[Effect of sample solvents on retention in packed column supercritical fluid chromatography]

Factors affecting the retention in supercritical fluid chromatography(SFC) on C18 and NH2 packed columns have been studied. These factors include the sampling solvent, continuous injection, stationary phase type and concentration of modifier. When supercritical carbon dioxide or low concentration(less than 0.5%) of methanol is employed as the mobile phase, retention time increases with the polarity of the solvent on NH2 column, while it keeps almost constant on C18 column. Methanol will cover the active sites on the surface of the bonded phase, forming the "pseudo" stationary phase, which will lower the interaction between the analytes and the stationary phase, hence the shorter will be the retention time. The hydrogen bond between stationary phase (including the amino group and the remaining hydroxyl group) and methanol is the most obvious. So it's quite more evident on NH2 column than on C18 column that the delaying effect of the solvent, with methanol, acetone and chloroform in decreasing sequence, encourages the elution of the analytes during continuous injections. The elution will recover to the first injection after adequate interval has passed, i.e., more than 10 column volumes. When the methanol concentration is greater than 1.0% in carbon dioxide, the effect decreases and becomes negligible. The solvent effect is significant to the reasonable injection and reproducible chromatographic data in packed column SFC.