As can be seen, at pH 5 and at concentration of salt 5 mM, theoretical plate
number decreased and peak asymmetry increased significantly, and it resulted in the peak broadening and decreased peak resolution.
In the first step, columns of different properties were investigated: Hypersil Gold C8, 5 [micro]m, 150 mm x 4.6 mm ID; Zorbax, 5 [micro]m, 150 mm x 4.6 mm ID; Knauer C8, 5 [micro]m, 250 mm x 4.6 mm ID; and Supelco C18, 5 [micro]m, 250 mm x 4.6 mm ID To choose the most appropriate column, the retention factor, peak asymmetry factor, and height equivalent to a theoretical plate
(HETP), as well as the retention time of ivabradine, were taken into consideration (Table 1).
The plate thickness ([t.sub.p]) in the prototype models was then determined for target theoretical plate
shear strength ([V.sup.*.sub.p]) of about 50% of the total theoretical capacity ([V.sup.*.sub.u]).
The [W.sub.avg] term in equation 1 can be defined in terms of time and theoretical plate
number (N), as shown in equation 2.
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Height equivalent to one theoretical plate
, H: The distance along the column occupied by one theoretical plate
; H = L/N.
In this paper, to obtain a good separation degree, higher theoretical plate
number, and good symmetry of chromatographic peak, different mobile phase conditions were investigated for system adaptability test.
The Mean theoretical plate
count, based on USP tangent calculations (USP-2014) for metronidazole peak was 9574 (Table-1).
Interestingly, this complex relationship is similar in some respects to the efficiency profile of gas and liquid chromatographic separations, where the maximum efficiency is related to the carrier gas velocity at which the theoretical plate
height is minimized (19).
For peak response, ZChrom chromatography manager software version 4.2.was used to calculate the statistical data of different parameters like, retention time (tR), peak area (A) symmetry factor of SFV and theoretical plates
(N) (Table 1).
In practice, to obtain a TBP curve, distillation columns with 15 or more theoretical plates
and high reflux ratios (often 5:1 or higher) are used.
Column efficiency was investigated by performing system suitability test including parameters like retention time (tR), tailing factor (T), theoretical plates
(N) and resolution (Res).