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
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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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