Development and validation of a HPLC method for 4,7-phenanthroline-5,6-dione I and identification of its major impurity by HPLC-MS-APCI.
In this study, the development and validation of an analytical method for the assay of 4,7-phenanthroline-5,6-dione I (dione I) using high-performance liquid chromatography (HPLC) and the determination of its synthetic impurities by employing the method in HPLC-mass spectrometry with atmospheric-pressure chemical ionization and photodiode-array UV detection is reported. The results show that dione I is eluted as a spectrally pure peak resolved from its impurities. 5-Bromo,4-7-phenanthroline is identified as the main impurity. This is supported by elemental analysis of the dione I, which demonstrated the presence of bromine. Validation parameters such as specificity and selectivity, linearity, accuracy, precision, limit of detection (LOD), limit of quantitation (LOQ), ruggedness, stability, and system suitability, which are evaluated for this method. The LOD and LOQ are 2.0 microg/mL and 50 microg/mL with a 0.50% relative standard deviation (%RSD), respectively. The calibration curves showed good linearity over the concentration range of 0.05-1.50 mg/mL. The correlation coefficient is > 0.9991 in each case. The %RSD values for intra- and interday precision studies are < 0.40%.
Shabir
Ghulam A
GA
Abbott Laboratories, R and D, Range Road, Witney, Oxon OX29 0YL, UK. ghulam.shabir@abbott.com
Forrow
Nigel J
NJ
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004340:Drug Contamination; D004355:Drug Stability; D013058:Mass Spectrometry / Q000379:methods; D010618:Phenanthrolines / Q000032:analysis* / Q000138:chemical synthesis / Q000302:isolation & purification / Q000592:standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
C503074:5-bromo-4,7-phenanthroline; D010618:Phenanthrolines; C004902:phanquinone