| ObjectiveCommonly used traditional Chinese medicine(TCM)sandalwood for sandalwood Santalum album L.plant trunk heartwood,have the effect of gas pain.Natural conditions,sandalwood heartwood form extremely slow,generally require more than 30 years to become useful for medicinal use.Reported from the artificial fragrance method can only make the sandalwood plant local node(form heartwood),hard to implement industrialization.To this end,the author has carried out biological candidated induced sandalwood overall knot technology research,and a preliminary evaluation and sample quality,in order to provide some commonly used for the production of modern scientific basis.Methods1 Experimental study of stimulate Method1.1 The effect of stimulation way in sandalwood heartwood formationRefer to previous induced junction way,select 5 years gave birth to healthy sandalwood plants as experimental object,design 1 hole injection fluid,dozen 2 hole fluid injection and infusion group,observation records 1 week,1 month,3 months,6 months of leaves,branches,trunk growth,at the same time,a detailed record experiment after 1 month,3 months,6 months,12 months sandalwood heartwood color,smell,and so on and so forth,induced to choose the best way.1.2 The effect of stimulation reagents in sandalwood heartwood formationSelect 5 years gave birth to healthy sandalwood plants as experimental object,design thiamethoxam benzene lung,green fresh vegetable,formic acid,acetic acid,hydrogen peroxide,biological inducers group,observation records 1 week,1 month,3 months,6 months of leaves,branches,trunk growth,at the same time,a detailed record 1 month,3 months,6 months,12 months sandalwood heartwood color,smell,etc.,choose the best inducer.1.3 Induced optimal concentration and dose of a-reagent experimentSelect 5 years gave birth to healthy sandalwood plants as experimental object,design respectively 1.0%,1.5%,2.0%,2.5%,3.0%,3.5%concentration of biological inducers category,dosage respectively 60,70,80,90,80 ml,experimental observation records after 1 week,1 month,3 months,6 months of the leaves,branches,the tree growth situation,detailed record experiment after 6 months,9 months,12 months sandalwood heartwood color,smell,and so on and so forth,to 12 months after the quality and production as a key evaluation index,select the best concentration and dosage.1.4 Verify and optimization the sandalwood optimal stimulation techniqueSelect 5 years gave birth to healthy sandalwood plants as experimental object,each set of 5 strains,were divided into group A(2.5%drip biological inducers 80 ml)and group B(80 ml)drip 1.5%biological inducers,group C(2.5%biological inducers 80 ml,drip to leaf),observation records 1 week,1 month,3 months,6 months of leaves,branches,tree growth,at the same time,A detailed record after the experiment 1 month,3 months,6 months sandalwood heartwood color,smell,etc.,at the same time in six months after the production as evaluation index,optimize the biological candidated knot incense technology and verification.2 Quality analysis of sandalwood heartwood sample by "Bio-physical stimulation method"2.1 Preliminary evaluation of sandalwood heartwood sample by "Bio-physical stimulation method"Sandalwood output induced knot using volume method,observed by the drilling method and record knot height,DBH,volume calculation formula of sweet = 3.14 x(diameter at breast height/2)2 x height.Will be commercially available different batch samples,induction and 6 months,12 months and the base of natural fragrant herbs samples quality analysis.According to the 2015 edition of China pharmacopoeia "(a)method under the sandalwood requirement,on the behavior of the experimental samples,microscopic,thin layer test.According to"Chinese pharmacopoeia" 2015 edition(four)the general principles of the 0832 "moisture determination method" under "the fourth method(toluene)" determination of moisture.Quenching method with ether ether extract content;According to the 2015 edition of"China pharmacopoeia"(four)determination of general principles of the 2204 volatile oil"armor" sandalwood essential oil content.2.2 Determining content of 9 kinds of inorganic elements in samples of sandalwood incenseUsing flame atomic absorption spectrophotometer method and graphite furnace method determination of 9 kinds of inorganic elements in samples 15 batches of sandalwood,Cu,Fe,zinc,Mn,Ca,Pb,As,Hg,Cr content,using SPSS20.0 content results are principal component analysis,correlation analysis and cluster analysis.2.3 Study on HPLC fingerprint of sandalwood incense samples.Kromasil C18 chromatographic column(5.0 mu m3 4.6 x 250 mm).Mobile phase:0.2%phosphoric acid aqueous solution(C,v/v)acetonitrile,gradient elution;Flow rate:1 ml/min;Detection wavelength of 254 nm;Column temperature 25 ?;Sample quantity:10 mu L.To establish the fingerprint of different batches of sandalwood samples by HPLC-UV method,the use of traditional Chinese medicine "refers to the State Pharmacopoeia Commission.Pattern similarity evaluation software 2004A version of the 12 batches of samples were analyzed and compared to determine the common peaks and the similarity,the use of SPSS20.0 software for cluster analysis.HPLC-UV method is adopted to establish the fingerprint of different batch samples of sandalwood,using the national pharmacopoeia committee "traditional Chinese medicine(TCM)fingerprint similarity evaluation software version 2004A" 12 batches of samples are compared,and spectrum analysis,identify common peak and similarity using SPSS20.0 software cluster analysis.2.4 Sandalwood incense knot samples analysis and GC-MS fingerprint of volatile oil componentsThe temperament-mass spectrometry analysis of 15 sandalwood chemical composition of the sample.HP-5 quartz capillary column,injection port temperature 270 ?;Temperature program:column temperature of 90?,5?.min-1 to 160?,and then to 0.5?.min-1 to 165 ?,and then to ?.min-1 to 180?,finally at a rate of about 10?.min-1 to 230?.Vaporizing chamber temperature:250?,the carrier for high purity helium;Before the column pressure:52.6kPa,carrier gas flow rate:1.0 mL.min-1;Sample quantity:1 mL;The split ratio is 20:1.Sandalwood essential oil for composition analysis and establish the fingerprint of the central south university "similarity evaluation software"similarity analysis,application SPSS20.0 statistical software for data clustering analysis,establish a method to control the quality of sandalwood essential oil.Results1 Experimental study of stimulate Method1.1 The effect of stimulation way in sandalwood heartwood formationThree ways of experimental phenomena have fallen leaves,a week after about a month after slowly recovering,after 3 months induced hole parts appear brown substancer,solid,burning with fragrance,about six months later,most of the injection agent of tree body around induced hole palm red,and the blank control group and the negative control group had no this phenomenon appears.6 months and 1 year after digging and truncated to look at the tree body,1 hole fluid injection method,dozen 2 hole fluid irjection method and infusion fluid injection method can form a palm red heartwood,three methods induced heartwood from the material in production have significant differences,infusion method were significantly influenced by area is larger than 1 hole and two hole injection fluid method,and liquid infusion method to fastest delivery.1.2 The effect of stimulation reagents in sandalwood heartwood formationGreen fresh vegetable,formic acid,acetic acid,biological inducers and thiamethoxam benzene lung group can make the sandalwood incense,hydrogen peroxide group did not knot fragrant effect,inductive substance of sandalwood incense production from high to low is:biological inducers benzene lung>green fresh vegetable>thiamethoxam formic acid>acetic acid>hydrogen peroxide,and medicinal materials produced by biological inducers induced sandalwood 1 year blond color,strong fragrance,heartwood diameter significantly greater than the other group,the best quality.1.3 Induced optimal concentration and dose of a-reagent experimentConcentration of 2.5%,sandalwood experiment plant heartwood length can reach 248.3 cm a year later,about 4.9cm in diameter and heartwood color scarlet,qualitative hard,strong aroma,is better than that of 1%,1.5%and 2%concentrations incense heartwood experiment group,the concentration of 3%and 3.5%,the tree wither or death occurs;2.5%concentration of biological inducers,80 ml of inducing category heartwood from the length of 258.3cm,4.6 cm width,dosage of heartwood quality is better than the other group,90ml and 100 ml dosage,the plants withered,decay and death.1.4 Verify and optimization the sandalwood optimal stimulation techniqueBiological inducers three groups of sandalwood,incense production of 2495.92 cm3 half A year,biological inducers is significantly higher than group A(annual output 1972.88cm3 half a year)and biological inducers of group B(the average yield of 624.98 cm3 half a year),biological inducers heartwood group C sample color,smell,have reached 10 years raw natural sweet sandalwood,advance the "sweet point for 10 years,and from the root to the trunk 191.70 cm overall knot.2 Analysis on the yield and quality of sweet stimuli induced by biological samples of sandalwood2.1 Sandalwood incense yield and the conventional evaluation stimulus node sample qualityIdentification results showed that six months are medicinal heartwood class material,heartwood from the 12 months quality better,yield an average of about 4243.12 cm3,heartwood palm red color and quality of a material is solid,is not quickly broken.Smells faint scent,fragrance thick.Microscopic identification found induction and incense are bordered pit containing pharmacopoeia rules catheter,calcium oxalate Fang Jing,crystal thick-walled cells such as material,but the amount of these basic characteristics between different samples or form exist certain differences,induction and perfume sample pigment block is relatively small,fiber,wood fiber and wood ray is light yellow,Fang Jing;TLC identification found induction knot samples of 6 months and 12 months and sandalwood oil reference substance in different Rf location have the same spots;Water content,ether extract content determination results show that the induction of 6,12 months and the samples in 11.75?11.96%moisture content,ether extract was 1.45%?1.45%;Induced to volatile oil content in six months was 1.14%,the induction of 12 months samples volatile oil content of 2.84%.Commercial samples of sandalwood essential oil content is between 1.49%-3.84%.2.2 Determining content of 9 kinds of inorganic elements in samples of sandalwood incenseThe average content of inorganic elements(?g/g)order from high to low is:Cu(3.5110-52.6808,average 18.7420)>Fe(0.1840-22.7640,average 6.4791)>Mn(1.1923-23.2843,average 5.0050)>Cr(0.4112-12.7130,average 1.7584)>Cd(0.0550-0.6045,average 0.2104)>Hg(0.0041-0.7431,average 0.1066)>As(0.0242-0.0856,average 0.0409)>Pb(0.0055-0.3405,average 0.0851)>Zn(0.0053-0.0915,average 0.0252).Principal component analysis,principal component 1 characteristic value was 3.521,eigenvector larger element Mn,As,Zn,Hg,Cr,were 0.946,0.922,0.790,0.759,0.602.2 principal component mainly element Fe,Pb,Cu,Cd,As,characteristic vector valued for 0.764,0.748,0.499,0.320,0.320.3 linear combination of principal component main content are as follows:Cr,Cd,zinc,Cu,Feelement,feature vector is respectively 0.594,0.584,0.545,0.292,0.101.4 linear combination of principal component content mainly include:Cd,Cr,Cu,Pb,zinc element,feature vector were 0.503,0.474,0.438,0.311 and 0.171.Combined with principal component analysis,correlation analysis and cluster analysis,Mn,As,Zn,Hg,Cr may is sandalwood inorganic elements characteristics.Inducing incense samples and most of the natural fragrant sample contents of inorganic elements in the same node.Determination of inorganic elements in the results showed that,Mn,Zn,As,Hg,Cr may sandalwood inorganic elements characteristics.Among them,Mn,Zn Trace elements,to further explore the Mn,the relationship between Zn and sandalwood efficacy,As,Hg,Cr for heavy metal elements,for the latter part of the development of sandalwood heavy metals limit standard provides a basis for the study.Correlation analysis results,Zn and Mn,Hg and Mn in 0.05 level was significantly positively related,Mn and Cr,Hg and Zn,Zn and the As,Mn,As,Hg and As at the 0.01 level was significantly positively related.The correlation between other elements did not reach significant level.Clustering analysis results,and 10 for distance,15 samples can be divided into three classes.Class 1:S2,S4,S6,S7,S8,S10,S11,S12and S15 samples,the samples of Cu element content is higher;Class 2,S13,S14 samples,each sample element content is relatively consistent;S1 and S3,S5 can get together for a class,the class samples containing Cu and Fe.On 9 kinds of element factor R type cluster analysis,with 20 for distance,9 kinds of elements can be divided into 4 classes.Category 1 Mn,As,Hg,Zn,Cr,combined with the correlation analysis showed that the correlation is larger,belong to the first principal component in the principal component analysis;Class 2 for Cu;Class 3 for Pb,Fe,both belong to the second principal component.Class 4 for Cd.The content of each element of different regions there are big differences in the variation coefficient.May be associated with producing area environment,climate,etc,to influence elements of absorption,transport,and the degree of enrichment is not the same.2.3 Study on HPLC fingerprint of sandalwood incense samples.12 batches of sandalwood a map on the control of the HPLC fingerprint and the sample of R similarity between 0.466 to 0.703.In 10,for distance,clustering analysis will be 12 batches of samples can be divided into four types,including inducing samples of 6 months and 12 months,most of the commercially available classified samples and reference substance.2.4 Sandalwood incense knot samples analysis and GC-MS fingerprint of volatile oil componentsSandalwood essential oil products is mainly containing alpha santalol,beta santalol,two component content as high as 75%,induction and fragrant 6 months sandal samples two component content is low,and the induction and sweet 12 months samples of two component content of 66%.15 batches of samples volatile oil(including commercial samples,sandalwood oil induced junction reference substance,sandalwood incense naphtha samples)similarity between 0.622?0.984,the difference is bigger,biological candidated knot incense heartwood naphtha similarity between 0.812 0.886,similarity is higher,the clustering analysis results 15 batches of samples can be divided into 2 groups,induction and fragrance samples of 6 months and 12 months with sandalwood essential oil as a kind of reference substance in the clustering analysis.Conclusion1 Experimental study of stimulate MethodBest method of induction and incense is:the infusion method;Induction knot agent:biological inducers;Induction knot agent concentration:2.5%;Sweet agent:80 ml;To the leaves.The law from the sandalwood incense quality solid heartwood,blond color;strong fragrance.Sweet time points ahead of the more than 10 years,the production and the comprehensive use of sandalwood is of great significance.And induce optimal season,heartwood harvest time of incense,incense agent in sandalwood plants such as conducting further studies are needed.2 Quality analysis of sandalwood heartwood sample by "Biological stimulation method"Biological candidated induction knot fragrant plant half year heartwood length of 191.7 cm,heartwood 4.3 cm in diameter,half year production 15767.1 cm3,volatile oil content of 2.84%.Induction and sweet after a year of the sample character,microscopic and thin layer,water content,content of ether extract and volatile oil content of the temporary not meet the requirements of pharmacopoeia,but has been higher than most commercially available sample quality.The GC-MS fingerprint and sandalwood essential reference substance.In conclusion,biological induction method is a kind of method can effectively induce sandalwood overall knot incense. |
PDF Full Text Request