Abstract��OBJECTIVE To compare the HPLC-PAD method and HPLC-ELSD method for determination of related substances in spectinomycin hydrochloride for injection, and determine 31 batches of samples by the two methods. METHODS The following conditions were used for both methods:the analytical column was Apollo C18 column (4.6 mm��250 mm,5 ��m), mobile phase was 0.1 mol��L-1 trifluoroacetic acid solution, the column temperature was maintained at 30 ��, and the sample volume was 20 ��L. For the PAD detection, gold electrode (3 mm)was used as the detection electrode, Ag-AgCl as reference electrode, titanium alloy as counter electrode, and integrated amperometric detector was employed with four waveform detection potential. For the ELSD detection, the drift tube temperature was set at 110 ��, and the carrier gas flow rate was 2.6 L��min-1, with gain of 1. RESULTS The two methods were consistent in the species and number of detected impurities. The detection limits of the PAD method and ELSD method were 2.4 and 72.8 ng, respectively (S/N=3). For the PAD method there existed a direct linear relationship in the concentration range of 0.15-150 ��g��mL-1, while a double logarithmic linear relationship was shown in the concentration range of 17-170 ��g��mL-1 for the ELSD method. When 31 batches of samples were determined for related substances by the two kinds of methods, the contents of impurity D and E were basically the same, but there were significant differences in the contents of impurity A and (4R)-dihydrospectinomycin. After correction, the contents of impurity A and total impurities determined by the PAD method were consistent with those by the ELSD method. CONCLUSION The two methods can both effectively detect the related substances of spectinomycin to achieve the purpose of controlling related substances. When using HPLC-PAD method, the impurity A and (4R)-dihydrospectinomycin should be adjusted by using response factor or the reference substances. The ELSD method needs to use double log linear regression.
����Ƽ, ���ٽ�, ��Ӣ. ע����������ù���й��������ֲⶨ�����ıȽ�[J]. �й�ҩѧ��־, 2017, 52(20): 1786-1791.
WANG Li-ping, LI Shao-jie, LIU Ying. Determination of Related Substances of Spectinomycin Hydrochloride for Injection by Two Analysis Methods. Chinese Pharmaceutical Journal, 2017, 52(20): 1786-1791.
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