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Methods For Detecting Reverse Triiodothyronine By Mass Spectrometry

A technique of mass spectrometry, trans-type, applied in methods using particle spectrometers, mass spectrometry, material analysis by electromagnetic means, etc.

Inactive Publication Date: 2014-09-10
QUEST DIAGNOSTICS INVESTMENTS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Thus, rT3 does not stimulate the metabolic activity of target cells and, in fact, blocks T3 from activating receptor sites

Method used

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  • Methods For Detecting Reverse Triiodothyronine By Mass Spectrometry
  • Methods For Detecting Reverse Triiodothyronine By Mass Spectrometry
  • Methods For Detecting Reverse Triiodothyronine By Mass Spectrometry

Examples

Experimental program
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Effect test

preparation example Construction

[0072] Protein precipitation is a preferred method of preparing samples, especially biological samples such as serum or plasma. Protein precipitation can be used to remove at least a portion of the protein present in the sample, leaving rT3 in the supernatant. The precipitated sample can be centrifuged to separate the liquid supernatant from the precipitated protein; alternatively the sample can be filtered, eg, through a glass fiber filter, to remove the precipitated protein. The resulting supernatant or filtrate can then be applied directly to mass spectrometry; or alternatively to liquid chromatography followed by mass spectrometry.

[0073] Various precipitating agents are known in the art, such as acetone, alcohols such as methanol, or various acidifying agents. In certain embodiments, the use of protein precipitation such as, for example, methanolic protein precipitation can eliminate the need for solid phase extraction (SPE) such as high turbulence liquid chromatograph...

Embodiment 1

[0099] Example 1 : Sample (serum) and reagent preparation

[0100] By collecting blood in standard Red Hat serum Serum samples were prepared in tubes and allowed to clot for 30 min at room temperature. The samples were then centrifuged and the serum was immediately separated from the cells. Optionally, blood is collected in double gel barrier tubes and allowed to clot at room temperature. The samples were then centrifuged and the serum was separated from the cells within 24 hours.

[0101] Also prepare plasma in EDTA tube and heparin Plasma samples collected in tubes for analysis.

[0102] Prepare three rT3 stock solutions. Prepare a 1 mg / mL initial rT3 stock solution in methanol / basic solution by dissolving rT3 in 40 mL of concentrated NaOH and diluting to 100 mL with methanol. Prepare an intermediate stock solution of 1,000,000 pg / mL rT3 by further diluting a portion of the initial stock solution with methanol. Finally, a working stock solution of 10,000 pg / mL...

Embodiment 2

[0104] Embodiment 2: rT3 in enrichment serum by protein precipitation

[0105] First add 100 μL of the sample to the wells of the 96-well plate. Then 500pg / mL in 300μL methanol solution 13 C 6 - rT3 (internal standard) was added to each well and each well was checked for precipitate formation. After visual confirmation of precipitation, the plate was mixed at about 1500 rpm for about 1 minute, allowed to stand, mixed again, refrigerated for about 30 minutes, and mixed one final time. After the final mixing, the plates were centrifuged at a minimum of 3000 xg for at least 30 minutes.

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PUM

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Abstract

Provided are methods for determining the amount of reverse T3 in a sample using mass spectrometry. The methods generally involve ionizing reverse T3 in a sample and detecting and quantifying the amount of the ion to determine the amount of reverse T3 in the sample.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit under 35 U.S.C. §120 of US Application Serial No. 13 / 311,412, filed December 5, 2011, the entire contents of which are incorporated by reference into this disclosure. technical field [0003] The invention relates to the detection of trans-triiodothyronine. In a specific aspect, the invention relates to a method of detecting trans-triiodothyronine by mass spectrometry. Background technique [0004] The following description of the background of the invention is provided simply to aid in the understanding of the invention and is not an admission that describes or constitutes prior art to the invention. [0005] Trans-triiodothyronine ((2S)-2-amino-3-[4-(4-hydroxy-3,5-diiodophenoxy)-3-iodophenyl]propionic acid) (rT3) It is an inactive isomer of triiodothyronine (T3). Both T3 and rT3 are derived from thyroxine (tetraiodothyronine, T4) through the action of the following deiodinases: ...

Claims

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Application Information

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IPC IPC(8): G01N33/50G01N27/62G01N30/02B01D15/08
CPCG01N33/78H01J49/0081G01N2560/00G01N33/6848H01J49/0431H01J49/0031H01J49/00G01N30/7233H01J49/0027H01J49/004H01J49/04H01J49/10
Inventor J·F·班克斯P·P·周N·M·马特
Owner QUEST DIAGNOSTICS INVESTMENTS INC
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