Development and validation of an HPLC assay for fentanyl and related substances in fentanyl citrate injection, USP

The stability indicating properties of the: USP method for the assay of fen tanyl in fentanyl citrate injection were evaluated [1] by analyzing fentany l drug substance and product after acid, hydrogen peroxide, heat, and light treatment. N-phenyl-N-(4-piperidinyl)propionamide (PPA), which is a known degradation product/process impurity of fentanyl, was not adequately resolv ed from the fentanyl peak, and mobile phase adjustments did not improve the resolution (Fig. I). Therefore, the USP method did not meet the requiremen ts for a stability-indicating assay. In addition, the wavelength in the USP method was too high (230 nm) to provide adequate levels for the quantitati on of the related substances of fentanyl and, in addition, the acetate ions in the mobile phase could interfere with a lower wavelength detection. An isocratic, reversed phase, stability indicating, high performance liquid ch romatographic (HPLC) method for the assay of fentanyl and related substance s in fentanyl citrate injection, USP has been developed and validated. The chromatographic conditions employed an Inertsil C8, 5 column (25 cm x 4.6 m m), a mobile phase of aqueous perchloric acid [0.23%. w/v]-acetonitrile [65 :35, v/v], and ultraviolet (UV) detection at 206 nm. Under the chromatograp hic conditions of the method, PPA and seven other known process impurities were separated from the active. Degradation studies showed that the active eluted as a spectrally pure peak resolved from its degradation products. (C ) 1999 Elsevier Science B.V. All rights reserved.