Determination Of Alliin,diallyl Disulfide And Diallyl Trisulfide In Garlic By High Performance Liquid Chromatrgraphy

Garlic is a kind of allium plants and always use as food and drug in common people’daily life.People pay attention to the quality and safety of food and drugs now.In this trend of eagering to return to nature,Garlic has so many adventages such as inhibiting and killing the growth of bacteria,preventing cancer defending against cancer,preventing and treating cardiovascular and cerebrovascular diseases and improving immunity that it becomes the hot topic of the reasearch.In America,There are dozens of health care products, medicines and food supplements which are made from garlic.Alliin is the specific endogenous amino acid in the fresh garlic.In the complete garlic cells,alliin exists in vacuole and the alliin enzyme that can decompose alliin is located in the cytoplasm.The two substances are separated by the cell membrane.When garlic cell is damaged,such as estrusion and comminution,alliin is cracked into a series of important sulfur-containing compounds under the action of alliin enzyme.Alliin and the series of sulfur-containing compounds that generated by alliin are the important bioactive substances in garlic.we should establish a simple, fast, sensitive and accurate method to detect the content of the important sulfur-containing compounds in garlic.That is beneficial to control the market’s quality effectually of pharmaceutical raw materials, cosmetic, feed and food additive,standardize garlic products and guarantee sustainable development of garlic industry steadily.If so,It can bring social acceptance value and great economic benefits and provide convenience and positive impace for multi-purpose development and deep processing of garlic to relevant enterprises.This paper is divided into four chapters:The first chapter:firstly a brief introduction about the study of garlic,the main ingredients and the benefits of garlic, the active ingredients and the relations between them was introduced;secondly the nature and benefits of alliin,diallyl disulfide and diallyl trisulfide were described briefly;thirdly the research progress of the alliin,diallyl disulfide and diallyl trisulfide’s detection methods was summarized.The second chapter:Establish a High performance liquid chromatography with photodiode array detector analytical method for determination of alliin in garlic.The alliin was extracted from the fresh garlic peeling after microwave out the enzyme by 0.01% phosphoric acid aqueous solution.The chromatographic separation and determination was carried out on an C18 column(4.6 x 250 mm, 5μm), using 0.01% phosphoric acid solution as mobile phase, at a flow rate of 0.8 m L/min.The chromatographic column temperature was kept at 30 ℃, The injection volume was 20μL.Photodiode array detector detected and the alliin was qualitated by retention time and quantitated by the peak area which detected under 214 nm.Optimized a series of factors of sample pretreatment conditions including the time of microwave treatment, extraction solvent, extraction time, extraction temperature,solid-liquid ratio and ultrasonic power when ultrasonic extraction, the pretreatment conditions showed that the microwave treatment method’s optimum conditions was 900 W,1min;solid-liquid ratio was 5:100(g/m L); under the temperature of 50℃,the ultrasonic extraction conditions was 300 W,15min.The linear range of the method was 5.0~500.0 u g/m L.The standard addition recovery was form 96.8% to 100.7%, and RSD was 1.1%~2.5%.The method is simple, rapid and high sensitivity.The third chapter:Establish a High performance liquid chromatography analytical method for determination of diallyl disulfide and diallyl trisulfide in garlic. The experiment determined the optimum conditions of enzymolysis,extraction and chromatography through comparing and screening.Linearity between the diallyl disulfide concentration and the peak area was good within the concentration ranges from 6 to 240 μg/ m L, R2 was 0.9989,and the diallyl trisulfide’s standard curve was linear within the range of 147.5~1475.1μg/ m L,R2 was 0.9991. The results were diallyl disulfide of recovery was 91.83%~100.04% and RSD was 2.74%~3.49% and diallyl trisulfide of recovery was from 91.26% to 99.92% and RSD was 0.46%~3.15% when we added the low, medium and high three levels of the standard solution to the garlic samples(every level repeated three times).The four chapter: The research work of this paper was summarized simply; the two kinds of detection methods were analyzed and compared in the paper;the content need to be completed in this study and the bottlenecks and problems should to be solved in garlic industry were proposed.