Studies On Structure Of Bark And Chemical Component Of Lacquer And The Bark In Toxicodendron Vernicifluum (Stokes) F.A. Barkley

Oriental lacquer which is wellknown used as the earliest excellent natural coatings, called as the king of paint,is one of the most important raw materials in mordern industries,angriculture,national defence,technology and civil use,having been used for more than 6900 years.The relic painted by oriental lacquer can keep their state for thousands of years,and it is shown that the durabilities of the synthesis paint can not reach those of oriental lacquer.Besides Oriental Lacquer,the flowers of lacquer tree not only contain abundant sweet liquid but also are important nectar source plant.The oil which is squeezed by fructification,can be edible and used as material of paint.The leaves,flowers, roots,skin,fruits,resina toxicodendri and heartwood of lacquer tree can all be used as medicine.Resina toxicodendri registered in the pharmacopoeia of the nation,has the character istics of symplectic.warm.poisonous.meridian,and breaking congestion. eliminauion integrable.disinsection,used for the treatment of amenorrhea.congestion. bollyache of integrable of insect.Toxicodendron vernicifluum(Stokes)F.A.Barkley originated in China and distributed in most provinces or region of China,such as Shaanxi,Hubei,Sichuan,Chongqing,Gansu, Guizhou and Yunnan,The maximal yield of Oriental Lacquer distributed in the seven provinces above.At present,although many studies on constituents in extracts of oriental Lacquer focus on the mensuration of their content and structure and the research of their antiseptic performances,little attention has been paid to system analysis and comprehensive estimate on the component and content of oriental Lacquer from different origins and species. Moreover,oriental Lacquer from different origins,varieties,altitude is greatly different in the aspects of appearance,quality,performance,being lack of some efficient ways of quality control of them.There is some problem further to research into biology,such as to differentiate varieties,confirm on relative of in Lacquer tree.Based on high performance liquid chromatography(HPLC)and high perfor- mance liquid chromatogrammphy-electrospray ionization trap mass spectrometry(HPLC--ESI-MSⁿ),constituents and main chemical compound have been analysed in Oriental Lacquer,and besides,way that authenticated the constituents of urushiol have been detected by HPLC.On this basis,the main chemical components of urushi and lacquer tree bark extract were analysed from different regions of Shaanxi and 13 varieties,the established extracts HPLC fingerprint of Oriental Lacquer and lacquer tree bark. Application of plant anatomy methods comparative have researched on different lacquer tree bark structure above 13 samples.Through the several aspects of the study,with a view to improving the quality of Oriental Lacquer current detection method to identify different regions of the Oriental Lacquer quality and lacquer tree varieties provide different chemical indicators.The main results acquired through these systematic studies as follows:1.Application of high performance liquid chromatography(HPLC)and high performance liquid chromatogrammphy-electrospray ionization trap mass spectrometry(HPLC-ESI-MSⁿ) analyse and determine the urushiol compounds in the Oriental Lacquer.Instrument model of HPLC-ESI-MSⁿ is 1100 series of Agilent Co.Ltd..The characters were as follows: Agilent ZORBAX C-18 columns(150×4.6 mm,5μn),flow rate:0.8 ml/min;mobile phase: the volume ratio between methanol and ammonmium acetate of aqueos solution:82:18, column temprature:35℃;uv detection at 225 nm;volume of injecting sample,10μL;ESI ionization source,negative ion mode,the press of evaporative gas:35.0 psi,the flow rate and temprature of drying gas:7.0 L/min,350℃;the range of scanning the electronic charge-mass ratio:50～1200m/z.According to the ion flow diagram of HPLC-ESI-MSⁿ identified nine kinds of urushiol compounds.This method not only can be separate and identify Oriental Lacquer urushiol compounds as effective and rapid but also provide information of compounds structural.2.Application of HPLC and HPLC-ESI-MSⁿ connected technology,urushi constituents which processed resina toxicodendri is analysed.Finnigan Surveyor-LCQ Advantage MAX (LC/MS)has been used.The characters were as follows:Hypersil ODS2 chromatography column(5μm,2.0×150 mm),the flow rate was 0.2 ml/min,detecting wavelength was at 225 nm and the mobile phase was that methanol compared with water is equal to 88:12(v/v),the injection volume was 10μl;ESI ionization source,negative ion mode,the press of evaporative gas:35.0 psi,the flow rate of drying gas:5.0 L/min,capillary temprture:350℃,capillary voltage:-47 V;power voltage:4.5 KV;source current:80.00μA,the range of scanning the electronic charge-mass ratio:50～1200 m/z.According to separating 11 components by liquid chromatography and identifying nine kinds of urushiol constituents by ionization trap mass spectrometry,urushiol constituents of oriental Lacquer which processed resina toxicodendri can be efficiently and fast separated in this way.3.The HPLC fingerprints of Oriental Lacquer and the extraction of lacquer tree bark were established.The main components of them were determined by HPLC-ESI-MSⁿ.Analysis by HPLC conditions were as follows:DIKMA C₁₈column(5μm,4.6 mm×250 mm)was used,the flow rate was 0.8 ml/min,detecting wavelength was at 225 nm and column temperature was at 35℃,Mobile phase:methanol and water ratio of 83:17(v/v),time for analysis was 130 min,the injection volume was 10 ul.The 13 raw lacquer samples and lacquer tree bark extract samples of Shaanxi Province were analyzed by HPLC.The fingerprint constructed by Oriental Lacquer and the chemical constituents from the petroleum ether extract of lacquer tree bark showed seven common peaks,and that of the chemical constituents from the methanol extract of lacquer tree bark showed five ones. Validating with methodology,RSD of precision,stability,reappearance were all less than 3%.Through similarity evaluation software(2004A)published by committee of codex, analyzing similarity of the fingerprints,whose similarity was all more than 0.990,it is revealed that the quality of 13 kinds of sample of lacquer was roughly the same,the average similarity of the chemical constituents from the methanol extract of lacquer tree bark was 0.974,it is revealed that the quality of 13 kinds of sample of the extraction of lacquer tree bark was roughly the same,belong to the same species,and that of the chemical constituents from the methanol extract of lacquer tree bark was low,whose average was 0.799,we can acquire that the chemical constituent of 13 kinds of the petroleum ether extract of lacquer tree bark present some diversity.4.Lacquer tree bark from the stem which is made up of secondary phloem,periderm and dead bark around it is main part cutting urushi..The secondary phloem consists of the axial system and the radial system,the axial system consists of sieve vessels,companion cells, parenchyma cells,stone cells and laticiferous canals;the radial system is ray.The bark structure of 13 lacquer tree samples which using safranine and fast green double dyeing were observed by the paraffin section method.The results show that the similar in structure. According to the statistical analysis the amount and area of laticiferous canals in per unit area and diameter of laticiferous canal existed certain difference in 13 kinds of samples of lacqer tree bark.5.Through the cluster analysis for indexes about urushiol content in Oriental lacquer of 13 kinds of samples of lacqer tree,that of the extraction of lacquer tree bark,amount and area laticiferous canals in per Unit Area by Statistica which is statistical software,the result show:the relationship between the first two samples analysed in chemical index was roughly the same,while that of analysed in anatomy existed certain difference.According to the result analysed from three aspects above,the samples whose relationship were close were:Da Hongpao and Jin Zhouhong,Longxian Da Mu and Taibai Da Mu,and those analysed from contents of chromatographic peaks of Oriental Lacquer sample and the chemical constituents from the methanol extract of lacquer tree bark were:Da Hongpao and Jin Zhouhong;Huang Ronggaobachi,Huo Yanzi and Qie Ketou;Longxian Da Mu, Nanzheng Da Mu and Tai Bai Da Mu.