UV SPECTROPHOTOMETRIC AND RP-HPLC ESTIMATION OF DRUG ASENAPINE IN TABLET DOSAGE FORM

UV SPECTROPHOTOMETRIC AND RP-HPLC ESTIMATION OF DRUG ASENAPINE IN TABLET DOSAGE FORM

1. A. Borkar ^* and N. J. Gaikwad

Department of Pharmaceutical Sciences, Rashtrasant Tukadoji Maharaj Nagpur University, Amravati Road, Nagpur - 440033, India

ABSTRACT: Asenapine in Tablet dosage form is used as antipsychotic agent. Literature revealed that there is no single method for the simultaneous estimation of this drug in tablet dosage form, which prompted us to develop a simple, rapid, accurate, economical and sensitive UV spectrophotometric and RP-HPLC methods. The simultaneous estimation of UV method is based on the principle of additivity of absorbance, for the determination of Asenapine in tablet formulation. The absorption maximum of the drug was found to be 270.0 nm in methanol. For RP-HPLC mobile phase is a mixture of acetonitrile and 0.1 M phosphate buffer (pH = 3.2) in the ratio of 65:35 v/v and detection wavelength is 270 nm. The accuracy and reproducibility of the proposed method was statistically validated by recovery studies

Asenapine, Simultaneous estimation, UV-spectrophotometry, RP-HPLC, Recovery study

INTRODUCTION: Asenapine (ASP) ¹ is antipsychotic agent whose IUPAC name is (3aRS,12bRS)-rel-5-Chloro-2,3,3a,12b-tetrahydro-2-methyl-1H-dibenz[2,3:6,7]oxepino[4,5-c]pyrrole. There are some repeated methods for estimation of ASP. Literature survey revealed that Liquid Chromatography–Tandem Mass Spectrometry method have been reported for the estimation of ASP ² and Formulation Development and Evaluation of Sublingual Film of Asenapine ³ However, no method is reported for the simultaneous estimation of this drug in tablet formulation.

This prompted us to develop simple, rapid, accurate, economical and sensitive UV spectrophotometric and RP-HPLC methods. The developed method was validated using ICH guidelines for validation ⁴.

MATERIALS AND METHODS: Shimadzu 1700 UV/Vis spectrophotometer with matched cuvettes and Shimadzu ULFC HPLC system with C₁₈ column(100 mm × 4.6 mm, 3.5 μ) equipped with UV visible detector SPD-M20A were used for the experimental work. The chemicals used were of analytical grade. Commercially available tablets (Asenapt 10^® and Sublingual Tablet Saphris^®) of ASP were procured from the local pharmacy. Standard ASP was received as gift sample from Lundbeck Pvt. Ltd., London.

 UV Estimation of ASP:

Standard stock solution of ASP was prepared by dissolving 25.0 mg of standard ASP in methanol and the volume adjusted to 50.0 ml. Standard solution (25 μg/ml and 10 μg/ml) ASP was further prepared by diluting 2.5 ml and 1.0 ml of stock solution of drug to 50.0 ml in volumetric flask with the methanol.

Spectra of standard solutions of ASP was obtained and scanned between 200- 400 nm (Fig. 1). ASP exhibited absorption maxima at 270.0 nm. Calibration curve for drug was prepared in the concentration range of 1.0-90.0μg/ml at corresponding wavelength. Amount of drug was determined using eqn. as C = A / a, where,   C = concentration of ASP in g/100 ml, A = absorbance of laboratory mixture at 270.0 nm, a = absorptivity of ASP at 270.0 nm. Percent estimation = (C×D)/W×100, where, C = concentration of ASP in g/100 ml, D= dilution factor and W= weight of drug in the laboratory mixture.

FIG. 1: SPECTRA OF ASENAPINE, SOLVENT METHANOL AND λ_max 270.0 nm

Marketed tablets Asenapt 10 and Sublingual tablet Saphris (Sun Pharma Pvt. Ltd., India) were used for the estimation of ASP. Twenty tablets were weighed and crushed to a fine powder. Powder equivalent to 25 mg of ASP (tablet contains 10 mg ASP) was dissolved in the solvent and volume was made up to 50 ml. Insoluble excipients were separated by filtration. The filtrate was further diluted to get final concentration of the drug in the linearity range. Absorbance was noted at the selected wavelength and percent label claim was determined by using the eqn. percent label claim= (C×D×W)/(Wm×L)×100 where, C = concentration of ASP in g/100 ml, W= average weight of tablet, Wm= weight of sample taken and L= label claim of sample taken.

For specificity study weigh tablet powder equivalent to 25.0 mg of ASP in four different volumetric flasks. The samples were exposed to stress conditions for 24 h. First flask at 50^oC sample with 1 ml 0.1 M NaOH (Alkali), second flask at 50^oC sample with 1 ml 0.1 M HCl (Acid), third flask at 50^oC sample with 1 ml 3% H₂O₂ (Oxide) and fourth flask at 50^oC only sample(Heat). After 24 h the flasks were cooled to room temperature and diluted with methanol upto 50.0 ml. Prepare 10 µg/ml concentration of each sample solution and analyzed.

RP-HPLC Estimation of ASP:

Preparation of mobile phase: The mobile phase used was a mixture of acetonitrile and 0.1 M phosphate buffer (pH = 3.2) in the ratio of 65:35 v/v and was filtered before use through membrane filter paper (0.4 μ). The elution was carried out at the flow rate of 1 ml/min. Detection was carried out at 270 nm at ambient temperature.

Preparation of standard stock solution:

Standard stock solution of the drug was prepared by dissolving 25 mg of ASP in mobile phase and volume was made up to 50 ml with the same solvent (500 μg/ml).

Preparation of working solution:

Working standard solution was prepared by diluting 2.5 ml of the standard stock solutions to 50 ml with mobile phase (25μg/ml).

Standard calibration curve: Chromatogram of standard solution of ASP was obtained and scanned between 200-400nm (Fig. 2) for RP-HPLC method. Various dilutions were prepared by taking 10-50μg/ml ASP solutions. Twenty microliters of the solution from the flasks was injected. Calibration curve was constructed by plotting peak areas against the corresponding drug concentrations.

FIG. 2: CHROMATOGRAM OF ASENAPINE. RETENTION TIME IS 4.0 MIN

Estimation of drug in commercial tablet formulation: For the estimation of drug from commercial formulation, twenty tablets, each containing 10 mg of ASP, was weighed and finely powdered. Accurately weighed tablet powder (25 mg) was suspended in the mobile phase and shaken for 15 min. The volume was made up to 50 ml with mobile phase and filtered through Whatman filter paper. Aliquot portion of this solution was diluted to produce 10μg/ml ASP and filtered through membrane filter paper (0.4 μ). Equal volumes of 20μl of standard and sample solutions were injected separately after the equilibrium of stationary phase and area under the curve noted. Amount of the drug in the tablet formulation was calculated using formula (A_U x W_S x W_AV x 100) / (A_S x L_C x W_T) where, A_U = area of unknown sample, W_S = weight of standard (mg), W_AV = average weight of tablet (mg), A_S = area of standard, L_C = labelled claim (mg/tablet), W_T = weight of sample (mg).

For specificity study same procedure was followed mentioned above in UV spectrophotometry method.

RESULTS AND DISCUSSION:

Reproducibility, repeatability and accuracy of the proposed methods by UV Spectrophotometry and RP-HPLC were found to be satisfactory which is evident from the low values of standard deviation (SD), relative standard deviation (RSD) and standard error (SE). Results for UV visible spectrophotometric method was discuss in Table 1, 2 and 3. Results for RP-HPLC method was discuss in Table 4, 5, 6 and 7.

The accuracy and reproducibility of the proposed methods were confirmed by recovery experiment, performed by adding known amount of the drug to the preanalyzed formulation and reanalyzing the mixture by proposed methods. Percent recovery obtained indicates non–interference from the excipients used in the formulation. Thus, the methods developed in the present investigation are found to be simple, sensitive, accurate and precise and can be successfully applied for the estimation of Asenapine in tablets.

TABLE 1: RESULTS OF STATISTICAL DATA OF MARKETED FORMULATION BY UV SPECTROPHOTOMETRIC METHOD

 TABLE 2: RESULTS OF DRUG RECOVERY STUDY BY UV SPECTROPHOTOMETRIC METHOD

 TABLE 3: RESULTS OF SPECIFICITY STUDY BY UV SPECTROPHOTOMETRIC METHOD

 TABLE 4: SYSTEM SUITABILITY PARAMETERS FOR RP-HPLC METHOD

 TABLE 5: RESULTS OF STATISTICAL DATA OF MARKETED FORMULATION BY RP-HPLC METHOD

 TABLE 6: RESULTS OF DRUG RECOVERY STUDY BY RP-HPLC METHOD

 TABLE 7: RESULTS OF SPECIFICITY STUDY BY RP-HPLC METHOD

CONCLUSION: The present study proposed an UV Spectrophotometry and RP-HPLC methods to determine ASP from tablet dosage form. The values of percent recovery and standard deviation indicate that the methods are accurate, reproducible and precise.

 ACKNOWLEDGEMENTS: The author wishes to thanks Head of the Department, Department of Pharmaceutical Sciences, RTM Nagpur University, Nagpur for providing necessary facilities. Also, thanks to Lundbeck Pvt. Ltd., London for providing the authentic sample of drug.

REFERENCES:

1. British Pharmacopoeia, Her Majesty's Stationary Office, London, UK, (2007).
2. Boer TD, Meulman E, Meijering H, Wieling J, Dogterom P and Lass H: Quantification of Asenapine and Three Metabolites in Human Plasma using Liquid Chromatography–Tandem Mass Spectrometry with Automated Solid-Phase Extraction: Application to a Phase I Clinical Trial with Asenapine in Healthy Male Subjects. Biomedical Chromatography 2012; 26(2):156 – 165.
3. Kapadia YD, Sodha HP and Patel VP: Formulation Development and Evaluation of Sublingual Film of Asenapine Maleate. Pharma Science Monitor 2013; 4(3):190 – 209.
4. International Conference on Harmonization (ICH), Q2 [R1], “Validation of analytical procedure- text and methodology” 2005
   

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How to cite this article:

Borkar AA and Gaikwad NJ: UV Spectrophotometric and RP-HPLC Estimation of Drug Asenapine in Tablet Dosage Form. Int J Pharm Sci Res 2016; 7(7): 3080-84.doi: 10.13040/IJPSR.0975-8232.7(7).3080-84.

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