Accurate Measurement of glutathione for disease diagnosis and drug metabolite screening
AssigneeDefinitek, Inc.
InventorsHoward M. Kingston, Mehmet Pamukcu
Life of this patent
Estimating the term…
Abstract
A method of measuring and calculating (preferably by a computer with output to a user) tGSH (total glutathione very particularly defined) with sample preparation and assay methods that have been confirmed to provide accurate and reliable tGSH and related diagnostic assays in blood or tissue from a patient.
Claims11
A method of accurate measurement and calculation of tGSH, GSH (reduced glutathione) and the ratio of GSH to oxidized glutathione (GSSG) in a sample of patient blood, urine, saliva, blood components or tissue, comprising collecting a blood or tissue sample;
- combining the sample with isotopically tagged spikes;
- and mass spectronometrically analyzing and computing tGSH, GSH and
- GSH/GSSG, where tGSH is the sum of GSHcorrected, GSSGcorrected and CGSH, and the ratio is given by using the formulas:
- GSH
- GSSG
- =
- z
- γ
- Where:
- z
- =
- GSH
- corrected
- =
- GSH
- SIDMS
- +
- α
- +
- β
- α
- =
- [
- GSH
-
-
- oxidation
-
- GSSG
- ]
- β
- =
- CGSH
- =
- ∑
- i
-
- CGSH
- 1
- +
- CGSH
- 2
- +
- …
-
-
- CGSH
- i
- γ
- =
- GSSG
- corrected
-
- (
- GSSG
- -
- [
- GSH
-
-
- oxidation
-
- GSSG
- ]
- )
- wherein the tGSH, GSH and GSH/GSSG data is calculated by a computer and output to a user according to the above calculations carried out on measurements performed by the mass spectrometer.
Glutathione screening of drug metabolites where CGSH is determined according to given by using the formulas:
- CGSH=ΣiCGSH1+CGSH2+ . . . CGSHi
- wherein the target drug and enzyme such as cytochrome P450 enzyme but not limited to it producing one or more data of chemically reactive metabolites of CGSH concentrations which are calculated by a computer and output to a user according to the above calculations carried out on measurements performed by the mass spectrometer.
A method of algorithmic and mathematical quantification of GSH, GSH-species, GSH-conjugates and GSH-metabolites, by following the principles and sample preparation protocols of D-IDMS and/or D-SIDMS.
- Wherein the algorithmic measurement involves and relies on properties of natural abundant and enriched molecules that includes but not limited to GSH, and related species, conjugates and species;
- Wherein in the purpose of measurement is toxicological screening of biologic drugs;
- Wherein the preferred need for GSH-screening is during biologic drug development, although it can also be done subsequent to a drug's release to the market;
- Wherein the measurement is done for GSH, GSH-conjugates, GSH-metabolites or any forms that will be generated after an established digestion protocol prior to mass spectrometric detection;
- and
- Wherein GSH-metabolites can be either or both soft-metabolites or hard metabolites.
Dependencies are inferred from the claim text.