A rational approach for HPLC calibration using mass spectrometer as detector

Abstract

A direct, accurate, precise, reproducible and cost-time efficient approach for calibration of HPLC using mass spectrometer as detector has been described which simulates the routine analysis in bioanalytical laboratories. Injector position check and injection volume repeatability test for the calibration of autosampler; and pump flow rate accuracy test for the calibration of pumps were illustrated using direct weighing method. Carryover test, pump repeatability test, linearity and precision were carried out for holistic (overall system) calibration of the instrument using a sensitive high throughput LC–MS/MS pre-validated method. The results of calibration experiments for individual modules suggested that pumps are yielding accurate flow rate, autosampler is aspirating the correct injection volume irrespective to position or volume, and column oven and autosampler chambers are maintaining steady temperatures. Holistic calibration verified the accuracy, precision, repeatability and reproducibility of the instrument with insignificant carryover.