Combination of High Performance Liquid Chromatography and Flame Atomic Absorption Spectrophotometry Using a Novel Nebulizer Interface Supported T Shaped Slotted Quartz Tube for the Determination of Vitamin B12

Abstract

A novel nebulizer interface (NI) was proposed to combine high performance liquid chromatography (HPLC) and flame atomic absorption spectrophotometer (FAAS). A glass concentric nebulizer was linked to T-shaped slotted quartz tube (T-SQT) using a tubing to transfer the liquid solution eluted from the chromatographic system into the atomization region of FAAS system. T-SQT was also used to intensify the interaction of atoms with the hollow cathode lamp light. Vitamin B12 was selected as an analyte to show the applicability of the new hyphenated system. After optimizing some parameters such as mobile phase flow rate and pH, nebulizer gas flow rate, T-SQT height and injection volume, linear range for the analyte was determined between 4.7 and 92 mg/kg as Co. Limit of detection (LOD) and limit of quantitation (LOQ) for the HPLC-NI-T-SQT-FAAS system were calculated to be 1.6 and 5.3 mg/kg as Co, respectively. Recovery studies were also conducted to verify the accuracy and applicability of the developed method for vitamin tablets and excellent percent recovery results (~ 100%) with low standard deviation values were obtained when matrix-matching calibration strategy was performed for each vitamin tablet. A successful separation and detection of the analyte was achieved within 3.0 min that offers high sample throughput. Two different vitamin tablets were analyzed by the optimized hyphenated system. The developed method also provides low usage of sample solution in contrast to conventional nebulizer in the FAAS system.