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A sensitive reverse phase-high performance liquid chromatography (RP-HPLC) method with fluorescent detector (FLD) was developed and optimized for salbutamol sulfate (SS) determination in human plasma. In this regard, mobile phase specifications, extraction procedures and excitation and emission wavelengths were optimized. The HPLC system consisted of a Lichrosorb RP-C18 analytical column (4.6 × 200 mm, 5 µm) with FLD operated at excitation 228 nm and emission 310 nm. Mobile phase {CH3OH/(NH4)H2PO4 (67 mM) (pH 3.0)/triethylamine (TEA), 50/50/0.02 (v/v/v %)} was run at a flow rate of 0.7 mL/min. To clean up the samples, a liquid-liquid extraction (LLE) procedure was selected and optimized. SS and tramadol hydrochloride (TH) eluted at 4.1 min and 5.2 min, respectively. Adequate extraction efficiency was achieved by DEHP (75.88-85.52 %). The standard curve was linear for the range tested (0.5-80 ng/mL) and the coefficient of determination was 0.9989. A detection limit of 0.17 ng/mL was achieved. The intra-and inter-day precision was less than 4 %. The present assay combines adequate accuracy and precision with sensitivity for SS determination in human plasma and can be applied to study pharmacokinetics of SS sustained release tablets after oral administration in human.

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Protease Inhibitors belong to class of drug which are used to cure or prevent infection by viruses like HIV and HCV. Protease Inhibitors from Bacillus subtilis-M15 prevent viral multiplication by inhibiting the activity of Trypsin and Pepsin. Pharmaceutically designed ion-exchange resins are proved as promising candidates to enhance the Inhibitory activities of Protease. The novel Biological anti-viral drugs are characterized by Hydrogen Peroxide, Dimethyl Sulfur, HCl, and Acidic-Alkaline Proteinase. When Protease Inhibitory activity of chemically formulated drug already available in market i.e. Lopinavir/Ritonavir are compared with novel anti-viral drug from Bacillus subtilis-M15 then Lopinavir/Ritonavir are proved as less effective than Biological drug due to their lower Inhibitory activity in terms of units per milliliter. Lopinavir/Ritonavir has 2446U/ml and 241U/ml Protease Inhibitory activities against trypsin and pepsin respectively whereas Biological drug from Bacillus subtilis-M15 has 2581U/ml and 3360U/ml Protease Inhibitory activities against Trypsin and Pepsin respectively. In this context Biological drug developing treatment has attracted much attention than chemically designed drug due to their high efficacy, least side effect, cost efficient and environment friendly nature. Hence we conclude the current application of novel anti-viral drug as a Protease Inhibitors from Bacillus subtilis-M15 and its future clinical prospects against targeted Protease based HIV and HCV therapy.