基于ITS基因条码的保健食品中植物源性成分的鉴定

Utilization of ITS Gene Barcodes for Identifying Plant-derived Functional Ingredients in Health Food Products

【目的】研究并建立运用ITS基因条码技术针对保健食品常见植物原料源性成分的分子生物学鉴定技术。【方法】通过采用回顾性调查方法,选取西洋参(Panax quinquefolius)、铁皮石斛(Dendrobium candidum)以及红景天(Rhodiola nobilis)、枸杞子(Lycium barbarum L.)与人参Ginseng(Radix Ginseng)作为研究对象。探索其作为生产原料的常见保健食品剂型中,目标DNA的最佳提取方法。运用ITS2内转录间隔区保守特性设计引物序列,并通过调整优化体系中各物质的组分,由此构建稳定高效准确的PCR反应体系。【结果】采用改良CTAB法可实现对所研究的保健食品中常见剂型目标DNA的提取,纯度及质量达到反应要求。所建立的PCR法扩增效果良好,对西洋参、铁皮石斛以及红景天的方法灵敏度均为1 ng·μL^(-1),枸杞子与人参的方法灵敏度为0.1 ng·μL^(-1)。【结论】运用建立的方法可实现对保健食品中常见植物性原料西洋参、人参、铁皮石斛、枸杞子、红景天成分的鉴定,适用于丸剂、片剂、口服液等不同剂型的保健食品中原料鉴别,可运用于对保健食品中植物源性的检测。

【Objective】A molecular biology identification method to detect common plant-derived functional ingredients in health food products using genetic barcodes was developed.【Method】By means of retrospective investigation,5 common health food raw materials in China,i.e.,American ginseng(Panax quinquefolius),Dendrobium candidum,rhodiola(Rhodiola nobilis),wolfberries(Lycium barbarum L.),and ginseng(Radix Ginseng),were firstly tested for functional ingredients using the best extraction method on the target DNA.A stable,efficient,and accurate PCR reaction system was constructed with primer sequence designed based on the conservative characteristics of ITS2 internal transcribed spacer and components of each substance adjusted and optimized.Molecular identification on health food products of various forms was tested using the developed method.【Result】The innovative application of a modified CTAB method afforded successful extraction of the target DNA in health food materials as well as products at commonly formulated dosages.The PCR amplified with the detection sensitivity on nucleic acids of American ginseng,D.candidum,and rhodiola reached 1 ng·μL^(-1),and of wolfberries and ginseng 0.1 ng·μL^(-1).【Conclusion】By using the ITS gene barcodes obtained through the DNA extraction and PCR amplification,the key functional ingredients from plant materials such as American ginseng,ginseng,D.candidum,wolfberries,and rhodiola rosea could be identified for authentication of health food products in the forms of pills,tablets,and concoctions.