SPECTROFLUORIMETRIC DETERMINATION OF TOLTERODINE TARTARATE IN PURE FORM AND PHARMACEUTICAL PREPARATION
HTML Full TextSPECTROFLUORIMETRIC DETERMINATION OF TOLTERODINE TARTARATE IN PURE FORM AND PHARMACEUTICAL PREPARATION
Mohammed W. Nassar, Khalid A. Attia, Hamed M. Abou-Seada and Ahmed El-Olemy Allam*
Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt
ABSTRACT: A Simple, rapid, sensitive, accurate and precise spectrofluorimetric method was developed for the determination of tolterodine tartrate (TOL) in bulk powder and in pharmaceutical preparation. This method is based on measuring the native fluorescence of TOL in methanol at 313 nm after excitation at 285 nm using 1 ml of phosphate buffer pH 6. All variables that affect fluorescence intensity such as diluting solvents, pH, buffer and buffer volume were studied and optimized .The fluorescence-concentration plot was rectilinear over the range of 0.5-8 µg/ml with a lower detection limit (LOD) of 0.090 µg/ml and lower quantification limit (LOQ) of 0.273 µg/ml. The method was validated and successfully applied to the determination of TOL tablets with an average percent recovery ± RSD% of 100.57 ± 0.76. The obtained results were statistically compared with those of the reported method by applying t-test and F-test at 95% confidence level and no significant difference was observed regarding accuracy and precision.
Keywords: |
Tolterodine, Spectrofluorimetric, pure form, Pharmaceutical preparation
INTRODUCTION:Tolterodine tartrate (Fig. 1) is 2-[(1R)-3-[Bis(1-methylethyl)amino]-1-phenyl propyl]-4-methylphenol 1. It is a tertiary anti-muscarinic with actions similar to those of atropine. It is used in the management of urinary frequency, urgency and incontinence in detrusor instability 2. Literature survey shows that several spectrophotometric 3-9 and chromatographic 10-18 methods for determination of tolterodine in pure form, pharmaceutical preparations and/or biological fluids have been reported.
FIGURE 1: STRUCTURAL FORMULA OF TOLTERODINE TARTARATE
MATERIALS AND METHOD:
Apparatus:
- Jasco FP-6200 Spectrofluorometer (Japan), equipped with 150 Watt Xenon lamp, holographic gratting excitation and emission monochromators for all measurements. Slit widths for both monochromators were set at 10 nm. A 1 cm quartz cell was used.
- Jenway, 3510 pH meter (Jenway, U.S.A.).
Materials and Reagents: All chemicals and reagents used throughout the work were of analytical grade.
- Tolterodine tartrate was kindly supplied by El-Kahira, Cairo, Egypt. The purity was assigned as 99.55 %.
- Detrusitol® tablets contain 2 mg tolterodine-L-tartrate. (B. No. - H885A, manufactured by Pharmacia & Upjohn) was obtained from commercial source.
- Water used throughout the procedures was freshly double distilled.
- Acetonitrile, methanol, ethanol, 1-propanol, chloroform, dichloroethane and tetrahydro-furan, all of HPLC grades [Sigma, Germany].
- Sodium hydroxide (El-Nasr Company, Egypt) prepared as 0.1 N aqueous solution.
- Hydrochloric acid (El-Nasr Company, Egypt) prepared as 0.1 N aqueous solution.
- Monobasic potassium phosphate, potassium chloride, boric acid, glacial acetic acid and sodium acetate trihydrate (El-Nasr Company, Egypt).
- Buffers of different pH values prepared as prescribed in US pharmacopeia 19:
- Acetate buffer pH range from 4.1 to 5.5.
- Phosphate buffer pH range from 6 to 8.
- Alkaline borate buffer pH range from 8 to 10.
Standard Solution: A stock solution of TOL (200 μg/ml) was prepared by dissolving 20 mg of Tol in 50 ml of water and complete to 100 ml with water and was further diluted with the same solvent as appropriate. The standard solution was stable for 2 weeks when kept in the refrigerator 6.
Procedure:
Construction of the Calibration Curve (General Procedure): Different aliquots of TOL stock solution ranging from (5-80) µg were transferred to a 10 ml volumetric flasks and 1 ml of phosphate buffer pH 6 was added. The solutions were diluted with methanol to 10 ml and mixed well. The fluorescence intensity was measured at 313 nm (ʎexcitation= 285 nm).The measured fluorescence intensity vs the final concentration in μg/ml were plotted to get the calibration graph. Alternatively, the regression equation was derived.
Analysis of Pharmaceutical Preparation:Weigh and pulverize 20 tablets. Transfer a weighed quantity of the powder equivalent to 20.0 mg of TOL into a small conical flask. Extract with 3 x 30 ml of water on three successive times each with 30 ml. Filter the extract into a 100 ml volumetric flask. Wash the conical flask with few mls of water. Pass the washings into the same conical flask and complete to the mark with the same solvent. Transfer aliquots covering the working concentration range into 10 ml volumetric flasks. Proceed as described under “General Procedure”, adopting the method. Determine the nominal content of the tablets either from the calibration curve or using the corresponding regression equation.
RESULTS AND DISCUSSION:
Spectral Characteristics: TOL exhibits a native fluorescence in methanol and its emission can be measured at 313 nm (λemission) after excitation at 285 nm (λexcitation). The emission and excitation spectra of TOL in methanol are shown in figure 2.
FIGURE 2: EXCITATION AND EMISSION SPECTRA OF TOLTERODINE (5 µg/ml) IN METHANOL USING 1 ml of PHOSPHATE BUFFER pH 6.
Optimization of Experimental Conditions:
(i) Effect of Diluting Solvents: The general procedure for the method was repeated using a fixed amount of TOL (50 µg) and different diluting solvents and found that; methanol is the best diluting solvent as shown in figure 3.
(ii) Effect of pH and Buffer: The general procedure for the method was repeated using a fixed amount of TOL (50 µg) and different buffers with different pH and found that; phosphate buffer pH 6 gives the best results as shown in figure 4.
(iii)Effect of Buffer Volume: The general procedure for the method was repeated using a fixed amount of TOL (50 µg)and different volumes of phosphate buffer pH 6 and found that; 1 ml gives the best results as shown in figure 5.
(iv)Effect of Time: The general procedure for the method was repeated using a fixed amount of TOL (50 µg)at different time interval and found that; it is stable at least for one hour as shown in figure 6.
FIGURE (3): EFFECT OF DIFFERENT DILUTING SOLVENTS ON FLUORESCENCE INTENSITY OF TOLTERODINE (5 µg/ ml).
FIGURE 4: EFFECT OF DIFFERENT BUFFERS (1 ml) ON FLUORESCENCE INTENSITY OF TOLTERODINE (5 µg/ ml).
FIGURE (5): EFFECT OF VOLUME OF PHOSPHATE BUFFER PH 6 ON FLOURESCENCE INTENSITY OF TOLTERODINE (5 µg/ ml)
FIGURE (6): EFFECT OF TIME ON FLUORESCENCE INTENSITY OF TOLTERODINE (5 µg/ ml)
Validation of the Method:
(i) Linearity: Under the described experimental conditions, the calibration graph for the method was constructed by plotting fluorescence intensity versus concentration in μg/ml. The regression plot was found to be linear over the range of 0.5-8 µg/ml. The linear regression equation for the graph is:
FI= 94.80C + 0.354 (r2 = 0.9998)
Where FI is the fluorescence intensity, C is the drug concentration in μg ml-1 and r2 is the correlation coefficient.
Linearity range, regression equation, intercept, slope and correlation coefficient for the calibration data were presented in table 1.
TABLE 1: SPECTRAL DATA FOR DETERMINATION OF TOL BY THE PROPOSED METHOD:
Parameters | Proposed Method |
λemission (nm) | 313 |
λexcitation (nm) | 285 |
Linearity range (µgml-1) | 0.5 — 8 |
LOD (µgml-1) | 0.090 |
LOQ (µgml-1) | 0.273 |
Regression equation* | |
Slope (b) | 94.80 |
Intercept (a) | 0.354 |
Correlation Coefficient (r2) | 0.9998 |
* y= a + bx where y is the fluorescence intensity and x is the concentration.
(ii) Sensitivity: The limit of detection (LOD) and the limit of quantitation (LOQ) were calculated according to ICH Q2 Recommendation20 from the following equations:
LOD = 3.3 Sa / slope
LOQ = 10 Sa / slope
Where Sa is the standard deviation of the intercept of regression line.
LOD was found to be 0.090 μg/ml, while LOQ was found to be 0.273 μg/ml. The small values of LOD and LOQ indicate good sensitivity.
(iii)Accuracy and Precision: Three replicate determinations of three different concentrations of TOL in pure form within linearity range were performed in the same day (intra-day) and in three successive days (inter-day). Accuracy as recovery percent (R%) and precision as percentage relative standard deviation (RSD%) were calculated and results are listed in table 2. The small value of RSD% indicates high precision of the method. Moreover, the good R% confirms excellent accuracy.
TABLE (2): INTRADAY AND INTERDAYS ACCURACY AND PRECISION FOR THE DETERMINATION OF TOL BY THE PROPOSED METHOD
Conc. mg.ml-1 | Intra-day | Inter-day | ||||
Found
Conc. + SD |
Accuracy
(R%) |
Precision
(RSD%) |
Found
Conc. + SD |
Accuracy
(R%) |
Precision
(RSD%) |
|
1
2 4 |
0.99 ± 0.008
1.98 ± 0.024 4.02 ± 0.033 |
99.44
99.23 100.40 |
0.798
1.215 0.033 |
1.00 ± 0.006
1.98 ± 0.020 4.02 ± 0.038 |
100.22
99.11 100.55 |
0.631
0.999 0.946 |
Pharmaceutical Applications: The proposed method was applied to the determination of the studied drug in its tablet preparation. The results were validated by comparison to a previously reported method 6. No significant difference was found by applying t-test and F-test at 95% confidence level, indicating good accuracy and precision of the proposed method for the analysis of the studied drug in its pharmaceutical dosage form (table 3).
TABLE 3: DETERMINATION OF TOLTERODINE IN DETRUSITOL® TABLETS (2 mg) BY THE PROPOSED AND REPORTED METHODS
Parameters | Proposed Method | Reported method (6) |
N* | 5 | 7 |
X‾ | 100.57 | 99.86 |
SD | 0.76 | 0.64 |
RSD% | 0.76 | 0.64 |
t** | 1.69 (2.23) | —— |
F** | 1.41 (6.16) | —— |
* No. of experimental. ** The values in the parenthesis are tabulated values of t and F at (p= 0.05).
CONCLUSION: The proposed method is simple, rapid and inexpensive. So, it is good alternative to the other few reported methods and to the high cost HPLC methods.
REFERENCES:
- The Merk Index, Fourteenth edition, Whitehouse Station, NJ: Merk & co., Inc., 2005, Electronic version.
- Sweetman S: Martindale. The Complete Drug Reference, 35th ed. The Pharmaceutical Press: London, 2007.
- Nanda RK, Gaikwad J and Prakash A: Simultaneous spectrophotometric estimation of tolterodine in pharmaceutical dosage form. Research J Pharm and Tech 2009; 2(2):312-314.
- Shetty SK and Shah A: New spectrophotometric method for estimation of tolterodine in bulk and pharmaceutical formulation. International Journal of Pharmaceutical Sciences and Research 2011; 2(6):1456-1458.
- Ishaq BM, Praksh KV, Manjula B, Kumar CH and Rani GU:New aurum coupling reaction for visible spectrophotometric determination of tolterodine tartrate in pharmaceutical preparations. International Journal of Chemical and Analytical Science 2010; 1(7):165-167.
- Walash MI, Belal F, El-Enany N and Elmansi H: Determination of tolterodine tartrate in pharmaceutical preparations using eosin, application to stability study. International Journal of Pharmaceutical Sciences and Research 2011; 2(11): 2849-2855.
- Ganesh M, Hemalatha P, Peng MM, Vinodh R, Sakthimanigandan K and Jang HT : A simple and reproducible estimation of tolterodine tartrate by ion-pair extractive colorimetric method using methyl orange as chromogen. Journal of Pharmacy Research 2013; 7:367-373.
- Bab MS, Prasad UV and Ramu BK: Assay of tolterodine tartrate using MBTH reagent in bulk and its pharmaceutical formulations. American Journal of PharmTech Research 2012; 2(4):395-404.
- Bab MS, Prasad UV and Ramu BK: Visible spectrophotometric determination of tolterodine tartrate from capsule formulations by oxidative coupling reaction. Journal of Scientific Research in Pharmacy 2012; 1(2):70-72.
- Krishna SR, Rao BM and Rao NS: A validated stability-indicating HPLC method for the determination of related substances and assay of tolterodine tartarate. Rasayan J Chem 2009; 2:144-150.
- Sinha VR, Jindal V, Kumar RV, Bhinge JR and Goel H: Development and validation of a simple, stability-indicating high-performance liquid chromatographic method for analysis of tolterodine tartarate in the bulk drug and in its tablet formulation. Acta Chromatogr 2011; 23:133-143.
- Shetty S K and Shah A:Development and validation of tolterodine by RP-HPLC method in bulk drug and pharmaceutical dosage forms. International Journal of PharmTech Research 2011; 3:1083-1087.
- Dwibhashyam VS, Keerthi P, Ratna JV and Nagappa AN: Reverse-phase, high performance liquid chromatographic method for the determination of tolterodine tartrate in routine quality control. PDA Journal of Pharmaceutical Science and Technology 2009; 63:234-239.
- Yanamandra R, Vadla CS, Puppala U, Patro B, Murthy YLN and Ramaiah PA: A new rapid and sensitive stability-indicating UPLC assay method for tolterodine tartrate: application in pharmaceuticals, human plasma and urine samples. Scientia Pharmaceutica 2012; 80:101-114.
- Vasantharaju SG, Mishra A, Arumugam K, Musmade PP, Udupa N and Bhat KM: Stability indicating RP-HPLC method for determination of tolterodine in solid dosage form. Journal of Pharmaceutical Research 2009; 8(4):184-186.
- Xia ZL, Chen ZY and Yao TW: An enantio specific HPLC method for the determination of (S)-enantiomer impurities in (R)-tolterodine tartarate. Pharmazie 2007; 62:170-173.
- Ramathilagam N, Meeradevi M, Solairaj P and Rajesh SC: Development and validation of HPLC method for the estimation of tolterodine tartarate in tablets. International Journal of Pharmacy and Biological Sciences 2012; 2(4):332-337.
- Madhavi A, Reddy GS, Suryanarayana MV and Naidu A: Development and validation of a new analytical method for the determination of related components in tolterodine tartarate using LC. Chromatographia 2008; 68:399-407.
- United States Pharmacopoeia 30 National formulary 24, United State Pharmacopoeia Convention, 2007.
- ICH Q2 (R1), Validation of analytical procedure, Text and methodology, Geneva, International conference on Harmonization, 2005.
How to cite this article:
Nassar MW, Attia KA, Abou-Seada HM and El-Olemy Allam A: Spectrofluorimetric determination of Tolterodine tartarate in pure form and Pharmaceutical preparation. Int J Pharm Sci Res 2013; 4(10): 3845-49. doi: 10.13040/IJPSR. 0975-8232.4(10).3845-49
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Article Information
15
3845-3849
492KB
978
English
IJPSR
Mohammed W. Nassar, Khalid A. Attia, Hamed M. Abou-Seada and Ahmed El-Olemy Allam*
Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt
ahmed_olemy2000@yahoo.com
19 May, 2013
01 August, 2013
26 September, 2013
http://dx.doi.org/10.13040/IJPSR.0975-8232.4(10).3845-49
01 October, 2013