Abstract��OBJECTIVE To establish an HPLC-PAD method to determine the related substances of sisomicin sulfate injection and compare with the statutory method. METHODS IonPac AMG C18(4.0 mm��150 mm, 3 ��m)chromatographic column was used with acetonitrile-0.1 mol��L-1 trifluoroacetic acid (containing 0.025% of pentafluoropropionic acid, 5 mL of 50% NaOH solution without carbonate, pH of the aqueous solution adjusted to 2.3 with 50% NaOH solution.)as mobile phase at a flow rate of 0.7 mL��min-1. NaOH solution of 0.76 mol��L-1 was added post column at a flow rate of 0.35 mL��min-1. The column temperature was maintaine at 30 ��. PAD detector was operated with the cell temperature set at 35 ��. The working electrode was a gold electrode (diameter of 3 mm)and a quadruple-potential waveform was selected as detection waveform. The reference electrode was Ag/AgCl, the detection potential was four potential. The determination result of the related substances of sisomicin sulfate injection was compared with that of the statutory method. RESULTS The peaks of sisomicin sulfate, gentamicin C1a and netilmicin could be completely separated, and other impurities could also be effectively separated. The blank sample had no interferences. The LOD and LOQ of etimicin were found to be 2 and 6 ng respectively, and the RSD of precision test (n=6) was 0.9%. Paired-samples t-test showed significance levels of P=0.034, P=0.364 and P=0.605 for total amount of impurities (%), the biggest single impurity (%)and content (%)respectively between the statutory method and the method of HPLC-PAD. CONCLUSION Compared with the statutory method, this HPLC-PAD method shows higher sensitivity, and is accurate and reliable. It can be applied to the determination of related substances in sisomicin sulfate injection.
�ƴ���, ����ϼ, ����, �΄���, �ܹ��, ��. ������������ע��Һ�й����ʷ��������ıȽ�[J]. �й�ҩѧ��־, 2017, 52(20): 1780-1785.
HUANG Chun-qing, HAN Chun-xia, LI Hong-di, HE Jie-yi, CAO Gui-hong, XU Bo. Comparison of Analytical Methods for Related Substances of Sisomicin Sulfate Injection. Chinese Pharmaceutical Journal, 2017, 52(20): 1780-1785.
WANG J, WANG Z J,CHEN G B,et al. Analysis of sisom icin sulfate and its related substances by HPLC-ELSD and HPLC-MSn[J]. Chin Pharm J(�й�ҩѧ��־),2007,42(1):62-66.
[2]
WANG W, PAN Q,REN W Z,et al. Determ ination of sisom icin sulfate injection by HPLC-ELSD[J]. West China J Pharm Sci(����ҩѧ��־),2009,24(3):306-307.
[3]
LU X H. Sisomicin combination of acute renal failure analysis in 3 cases [J]. Chin J Pharmacoepidemiol(ҩ�����в�ѧ��־),2007,16(1):53-54
[4]
YUAN Y Z,ZHANG X,ZHANG M,et al. HPLC-ESI-IT-MSn characterization of related substances in sisomicin sulfate [J]. China J Pharm Anal(ҩ�������־),2011,31(7):1288-1295.
[5]
ZHOU H P. Resarch of the defferential refraction-HPLC mothod about assay testing of sisomicin sulate ingredient product[D]. Zhejiang:Zhejiang University of Technology,2007.
CHEN W D. Aminoglycoside antibiotics related substances test method research[J]. Strait Pharm J(��Ͽҩѧ),2008,20(9):128-130.
[9]
WANG Y,WANG M J,YAO S C, et al. The application and outlook of electrochemical detector (ECD)in the Chinese Pharmacopeia, the US Pharmacopeia and the European Pharmacopeia [J]. Chin J Antibiot(�й���������־),2012,37(11):801-805.
[10]
WU Y N,ZHAO W,ZHU X Y,et al. Improved liquid chromatography combined with pulsed electrochemical detection for the analysis of etimicin sulfate[J]. Chin Pharm J(�й�ҩѧ��־),2015,50(20):1797-1782.
[11]
ZHANG Y H,ZHEN H S, CHENG L. Comparison of determination of the components and related substances in gentamicin by two HPLC methods[J]. Chin Archives Tradit Chin Med(�л���ҽҩѧ��),2007,25(4):831-833.
WU Y N,ZHAO W,ZHU X Y,et al. Determination of related substances in etimicin sulfate injection and etimicin and sodium chloride injection by LC-PED [J]. Chin J Antibiot(�й���������־),2016,41(8):618-623.
[14]
YANG H,YUAN Y Z,ZHAO X,et al. Study on impurity profile of C1a by HPLC-ESI-IT-MSn [J]. Pharm Clin Res(ҩѧ���ٴ��о�),2012,20(5):413-418.
2017, Vol. 52 
