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AHP (2-Amino-3-hydroxyphenazine) and DAP (2, 3-Diaminophenazine) are process relevant impurities in Carbendazim Technical (fungicide), but as they are classified as toxic impurities these should be monitored at trace level. As their quantification limit are too low (0.5 mg Kg-1 for AHP and 3.0 mg Kg-1 for DAP), the research work is carried on mass spectrometer (MS) as detector by using HPLC. The chromatographic condition was optimized using gradient elution program on C18 stationary phase in reverse phase HPLC using selective ion monitoring (SIM) mode in MS detector. 2.0 mM aqueous Ammonium acetate and Acetonitrile (organic modifier) were used in mobile phase. The detector response was found linear over the concentration range of 0.0001 to 0.0024 mg L-1 (equivalent to 0.049 mg Kg-1 to 1.184 mg Kg-1) for AHP with coefficient of correlation 0.997 and at a concentration of 0.0001 to 0.0123 mg L-1 (equivalent to 0.051 mg Kg-1 to 6.129 mg Kg-1) for DAP with coefficient of correlation 0.997. The detection limit (LOD) was 0.02 mg Kg-1 for AHP and 0.02 mg Kg-1 for DAP, where as the quantification limit (LOQ) was 0.051 mg Kg-1 for AHP and 0.049 mg Kg-1 for DAP. The% RSD for method precision was 1.92% and 2.21% for AHP and DAP, respectively. The accuracy was found to be between 97.32% 104.47% for AHP and 96.57% 98.87 % for DAP. This analytical method can be applied for quantification of AHP and DAP in Carbendazim Technical.

oriental journal of chemistry/oriental journal of chemistry

Determination of 2-Amino-3-Hydroxyphenazine (AHP) and 2, 3-Diaminophenazine (Toxicologically Relevant Impurities) by RPHPLC-MS Method in Carbendazim Technical