Balanced-density method

Slurry packing techniques are required for the preparation of efficient columns with rigid particles of less than 20 micrometers in diameter. The same general packing apparatus. Figure 4.8, can be used to pack columns by the balanced-density slurry, liquid slurry, or the viscous slurry techniques. Down-fill slurry packing is the method of choice for small bore columns and packed capillary columns. 

The two main methods of packing columns for HPLC are dry packing, which is suitable for particles of diameter > 30 jim, and balanced-density slurry packing which is best for small particles of diameter 5-30 jum. 

Columns consisting of particles of less than 30-50 jum in diameter are prepared most efAciently by slurry packing. Balanced-density slurry packing [11,12] is the most successful of such methods. In this technique, a supporting liquid is used which has the same density as that of the particles. This eliminates sedimentation problems. A typical balanced-density slurry-packing apparatus is shown in Fig.3.42. For the preparation of a... 

Chromatography. I. Packing of columns Balanced density slurry method was employed and the quality of packed columns was tested chromatographically in methylene chloride. Benzene was injected as testing solute. Columns which gave satisfactory results [l.e., symmetrical elution curve, relative band broadening of 50 ym at linear velocity of 5 mm/sec, and permeability better than 1.10 -9 cm 2] were used for further experiments. 

An HPLC (39) method was developed for determination of the drug and its metabolites in human and rat bile. A stainless-steel column (15 cm X 4.6 mm I.D.) packed with UChrosorb RP-8 (Pore size 5pm) or Nucleasil Ci8 (pore size 5pm) was used. The columns were packed by means of a balanced density slurry method specially developed for the ammonia elution system. Gradient elution was performed with water (0.005 M ammonia) to which methanol was added, according to the desired programme. The final elution was usually effected with 100% methanol. Flow rate was Iml/min. A wavelength of 235 nm was found suitable for the detection of drug and its metabolites. 

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