Can you please suggest me the mobile phase & hplc column for calcitriol in calcitrial capsulescontains calcium
Answer:
Carry out the method for liquid chromatography, in subdued light using the following solutions.
For capsules containing 0.25 µg or less, use the mixed contents of 10 capsules as solution (1). For capsules containing more than 0.25 µg, prepare solution (1) by diluting a quantity of the mixed contents of 10 capsules containing 1.5 µg of Calcitriol to 1 ml with the mobile phase.
Solution (2) contains 0.00015% w/v of calcitriol EPCRS in the mobile phase.
The chromatographic procedure may be carried out using
(a) a stainless steel column (25 cm × 4.6 mm) packed with stationary phase A (5 µm) (Lichrosorb Si60 is suitable),
(b) a mixture of 1 volume of propan-1-ol , 2 volumes of methanol , 40 volumes of hexane and 60 volumes of ethyl acetate as the mobile phase with a flow rate of 1.2 ml per minute and
(c) a detection wavelength of 265 nm.
If necessary, adjust the composition of the mobile phase so that the principal peak in the chromatogram obtained with solution (1) is clearly separated from the tail of the peak due to the excipient.
Calculate the content of calcitriol, C27H44O3, using the declared content of C27H44O3 in calcitriol EPCRS.
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