Analyte mass spectrometry quantitation using a universal reporter
Quantitation of analytes, including but not limited to peptides, polypeptides, and proteins, in mass spectrometry using a labeled peptide coupled to a reporter, and a universal reporter.
1. A method of generating a dilution curve in a single injection into a liquid chromatograph/mass spectrometer, the method comprising
injecting into a first liquid chromatography (LC) column coupled to a mass spectroscopy detection system a single peptide composition, each peptide in the composition having the identical amino acid sequence A whereby each amino acid is present in the composition as a heavy labeled isomer resulting in the composition being a sequence A blend of all heavy labeled isomers, each isomer in the blend of sequence A having a mass shift and each isomer in the blend of sequence A present in a different concentration,
analyzing each peptide in the co-eluted peptide composition by mass spectroscopy, and
generating from the single peptide composition injection of sequence A into the LC column a dilution curve from the analysis.
2. The method of claim 1 resulting in a decreased loss of the less abundant isomer by non-specific binding to a component of the LC column due to the concentration of the most abundant isomer being higher than the concentration of the most diluted isomer.
3. The method of claim 1 further comprising
injecting at least an additional sequence B and repeating the method, and
comparing the result of analysis of each of the sequence A blend and sequence B blend to compare limits of detection (LOD), and limits of quantitation (LOQ) between different column injections.
4. The method of claim 1 further comprising
injecting the peptide composition from claim 1 into a second liquid chromatography (LC) column coupled to a mass spectroscopy detection system apparatus and repeating the method, and
comparing the result of analysis from each of the first column and second column to compare limits of detection (LOD), and limits of quantitation (LOQ) between different systems.
5. The method of claim 4 wherein the systems are the same brand or are different brands.