Vijayageetha, R (2012) New Analytical Method Development, Optimization and Efficient Validation by Chemometric Approach for Marketed Formulations. Doctoral thesis, The Tamil Nadu Dr. M.G.R. Medical University, Chennai.
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Abstract
SPECTROPHOTOMETRIC METHODS: Spectrophotometric methods are the oldest technique for the analysis of components. It have advantages of being simple, reproducible and fast; the apparatus is relatively cheap, still make the methods more attractive. The main disadvantage of spectrophotometric method is the simultaneous analysis of two or more active components in the same mixture was quite tough in resolving the overlapped bands. But this was overcome by new technique called spectrophotometric coupled with chemometrics. CHEMOMETRIC METHODS: Multivariate calibrations methods are playing a very important role in the multi component analysis of mixtures. The approach is useful in the resolution of band overlapping in quantitative analysis. The multivariate calibration has been found to be the method of choice for complexed mixtures. The advantage of multi component analysis using multivariate calibration is the speed of the determination of the components in a mixture, avoiding a preliminary separation step. Control analyses on pharmaceutical preparations using multivariate calibration method, has been proved to be a valid alternative to HPLC. The application of quantitative chemometric methods, particularly principal component regression (PCR) and partial least square (PLS) to multivariate methods. PLS and PCR are calibration methods more effective for the resolution of mixtures having serious spectral overlapping and non-linear absorbance. The main advantage of a multivariate calibration when applied on spectral data is the high speed in processing both absorbance and concentration values. At the same time errors of calibration model are minimized by measuring the absorbances at many points in the wavelength range of the zero order and derivative spectra. The following combinations were analysed by PLS and PCR: 1. Drotaverine Hcl and Mefenamic acid : 181 Data points. 2. Etoricoxib and Paracetamol : 101 Data points. 3. Tolperisone Hcl and Paracetamol : 70 Data points. 4. Thiocholchicoside and Etoricoxib : 108 Data points. 5. Thiocholchicoside, Aceclofenac sodium and Paracetamol : 91 Data points. 6. Diacerein and Aceclofenac sodium : 67 Data points. 7. Diacerein and Celecoxib : 64 Data points. 8. Rabeprazole sodium and Domperidone maleate : 89 Data points. 9. Lafutidine and Domperidone maleate : 51 Data points. 10. Famotidine, Diclofenac potassium, Paracetamol and Chloroxazone : 101 Data points. 11. Phenylephrine Hcl, Paracetamol, Caffeine and Cetrizine Di Hcl : 101 Data points. 12. Phenylephrine Hcl, Paracetamol, Caffeine. and Chlorpheniramine maleate. : 101 Data points The PLS and PCR models were developed using above mentioned data points. The pre-processing and validation steps were carried out for all the mixtures to get the optimum results and to get low values of RMSEC and RMSEP. The results obtained confirm the suitability of the proposed method for accurate and precise analysis of studied drugs. These methods were applied directly to the commercial mixture preparations without previous treatment. According to these studies, multivariate calibration techniques coupled with spectrophotometric can be recommended as a very suitable choice to resolve band overlapping in quantitative analysis Also no expensive laboratory technique is needed. In addition the proposed methods are suitable for application without interference of the excipients. CHROMATOGRAPHIC METHODS: History of chromatography can give an idea about improvement in technology from conventional column chromatography to high performance liquid chromatography and finally at this stage an ultra performance liquid chromatography. The most characteristic feature of the development in the pharmaceutical analysis in the past 25 years is that various forms of HPLC became undoubtedly the most important method. This created an interest in developing a separation technique using UHPLC for quantitative estimation of studied drugs with minimal use of solvents in less time. Another effort was made to utilise the advantage of HPTLC, that a large number samples can be analysed in a shorter time period. This method also utilises less consumption of solvents. REVERSE PHASE ULTRA HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (RP-UHPLC): Thermo scientific Ultra High Performance Liquid Chromatography equipped with Class Accela 600 pump quaternary solvent delivery system, Accela auto sampler and Accela PDA detector. The data acquisition was made with CHROMQUEST software.Separation and quantification of selected drugs was made with Agilent C18 ZORBAX column eclipse plus Rapid Resolution HT (50 x 2.1, 1.8 μm particle size) and Thermo scientific Hypersil Gold C18 (50 x 2.1, 1.9μm particle size) Optimized chromatographic conditions for studied drugs 1. The isocratic technique is used to elute two drugs in 10 mM O-phosphoric acid buffer: Acetonitrile: methanol (pH adjusted to 3.5) 20:50:30 detection at 230 nm for Drotaverine hcl and Mefenamic acid. 2. The gradient profile consisted of 10mM potassium dihydrogen phosphate buffer, acetonitrile and methanol (pH adjusted to 3.00) and the transition in the composition was made by a short gradient programme. This system was used for the elution of four combinations in one system. The combinations are Etoricoxib and paracetamol, Tolperisone Hcl and paracetamol, Thiocholchicoside and etoricoxib, & Thiocholchicoside, Aceclofenac sodium and paracetamol. Detection at 274 nm. 3. The isocratic technique is utilized here to elute two combinations in one system. 10mM potassium dihydrogen phosphate buffer and acetonitrile. (pH adjusted to 3.00), 45:55 The combinations are Diacerein and Aceclofenac & Diacerein and Celecoxib. Detection at 254nm. 4. The isocratic technique is utilized here to elute two combinations in one system. 0.5% Ammonium acetate buffer, water and methanol.50:40:10 (pH adjusted to 7.00) -Rabeprazole sodium and Domperidone Maleate & Lafutidine and Domperidone Maleate. Detection at 220nm. 5. 0.1% Triethylamine buffer and acetonitrile 55: 45 (pH adjusted to 2.5) - Famotidine, paracetamol, diclofenac potassium and chloroxazone in isocratic technique. Detection at 230nm. 6. 0.1% Triethylamine buffer and acetonitrile, 80:20 (pH adjusted to 2.5) – Phenylephrine hcl, Paracetamol, Caffeine and Cetrizine Di hcl. Detection at 220nm. 7. 0.1% Triethylamine buffer, acetonitrile and water in isogradient profile ( pH adjusted to 2.5)– Phenylephrine hcl, Paracetamol, Caffeine and Chlorpheniramine maleate. Detection at 220nm. Based on the results obtained from analysis using optimized methods, it can be concluded that there is no co-eluting peak with the main peaks and the methods are specific for their particular combinations. The methods have linear response in stated range and are simple, precise and accurate. Recovery was in the range with the % RSD of less than 2% for all the drugs. The high percentage recovery and low coefficient of variation confirm the suitability of the method for simultaneous analysis of all the combinations. 7.2.2 High Performance Thin Layer Chromatography (HPTLC) Analysis was performed on 20 x 20 cm aluminium plates pre-coated with 0.2mm layers of silica gel 60 F254 (Merck Mumbai). Standard solutions were applied to pre-washed activated plates, as 6mm bands under a stream of nitrogen, by means of a Camag (Switzerland) Linomat V automated spray-on band applicator with a Hamilton 100μl syringe. Linear ascending development of the plates to a distance of 8cm was performed with optimized mobile phase in a Camag twin trough chamber previously saturated with mobile phase for 20 min at room temperature. After development the plates were scanned at the selected wavelength by means of Camag TLC scanner 3 using WinCATS (version 1.4.4) software incorporating track position optimization .the slit dimension were 4.00 x 0.30 mm with scanning speed 20 mm s-1. System 1 Ethyl acetate: hexane: toluene: methanol (5: 2: 2: 3) - Drotaverine hcl and Mefenamic acid. Detection was at 230nm. System 2 Toluene: ethyl acetate: glacial acetic acid (5:5: 0.02) – Diacerein and Celecoxib. Detection was at 254nm. System 3 Toluene: ethyl acetate: Triethylamine (3: 7: 0.1) – Tolperisone Hcl and Paracetamol. Detection was at 274nm. System 4 Chloroform: n-Hexane: methanol (6: 3: 1) –Lafutidine and Domiperidone. Detection was at 220nm. System 5 Ethyl acetate: methanol: triethylamine (5: 5: 0.02) – Thiocholchicoside, Aceclofenac sodium and Paracetamol. Detection was at 274 nm. The proposed HPTLC method offers simultaneous analysis of studied drugs. The advantage of this method includes small amount of mobile phase requirement, speed of analysis and reduction in the cost. HPTLC does not suffer from pH restrictions and sample cleanup procedures. Several samples can be analysed simultaneously and the plates can be scanned at different wavelengths for several times. No chromatographic interference from the tablet excipients was found. Statistical analysis showed that the method was repeatable and selective for the simultaneous quantitation of the above studied drugs in tablet formulation. and for routine quality control of the drugs.
| Item Type: | Thesis (Doctoral) |
|---|---|
| Uncontrolled Keywords: | analytical method, development, optimization, validation, chemometric approach, marketed formulations. |
| Subjects: | PHARMACY > Pharmaceutical Analysis |
| Depositing User: | Devi S |
| Date Deposited: | 04 Jul 2017 10:40 |
| Last Modified: | 17 Sep 2022 11:19 |
| URI: | http://repository-tnmgrmu.ac.in/id/eprint/913 |
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