UHPLC-Q/TOF MS结合HPLC-DAD定性和定量分析硫酸黏菌素可溶性粉中的未知添加物

Qualitative and Quantitative Analysis of Unknown Additives in Colistin Sulphate Soluble Powder by UHPLC-Q/TOF MS and HPLC-DAD

定性和定量分析一批兽药硫酸黏菌素可溶性粉中的未知添加物。照《中国兽药典》2010年版一部对该批检品采用微生物检定法进行含量测定时,发现该样品的抑菌圈为虚圈,薄层色谱鉴别该样品,未显示与标准品溶液一致的主斑点,怀疑该样品中有处方外非法添加物。采用超高效液相色谱-四级杆-飞行时间质谱(UHPLC-Q/TOF MS)对该样品进行筛查,发现疑似添加物,并使用液相色谱-二极管阵列检测(HPLC-DAD)法进行了双重确证和含量测定。该样品中非法添加物确证为磺胺氯达嗪和甲氧苄啶,添加量分别为58.5 mg/g和13.4 mg/g。

A qualitative and quantitative analysis of unknown additives in a batch of veterinary colistin sulphate soluble powder had been implemented.During the content determination of the sample with the method of microbiological assay according to Chinese Veterinary Pharmacopoeia2010,the inhibition zones were virtual.And the spot of the sample did not accord with the spot of the reference substance in the identification by the thin-layer chromatography(TLC).It revealed that some unkown compounds may be doped in the sample.Then the screening of the sample through ultra high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(UHPLC-Q/TOF MS)was carried out.Furthermore,double confirmation and content determination were accomplished by high performance liquid chromatography-diode array detection(HPLC-DAD).Sulfachloropyridazine and trimethoprim as the illegal additives were detected at58.5mg/g and13.4mg/g respectively.The research provided the technique references for the inspection of illegal adulterations in veterinary drug.