Serratiopeptidase Essay

Development and Validation of Analytical Method for Simultaneous Estimation of Diclofenac potassium and Serratiopeptidase in Pharmaceutical Formulation

Dhara Parekh*, Alpesh B. Limbasiya, Pankaj P. Kapupara, Ketan V. Shah

Department of Quality Assurance, School of Pharmacy, RK University, Rajkot-360 020, Gujarat, India

“*Address for correspondence”:

School of Pharmacy, RK University,

C/O: R.K. College of Physiotherapy Center,

Near: Bhaktinagar Circle, Rajkot-360002, Gujarat, India.

Email Address: [e-mail]

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ABSTRACT:

A simple and novel method has been developed for simultaneous estimation of Serratiopeptidase (SER) and Diclofenac potassium (DCL) by spectrophotometry. The method

This was done by dissolving 10 mg of drug in distilled water distinctively. SER had λmax at 195 nm and DCL’s λmax was at 222 nm and 276 nm. To carry out simultaneous determination of SER and DCL, the two wavelengths selected should be such that at one wavelength the absorbance of one drug should be zero, whereas other drug shows significant absorbance value. At other wavelength, both the drugs should have significant absorbance value. So the two wavelength chosen were 210.0 nm and 300.0 nm. At 210.0 nm Serratiopeptidase and Diclofenac potassium both showed significant absorbance and at 300.0 nm Serratiopeptidase does not show any absorbance, but Diclofenac potassium showed significant

The samples were put into series of 10 ml volumetric flask. The volume was made upto the mark with distilled water. Sets of various concentration of solutions were obtained like 1, 2, 5, 10, 20, 30, 40, µg/ml for Serratiopeptidase and 1, 2, 5, 10, 20, 30, 40, 50, 60, 70 µg/ml for Diclofenac potassium. The absorbance of all above solutions were measured at the specific wavelengths (i.e., 210 nm and 300 nm).Absorbances of each solution were plotted against respective concentrations. Drug concentration range which obeyed Beer’s law was chosen. SER and DCL were linear with absorbances within range of 1-40 µg /ml and 1-70 µg /ml at their selected wavelengths respectively. Correlation coefficients obtained were 0.9996 for SER and 0.9933 for DCL. The optical characteristics and regression values for the calibration curve are presented in Table 1.

Solutions of 10µg /ml SER and 50µg/ml of DCL were prepared. This preparation was done by dilution of appropriate volumes of the standard stock solutions. The solutions of SER and DCL were scanned between 190 to 400 nm to obtain overlay spectra(figure 1). Absorbance and absorptivity of both standards was checked at selected wavelengths λ1(300nm) and λ2(210 nm). The mixed standard for the pure drug was prepared from the stock solutions. Mixed standards that were prepared contained 10 µg/ml SER and 50 µg/ml DCL . The absorbances were recorded at selected