Specific analysis of ketone and aldehyde analytes using reagent compounds, labeling strategies, and mass spectrometry workflow
View Patent ↗Labeling reagents, sets of labeling reagents, and labeling techniques are provided for the relative quantitation, absolute quantitation, or both, of ketone or aldehyde compounds including, but not limited to, analytes comprising steroids or ketosteroids. The analytes can be medical or pharmaceutical compounds in biological samples. Methods for labeling, analyzing, and quantifying ketone or aldehyde compounds are also disclosed as are methods that also use mass spectrometry.
1. A method for mass analysis of an analyte in a sample comprising:
derivatizing an analyte comprising an aldehyde or ketone functional group, with a labeling reagent of formula:
Y—(CH 2 ) n -ONH 2
wherein n is an integer from 2 to 10 and Y is a group selected from the group consisting of:
subjecting the labeled analyte to ionization; and
detecting the analyte by mass analysis.
2. The method of claim 1 , further comprising determining a concentration of the analyte in a sample.
3. The method of claim 1 , further comprising the step of subjecting the labeled analyte to ion fragmentation to yield an ionized reporter group.
4. The method of claim 1 , wherein the step of detecting the analyte is comprised of detecting a first transmitted parent ion and a daughter ion fragment by parent-daughter ion transition monitoring.
5. The method of claim 1 , wherein the labeling reagent contains one more heavy atom isotopes, the method further comprising derivatizing an aldehyde or ketone functional group of a standard compound with a labeling compound that is isobaric to the labeling compound used to form the labeled analyte and measuring the relative concentration of the analyte.
6. The method of claim 1 , wherein the labeling reagent is isotopically enriched with two or more heavy atoms.
7. The method of claim 1 , wherein at least two analyte compounds are derivatized with the labeling reagent and the method further comprises the step of determining a relative concentration between at least two analytes.
8. The method of claim 1 , wherein at least two analyte compounds are derivatized with the labeling reagent and the method further comprises the step of determining an absolute concentration of at least one analyte.
9. The method of claim 1 , wherein the ionization produces structurally specific fragment ions and Q3 MRM ions, the labeling reagent is wholly or partly contained in the structurally specific fragment ions, and the method provides both sensitivity and specificity for the Q3 MRM ions.