Study On The Characteristics Active Substances Analysis And Quality For Garlic

Objective：Garlic（Allium Sativum L.）is used as both food and medicine forthousands of years, which has been listed in Pharmacopeia as traditional medicinalmaterials, dietary supplements and herbal medicine in many countries. But themonography are quite different, especially in Chinese Pharmacopeia. The purpose of thisstudy was to establish the TLC and HPLC methods and fingerprint for identifying aminoacids and sulfur-containing compounds in garlic. Utilizing mathematics analysis to extractand express information from fingerprint and screening the effective method ofdistinguishing garlic from other Allium L. species and garlic from different regions.According to foreign pharmacopoeia standards on garlic methods for the revision ofspecification of garlic in Chinese pharmacopoeia (2010) will be suggested. Microemulsionthin layer chromatography for grlic TLC identification will be studied and applied totwo-dimension thin layer chromatography. Methods:1.Established the fingerprint ofamino acids and sulfur compounds active substances for different Allium L. species andgarlic from different regions by TLC, respectively. similarity analysis, cluster, principalcomponent and numerical taxonomy were used to extract and express the TLC fingerprintfeatures of active substances of Garlic.2. Established the fingerprint of amino acids andsulfur compounds active substances for different Allium L plants and garlic from differentregions by HPLC, respectively. Several same mathematical models were used to analyzethe fingerprint of the two types of active substances.3. Established the TLC identificationand determination for Garlic, according to foreign pharmacopoeias. Water content，totalash and acid insoluble ash were tested additionally and the quality standard ofGarlic (draft) and drafting instructions were wrote.4. Microemulsion TLC was used toidentify the amino acids of garlic and applied it in the two-dimensional TLC for Garlic.Results：1.The TLC fingerprint conditions for amino acids and sulfur compounds in Garlicwere n-butanol：glacial acetic acid：water=4∶1∶1and toluene：ethyl acetate(10:3), respectively. Each compound was effectively separated under these conditions. However,the feature components were not effectively classified by extraction with differentmathematical methods.2.18kinds of amino acids were effectively separated by thephenylisothiocyanate derivatives methods for amino acids in Garlic.10chromatographicpeaks of sulfur compounds were effectively separated under the gradient elution with1%formic acid-methanol. Data analysis with various mathematical methods shows that themathematical expressions of the TLC and HPLC fingerprint of amino acids and the TLCfingerprint of sulfur compounds could not distinguish Garlic from other Allium L Plants,however, the mathematical expressions of the HPLC fingerprint of sulfur compoundscould effectively distinguish Garlic from other Allium L Plants, the mathematical model ofnumerical taxonomy could be used in the quality comparison among Garlic.3. Establishedthe thin layer identification and determination for Garlic with potential allicin, alliin andarginine as index components. The water determination of Garlic was established and thelimitation was less than65.0%. The content of alliin, arginine, allicin is not less than1.5%、1.0%、0.60%(dry weight).4. The optimal microemulsion TLC identification methodof Garlic was20%SDS-n-butanol-n-hexane (3.0g-17.3ml-3.0ml)-water microemulsionsystem.The two dimensional TLC for garlic, which was effectively separated by organicsolvent–microemulsion development. Conclusion:1.For rich sulfide active substancesand the particular enzyme system in garlic and research status at home and abroad, aminoacids of enzyme inactivation system and sulfur compounds of enzyme active system as theactive substances to research are reasonable. TLC and HPLC conditions established can beused to separate active substances of the amino acids and sulfur compounds in garlic, andthe method validation of fingerprint meets the guiding principles.2. Several mathematicalmodel sused to extract and express the fingerprint of active substances of aminoacids,There are have a high similarity among garlic from different regions and a certainsimilarity with same generic plants. But the active substances of amino acids fingerprintcharacteristics are not sufficient to distinguish garlic.3. The reaction which characteristicalliinase participated in could generate the sulfur compounds, whose HPLC fingerprintcould effectively distinguish garlic from other Allium species, even the quality evaluationfor garlic of different origins. HPLC fingerprint of sulfur compounds in the alliinaseactivation system can be uesd to effectively distinguish the garlic from other plants.4. Toeffectively distinguish between the fingerprint of the active substances of amino acids,which shuld extract the characteristic information from fingerprints, using appropriatemathematical models, or creating a viable mathematical model to extract and digital express the different information.5. Garlic quality standards established in this study hadconnected with international standards, which could provide basis and reference foramending the garlic quality standards in the Chinese Pharmacopoeia.