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目的分析比较注射用盐酸大观霉素有关物质测定的高效液相色谱脉冲安培检测法(HPLC-PAD)和高效液相色谱蒸发光散射检测器法(HPLC-ELSD),并采用两种方法测定了31批次样品。方法两法色谱柱均为Apollo C_(18)(4.6 mm×250 mm,5μm),流动相均为0.1 mol·L~(-1)三氟乙酸溶液,柱温均为30℃,进样量均为20μL。PAD检测:检测电极为金电极(3 mm),Ag-AgCl参比电极,钛合金对电极,积分安培检测器,四波形检测电位。ELSD检测:漂移管温度110℃,载气流速2.6 L·min~(-1),增益1。结果两法专属性一致,检出有关物质种类和个数一致;PAD检出限为2.4 ng,ELSD检出限为72.8 ng(以S/N=3计);PAD为直接线性关系,质量浓度范围为0.15~150μg·mL~(-1);ELSD为双对数线性关系,浓度范围为17~170μg·mL~(-1)。两法测定31批次样品有关物质,杂质D和杂质E结果一致,杂质A、4R-双氢大观霉素和总杂差异较大;PAD中杂质A采用校正因子校正后,杂质A和总杂结果均与ELSD一致。结论两种方法均可有效检出有关物质,达到控制大观霉素有关物质的目的。HPLC-PAD需对杂质A和4R-双氢大观霉素进行响应因子校正,ELSD需采用双对数线性回归。
OBJECTIVE: To compare the HPLC-PAD and HPLC-ELSD methods for the determination of spectinomycin hydrochloride for injection, and to determine 31 batches of samples. Methods The two chromatographic columns were Apollo C_ (18) (4.6 mm × 250 mm, 5 μm). The mobile phase was 0.1 mol·L -1 trifluoroacetic acid and the column temperature was 30 ℃. The injection volume Both are 20 μL. PAD test: the detection electrode is a gold electrode (3 mm), an Ag-AgCl reference electrode, a titanium alloy counter electrode, an integral amperometric detector, and a four-wave detection potential. ELSD detection: drift tube temperature 110 ℃, carrier gas flow rate 2.6 L · min -1, gain 1. Results The specificity of the two methods was the same. The detection limit was 2.4 ng for PAD and 72.8 ng for ELSD (S / N = 3). The PAD was directly related to the mass The concentration range was 0.15-150 μg · mL -1. ELSD had a logarithmic linear relationship with a concentration range of 17-170 μg · mL -1. Two methods for the determination of 31 batches of samples of related substances, impurities D and impurities E results are consistent, impurities A, 4R-dihydropermethacin and total heterodyne differences are larger; PAD impurity A correction factor correction, impurity A and total miscellaneous The results are consistent with the ELSD. Conclusion Both methods can effectively detect the related substances and achieve the purpose of controlling the related substances of spectinomycin. HPLC-PAD requires impurity factor A and 4R-dihydronaphthalene response factor correction, ELSD dual logarithmic linear regression.