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Blank biological sample preparation method

A biological sample, blank technology, applied in the field of blank biological sample preparation, can solve problems such as interference

Pending Publication Date: 2019-02-26
上海可力梅塔生物医药科技有限公司 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When the sample to be tested is serum, serum or similar matrix should also be used to establish a standard curve for the test, and vitamins with high endogenous content will cause great interference to this type of detection method, and cannot be used at the detection concentration that needs to be covered. Accurate measurement results within the range

Method used

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preparation example Construction

[0034] Preparation of blank biological samples

[0035] In the existing mass spectrometry or chromatographic analysis methods, the establishment of the standard curve is often interfered by endogenous vitamins in the blank sample. Since vitamins in biological samples not only exist in free form, but also partly in endogenous form, the existing treatment methods cannot completely remove endogenous vitamins. In this regard, the present invention provides a process for removing endogenous analytes from biological samples while retaining the biological characteristics of the original biological samples, which can be used for the removal of vitamin compounds in biological samples. The basic flow of this method is as follows:

[0036] a) hydrolytic release of vitamin compounds in biological samples;

[0037] b) Utilize the adsorption material to absorb the dissociated vitamin compounds;

[0038] c) centrifuging the adsorption material and the biological sample after adsorption to...

Embodiment 1

[0063] Example 1 The preparation process of the blank human serum sample not containing vitamin compounds

[0064] (1) Take 10 mL of serum to be treated, and add 0.5% formic acid by its volume as a hydrolysis reagent for vitamin-binding protein. The hydrolysis reagent needs to be evenly mixed with the serum, and incubated at 37°C for 10 minutes to obtain pretreated serum, in which all endogenous vitamins exist in free form.

[0065] (2) Add 5% activated carbon by volume to the pretreated serum obtained in (1). Activated charcoal needs to be fully mixed with serum and incubated at 37°C for 1 hour to obtain a solid-liquid mixed serum with all free vitamins removed.

[0066] (3) The solid-liquid mixed serum obtained from (2) can be centrifuged at a high speed to remove the solid activated carbon. The centrifugation speed is 15,000 rpm. At this centrifugation speed, it is necessary to take out the supernatant after one centrifugation and centrifuge again. Repeat this step 2 The ...

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Abstract

The invention relates to a blank biological sample preparation method, which specifically comprises: (1) separating vitamins bound to proteins under an acid / alkali condition to release endogenous vitamins; (2) adsorbing the released vitamins with an adsorption material; and (3) separating the adsorption material from a biological sample through high-speed centrifugation to obtain a vitamin-free blank biological sample. According to the present invention, the preparation process can effectively remove the endogenous vitamin compounds in the biological sample, can retain the characteristics of the biological sample to the greatest extent, and can provide the matrix environment similar to the to-be-detected biological sample for the clinical quantitative detection of vitamin compounds.

Description

technical field [0001] The invention relates to the field of biological sample preparation technology, in particular to a method for preparing a blank biological sample that removes endogenous vitamins from biological samples and retains the characteristics of the original organism. [0002] The invention can quickly and efficiently remove the endogenous analyte that interferes with the detection of the analyte, and maintain the matrix environment of the biological sample itself, and provide a method that does not contain the analyte and has biological characteristics for the establishment of a standard curve in the quantitative analysis process. A blank biological sample close to the sample to be tested. Background technique [0003] Vitamin compounds, including fat-soluble vitamins such as vitamins A, D, E, and K, and water-soluble vitamins such as vitamin B group and vitamin C. Although different vitamins have different chemical structures and biological activities, they...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/34
CPCG01N1/34
Inventor 蔡婷婷官培龙李仁杰
Owner 上海可力梅塔生物医药科技有限公司
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