Barnidipine Hydrochloride Lab Report

Barnidipine hydrochloride is ([3S]-l-benzyl-3-pyrrolidinyl methyl [4S]-2,6-dimethyl-4-[m-nitrophenyl]-l,4-dihydropyridine-3,5-dicarboxylate hydrochloride) has been synthesized and developed as an antihypertensive drug by Yamanouchi Pharmaceutical Co Ltd. (Tokyo, Japan). Physicochemically, Barnidipine hydrochloride differs from other dihydropyridines, such as nifedipine or nisoldipine, in its water solubility. The solubility of Barnidipine at room temperature is 2.89 mg/ml [1]. Literature review reveals that several analytical methods by HPLC and LC–MS–MS have been developed for the detection of Barnidipine in formulations and biological fluids [2,3]. Hence, authors propose a simple LCMS method that is developed and validated in achieving good
Initially, a stock solution of 1 mg/ml was dissolved and used to prepare desired dilutions with LC grade milli-Q water. Prepare 10 ml each of 1000.0, 1200.0, 1500.0, 2500.0, 5000.0, 9000.0, 18000.0 and 20000.0 ng/ml of Barnidipine standard solutions using the Barnidipine standard stock solution and mobile phase and store at –20 ± 2ºC until analysis. Prepare 10.0 ml each of 50.0, 60.0, 75.0 ,125.0, 250.0, 450.0, 900.0 ,1000.0 ng/ml of Barnidipine Calibration curve samples using 0.5 ml of Barnidipine standard stock solution and make up the volume with blank plasma, transfer in to different 2 ml centrifuge tubes and store at –70 ± 2ºC until
The standard and sample peaks were recorded and compared. The sample peaks were seen to match with corresponding standard drug peaks. Some additional peaks observed in the sample chromatograms. These peaks, however, did not interfere with the Barnidipine and internal standard peak (Figure 1 and 2).

4.5.4 Accuracy: The absolute recovery of Barnidipine was determined by comparing the response factor of the drug obtained from the plasma with response factor obtained by the direct injection of Barnidipine in mobile phase at three different levels. Recovery studies were carried out for three levels at six times and the % recovery, mean, standard deviation and % CV was calculated.

4.5.5 Stability: The stability studies of plasma samples spiked with Barnidipine were subjected to three Freeze - thaw cycles, Short term stability at room temperature for 3 hrs and Long term stability at – 70ºC over four weeks. In addition, stability of standard solutions was performed at room temperature for 6 hr and freeze condition for four weeks. The stability of triplicate spiked human plasma samples following three freeze thaw cycles was analyzed. The mean concentrations of the stability samples were compared to the theoretical