Multi-Analyte Calibration Curve for High-Performance Liquid Chromatography with an Inductively Coupled Plasma Carbon Emission Detector

Abstract

A liquid chromatography system with an inductively coupled plasma detector is used to prepare a single calibration curve that is useful for multiple analytes. The detector monitors the atomic emission from carbon at 193.09 nm. Hence, the analytes need not exhibit appreciable molar absorptivity or native fluorescence. Since the carbon signal is independent of molecular structure, the sensitivities for different compounds are similar as long as nebulization efficiencies are comparable. In fact, with a suitable internal standard, no calibration curve is necessary. The capability of the system is demonstrated with a test mixture of nine amino acids separated with a C30 reversed-phase column and a 20 mM phosphate buffered mobile phase. The system provides a detection limit of 30 ng carbon. A multi-analyte calibration curve is prepared with 135 distinct measurements: each of nine analytes, at five different concentrations, repeated in triplicate. The average relative standard deviation for 27 measurements of different amino acids at a given concentration is 2.5%. Clearly, a single analyte will suffice for the calibration of all nine test compounds. Similarly, the internal standard method provides an average percent error of 2.0% for the determination of 45 different amino acid concentrations using only a single replicate for each sample.