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.

中文翻译：

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RPHPLC-MS法测定多菌灵技术中的2-Amino-3-Hydroxyphenazine（AHP）和2,3-Diaminophenazine（毒理学相关杂质）

AHP（2-Amino-3-hydroxyphenazine）和DAP（2，3-Diaminophenazine）是多菌灵工业（杀菌剂）中与工艺相关的杂质，但是由于它们被分类为有毒杂质，因此应进行痕量监测。由于它们的定量限太低（AHP为0.5 mg Kg-1，DAP为3.0 mg Kg-1），因此研究工作是通过HPLC作为检测器在质谱仪（MS）上进行的。色谱条件通过在反相HPLC中使用C18固定相的梯度洗脱程序在MS检测器中使用选择性离子监测（SIM）模式进行优化。在流动相中使用2.0 mM醋酸铵水溶液和乙腈（有机改性剂）。发现在AHP的浓度范围为0.0001至0.0024 mg L-1（相当于0.049 mg Kg-1至1.184 mg Kg-1）范围内，检测器响应呈线性关系，相关系数为0。997，DAP的浓度为0.0001至0.0123 mg L-1（相当于0.051 mg Kg-1至6.129 mg Kg-1），相关系数为0.997。AHP的检出限（LOD）为0.02 mg Kg-1，DAP的检出限（LOQ）为0.051 mg Kg-1，DAP的检出限（LOQ）为0.051 mg Kg-1。AHP和DAP的方法精密度的RSD％分别为1.92％和2.21％。发现准确度在AHP的97.32％104.47％和DAP的96.57％98.87％之间。该分析方法可用于多菌灵技术中AHP和DAP的定量分析。对于AHP为051 mg Kg-1，对于DAP为0.049 mg Kg-1。AHP和DAP的方法精密度的RSD％分别为1.92％和2.21％。发现准确度在AHP的97.32％104.47％和DAP的96.57％98.87％之间。该分析方法可用于多菌灵技术中AHP和DAP的定量分析。对于AHP为051 mg Kg-1，对于DAP为0.049 mg Kg-1。AHP和DAP的方法精密度的RSD％分别为1.92％和2.21％。发现准确度在AHP的97.32％104.47％和DAP的96.57％98.87％之间。该分析方法可用于多菌灵技术中AHP和DAP的定量分析。