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US 2022/0178951 A1Published application
Method for quantitative measurement of catechol estrogen bound protein in blood sample
AssigneeNational Cheng Kung University
InventorsShu-Hui CHEN, Yu-Shan HUANG, Hung-Hsiang JEN, Yu-Min LIN
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Abstract
The present invention relates to a method for quantitative measurement of catechol estrogen bound protein in blood sample. By detecting adduction levels of binding sites of the catechol estrogen on the protein in blood sample, the catechol estrogen bound protein in the blood sample can be detected quantitatively and a limit of quantitation can be decreased.
Claims14
A method for quantitative measurement of catechol estrogen bound protein in a blood sample, comprising:
- providing the blood sample, wherein the blood sample comprises a first amount of protein, the catechol estrogen binds to at least one site of the protein or not;
- performing an analytical procedure of a mass spectrometry that the blood sample is subjected to an analytical method of a tandem mass spectrometry for obtaining mass spectrometric data, wherein the mass spectrometric data comprise a plurality of unbound mass peaks and a plurality of bound mass peaks corresponding to the plurality of the unbound mass peaks, the plurality of the unbound mass peaks corresponds to catechol estrogen unbound protein, and the plurality of the bound mass peaks corresponds to catechol estrogen bound protein;
- and
- performing a determination procedure to obtain a second amount from the mass spectrometric data according to a formula (I) as follows, wherein the catechol estrogen bound protein has the second amount:
- B1=A1×Y1 (I)
- in the formula (I), B1 represents the second amount of the catechol estrogen bound protein, Y1 represents an adduction level of the catechol estrogen bound to the protein, A1 represents the first amount of the protein, and Y1 is obtained according to a formula (II) as follows:
- Y
-
-
- 1
- =
- f
- ×
- P
-
- 2
- P
-
- 1
- +
- P
-
- 2
- (
- II
- )
- in the formula (II), P1 represents a sum of areas of the plurality of the unbound mass peaks, P2 represents a sum of areas of the plurality of the bound mass peaks, and f represents a conversion factor.
24 more elements
A method for quantitative measurement of catechol estrogen bound hemoglobin in a blood sample, comprising:
- providing the blood sample, wherein the blood sample comprises a first amount of the hemoglobin, the hemoglobin has an amino acid sequence as shown by SEQ ID NO:
- 2, the catechol estrogen binds to at least one site of the hemoglobin or not;
- enzymatically digesting the hemoglobin with trypsin or chymotrypsin to obtain a plurality of amino acid fragments;
- performing an analytical procedure of a mass spectrometry by using a parallel reaction monitoring mode of an analytical method of a tandem mass spectrometry for obtaining mass spectrometric data, wherein the mass spectrometric data comprise a plurality of unbound mass peaks and a plurality of bound mass peaks corresponding to the plurality of the unbound mass peaks, the plurality of the unbound mass peaks corresponds to the catechol estrogen unbound hemoglobin, and the plurality of the bound mass peaks corresponds to the catechol estrogen bound hemoglobin;
- and
- performing a determination procedure to obtain a second amount from the mass spectrometric data according to a formula (I) as follows, wherein the catechol estrogen bound hemoglobin has the second amount;
- B1=A1×Y1 (I)
- in the formula (I), B1 represents the second amount of the catechol estrogen bound hemoglobin, Y1 represents an adduction level of the catechol estrogen bound to the hemoglobin, A1 represents the first amount of the hemoglobin, and Y1 is obtained according to a formula (II) as follows:
- Y
-
-
- 1
- =
- f
- ×
- P
-
- 2
- P
-
- 1
- +
- P
-
- 2
- (
- II
- )
- in the formula (II), P1 represents a sum of areas of the plurality of the unbound mass peaks, P2 represents a sum of areas of the plurality of the bound mass peaks, and f represents a conversion factor.
26 more elements
Dependencies are inferred from the claim text.