Efficient liquid-phase chromatographic detection method for content of polyamines in goose tissue as well as application thereof
A technology of high performance liquid chromatography and detection method, which is applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of inseparable peak time, cumbersome methods, and unsuccessful extraction.
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Embodiment 1
[0050] Example 1 Solid phase extraction and high performance liquid chromatography detection of polyamines in Sichuan white goose grade follicles.
[0051] (1) The grade follicle samples of goose were taken out from the -80 ℃ refrigerator, ground in liquid nitrogen, and 0.1 g of tissue samples were weighed and placed in Eppendorf tubes.
[0052] (2) Homogenize the largest preovulatory follicle of goose twice, extract the supernatant, add 2 mL of 2.5 mol / mL NaOH, and 1.4 μL of benzoyl chloride, vortex and derivatize in a water bath at 40°C 30 min.
[0053] (3) Take out the derivative solution, adjust the pH value to 7.0 with 6 mol / mL HCl, then add the derivative solution to an activated C18 solid-phase extraction column for filtration, and then use 15 mL of ultrapure water and 15 mL of 15% methanol was used to wash away impurities.
[0054] (4) Elute the sample with 0.5 mL of chromatographic methanol, and then filter the eluate with a 0.22 μm syringe filter. Take 20 μL of ...
Embodiment 2
[0056] Example 2 Detection of polyamines in atretic follicles of Sichuan white goose by high performance liquid chromatography.
[0057] (1) The goose atretic follicle samples were taken out from the -80°C refrigerator, ground in liquid nitrogen, and 0.1 g of tissue samples were weighed and placed in Eppendorf tubes.
[0058] (2) Goose atretic follicles were homogenized twice, and the supernatant was extracted, then 2 mL of 2.5 mol / mL NaOH and 1.4 μL of benzoyl chloride were added, vortexed and derivatized in a water bath at 40°C for 30 min.
[0059] (3) Take out the derivative solution, adjust the pH value to 7.0 with 6 mol / mL HCl, then add the derivative solution to an activated C18 solid-phase extraction column for filtration, and then use 15 mL of ultrapure water and 15 mL of 15% methanol was used to wash away impurities.
[0060] (4) Elute the sample with 0.5 mL of chromatographic methanol, and then filter the eluate with a 0.22 μm syringe filter. Take 20 μL of the el...
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