David m bliesner validating chromatographic methods
Visit for more related articles at Pharmaceutical Analytical Chemistry: Open Access A novel and efficient stability indicating reverse phase High performance liquid chromatography with diode array detector (RP-HPLC-DAD) related substances analytical method has been developed, optimized and validated for the determination of degradation products and process impurities of Norethindrone in Norethindrone tablets: Noreandrostenedione (Imp-A), Norethindrone enolether (Imp-B), Delta-5(6)Norethindrone (Imp-C), Delta-5(10) Norethindrone (Imp-D).
This is done by RP-HPLC equipped with Diode array detector (DAD).All other peaks will be considered as unknown impurities.In this work, impurities will be injected in specificity experiment to determine their retention times .Gradient programme is 0/35, 18/35, 40/80, 45/35 and 55/35 (time (min)/%B mobile phase).Water is taken as mobile phase-A and acetonitrile as mobile phase-B.Norethindrone; HPLC; Degradation products; Related substances; Method validation RP-HPLC-DAD: Reverse Phase-High Performance Liquid Chromatography-Diod Array Detector; UV: Ultra Violet; PDA: Photo Diode Array; HCL: Hydrochloric acid; Na OH: Sodium hyderoxide; H: Hyderogen Peroxide; Acc: Accuracy; nm: Nanometer; RSD: Relative Standard Deviation; SD: Standard Deviation; μ: Micrometer; LOQ: Limit of Quantitation; LOD: Limit of Detection; Imp-A: Noreandrostenedione; Imp-B: Norethindrone enolether; Imp-C: Delta-5(6)Norethindrone; Imp-D: Delta-5(10)Norethindrone and RRT: Relative Retention Time.
Norethindrone is form of progesterone, a female hormone.
However, UPLC is useful for more sample quantitation in less time.
There are some analytical assay methods available for determination of steroidal hormones .
Chemical structures of Norethindrone and its impurities Currently there is one UPLC and one HPLC method available to separate and quantify the Norethindrone and its impurities in Norethindrone tablets [2,3].
More than 24 hours solution stability of impurities, reference solution and test solution is not yet reported [2- 4]. Reference standard and impurity standard of Norethindrone and Norethindrone tablets 0.35 mg were obtained as gift sample from Bayer, India.
Limit of detection (LOD) and limit of quantification (LOQ) were also obtained for all impurities.