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.

Inactive Publication Date: 2014-02-12
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The solid-phase extraction method is cumbersome and it is easy to cause unsuccessful or incomplete extraction due to operational errors. This program provides a complete and feasible method for extracting polyamines from goose tissues. The method is simple and feasible
[0005] High-performance liquid chromatography detection has the characteristics of fast analysis speed, high separation efficiency, high sensitivity, and wide application range. However, the detection conditions must be optimized during high-performance liquid chromatography detection. The peak time cannot be separated, and the detection repeatability is not good, etc.

Method used

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  • Efficient liquid-phase chromatographic detection method for content of polyamines in goose tissue as well as application thereof
  • Efficient liquid-phase chromatographic detection method for content of polyamines in goose tissue as well as application thereof
  • Efficient liquid-phase chromatographic detection method for content of polyamines in goose tissue as well as application thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses an efficient liquid-phase chromatographic detection method for content of polyamines (putrescine, spermidine and spermine) in a goose tissue. Chromatographic conditions of the method are as follows: a C18 chromatographic column, 5 mu m, 4.6*250mm; a volume ratio of mobile phase methanol: water of 62:38; flow velocity of 1.0 mL / minute; ultraviolet detection wavelength of 229 nm; and a column temperature of 25 DEG C. A tissue solid-phase extracting method comprises the following steps: adding 0.1g of tissue into 1 mL of 5% HClO4 and 10 mu L of an internal standard substance for homogenizing and extracting supernatant; adding 2 mL of 2.5 mol / mL NaOH and 1.4 mu L of benzoyl chloride, carrying out water-bath for 30 minutes at a temperature of 40 DEG C; adjusting pH value to 7.0, then, adding into the C18 column, and flushing by 15 mL of water and 15 mL of 15% methanol; and eluting by 0.5 mu L of methanol. The scheme can effectively separate putrescine, spermidine and spermine in the tissue, and can quickly and accurately detect content of the polyamines in the goose tissue, is simple and convenient to operate, high in accuracy and good in repeatability.

Description

technical field [0001] The invention belongs to a high-performance liquid chromatography detection method. The invention relates to a high-performance liquid chromatography detection method for polyamine content in goose tissues and its application, specifically including a solid-phase extraction method and a high-performance liquid chromatography method for polyamines in goose tissues. Chromatography was used to detect the content of putrescine, spermidine and spermine in the extracted samples. Background technique [0002] Polyamines (including putrescine, spermidine and spermine) are a class of low-molecular-weight aliphatic compounds with two or more amino groups, which exist in almost all eukaryotic organisms. This evolution The conservation suggests that polyamines are extremely important for cell survival. At physiological pH, polyamines have a high density of positive charges, and they can bind to biological macromolecules such as DNA, RNA, and proteins with a large...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 康波姜冬梅何珲王继文马容李亮王迅韩春春刘贺贺许恒勇
Owner SICHUAN AGRI UNIV
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