Quantitative Analysis Method Of Garlic, Alliin And Alliinase And Quality Standard Of Alliin Research

Objects Garlic (Allium Sativum L.) contains large numbers of biology activity matter an nutrition composition, In recent years, many studies have found that garlic possess a vast variety of biological effects such as antimicrobial, anti-inflammatory, antithrombotic and anticancer as well as for the the prevention of stroke and arteriosclerosis. More recently it has been proved that the active principle of garlic is assumed to be related to allicin (allyl-2-propenethiosylfinate). In the presence of alliinase, allicin and ajoene are formed from the precursor alliin (S-(+)– allyl–L–cysteine sulf oxide ). Because that many garlic preparations have lower content ,curative effect and easier preparation technics, and the large numbers of garlic preparations are selled as health protection, accoding to design of our gourp, relatively steadily Alliin and Allinase are distilled from garlic bg the technic of modern biology pharmacy, and then"high effect garlic preparation"will be executed by this Alliin and Alliinase. Determination the content of Alliin which as the uppermost activity composition and origination of flavor substance in garlic can provides credible gist for medium-size examination for the future. And analysis of each examine point can provides quality control for medium-size examination for the future in course of medium-size examination of Alliin and Alliinase. Finally , the quality standard (draft) and explain of standard of raw material Alliin which was gained from medium-size examination production was established. Methods Quantitative analysis of garlic, alliin, alliinase are researched using HPLC, and the examine point in medium-size examination of Alliin and Alliinase are determined. finaly, the quality standard (draft) and explain of standard of raw material Alliin which was gained from medium-size examination production was established. Results Analysis methods of Garlic , Alliin and Alliinase are established in this text. Outer-standard method and inner-standard method are introduced in determination of Alliin of Garlic, and are contrasted; And the two method are contrasted with the method of USP ; HPLC determination the activity of Alliinase is established. Upwards condition of determination as follow: 1.Analysis methods of Garlic, Alliin and Alliinase1.1 Determination the content of garlicOuter-standard method: column Hypersil BDS C18 4.6mm×250mm, 5 m, mobile phase: 0.04% CF3COOH, detection: UV 214 nm, flow rate 0.5 ml min-1. Inner-standard method: column Waters symmetry shield RP C18 4.6mm×250mm, 5m, mobile phase: water, detection: UV 214 nm, flow rate 0.5 ml min-1.1.2 Determination the content of Alliin(50%) Column Hypersil BDS C18 4.6mm×250mm, 5 m, mobile phase: methanol:0.02% CF3COOH(5:95), detection: UV 214 nm, flow rate 0.5 ml min-1.1.3 Contrastion with USP methodColumn Thermo ODS 4.0 mm×100 mm, 3μm, mobile phase: 0.045mol·mL-1 phosphate buffer: acetonitrile: 1,4-dioxane: Tetrahydrofuran(69.9 :25: 2.9: 2.2), detection: UV 337 nm, flow rate 1.0 ml min-1.1.4 Determination the activity of Alliin(50%)Column Hypersil BDS C18 4.6mm×150mm, 5 m, mobile phase: methanol: water (55 : 45), detection: UV 214 nm, flow rate 0.5 ml min-1.2.Examine point determination in medium-size examination of Alliin and AlliinaseThis text have confirmed that the determination method of Alliin medium-size examination is HPLC, and the condition is column Hypersil BDS C18 4.6mm×250mm, 5 m , mobile phase: 0.04% CF3COOH, detection: UV 214 nm, flow rate 0.5 ml min-1; And the determination method of Alliinase medium-size examination is UV, determining the activity of Alliinase with pyruvic acid method, determining the content of Alliinase protein with Bradford method, thereby activity of Alliinase will be gained.3.Study of quality standard of AlliinQuality standard of Alliin(≥50%) and the explain are established. Conclusion quantitative analysis method of garlic,alliin,alliinase have be established in this text , The method proved to be simple, nicely, rapid and suitable; the examine point in medium-size examination of Alliin and Alliinase are determined; Quality standard of Alliin(≥50%) and the explain are established.