反相色谱峰形拖尾的原因与改善方法(英文).docVIP

Correcting Peak Tailing Problems in Reversed Phase HPLC Peak tailing in reversed phase HPLC continues to be a common complaint. It is particularly prevalent when separating basic compounds and, therefore, a source of constant problems to those analyzing pharmaceutical compounds by HPLC. Peak tailing causes a number of problems, including lower resolution, reduced sensitivity, and poorer precision and quantitation. Figure 1 illustrates how resolution between peaks and sample sensitivity is negatively affected by peak tailing.? Figure 2 illustrates how accuracy and precision of an analysis can suffer because of the inability of data systems to identify exactly where a tailing peak begins and ends. FIGURE 1Effect of Peak Tailing on Resolution and Sensitivity As peak tailing (T, Tailing factor) increases from 1.0 to 2.0, resolution (R s ) decreases from 1.5 to 1.0. Sensitivity (peak height) also decreases with peak tailing since the peak volume increases and the sample concentration decreases. ????? FIGURE 2Accuracy and Precision isAdversely Affected by Peak Tailing Tailing peaks make it more difficult for data systems to identify exactly where a peak ends. Because of this, accuracy and precision can suffer. In this example, the peak area measured at point B is 3% less than the peak area measured at point A. Causes of Peak TailingThe major causes of peak tailing include:?injecting sample in a solvent that is significantly stronger than the mobile phase,?sample mass overload,?stationary phase silanol interactions with amines,?adsorption of acidic compounds onto silica, and?a void in the column抯 packing bed. Once you have identified the cause of the tailing, you can take action to reduce or eliminate it. (See Table 1) TABLE 1Peak Tailing: Causes and Cures Causes Cure Sample solvent stronger than the mobile phase Dissolve sample in mobile phase or at least reduce the strength of the sample solvent as much as possible Sample mass overload Reduce the amount