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Determination method for converting content of free substances under equilibrium dialysis by using ultrafiltration method

A technology of free substance and equilibrium dialysis, which is applied in the direction of measuring devices, material separation, and analysis materials, etc., can solve the problems of inability to meet the rapid analysis results of clinical biological samples, large deviations of measurement results, and difficult temperature control, etc., to achieve reliable detection results, The effect of lower detection limit and easy temperature control

Pending Publication Date: 2022-06-07
合肥歆智医疗器械有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] For the pre-treatment of the existing high-performance liquid chromatography tandem mass spectrometry technology to detect free substances, the ultrafiltration method is efficient and fast, but the temperature is not easy to control during the treatment process, resulting in large deviations in the measurement results, while the equilibrium dialysis method is more accurate. Accurate, but time-consuming, unable to meet the requirements of rapid analysis results of clinical biological samples, this application provides a method for determining the content of free substances in equilibrium dialysis using ultrafiltration, which can take into account the high detection accuracy and short time-consuming Require

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  • Determination method for converting content of free substances under equilibrium dialysis by using ultrafiltration method
  • Determination method for converting content of free substances under equilibrium dialysis by using ultrafiltration method
  • Determination method for converting content of free substances under equilibrium dialysis by using ultrafiltration method

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Embodiment 1

[0060] One of the present embodiments using an ultrafiltration method converts a method for determining the content of ionizing substances downstream of equilibrium dialysis, the sample is free testosterone.

[0061] The method of the present embodiment, first select 20 ° C, 25 ° C, 30 ° C and 34 ° C under the four temperature values, respectively, ultrafiltration method and equilibrium dialysis method to separate free testosterone, in fact, other temperature values may be used. It was then detected by high performance liquid chromatography tandem mass spectrometry, and the results are shown in Table 1. thereinto

[0062] The specific schemes used in the ultrafiltration method are:

[0063]Ultrafiltration: Take 300 μL of serum samples, add 900 μL of 4-hydroxyethylpiperazine ethsulfonic acid buffer, blow and mix well with a pipette, and then transfer to the activated ultrafiltration tube, at 20 °C, 25 °C, 30 °C and 34 °C, respectively, 2000 g centrifugation for 1 h;

[0064] Solid ...

Embodiment 2

[0088] The method of the present embodiment, first selected 20 ° C, 25 ° C, 30 ° C and 34 ° C under the four temperature values, respectively, ultrafiltration method and equilibrium dialysis method to separate the free T3 (triiodothyronine), in practice can be used other temperature values. It was then detected by high performance liquid chromatography tandem mass spectrometry, and the results are shown in Table 3. thereinto

[0089] The specific schemes used in the ultrafiltration method are:

[0090] Ultrafiltration: take 100 μL of serum samples, add 100 μL of 4-hydroxyethylpiperazine ethsulfonic acid buffer, blow and mix well with a pipette, and then transfer to the activated ultrafiltration tube, at 20 °C, 25 °C, 30 °C and 34 °C, respectively, centrifuge at 2000 g for 1 h;

[0091] Internal standard precipitation: Take 100 μL ultrafilter and add 100 μL of internal standard solution, vortex for 3 min, 4 °C, centrifuge at 12000 rpm for 10 min.

[0092] The specific schemes used ...

Embodiment 3

[0114] The method of the present embodiment, first of all, 20 ° C, 25 ° C, 30 ° C and 34 ° C under the four temperature values, respectively, ultrafiltration method and equilibrium dialysis method to separate the free T4 (thyroxine), in practice can be used other temperature values. It was then detected by high performance liquid chromatography tandem mass spectrometry, and the results are shown in Table 5. thereinto

[0115] The specific schemes used in the ultrafiltration method are:

[0116] Ultrafiltration: take 100 μL of serum samples, add 100 μL of 4-hydroxyethylpiperazine ethsulfonic acid buffer, blow and mix well with a pipette, and then transfer to the activated ultrafiltration tube, at 20 °C, 25 °C, 30 °C and 34 °C, respectively, centrifuge at 2000 g for 1 h;

[0117] Internal standard precipitation: Take 100 μL ultrafilter and add 100 μL of internal standard solution, vortex for 3 min, 4 °C, centrifuge at 12000 rpm for 10 min.

[0118] The specific schemes used by the b...

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Abstract

The invention belongs to the technical field of free substance detection, and relates to a determination method for converting the content of free substances under equilibrium dialysis by using an ultrafiltration method. The invention aims to solve the problems that when the existing high performance liquid chromatography-tandem mass spectrometry technology is used for detecting the pretreatment of free substances, an ultrafiltration method is efficient and rapid, but the temperature is not easy to control in the treatment process, so that the deviation of a measurement result is large, and an equilibrium dialysis method is easy to control the temperature, more accurate in measurement result, long in time consumption and low in cost. According to the determination method for converting the content of the free substances under equilibrium dialysis by using the ultrafiltration method, the free substances are separated from the sample by using the ultrafiltration method to obtain first concentration data, the free substances are separated from the sample by using the equilibrium dialysis method to obtain second concentration data, and the first concentration data and the second concentration data are obtained. The linear equation is established by using the first concentration data and the second concentration data, and the requirements of high detection accuracy and short time consumption can be met.

Description

Technical field [0001] The present invention belongs to the field of free matter detection technology, specifically, relates to a method of measuring the content of ionized substances downstream of equilibrium dialysis using ultrafiltration conversion. Background [0002] Free hormones are more biologically active in the human body, is the main part of the realization of the biological effects of the hormone, the current method of isolating the free analyte is mainly balanced dialysis and ultrafiltration. Among them, equilibrium dialysis is considered to be the gold standard for measuring free analytes. [0003] For example, China Patent Application Publication No. CN111398490A, the name of the invention is "Mass Spectrometry For the Detection of Free Triiodothyronine and Free Thyroxine Kit", the disclosed kits include: reagents for balancing dialysis to isolate the binding state and free state of Triiodothyronine and thyroxine; Also includes dialysate; and reagents for the extra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/72G01N30/06G01N30/08G01N30/14
CPCG01N30/02G01N30/72G01N30/06G01N30/08G01N30/14G01N2030/146G01N2030/062
Inventor 彭军江振作刘洋谢沙莎
Owner 合肥歆智医疗器械有限公司
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