|Year : 2010 | Volume
| Issue : 2 | Page : 172-179
HPTLC method development and validation of trandolapril in bulk and pharmaceutical dosage forms
N Sreekanth1, Bahlul Z Awen2, Ch. Babu Rao2
1 Department of Pharmacy, College of Public Health and Medical sciences, Jimma University, Jimma, Ethiopia
2 Faculty of Pharmacy, 7th April University, Zawia, Libya
|Date of Submission||01-Apr-2010|
|Date of Decision||28-May-2010|
|Date of Acceptance||06-Jun-2010|
|Date of Web Publication||2-Nov-2010|
Department of Pharmacy, College of Public Health and Medical sciences, Jimma University, Jimma
Source of Support: None, Conflict of Interest: None
| Abstract|| |
A simple, precise, accurate and rapid high performance thin layer chromatographic method has been developed and completely validated for the estimation of trandolapril in bulk and pharmaceutical dosage forms. Quantification of trandolapril was carried out with percolated silica gel 60F254 as stationary phase using mobile phase consisting of Chloroform: Methanol: Acetic acid (8:1.5:0.5 v/ v/ v) and scanned in Absorbancei Reflectance mode at 212 nm using Camag TLC scanner 3 with WinCAT software. The Rf value of trandolapril was found to be 0.54 (±0.03). The proposed method has permitted the quantification of trandolapril over the linearity range of 25150 ng/spot and its percentage recovery was found to 99.7%. The intraday and inter day precision were found to be 1.26% and 1.4%, respectively. The limit of detection and the limit of quantification were found to be 18 ng/ spot and 54 ng/ spot, respectively. The proposed method can be successfully applied for the estimation of drug content of different marketed formulations simultaneously on a single plate and provides a faster and cost effective quality control tool for routine analysis of trandolapril as bulk drug and in tablet dosage forms.
Keywords: HPTLC, Validation, Trandolapril
|How to cite this article:|
Sreekanth N, Awen BZ, Rao C. HPTLC method development and validation of trandolapril in bulk and pharmaceutical dosage forms. J Adv Pharm Technol Res 2010;1:172-9
|How to cite this URL:|
Sreekanth N, Awen BZ, Rao C. HPTLC method development and validation of trandolapril in bulk and pharmaceutical dosage forms. J Adv Pharm Technol Res [serial online] 2010 [cited 2023 Feb 5];1:172-9. Available from: https://www.japtr.org/text.asp?2010/1/2/172/72255
| Introduction|| |
Trandolapril, chemically, it is (2S, 3aR, 7aS)-1-[(S)-N-[(S)-1-carboxy-3phenylpropyl] alanyl] hexahydro-2indolinecarboxylic acid, 1-ethyl ester  and is not official in any pharmacopoeia The chemical structure of trandolapril was shown in [Figure 1]. Trandolapril is a nonsulphydryl prodrug that is hydrolysed to the active diacid trandolaprila. Trandolapril is an orally administered angiotensin converting enzyme inhibitor that has been used in the treatment of patients with hypertension and congestive heart failure, and myocardial infarction , . Literature survey revealed that few HPLC methods were reported for the estimation of trandolapril in the biological fluids ,,,,,, . The present study illustrates development and validation  of a simple, accurate, precise and specific HPTLC method for the estimation of trandolapril tablet dosage forms.
| Experimental|| |
Pure working standard of trandolapril was procured as a gift sample from Ranbaxy Ltd., Himachal Pradesh. All chemicals and reagents used were of analytical grade. A Silica gel 60F 254 TLC pre coated aluminum plates (10 x 10 cm, layer thickness 0.2 mm, E. Merck, Mumbai) were used as a stationary phase. Chloroform: Methanol: Acetic acid (8:1.5:0.5 v/ v/ v) was used as mobile phase and methanol was used as solvent. Commercially available Tablet formulations with labeled amount 2.0 mg of trandolapril (Zetpril-2, Hetero Drugs Pvt. Ltd. And Mavik-2, Abott Pharmaceuticals Ltd) were purchased from the local market.
A CAMAG HPTLC system (Switzerland) comprising a CAMAG Linomat IV semiautomatic sample applicator, a CAMAG TLC Scanner 3, A CAMAG twintrough chamber (10 x 10 cm), CAMAG CATS 4 software, A Hamilton syringe (100 μl), A Shimadzu libror AEG-220 weighing balance and A ultra sonicator (Frontline FS-4, Mumbai) was used during the study.
The chromatographic conditions were optimized and estimations were performed on a stationary phase, pre coated silica gel 60 F254 aluminum sheets (10 x 10 cm) which were prewashed with methanol and dried in air, with mobile phase of Chloroform: Methanol: Acetic acid (8:1.5:0.5 v/v/v) . The chromatographic chamber and plate was allowed to saturate for about 30 min and the migration distance allowed was 72 mm. The wavelength scanning was performed at 212 nm keeping the slit dimension 5 x 0.45 mm. The source of radiation was deuterium lamp emitting a continuous UV spectrum between 190-400nm. The standard solutions of trandolapril was spotted and developed at constant temperature of 25 ± 2°C.
Preparation of mobile phase
Chloroform: Methanol: Acetic acid (8:1.5:0.5 v/ v/ v) was employed as mobile phase.
Preparation of standard solution of trandolapril
A working standard of trandolapril about 2.5 mg was accurately weighed and transferred in to 100 ml volumetric flask. A volume of methanol about 25 ml was added and sonicated for about 20 min; finally the volume was made up to 100ml with methanol to obtain the concentration about 25 μg/ml. From this stock solution 0.1 ml was taken and the volume made up to 100m1 to get concentration about 25ng/ ml.
Preparation of calibration curve
Aliquots (1, 2, 3, 4, 5 and 6 μl) of standard solution of trandolapril were spotted on pre coated TLC plates using semi automatic spotter under nitrogen stream. The plate was dried in air and developed up to 72 mm at constant temperature with a mixture of Chloroform: Methanol: Acetic acid (8:1.5:0.5 v/ v/ v) as mobile phase in a CAMAG twin through chamber which was previously saturated with mobile phase for about 30 min. the plate was removed from the chamber and dried in air. Photometric measurements were performed at 212 nm in absorbance/reflectance mode with the CAMAG TLC scanner 3 using CATS 4 software incorporating track optimizing option. The standard plot of trandolapril was established by plotting the peak area Vs concentration (ng/ml) corresponding to each spot.
Estimation of trandolapril in marketed tablet formulation
Twenty tablets were accurately weighed and finely powdered. The powder which is equivalent to 2.5 mg of trandolapril was weighed, mixed with 25 ml of methanol and sonicated for 15 min. The solution of tablet was filtered through Whatman filter paper No. 41 and the residue was thoroughly washed with methanol. The filtrate and washings were combined in a 100 ml volumetric flask and diluted to the mark with the methanol to get the final concentration of 25 μg/ml of trandolapril. From this stock solution 0.1 ml was taken and the volume made up to 100ml to get concentration about 25ng/ ml. Three micro liters of sample solution was applied on a TLC plate under a nitrogen stream using a semi automatic spotter. The amount of trandolapril present in the sample solution was determined by fitting the area values of peaks corresponding to trandolapril into the equation of the line representing the calibration curve of trandolapril. All determinations were performed in triplicate.
| Results and Discussion|| |
Trandolapril was soluble in methanol, there fore methanol was selected as the solvent. A solvent system consisting of Chloroform: Methanol: Acetic acid (8:1.5:0.5 v/v/v) was selected as mobile phase, that would give dense and compact spot with appropriate R f values was selected for quantification of Trandolapril in pharmaceutical formulations. The present HPTLC method for the quantification trandolapril in bulk and pharmaceutical dosage, revealed as simple, accurate and precise with Rf value of 0.54. The typical densitogram of trandolapril was shown in [Figure 2].
Validation of method
The Linearity for the detection of trandolapril was 25-150 ng/ml with R 2 = 0.998; Y=21.07x + 21.71. The results were shown in the [Table 1]. The precision of the method (System reproducibility) was assessed by spotting 3 μl of drug solution six times on a TLC plate, followed by development of plate and recording the peak area for 6 spots. The % RSD for peak area values of trandolapril was found to be 1.04%. The results were shown in [Table 2]a. The method reproducibility (The intra-day precision) was determined by analyzing standard solutions in the concentration range of 75 ng/spot to 100 ng/spot of drug for 3 times on the same day and inter-day precision was determined by analyzing corresponding standards daily for 3 day over a period of one week. The intra-day and inter-day coefficients of variation (%RSD) are in range of 0.39 to 1.26 and 0.17 to 1.4, respectively. The results were shown in [Table 2]b, c. Recovery studies were carried out to assess accuracy of the method. These studies were carried out at three levels. The percentage recovery was found to be within the limits and shown in [Table 3]. The assay for the marketed formulation was established with the present chromatographic conditions developed and it was found to be more accurate and reliable. The average drug content was found to be 99.15% of the labeled claim. The results were shown in [Table 4]. Limits of Detection (LOD) and Quantification (LOQ), the limits of detection and quantitation were calculated by the method based on the standard deviation of response (a) and the slope of calibration plot (S), using the formulae LOD = 3.3σ/S and LOQ 10σ/S. The LOD and LOQ were calculated and found to be 18 ng/Spot and 54 ng/Spot, respectively. Robustness was determined by altering chromatographic conditions like mobile phase composition, Amount of mobile phase, Plate treatment, Time from spotting to chromatography and time from chromatography. The low value of RSD indicates robustness of the method. The results were shown in the [Table 5]. Specificity test of the proposed method demonstrated that there were no interference form excipients. Furthermore, well shaped peaks indicate the specificity of the method.
|Table 2: a: Precision of Trandolapril|
b: Intra-day precision of Trandolapril
c: Inter-day precision of Trandolapril
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| Intra-Day Precision|| |
| Conclusion|| |
The developed HPTLC technique is simple, precise, specific and accurate and the statistical analysis proved that method is reproducible and selective for the analysis of trandolapril in bulk and pharmaceutical formulations.
| Acknowledgement|| |
The authors are thankful to Department of Pharmacy, College of Public Health and Medical sciences, Jimma University, Jimma, Ethiopia and authors greatly acknowledge Ranbaxy Ltd., Himachal Pradesh for providing the gift sample of Trandolapril.
| References|| |
|1.||The Merck Index, An encyclopedia of chemicals, drugs, & , 13 th ed. Merck Research laboratories, New York, 9644. |
|2.||Peters D.C, Noble S, Plosker G.L. Trandolapril: An update of its pharmacology and therapeutic use in cardiovascular disorders, Drugs. 1998; 56: 871-893. |
|3.||Pedersen O.D, Bagger H, Kober L, Pedersen C. Trandolapril reduces the incidence of atrial fibrillation after acute myocardial infarction in patients with left ventricular dysfunction, Circulation. 1999; 100:376-380. |
|4.||Gumeniczek A, Hopklah H. Development and Validation of a liquid chromatographic method for the determination of trandolapril and verapamil in capsules, J. Liq. Chromatogr. Relat. Technol. 2001; 24:393-400. |
|5.||Constantinos P, Maria K, Irene P. Liquid chromatographic tandem mass spectrometric determination of trandolapril in human plasma, Anal. Chemica. Acta. 2005; 540:375-382. |
|6.||Cendrowska I, Bankowski K, Jopa J. A study on the stereochemical purity of trandolapril and octahydro-1 H-indole-2-carboxylic acid by HPLC method, Acta. Pol. Pharm. 2003; 60: 141-144. |
|7.||Harlikar J. N, Amlani A. M. Simultaneous Determination of Perindopril, Indapamide, Ramipril, Trandolapril in Pharmaceutical formulations using Reverse Phase Liquid Chromatography, Re. J. Chem. And Environ. 2003; 7:144-154. |
|8.||Ramakrishna V.S.N, Vishwottam N. K, Shrivastava W, Koteshwara M. Quantification of trandolapril and its metabolite trandolaprilat in human plasma by liquid chromatography! Tandem mass spectrometry using solid-phase extraction, Rap. Comm. Mas. Spec. 2006; 20:3709 -3716. |
|9.||Gumeniczek A, Hopklah H. Highperformance liquid chromatographic assay of trandolapril in capsules, Acta. Pol. Pharm. 2000; 57:253-265. |
|10.||Bonazzi D, Gotti R, Andrisano V, Cavrini V. Analysis of ACE inhibitors in pharmaceutical dosage forms by derivative UV spectroscopy and liquid chromatography (HPLC), J. Pharm. Biomed. Anal. 1997; 16:431-438. |
|11.||ICH steering committee, International Conference on Harmonization of Technical Requirements for Registration of Pharmaceutical for Human Use. Validation of Analytical Procedures: Text and Methodology, Q2 (R 1), Geneva, Switzerland, 2001. |
[Figure 1], [Figure 2]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5]