高效液相色谱法测定庆大霉素C组分的不同检测方式测定结果的比较

Comparison of the Determination Results of Gentamycin C Components by High Performance Liquid Chromatography with Different Detectors

各国药典中关于庆大霉素C组分的测定方法均为高效液相色谱法,但检测方式及分离效果不同。为此采用直观推导式演进特征投影法(HELP),对高效液相色谱-二极管阵列检测采集的庆大霉素C组分色谱数据进行解析,分辨出各物质的色谱曲线,在扣除了未分离的杂质峰对庆大霉素C1组分的干扰后,对柱前邻苯二醛衍生化-二极管阵列检测法及目前中国药典2005版收载的高效液相色谱-蒸发光散射检测方法测定庆大霉素C组分的准确性进行了比较,并运用柱切换技术,证明二者测定结果的一致性。

The results were compared for determining gentamycin C components by high performance liquid chromatography with different detectors in pharmacopoeias. Evaporative lightscattering detector （ELSD） was first used in Pharmacopoeia of People＇ s Republic of China （ChP） 2005, while British Pharmacopoeia （BP） 2005 adopted pulsed amperometric detector in quantitative analysis of gentamycin. As gentamycin C components could not produce signals using UV detector, pre-column derivatization method made the detection possible, referring to United State Pharmacopoeia （USP） 29th Ed, etc. However, the absorbance of by-products and reagents overlapped with gentamycin C components, which caused much trouble in integrating peaks. That is the reason that the quantitative results were calculated based on area and height in different editions of ChP. To make the determination more accurate, ＂pure＂ chromatogram was resolved by heuristic evolving latent projection （HELP） method, based on calculating the data obtained with high performance liquid chromatography-diode array detection （HPLC-DAD） using pre-column derivatization with o-phthalic aldehyde （OPA）. After subtracting the interference of impurities, gentamycin C, component in samples was quantitatively determined accurately. The results were comparable to what achieved with HPLC-evaporative light-scattering detector and column switching. At the same time, it was proved that the HPLC-ELSD method in ChP 2005 was more reliable than the pre-column derivatization method.