Sample pretreatment method for determining endogenous hormones in trace plant sample

A sample pretreatment and endogenous hormone technology, applied in the field of analytical chemistry, can solve the problems of large sample volume, numerous steps, and lengthy process, and achieve the effects of mild extraction conditions, improved detection sensitivity, and simple operation

Inactive Publication Date: 2019-07-09
INST OF CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This technology uses ultraviolet light waves that help break down plants into smaller pieces for better analysis by chemistry techniques like sonochemery (ultrasound). It also involves direct extractions from living cells without damaging them during processing steps such as cutting or polishing. Additionally, it allows for easy handling and disposal afterward while maintaining environmental benefits compared to traditional methods used beforehand. Overall, this process provides improved efficiency and accuracy in analyzing biological materials.

Problems solved by technology

This patented technical problem addressed by the present patents relates to improving methods for analyzing small amounts (<100 mL) of chemicals from plants without losing valuable material during processing procedures such as extractive distillations.

Method used

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  • Sample pretreatment method for determining endogenous hormones in trace plant sample
  • Sample pretreatment method for determining endogenous hormones in trace plant sample
  • Sample pretreatment method for determining endogenous hormones in trace plant sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1, determine the content of nine kinds of endogenous GAs in an Arabidopsis flower

[0030] Flowchart such as figure 1 shown.

[0031] 1) Accurately weigh three Arabidopsis flowers with a mass between 0.65 and 0.83 mg, transfer them to three centrifuge tubes respectively, and add nine kinds of 5 fmol [ 2 h 2 ]GAs (including [ 2 h 2 ]GA 4 , [ 2 h 2 ]GA 8 , [ 2 h 2 ]GA 9 , [ 2 h 2 ]GA 12 , [ 2 h 2 ]GA 19 , [ 2 h 2 ]GA 24 , [ 2 h 2 ]GA 29 , [ 2 h 2 ]GA 34 and[ 2 h 2 ]GA 51 );

[0032] 2) Weigh a certain amount of cellulase R-10 solid powder and dissolve it in water with a pH of 4.5 (adjusted with 0.12 mol / L hydrochloric acid diluted with concentrated hydrochloric acid);

[0033] 3) to the Arabidopsis flowers and [ 2 h 2 ] Add the above-mentioned cellulase solution (the amount added is 5% of the quality of the Arabidopsis thaliana flower) to the centrifuge tubes of GAs, and then place these centrifuge tubes in an ultrasonic cleaner ...

Embodiment 2

[0041] Embodiment 2, measure the content of endogenous indole acetic acid (IAA), cinnamic acid (CA) in 3mm long malt tip

[0042] Flowchart such as figure 1 shown.

[0043] 1) Accurately weigh five malt tips with a length of 3 mm, with a mass between 1.02 and 1.25 mg, transfer them to five centrifuge tubes respectively, add 100 fmol of IAA and CA deuterated internal standard ([ 2 h 5 ]IAA and [ 2 h 7 ]CA);

[0044] 2) Weigh a certain amount of cellulase R-10 solid powder and dissolve it in water with a pH of 4.7 (adjusted with 0.12 mol / L hydrochloric acid diluted with concentrated hydrochloric acid);

[0045] 3) Add the above-mentioned cellulase solution (the amount added is 6% of the mass of the malt tip) to the centrifuge tube containing the malt tip and the deuterated internal standard, and then place these centrifuge tubes in an ultrasonic cleaner for extraction for 10 min at a temperature of 50 ℃. After the extraction, centrifuge to collect the supernatant; the pla...

Embodiment 3

[0054] Embodiment 3, determine the content of six kinds of endogenous GAs in a grain of rice seed

[0055] Flowchart such as figure 1 shown.

[0056] 1) Accurately weigh five rice seeds with a mass between 25.3 and 25.9 mg, transfer them to five centrifuge tubes respectively, and add 15 fmol of the six kinds of [ 2 h 2 ]GAs (including [ 2 h 2 ]GA 4 , [ 2 h 2 ]GA 8 , [ 2 h 2 ]GA 19 , [ 2 h 2 ]GA 20 , [ 2 h 2 ]GA 29 and[ 2 h 2 ]GA 53 );

[0057] 2) Weigh a certain amount of cellulase R-10 solid powder and dissolve it in water with a pH of 5.0 (adjusted with 0.12 mol / L hydrochloric acid diluted with concentrated hydrochloric acid);

[0058] 3) Fill rice seeds and [ 2 h 2 ] Add the above-mentioned cellulase solution (the addition amount is 5% of the mass of rice seeds) to the centrifuge tubes of GAs, and then place these centrifuge tubes in an ultrasonic cleaner for extraction for 50min at a temperature of 40°C. After the extraction, centrifuge to collect t...

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Abstract

The invention discloses a sample pretreatment method for determining endogenous hormones in a trace plant sample. The sample pretreatment method for determining the endogenous hormones in the trace plant sample comprises the step of extracting the endogenous hormones in the plant sample by using a lyticase under ultrasonic irradiation, wherein the plant sample is fresh or dry plant tissues, organsor their sections; the lyticase is at least one of a cellulase, a pectinase, a hemicellulase, and a macerozyme; and the endogenous hormone is at least one of auxin, cytokinin, gibberellin, abscisic acid, jasmonic acid and derivatives thereof, and salicylic acid and derivatives thereof. The invention has the following advantages: 1, the principle of ultrasonic-assisted enzymatic hydrolysis is used, so that the endogenous hormones are directly extracted without pulverizing or grinding the plant sample, therefore, the extraction condition is mild, and the operation is simple and rapid; 2, the extractant is an aqueous solution, which avoids the use of organic solvents and is more environmentally friendly; and 3, chemical derivatization and efficient chromatographic separation are combined, sothat the detection sensitivity can be significantly improved, and the amount of the plant sample can be reduced to milligram or even sub-milligram levels.

Description

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Claims

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Application Information

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Owner INST OF CHEM CHINESE ACAD OF SCI
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