LC-MS/MS method for detecting IPP, GPP, FPP and GGPP in fresh tobacco leaf

A LC-MS and fresh tobacco leaf technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of complicated and time-consuming processing, and achieve the effects of simplifying sample pretreatment steps, improving analysis efficiency, and high sensitivity

Active Publication Date: 2015-01-21
ZHENGZHOU TOBACCO RES INST OF CNTC
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Problems solved by technology

[0004] The purpose of the present invention is aimed at the tedious and time-consuming problem of the pretreatment of IPP, GPP, FPP and GGPP assay methods in the prior art, and provides a simple liquid chromatography-tandem mass spectrometry method for detecting IPP, GPP, GPP, Methods of FPP and GGPP

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  • LC-MS/MS method for detecting IPP, GPP, FPP and GGPP in fresh tobacco leaf
  • LC-MS/MS method for detecting IPP, GPP, FPP and GGPP in fresh tobacco leaf
  • LC-MS/MS method for detecting IPP, GPP, FPP and GGPP in fresh tobacco leaf

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Embodiment Construction

[0026] The present invention is further described by the following specific examples, but does not limit the present invention.

[0027] Weigh 45-50 mg of freeze-dried fresh tobacco leaves into a glass test tube, add 1 mL of extraction solvent into the glass tube, and the internal standard is 20 ng ml -1 ABA-d6, capped and sealed, placed in an ultrasonic cleaner, ultrasonically extracted in an ice bath for 30 min, with an ultrasonic power of 100 Hz, and then centrifuged at 8000 rpm for 5 min, the supernatant was taken and filtered, and carried out LC-MS / MS analysis.

[0028] Using this method to extract flue-cured tobacco safflower Dajinyuan tobacco leaf samples, through LC-MS / MS analysis, the obtained total ion current TIC chromatogram and extracted ion current MRM are as follows: image 3 As shown, each quantitative ion pair ABA-d6, 269>159; IPP, 245>79; GPP, 313>79; FPP, 381>79; GGPP, 449>79 can achieve baseline separation and quantify IPP, GPP, FPP and GGPP. After test...

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Abstract

The invention provides an LC-MS/MS method for detecting IPP, GPP, FPP and GGPP in a fresh tobacco leaf. The LC-MS/MS method comprises the following steps: 1) preparing an extraction solvent; 2) conducting liquid nitrogen flash freezing on the fresh tobacco leaf, and conducting freeze drying and crushing; 3) adding the extraction solvent to the crushed sample, and conducting ultrasonic extraction at low temperature; 4) carrying out refrigerated centrifugation, taking supernatant, and filtering; 5) conducting LC-MS/MS method to detect IPP, GPP, FPP and GGPP in the detected sample. The LC-MS/MS method has the advantages that the pretreatment method of the sample is simple and only comprises ultrasonic extraction and filtration; compared with the existing method, the method is high in sensitivity, good in precision, and high in recovery of standard addition; the sample can be directly fed and analyzed through filtration, but according to the methods in the existing documents, the pretreatment method is complicated and time-consuming, and requires pyrophosphatase and alkaline phosphatase for carrying out biochemical reaction on the sample. Therefore, the LC-MS/MS method disclosed by the invention can effectively increase the analysis efficiency, and is suitable for analysis of the ultralow content target in the complex substrate.

Description

technical field [0001] The present invention relates to the research on terpenoids in fresh tobacco leaf samples of tobacco metabolomics, specifically a liquid chromatography-tandem mass spectrometry method for detecting isopentenyl pyrophosphate IPP, geranyl pyrophosphate GPP and farnesyl in fresh tobacco leaves Methods of FPP pyrophosphate and GGPP geranylgeranyl pyrophosphate. Background technique [0002] Terpenoids are the most numerous class of compounds in plant metabolites, which are composed of isoprene as a structural unit. It plays an important role in plant respiration, photosynthesis, growth, development, reproduction, signal transduction and defense. Some terpenoids have important economic value, and some can be used as natural fragrances and aroma compounds or anti-tumor chemotherapy drugs. In plants, the biosynthesis of terpenoids occurs in the cytoplasm and plastids, and its precursor is C 5 Structured IPP. IPP synthesized via the mevalonate pathway is a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 陈霞李锋周会娜翟妞金立锋陈千思刘萍萍郑庆霞王晨林福呈
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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