Analysis Of Trace Elements,Plant Hormone And Sterol In Different Cultivation Patterns Of Anoectochilus Roxburghii

Anoectochilus roxburghii(A.roxburghii)is Orchidaceae Anoectochilus plants,is the common folk rare herbsperennial,used in the treatment of cancer,diabetes,high blood lipids,hepatitis B and other diseases,the general distribution of 300~1200m in altitudehilly areas,Fujian province is one of the main producing areas of A.roxburghii.at present,because of the deterioration of the natural environment,human wanton excavating and so on,the artificial cultivation of A.roxburghii is main.The artificial cultivation of A.roxburghii is divided into tissue culture and Cultivation under forest.In order to improve the quality of the artificial cultivation of A.roxburghii,In this article carries,trace elements,plant hormone and sterol in the artificial cultivated and ferine A.roxburghii was analysed.These investigations would provide the theory reference to the artificial cultivation of A.roxburghii.Mainly This thesis were divided into the following four chapters.Chapters 1.This paper used Inductively Coupled Plasma Mass Spectrometry(ICP-MS)for the quantitative analysis of 26 elements in different cultivation patterns sample of A.roxburghii.,inclused Ca,Fe,K,Mg,Na,Ag,Al,As,Ba,Hg,Cd,Co,Cr,Cu,Mn,Mo,Ni,Pb,Sb,Th,Se,Tl,U,V,Zn,Hg.And pre-processing sample was optimized.Microwave digestion in the sealing PTFE digestion tank,less interference,complete digestion,the measurement results more believable.The analysis method,ICP-MS method was established for the simultaneous determination of 26 trace elements in certified,each element of a good linear relationship,repetitive RSD is below 8.30%,the recoveries were 84.43 ~ 97.96%(n=9).Chapter 2.Nine kinds of phytohormone compounds in Anoectochilus Roxburghii cultivated with different cultivation patterns were analysed by HPLC-Qqq-MS-MS,including indoleacetic acid,indolebutyric acid,gibberellin,kinetin,trans-zeatin,trans-zeatin riboside,6-benzyl amino purine phen,isopentenyl adenosine and isopentennyl adenine.The samples were grinded by liquid nitrogen in low temperature and extracted by methanol-H2O-formic acid mixture.Use C18(2.1×50mm,2.6?m)chromatographic column to separate the samples with gradient elution.The velocity of flow was 0.3 mL/min.The column temperature was 30 ?.Nine kinds of phytohormone compounds were quantitative by ESI and MRM.The concentrations of phytohormones between 2 ~ 200ng/mL indicated a good linear.The detection limit was 0.01016 ~0.09970ng/Ml,the rate of recovery was 69.6%~77.6% and RSD<15.2%(n = 9)?It showed that the method with simple operation,high sensitive and accuration was also suitable to detect phytohormone compounds in other plants.According to results,the main phytohormones in 12 batches of samples were gibberellin,indoleacetic acid,trans-zeatin riboside,isopentennyl adenine and isopentenyl adenosine.Other phytohormones were detected in part of the samples,but the contents were close to quantification limit.Chapter 3.Three kinds of plant sterols compounds in Anoectochilus Roxburghii cultivated with different cultivation patterns were analysed by GC-MS,(including: ergosterol,stigmasterol and beta-sitosterol).The method of ultrasonic extraction is using chloroform solution with internal standard,direct injection analysis after centrifugal filtration,in DB-5MS(30m × 2.5mm,0.25 ? m)column using gradient temperature to realize the separation of reference standards,and the NIST08 spectral library of qualitative,and quantitative with standard.The sterol concentration in 1 ~ 200 ? g/mL linear relation in the range of good,the detection limit is 5.08 ~ 10.23ng/mL,recovery between 93.6% ~ 97.2%,RSD < 1.1%(n = 9).The method is simple,sensitive and accurate,and is suitable for detecting thecontent of sterols.We know from the test results,12 batches of samples were detected in beta-sitosterol,ergosterol and stigmasterol,total sterol content was more than 1mg/g,the sterols have anti-inflammatory,anti ulcer,reducing blood fat,lowering cholesterol and preventing tumor and so on,also provides a theoretical and experimental basis for the pharmacological effects of Anoectochilus formosanus isforce,also provides methods and reference for further research in Anoectochilus roxburghii.