反相高效液相色谱负峰测定法研究以及酮类的测定

Negative Peaks Ultraviolet Detection in Reversed Phase High Performance Liquid Chromatography and Its Application to Determination of Ketones

研究了反相液相色谱中负峰现象及产生的原理,应用计量置换作用原理解释了样品在色谱柱中的计量置换保留行为;应用负峰法测试了甲基脂肪酮各组分的含量。本研究采用AgilentC18色谱柱(150mm×4.6mm,5μm),流动相为V(甲醇)∶V(水,含0.01mmol/L磺基水杨酸)=60∶40,紫外检测器(λsig=310nm,λref=275nm)。本方法简便,线性相关系数r均大于0.9993;甲基酮的定量下限为0.041μg;相对标准偏差为0.52%～0.80%。

The phenomenon of negative peaks ultraviolet detection （UV） was reported and the principle was explained. In liquid chromatography, solute molecules in the mobile phase undergo many successive adsorptions and desorptions. It was proved that there was a relationship called stoichiometric displacement. The stoichiometric displacement retention behavior of samples in the column was explained using the theory. Based on the theory, the content of each component in the mixed ketones was detected. From the study, UV detector can be used to detect the sample without UV absorbance. As the result of the study, liquid chromatography separation was performed on Agilent C18 column with methanol-water （60∶40, V/V） and sulfosalicyclic acid concentration of 0.01 mmol/L as mobile phase and with UV detection （λsig=310 nm, λref=275 nm ）. The method was simple and the correlation coefficients were above 0.9993. The limit of quantitative detection was 0.041 μg for methyl ketones and their relative standard deviations were between 0.5% and 0.8%.