Hi well the HPLC has the following four critical aspects of method development you can have a look and also I have given you the PDF below you can download it

Development and Analysis

Rapid Sample Preparation – minimum steps for maximum effectiveness, use updated tools (combination filters) and multi- sample preparation equipment (SPE 96-well plates)
Choose best bonded phases for high resolution – selecting from typical C18 and C8 bonded phases or those targeted to special sample types

Choose the best column configuration for minimum analysis time with high efficiency and resolution – best column length, internal diameter, particle size

Using your HPLC instrument to further reduce analysis time and increase sample throughput – optimizing the HPLC and using new features

Resolution is driven by the Bonded Phase Choice

Changing selectivity (α) influences resolution the most

Bonded phase is the column choice that controls selectivity


Bonded Phase Choosing

Select the columns known to have long lifetimes at operating mobile phase pH.

Choose bonded phases on high purity, low acidity silica for best peak shape.

Select a C18 or C8 bonded phase first for good retention and resolution with typical acidic, basic and neutral samples.

If sample is expected to be difficult (i.e. very polar, difficult to retain, very basic) then select targeted bonded phases available for these types of samples.