌、粪小短杆菌、冻土毛霉、粉红粘帚菌和腐皮镰孢菌的抑菌率达50％时的浓度分别为15.5μL/L、18.8μL/L、30.5μL/L、15.5μL/L、40.5μL/L；单一的百里香精油对枯草芽孢杆菌、粪小短杆菌、冻土毛霉、粉红粘帚菌和腐皮镰孢菌的抑菌率达50％时的浓度分别为6.4μL/L、6.7μL/L、14.4μL/L、67.2μL/L、58.3μL/L；并且，单一柠檬草精油10μL/L时，对枯草芽孢杆菌和粪小短杆菌的抑制率分别为35.5和34.3％；单一百里香精油10μL/L时，对枯草芽孢杆菌和粪小短杆菌的抑制率分别为68％、68.3％；而V柠檬草精油:V百里香精油＝1:3的复合精油浓度为10μL/L时，对枯草芽孢杆菌、粪小短杆菌、冻土毛霉、粉红粘帚菌和腐皮镰孢菌的抑菌率分别达到86.7％、76.7％、73.9％、67.3％和87.1％，因此，与单一精油相比，复合精油之间协同抑菌的效果明显优于单一精油。
实施例2
制备复合精油微胶囊
a.分别量取柠檬草精油1mL、百里香精油3mL，两者按1:3的体积比混合，得到复合精油；
b.将复合精油与95％乙醇按1:25的体积比混合，得到精油乙醇混合溶液。
c.称取β-环糊精5g至于烧杯中，加入100mL去离子水，70℃水浴加热，搅拌，溶解形成β-环糊精过饱和溶液；
d.将β-环糊精过饱和溶液冷却至40℃后，放置于恒温加热磁力搅拌器上，匀速缓慢滴加精油乙醇混合溶液1:10至β-环糊精过饱和溶液中，并将温度控制在40℃，搅拌包埋2小时，待温度降低至25℃，将样品置于4℃环境中，冷藏24h后进行真空抽滤，得到柠檬草-百里香复合精油微胶囊初品；
e.使用95％乙醇反复清洗微胶囊初品3次，再用纯水反复清洗样品3次，使粘附在胶囊表面的精油完全洗净，真空抽滤，将得到的样品转移至真空冷冻干燥机中，-40℃下干燥12h，得到柠檬草-百里香复合精油微胶囊。
测定复合精油最大吸收波长：将步骤a得到的柠檬草-百里香复合精油，在190～1100nm波长条件下进行扫描。扫描结果如图2所示，柠檬草-百里香复合精油在295nm处有最大吸收峰。
实施例3
复合精油微胶囊的制备工艺优化
以复合精油作为芯材，β-环糊精作为壁材，通过测定包埋率探讨包埋温度、包埋时间、壁芯比等对复合精油微胶囊包埋效果的影响(实验过程同实施例2)。结果如图3所示。
采用干燥失重法测定精油包埋率：干燥失重法测定的包埋率(％)＝m总×(m1-m2)/m1/m精油×100％。式中：m总为制备所得复合精油微胶囊的质量(g)；m1为振荡干燥前的微胶囊质量(g)；m2为振荡干燥后的微胶囊质量(g)；m精油为制备所使用的精油的质量(g)。
图3为单因素试验的最佳包埋温度、包埋时间、壁芯比结果，经过对各因素的极差大小排列，最后得出最优的组合为包埋温度为40℃、时间2h、壁芯比10:1，此条件下包埋效果最好，包埋率可达85.11％。
实施例4
对实施例2制备的柠檬草-百里香复合精油微胶囊以及β-环糊精进行结构表征及缓释性能研究：(1)扫描电镜分析：用场发射扫描电子显微镜观察β-环糊精、精油微胶囊的微观结构及表观形貌。结果如图4所示，β-环糊精整体分布不均匀，质地粗糙，多为块状分布，表面裂隙明显。包埋率较低的复合精油微胶囊(同实施例2的制备方法一样，改变了包埋温度或包埋时间、壁芯比等得到的)，虽有成功包埋精油但微胶囊表面裂隙仍较多，质地粗糙且稍有塌陷。实施例2得到的柠檬草-百里香复合精油微胶囊表面无塌陷或褶皱现象，颗粒大小均匀，饱满充实，表面光滑连续性好。
(2)热重分析：采用热重分析仪对实施例1得到的柠檬草-百里香复合精油微胶囊的热重进行分析，测得TG曲线。其中，红色线是失重率；深蓝色线是热力学效应；浅蓝色线是对深蓝色线的一种标记。结果如图5所示，微胶囊在311℃之前的热失重较为缓和，可能是由于β-环糊精腔内结合水已被柠檬草、百里香复合精油取代，而复合精油与环糊精通过疏水键相结合，挥发受扩散机理影响，逐渐释放，使得失重和缓。从305.39℃开始微胶囊发生热分解，失重率急剧上升，直到321.42℃。此外，通过曲线可以看出微胶囊在43.36℃时出现了吸热峰，可能是因为糊精型开始转变，与精油两相分离，在约为270℃时因为热力学效应，胶囊结构开始被破坏，环糊精受热开始分解。而精油含有碳氧双键，结构较环糊精稍显稳定，在333.79℃后才开始热分解。上述规律说明，微胶囊化可提高精油的热稳定性，延缓柠檬草-百里香复合精油的挥发，同时也证明了复合精油微胶囊的成功制备。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
Claims (2)
1.一种柠檬草-百里香复合精油在块菌保鲜中的应用，其特征在于，所述柠檬草-百里香复合精油中柠檬草精油和百里香精油的体积比为1～3:1～3。
2.如权利要求1所述的应用，其特征在于，所述复合精油抑制块菌贮藏中的枯草芽孢杆菌、粪小短杆、冻土毛霉、粉红粘帚菌和腐皮镰孢菌。
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