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Kit and method for measuring content of free fatty acid

A free fatty acid and kit technology, applied in the field of life sciences, can solve the problems of long detection process, poor repeatability of detection results, expensive instruments, etc., and achieve improved extraction methods and operating steps, sample pretreatment process is simple, and the process is simple and fast. Effect

Inactive Publication Date: 2016-05-11
SUZHOU COMIN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] But above-mentioned method mainly has following shortcoming: 1. the extract liquid composition of gas chromatography and liquid chromatography is more complicated, and processing process is loaded down with trivial details, and detection process takes a long time, and detection cost is higher, and instrument is expensive, and reagent consumption is bigger , cannot be widely used; 2. The operation time of diphenylcarbazide colorimetric method is shorter than that of gas chromatography and liquid chromatography in colleges and universities, but the reagent components used are complex and the repeatability of test results is poor

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Determination of the content of free fatty acids in the blood, the specific steps are as follows:

[0057] Step 1 preparation of instruments and supplies;

[0058] Mortar, desktop centrifuge, shaking instrument, visible spectrophotometer, 1mL glass cuvette;

[0059] Extraction of free fatty acids in the blood sample in step 2;

[0060]Let the collected blood stand at room temperature for 1 hour, then centrifuge at 3500rpm, 4°C for 15 minutes, take 0.1mL of the supernatant, add 1.2mL of the reagent 1, shake and extract for 3h, centrifuge at 8000rpm, 4°C for 10 minutes, take the supernatant Serum to be tested;

[0061] Step 3 Determination of free fatty acid content in blood samples;

[0062] 1) Preheat the spectrophotometer for more than 30 minutes, and adjust the wavelength to 715nm;

[0063] 2) Control tube: take 1mL of the supernatant of the sample, add 0.5mL of the reagent 2, shake fully for 5 minutes, let stand at room temperature for 5 minutes, take 0.8mL of th...

Embodiment 2

[0070] Determination of the content of free fatty acids in tissues, the specific steps are as follows:

[0071] Step 1 preparation of instruments and supplies;

[0072] Mortar, desktop centrifuge, shaking instrument, visible spectrophotometer, 1mL glass cuvette;

[0073] Extraction of free fatty acids in the tissue sample in step 2;

[0074] After the tissue was rinsed with distilled water, the surface moisture was absorbed with absorbent paper, crushed, weighed about 0.1 g, and transferred to a centrifuge tube. Reagent 1 was extracted by shaking for 3 hours, centrifuged at 8000 rpm, 4°C for 10 minutes, and the supernatant was taken for testing;

[0075] Step 3 Determination of free fatty acid content in tissue samples;

[0076] 1) Preheat the spectrophotometer for more than 30 minutes, and adjust the wavelength to 715nm;

[0077] 2) Control tube: take 1mL of the supernatant of the sample, add 0.5mL of the reagent 2, shake fully for 5 minutes, let stand at room temperature...

Embodiment 3

[0090] To determine the content of free fatty acids in bacteria and fungi, the specific steps are as follows:

[0091] Step 1 preparation of instruments and supplies;

[0092] Mortar, desktop centrifuge, shaking instrument, visible spectrophotometer, 1mL glass cuvette;

[0093] Extraction of free fatty acids in step 2 bacteria and fungi samples;

[0094] According to the number of cells (10 4 pcs): the volume (mL) of the reagent 1 is (500~1000): 1.2 ratio (it is recommended to add 1.2mL reagent 1 to 5 million cells), add the reagent 1, break the cells in an ice bath ultrasonically at a power of 300w, and ultrasonically 2 seconds, with an interval of 3 seconds, and a total time of 3 minutes; shake and extract for 3 hours, then centrifuge at 8000 rpm and 4°C for 10 minutes, and take the supernatant for testing;

[0095] Step 3 Determination of free fatty acid content in bacteria and fungi samples;

[0096] 1) Preheat the spectrophotometer for more than 30 minutes, and adjust...

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Abstract

The invention discloses a kit and method for measuring the content of free fatty acid. Reagents of the kit comprise the first reagent composed of isooctane, the second reagent formed by dissolving sodium acetate in distilled water, then adding pyridine and mixing the constituents uniformly and the third reagent formed by dissolving copper acetate in distilled water, then adding pyridine and mixing the constituents uniformly. The method comprises the steps that free fatty acid in a sample is extracted through the isooctane, then the free fatty acid reacts with copper salt to generate copper soap under the weak acidic condition, and the content of free fatty acid in the reaction sample at the light absorption value of 715 nm is measured. According to the method, based on measurement conducted through an existing colorimetric method, the extraction method and operation steps are improved, and the effects of simple extraction and fast, simple and convenient measurement are achieved. According to the method, the sample pre-treatment process is simple and convenient, and an extracting solution is singular in constituent and low in cost. According to the measurement steps of the method, only the three reagents are needed for completing measurement, and the process is simple, convenient and fast.

Description

technical field [0001] The invention belongs to the field of life sciences and relates to a kit, in particular to a free fatty acid content determination kit and a method thereof. Background technique [0002] Free fatty acids (FFA) are both products of fat hydrolysis and substrates for fat synthesis. The concentration of free fatty acids is related to lipid metabolism, glucose metabolism, and endocrine function, and can also reflect the quality changes in food storage. [0003] At present, the existing methods for detecting the content of free fatty acids in the market are mainly gas chromatography, high performance liquid chromatography and colorimetry. Gas chromatography uses organic solvent mixed solution and sulfuric acid to treat the sample, and uses organic solvent to extract free fatty acids at high temperature, and then conducts gas chromatography analysis. High-performance liquid chromatography uses organic solvents to extract free fatty acids in samples, followe...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/78
CPCG01N21/3103G01N21/78G01N2021/3122
Inventor 赵林川
Owner SUZHOU COMIN BIOTECH
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