Procedure
Introduce separately 20-µL portions of the
Assay preparation and the
Standard preparation into a high-pressure liquid chromatograph (see
Chromatography
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) operated at 25
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, by means of a suitable microsyringe or sampling valve, adjusting the operating parameters such that the peak obtained with the
Standard preparation is full-scale. Typically, the apparatus is fitted with a 4-mm × 30-cm column, packed with packing L1, is equipped with an UV detector capable of monitoring absorption at 280 nm and a suitable recorder, and is capable of operating at a column pressure of up to 3500 psi. Five replicate injections of the
Standard preparation show a relative standard deviation of not more than 1.5%. Calculate the quantity, in mg, of C
6H
6Cl
2N
2O
4S
2 in the portion of Dichlorphenamide taken by the formula:
50C(AU / AS),
in which
C is the concentration, in mg per mL, of
USP Dichlorphenamide RS in the
Standard preparation; and
AU and
AS are the area responses obtained at equivalent retention times from the
Assay preparation and the
Standard preparation, respectively.