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基于蜂王浆与盐酸显色反应和脂肪酸组成的蜂王浆质量控制研究
Studies on Quality Control of Royal Jelly Based on Chromogenic Reaction between Royal Jelly and HCI and Fatty Acid Compostion
【作者】 魏文挺;
【导师】 胡福良;
【作者基本信息】 浙江大学 , 特种经济动物饲养, 2014, 博士
【摘要】 长期实践和研究表明,蜂王浆在常温下容易变质,新鲜度评价是蜂王浆质量控制的重要方面。然而,由于缺乏简单、有效且被广泛接受的评价方法,目前蜂王浆国家标准中没有涉及新鲜度相关评价指标及检测方法,有关蜂王浆质量评价主要以反式10-羟基-2-癸烯酸(10-HDA)为技术指标,是影响蜂王浆价格的决定性因素。然而10-HDA含量却逐年下降,导致出口贸易和内需消费均受到严重影响,已经引起国内科研工作者和相关企业的高度重视。因此,亟需开展蜂王浆新鲜度及质量控制相关研究,为完善蜂王浆国家国际标准和整个产业的健康发展提供技术支持和理论依据。本研究一方面基于蜂王浆与盐酸显色反应,试图寻找一种简单、有效、快速、实用的蜂王浆新鲜度指标和相应检测方法;另一方面根据脂肪酸合成路径,采用饲喂有机酸方式提高蜂王浆中10-HDA等多种脂肪酸的含量,以提升蜂王浆品质;同时,对影响10-HDA含量的多种因素进行系统分析,获得可靠的证据。具体结果如下:1.基于蜂王浆与盐酸显色反应建立了一种全新的蜂王浆检测方法。显色反应a木、b*值随新鲜度降低会发生规律性变化,不同类型蜂王浆变化略有不同。杂花期蜂王浆:利用显色反应Y(Y=a*-b*)值,可以有效区分室温下贮存7d前后、9d前后、21d前后的蜂王浆样品。判别分析结果显示将新鲜度分为0-5、7-9、>11d等集合对不同温度下贮存样品进行判定可以获得良好的区分效果。油菜花期蜂王浆:根据a*值统计分析结果,室温下样品可以分为0-7、9、11~17、21、28和35~57d等区间,根据b木值则可分为0-11、14~17和21~57d等区间,区间内样品差异不显著,但与区间外差异显著。判别分析结果显示,将新鲜度分为0~11、14~17、>21d等集合对不同温度下贮存样品进行判定可以获得理想的判定结果。2.首次建立一种同时测定蜂王浆中10-羟基癸酸(10-HDAA)、3-羟基癸酸(3-HDAA)和癸二酸的气相色谱检测方法。利用该方法结合10-HDA国标法测定了蜂群饲喂有机酸前后生产的蜂王浆中10-HDA、10-HDAA、3-HDAA和癸二酸含量。结果显示,饲喂柠檬酸和硬脂酸可以显著提高10-HDA和癸二酸含量,提升效果分别可达5.6%-6.4%和22%-46%。柠檬酸还对10-HDAA和3-HDAA含量有显著提升效果,增产幅度分别为14.4%-20%和10.6%-23.4%。3.首次对10-HDA含量的影响因素进行了系统性、大样本量的研究。地理和蜂种均可显著影响10-HDA含量。在中国,10-HDA含量从高到低依次为西部地区(2.01%)>东北地区(1.87%)>东部地区(1.75%),意卡杂交蜂(Apis mellifera ligustica×A. m. carnica)(1.89%)>意大利蜜蜂(A. m. ligustica)(1.79%),其中地理为主要因素。油菜花期蜂王浆10-HDA含量显著高于荆条花期样品。蜂王浆产量与10-HDA含量呈显著负相关,油菜花期内两者相关系数为-0.362,杂花期内为-0.432。
【Abstract】 Long-term research and practice showed that royal jelly (RJ) spoils easily under room temperature, which indicates that freshness is closely related to its quality. However, freshness is not involved in national standard of RJ because of lacking of simple, effective and widely accepted detection methods. The content of trans-10-hydroxy-2-decenoic acid (10-HDA) is the most important marker in RJ, which directly determines the price of RJ in commercial trade. However, the10-HDA content declines a lotcompared to several years ago, which affects the export and native consumption. As a result, researches on RJ freshness and quality control that providing theoretical support and basic data is urgently necessary.In this study, a simple, fast and effective freshness assessment method was established based on a chromogenic reaction between RJ and HC1. Elevation of10-HDA content in RJ was achieved by feeding organic acids. And also, the factors which could influence10-HDA content were analyzed based on a large amount of samples with different origins. The results were showed as follow:1. A new RJ freshness measurement method was established based on a chromogenic reaction between RJ and HC1. The a*and b*value change as the freshness, the varying patterns differ a little between RJ produced on various kinds of pollen and on rape pollen. For RJ produced on various kinds of pollen: According to Y (Y=a*-b*) value, samples stored at RT got a good separation before and after7d,9d and21d. Discrimination analysis results showed that samples stored at different temperature for different storage could be correctly classified into proper freshness intervals based on RT samples.For RJ produced on rape pollen: according to a*value, samples at RT could be divided into0-7,9,11-17,21,28and35-57d intervals; While using b*value,0-11,14-17and21-57d intervals could be set. Based on a*and b*value, discrimination analysis showed a good classification of RT samples into0-11,14-17and21-57d intervals, and samples stored at different conditions could be correctly classified into proper freshness intervals based on RT samples.2. A gas chromatography (GC) method for detection of10-hydroxydecanoic acid (10-HDAA),3-hydroxydecanoic acid (3-HDAA) and sebacic acid was developed for the first time. Using this method combined with10-HDA detection method in national RJ standard,10-HDA,10-HDAA,3-HDAA and sebacic acid content in RJ produced before and after feeding with organic acids were detected. The results indicated that,10-HDA and sebacic acid could be elevated by5.6%-6.4%and22%-46%, respectively, by feeding citric or stearic acids. The content of10-HDAA and3-HDAA arised by14.4%-20.0%and10.6%-23.4%, respectively, after feeding citric acid.3. Factors influencing10-HDA content were studied systematically based on a large number of samples. The content of10-HDA could be affected by both geography and bee breed, in China, Western Group (2.01%)> Northeastern Group (1.87%)> Eastern Group (1.75%), Apis mellifera ligustica×A. m. carnica (1.89%)> A. m. ligustica (1.79%), geography is the dominant factor. The content of10-HDA in RJ produced on rape pollen is significantly higher than that in RJ on vitex pollen. A significant negative correlation between yield and10-HDA content of RJ has been observed, the correlation coefficient of these2parameters is-0.362for RJ produced during rape flowering season and-0.432for that produced on various kinds of pollen.
【Key words】 royal jelly; freshness; chromogenic reaction; 10-HDA; fatty acid; influencing factor;