A chromatographic mass spectrometry method for the detection of juvenile hormone jh II in insects
A detection method, juvenile hormone technology, applied in the field of analytical chemistry, can solve the problem of low detection sensitivity of juvenile hormone JHII, and achieve the effect of ensuring quantitative accuracy, good repeatability and high sensitivity
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Embodiment 1
[0015] Example 1 Evaluation of the method for detecting juvenile hormone JH II in insects based on liquid chromatography-mass spectrometry
[0016] Concentration is 1mg / mL juvenile hormone JH II standard sample is diluted into a series of different concentration solutions (20ng / mL, 10ng / mL, 5ng / mL, 1ng / mL, 0.5ng / mL, 0.1ng / mL with volume concentration 60% acetonitrile / mL, 0.05ng / mL, 0.025ng / mL, 0.01ng / mL). Then take 200 μL of standard sample solutions with different concentrations, add 20 μL of internal standard methoprene solution (concentration: 50 ng / mL) and mix well, and wait for instrumental analysis. Adopt the above-mentioned standard solution of different concentrations that adds internal standard to carry out liquid chromatography mass spectrometry separation and analysis, investigate the linearity of instrument method, take the concentration of juvenile hormone JH II and the relative area of peak as coordinates to draw linear standard curve, linear correlation coeff...
Embodiment 2
[0018] Embodiment two 4 instars cotton bollworm sample detection
[0019] Take 4 samples of the 4th instar cotton bollworm, put them into a 2mL Eppendorf tube, weigh 191.8mg, add grinding balls, add 250μL of acetonitrile, 250μL of 2% sodium chloride aqueous solution, 500μL of 1ng / mL internal standard Grind the n-hexane of methoprene for 2 minutes, centrifuge at 12000g for 5 minutes at 4°C, extract the supernatant n-hexane layer, then add 500 μL of pure n-hexane to the residue, grind, extract the supernatant, and repeat the extraction once with pure n-hexane , put the crude extract into a 2mL Eppendorf tube, vortex for 1min, centrifuge at low speed for 15s, blow dry slowly with nitrogen to obtain the crude extract; The solid-phase extraction column was pre-washed twice with 1 mL of methanol in advance, and eluted with 200 μL of acetonitrile for 3 times. The eluate in the collection box was taken out, and the collection box was washed once with 200 μL of acetonitrile. Centrifug...
Embodiment 3
[0022] Embodiment three 3 instars cotton bollworm sample detection
[0023] Take 6 samples of the 3rd instar cotton bollworm, weigh 127.2 mg, add grinding balls, 250 μL of acetonitrile, 250 μL of 2% sodium chloride aqueous solution, 500 μL of n-hexane containing 1 ng / mL internal standard methoprene, and grind for 2 minutes , centrifuge at 12000g for 5min at 4°C, extract the supernatant n-hexane layer, repeat the extraction twice with 500μL pure n-hexane, combine the crude extract into a 2mL Eppendorf tube, vortex for 1min, centrifuge at low speed for 15s, blow dry with nitrogen; Add 200 μL of 60% acetonitrile to the dried crude extract, vortex for 1 min, then pass the reconstituted solution through the solid phase extraction column into the collection box, and elute with 200 μL acetonitrile for 3 times, take out the eluate in the collection box, and use 200 μL Rinse the collection box once with acetonitrile, put it into a 2mL Eppendorf tube together with the eluent, centrifuge a...
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