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Method for detecting abscisic acid and jasmonic acid in fresh tobacco leaves through liquid chromatogram-tandem mass spectrometry

A technology of liquid chromatography and tandem mass spectrometry, which is applied in the field of liquid chromatography-tandem mass spectrometry for detecting abscisic acid and jasmonic acid in fresh tobacco leaves, can solve the problems of cumbersome and time-consuming processing, and achieves simplified sample pretreatment steps, good selectivity, The effect of improving analytical efficiency

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 to provide a simple method for detecting abscisic acid and jasmonic acid in fresh tobacco leaves by liquid chromatography-tandem mass spectrometry for the cumbersome and time-consuming pretreatment of the determination methods of abscisic acid and jasmonic acid in the prior art

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  • Method for detecting abscisic acid and jasmonic acid in fresh tobacco leaves through liquid chromatogram-tandem mass spectrometry
  • Method for detecting abscisic acid and jasmonic acid in fresh tobacco leaves through liquid chromatogram-tandem mass spectrometry
  • Method for detecting abscisic acid and jasmonic acid in fresh tobacco leaves through liquid chromatogram-tandem mass spectrometry

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Effect test

example 1

[0029] 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.

[0030] Using this method to extract flue-cured tobacco safflower Dajinyuan tobacco leaf samples, through LC-MS / MS analysis, the obtained total ion current TIC and extracted ion current MRM are as follows: image 3 As shown, each quantitative ion pair ABA-d4, 269>159; ABA, 263>153; JA, 209>59 can achieve baseline separation and accurately quantify abscisic acid and jasmonic acid. Abscisic acid, jasmonic acid 7.37 ng mL -1 and 0.53 ng mL -1 .

example 2

[0032] Weigh 20-25 mg of freeze-dried fresh tobacco leaves into a glass test tube, add 0.5 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.

[0033] Burley tobacco TN90 sample was extracted by this method, analyzed by LC-MS / MS, the obtained total ion current TIC and extracted ion current MRM were as follows: Figure 4 As shown, each quantitative ion pair ABA-d4, 269>159; ABA, 263>153; JA, 209>59 can achieve baseline separation and accurate quantification of abscisic acid and jasmonic acid. Abscisic acid, jasmonic acid 10.21 ng mL -1 and 0.67 ng mL -1 .

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Abstract

The invention provides a method for detecting abscisic acid and jasmonic acid in fresh tobacco leaves through liquid chromatogram-tandem mass spectrometry. The method comprises the following steps: (1), preparing an extraction solvent; (2), carrying out liquid nitrogen quick freezing on fresh tobacco leaves, and freezing, drying and smashing; (3), adding the extraction solvent into the well smashed samples, and carrying out low temperature extracting; (4), carrying out refrigerated centrifugation, taking supernate and filtering; (5), carrying out LC-MS / MS detection. The method provided by the invention has the following advantages: a pretreatment method is simple, only ultrasound and filtering are required, and solid phase extraction column purification treatment is not required. Compared with the conventional method, the method provided by the invention adopts LC-MS / MS, the sensibility is high, the accuracy is high, the recovery of standard addition is high, and pretreatment for 100 samples consumes less than 2 hours while methods in existing documents require tedious and time-consuming pretreatments, and about three days are required for 100 samples. Therefore, the method provided by the invention can effectively improve analysis efficiency, and is suitable for analyzing low content objects in a complex matrix.

Description

technical field [0001] The invention relates to the research on hormones in fresh tobacco leaf samples of tobacco metabolomics, in particular to a liquid chromatography-tandem mass spectrometry method for detecting abscisic acid and jasmonic acid in fresh tobacco leaves. Background technique [0002] Abscisic acid and jasmonic acid are two important physiologically active substances in plants, which perform functions in secondary metabolism, stress tolerance and signal transduction of tobacco plants, thereby affecting the growth and development of tobacco and the quality of tobacco leaves. Therefore, accurate and reliable qualitative and quantitative analysis of trace amounts of abscisic acid and jasmonic acid is of great significance. [0003] The simultaneous determination of abscisic acid and jasmonic acid in fresh plant leaves by solid-phase extraction-liquid chromatography / mass spectrometry has been reported in the literature. (1) 50 mg of plant samples were ground int...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/06
Inventor 陈霞周会娜刘萍萍陈千思金立锋翟妞郑庆霞卢鹏王燃林福呈
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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