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Preparation method of free fatty acid calibrator

A free fatty acid and calibrator technology, applied in the field of medical testing, can solve problems such as wrong results, result deviations, and unstable calibrator, and achieve high accuracy and good stability

Pending Publication Date: 2021-10-01
WENZHOU PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the fatty acid species involved in the preparation method of the ACS-ACOD coupled enzymatic detection reagent calibrator in the prior art is only one or a combination of a small number of free fatty acids, and the calibrator and serum / plasma reactions are not synchronized during the test. , when applied to the determination of biochemical analyzers of different models, there are large deviations in the results and even wrong results
In addition, if the free fatty acid calibrator is prepared directly with the mixed serum as the matrix solution, there will be a problem that the neutral fat, lipoprotein and other macromolecules in the matrix solution will continuously decompose to produce free fatty acid, which will make the calibration product unstable.

Method used

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  • Preparation method of free fatty acid calibrator
  • Preparation method of free fatty acid calibrator
  • Preparation method of free fatty acid calibrator

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1 A kind of preparation method of free fatty acid calibrator

[0051] Include the following steps:

[0052](1) Add 0.1 mol / L guanidine hydrochloride, 10 g / L trehalose, 9 g / L sodium chloride, and 0.2 g / L sodium azide to 10.0 mL of mixed serum, and age at 37°C for 24 hours.

[0053] (2) Place the solution prepared in step (1) in a refrigerated centrifuge, select a centrifugation temperature of 2° C., and a centrifugation speed of 3000 rpm, and take 9.8 mL of the supernatant solution after centrifugation for 10 minutes.

[0054] (3) Transfer the 9.8mL supernatant solution prepared in step (2) to the sample container of the tangential flow filtration system, set the ultrafiltration membrane inside the hollow fiber membrane filter, and the ultrafiltration membrane is modified polyethersulfone Membrane, molecular weight cut off is 1.0KD, membrane area is 20cm 2 , set the target pump speed to 30mL / min, and the shear force to 3000s -1 . Close the filtrate clamp o...

Embodiment 2

[0063] Embodiment 2 A kind of preparation method of free fatty acid calibrator

[0064] Include the following steps:

[0065] (1) Add 0.5mol / L urea, 50g / L trehalose, 9g / L sodium chloride, 1.0g / L sodium azide to 25.0mL mixed serum, and age at 37°C for 48 hours;

[0066] (2) Place the solution prepared in step (1) in a refrigerated centrifuge, select the centrifugation temperature to be 5° C., and the centrifugation speed to be 7500 rpm, and take 24.5 mL of the supernatant solution after centrifugation for 10 minutes;

[0067] (3) Transfer the 24.5mL supernatant solution prepared in step (2) to the sample container of the tangential flow filtration system, set the ultrafiltration membrane inside the hollow fiber membrane filter, and the ultrafiltration membrane is modified polyethersulfone Membrane, molecular weight cut off is 1.0KD, membrane area is 60cm 2 , set the target pump speed to 60mL / min, and the shear force to 4500s -1 . Close the filtrate clamp of the filtration c...

Embodiment 3

[0075] Embodiment 3 A kind of preparation method of free fatty acid calibrator

[0076] Include the following steps:

[0077] (1) Add 1 mol / L sodium lauryl sulfate, 100 g / L trehalose, 9 g / L sodium chloride, and 2.0 g / L sodium azide to 50.0 mL of mixed serum, and age at 37°C for 72 hours.

[0078] (2) Place the solution prepared in step (1) in a refrigerated centrifuge, select a centrifugation temperature of 8° C., and a centrifugation speed of 12000 rpm, and take 49.0 mL of the supernatant solution after centrifugation for 10 minutes.

[0079] (3) Transfer the 49.0mL supernatant solution prepared in step (2) to the sample container of the tangential flow filtration system, set the ultrafiltration membrane inside the hollow fiber membrane filter, and the ultrafiltration membrane is modified polyethersulfone Membrane, the molecular weight cut off is 1.0KD, and the membrane area is 115cm 2 , set the target pump speed to 80mL / min, and the shear force to 6000s -1 . Close the fi...

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Abstract

The invention provides a preparation method of a free fatty acid calibrator, which comprises the following steps: adding an aging accelerator into mixed serum to quickly release free fatty acid in the serum, centrifuging at low temperature, taking a supernatant solution, enabling the supernatant solution to pass through a tangential flow filtering system, selecting a hollow fiber membrane module, pump speed and shearing force, and performing purification and concentration operation, and diluting after concentration. The free fatty acid calibrator prepared by the method contains humanized free fatty acid types, does not contain serum neutral fat, lipoprotein, polypeptide and other substances with the molecular weight of more than 1.0 KD, has stable performance after being redissolved, and is suitable for batch production of the free fatty acid calibrator.

Description

technical field [0001] The invention belongs to the technical field of medical testing, and in particular relates to a preparation method of a free fatty acid calibrator. Background technique [0002] Free fatty acids, also known as non-esterified fatty acids (NEFA), refer to fatty acids that have not been esterified with glycerol, cholesterol, etc. in serum, and have a half-life of 2-3 minutes in plasma. Under normal circumstances, free fatty acids are extremely high in plasma. few. NEFA has strong cytotoxicity, can damage cell membrane, mitochondria and lysosome membrane, etc., cause intracellular damage, and can enhance cytokine toxicity, playing an important role in the pathophysiology of many diseases. Acetyl-CoA synthetase (ACS)-acetyl-CoA oxidase (ACOD) coupled enzymatic assay is currently the main method for clinical testing of NEFA. Nearly 20 free fatty acids including oleic acid are known to exist in human serum / plasma samples [1] , ACS and ACOD can also act on ...

Claims

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Application Information

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IPC IPC(8): G01N1/28G01N1/34G01N1/40G01N30/02G01N30/06G01N30/14C12Q1/25C12Q1/26
CPCG01N1/28G01N1/34G01N1/40G01N1/4044G01N1/4077G01N30/06G01N30/14G01N30/02C12Q1/26C12Q1/25G01N2001/4088
Inventor 潘利琴
Owner WENZHOU PEOPLES HOSPITAL