Sil 20a prominence

Chromatographic analysis of TSN was performed on a Shimadzu HPLC equipped with a SPD-20 AV Prominence UV/visible detector set at 280 nm, an LC-20AT pump, and SIL-20A Prominence autosampler. Separation of TSN was performed using a KinetexVR pentaflurophenyl 5 μm column (250 4.6 mm) (Phenomenex, Macclesfield, UK) maintained at 30 °C and TSN was eluted with a mobile phase consisting of acetonitrile and water (60:40 v/v) at a flow rate of 1 ml/min. Injection volume was 5 μl with retention time of 7.34 min (Kakadia and Conway 2018) .

The HPLC method for diclofenac sodium analysis was based on the method reported by Khan et al., (2016) with some modification. Chromatographic separation was performed on a Shimadzu HPLC system equipped with a SPD-20 AV Prominence UV/VIS detector, a LC 20 AT pump, and SIL-20A Prominence auto sampler. The data acquisition was carried out on a LC solution software integrator. The separation was performed using C18 L1, pH resistant (4.5 mm x 150 nm: 3.5μm) column (Waters, UK).The mobile phase composition was acetonitrile/water (60:40) (v/v). Stock solutions of diclofenac sodium were prepared by dissolving 100 mg in 100 mL of the mobile phase. The mobile phase was filtered (0.22 m filter) and degassed prior to use. Standard solutions were prepared in concentrations between 0.5 μg mL -1 and 200 μg mL -1 by diluting the stock solution with the mobile phase and were analysed in triplicate. Calibration standards were prepared by plotting the area under the curve (AUC) versus concentration. The limit of detection (LOD) and the limit of quantification (LOQ) were determined as 0.045 and 2.3 g/mL respectively.

Chromatographic analysis of TSN was performed on a Shimadzu HPLC equipped with a SPD-20 AV Prominence UV/Visible detector set at 280 nm, an LC-20AT pump, and SIL-20A Prominence auto sampler. Separation of TSN was performed using a Kinetex ® pentaflurophenyl 5 m column (250 x 4.6 mm) (Phenomenex, UK) maintained at 30°C and TSN was eluted with a mobile phase consisting of acetonitrile and water (60:40 v/v) at a flow rate of 1 ml/min. Injection volume was 5 L with retention time of 7.34 min. Standard solutions of TSN were prepared at concentration of 1, 5, 10, 20, 30, 40 and 50 g/ml and these standards were used to determine the linearity (R 2 = 0.999) and the limit of detection (0.60 g/ml) during HPLC method validation.