Method for the detection and / or direct quantification of at least one compound with a molecular weight of at least 200

摘要

Method for the direct detection and quantification of at least one compound with a molecular weight of at least 200, wherein the compound is a chemically complex molecule, wherein said chemically complex molecule is an inositol polyphosphate selected from the group containing 2 to 6 phosphate groups, including ions or salts thereof, wherein the compound or compounds are within a biological matrix, wherein said biological matrix is a fluid selected from the list consisting of blood, plasma, serum, urine, saliva, liquid lymphatic, cerebrospinal fluid and mixtures thereof, wherein the method comprises at least the following steps: i) preparing at least one standard sample of the chemically complex molecule to be detected and quantified; ii) introduction of the standard sample into the solvent system stream; wherein the solvent system consists of an aqueous solution of an amine; wherein the pH of the solvent has been adjusted, between 8 and 13; iii) passing the sample and the solvent system through at least one column in which the column is a stationary phase of inverse phase, the particle size being between 1.5-15 microns and the pore size between 60-500 Angstroms, while maintaining system pressure between 5 and 1500 atm; iv) identify the retention time and quantify the signal strength of when the chemically complex molecule is eluted, by a mass detector; v) preparing a chemically complex molecule sample from a biological matrix comprising the chemically complex molecule to be detected and quantified, wherein the process for preparing the sample comprises at least totally or partially dissolving the biological matrix to form a solution or a suspension and, if necessary, treat the solution or suspension to remove suspended particles; vi) repeat steps (ii) and (iii) with the sample prepared in step (v); together or sequentially with the standard; and vii) detecting and quantifying the presence of the chemically complex molecule by comparing the retention time and intensity of the signal of the standard sample or the standard samples, or by comparing the retention time and intensity of the signal obtained from previous studies or from the literature, where the detector capable of detecting the elution of the chemically complex molecule is a mass spectrometer; a tandem mass spectrometer or a single quadrupole; working under any of the following modes of operation: monitoring of selected ions (SIM); multiple reaction monitoring (MRM); Selected reaction monitoring (SRM) and SCAN (negative / positive); or a combination of them.