A REVIEW ON ETORICOXIB AND PREGABALIN IN METHOD VALIDATION BY RP-HPLC
- 1. GUIDED BY DHIRENDRA KUMAR TARAI PREPARED BY: UPEKSHA J. BAVADIYA Department of Pharmaceutical Quality Assurance GUJARAT TECHNOLOGICAL UNIVERSITY Chandkheda, Ahmadabad – 382424 - Gujarat “A REVIEW ON ETORICOXIB AND PREGABALIN IN METHOD VALIDATION BY RP-HPLC”
- 3. INTRODUCTION OF DRUG (ETORICOXIB)(1) 3 Etoricoxib, sold under the trade name Arcoxia, is a selective COX-2 inhibitor from McOLSON Research Laboratories. Currently it is approved in more than 80 countries worldwide but not in the US, where the Food and Drug Administration (FDA) has required additional safety and efficacy data for Etoricoxib before it will issue approval Etoricoxib is indicated for the treatment of rheumatoid arthritis, psoriatic arthritis, osteoarthritis, ankylosing spondylitis, chronic low back pain, acute pain, and gout. Approved indications differ by country. In the U.K., it is also "used for the short term treatment of moderate pain after dental surgery" of adults. MOA : Like any other COX-2 selective inhibitor Etoricoxib selectively inhibits isoform 2 of cyclo-oxigenase enzyme (COX-2), preventing production of prostaglandins (PGs) from arachidonic acid.
- 4. INTRODUCTION OF DRUG (PREGABALIN)(2) Marketed under the brand name Lyrica among others, is a medication used to treat epilepsy, neuropathic pain, fibromyalgia, restless leg syndrome, and generalized anxiety disorder Its use in epilepsy is as an add-on therapy for partial seizures. When used before surgery, it reduces pain but results in greater sedation and visual disturbances It is taken by mouth Common side effects include headache, dizziness, sleepiness, confusion, trouble with memory, poor coordination, dry mouth, problem with vision, and weight gain. Serious side effects may include angioedema, drug misuse, and an increased suicide risk. MOA : Act as a ligand of the alpha2-delta subunit of calcium channels. Decreased calcium entry into nerve endings. less glutamate released from nerve endings .than relief of neuropathic pain. 4
- 5. Introduction to Dosage form Brand Name Contents Manufacturer Formulation Etoshine NP Etoricoxib + Pregabalin Sun Pharma laboratory pvt. Ltd Tablet 5
- 6. TECHNIQUES OF METHOD DEVLOPMENT 6
- 7. INTRODUCTION OF ANALYTICAL METHOD (3) Analytical chemistry is divided into two branches: Qualitative: A qualitative analysis provides information about the identity of atomic or molecular species or functional groups in sample. Quantitative: A quantitative analysis provides numerical information as to the relative amount of one or more of these components. Analytical methods development and validation play important roles in the discovery, development and manufacture of pharmaceuticals with the objectives. To qualify and quantify the active pharmaceutical ingredients in bulk as well as dosage form To establish impurity profile and limit of allowable impurities in dosage form. 7
- 8. 8 INTRODUCTION OF HPLC METHOD (4) HPLC is an analytical technique widely used for identification, separation, detection and quantification of various drugs and its related degradents. High performance liquid chromatography (HPLC), is a separation technique based on a solid stationary phase and a liquid mobile phase. Most of the drugs in multi- component dosage forms can be analyzed by HPLC method because of the several advantages like rapidity, specificity, accuracy, precision and ease of automation in this method. HPLC method eliminates tedious extraction and isolation procedures. Principle of separation: The principle of separation in normal phase mode and reverse phase mode is adsorption. When mixtures of components are introduced in to a HPLC column, they travel according to their relative affinities towards the stationary phase. The component which has more affinity towards the adsorbent travels slower. The component which has less affinity towards the stationary phase travels faster. Since no two components have the same affinity towards the stationary phase, the components are separated.
- 9. CONT….. Different modes of separation in HPLC: 1) Normal phase mode. 2) Reverse phase ion pair chromatography. 3) Reversed phase mode. 4) Size exclusion chromatography. 9
- 10. Method development by RP-HPLC (5) 10 Reversed-phase chromatography is the mainly used in chromatographic mode, it is used to separate neutral molecules in solution based on their hydrophobicity. As the name suggested that, reversed-phase chromatography is the reverse of normal-phase chromatography in the intelligence that it involves the employ of a polar mobile phase and a non-polar stationary phase. It ensures that a decrease in the polarity of the mobile phase results in a decreases in solute retention.
- 11. 11 HPLC Techniques Normal phase Mode Stationary phase : Polar Mobile phase : Non polar Reverse phase Mode Stationary phase : Non polar Mobile phase : Polar
- 12. 12 Fig. A schematic diagram of HPLC equipment.
- 13. VALIDATION OF ANALYTICAL METHOD (14) 13 Validation Validation is a process of establishing documented evidence, which provides a high degree of assurance that a specific activity will consistently produce a desired result or product meeting its predetermined specifications and quality characteristics. Method Validation Method validation is the process used to confirm that the analytical procedure employed for a specific test is suitable for its intended use. Results from method validation can be used to judge the quality, reliability and consistency of analytical results.
- 14. Validation of Analytical method 14
- 15. 2. DRUG PROFILE (15-17) 15
- 16. 2. Drug profile of Etoricoxib (17) INTRODUCTION CAS Number 202409-33-4 Description Etoricoxib is a synthetic, nonsteroidal anti-inflammatory drug (NSAID) with antipyretic, analgesic, and potential antineoplastic properties. Etoricoxib specifically binds to and inhibits the enzyme cyclooxygenase-2 (COX-2), resulting in inhibition of the conversion of arachidonic acid into prostaglandins. Structure Chemical Formula C18H15ClN2O2S 16
- 17. Mol. Weight 358.84 gm/mol IUPAC Name 5-chloro-2-(6-methylpyridin-3-yl)-3-(4-methylsulfonyl) phenyl) pyridine. Categories COX-2 Inhibitors Anti-Inflammatory Agents Solubility Slightly Soluble in water, freely soluble in methanol Mechanism Of Action Like any other COX-2 selective inhibitor Etoricoxib selectively inhibits isoform 2 of cyclo-oxigenase enzyme (COX-2), preventing production of prostaglandins (PGs) from arachidonic acid. LogP 3.7 pKa 4.96 Melting point 134-135°C 17
- 18. 2. Drug profile of Pregabalin (15-16) INTRODUCTION CAS Number 148553-50-8 Description Pregabalin is structurally similar to gamma- aminobutyric acid (GABA) - an inhibitory neurotransmitter. It may be used to manage neuropathic pain, postherpetic neuralgia, and fibromyalgia among other conditions Structure Chemical Formula C8H17NO2 18
- 19. Mol. Weight 259.22 gm/mol IUPAC Name 3-isobutyl GABA, (S)-3-isobutyl-γ-aminobutyric acid Categories Anticonvulsants Analgesics Solubility Freely Soluble in water Mechanism Of Action Act as a ligand of the alpha2-delta subunit of calcium channels. Decreased calcium entry into nerve endings. less glutamate released from nerve endings .than relief of neuropathic pain. LogP -1.3 pKa 4.2 Melting point 176 - 178ºC 19
- 20. 3. REVIEW OF LITERATURE 20
- 21. 3. Review of Literature Official Drugs Method Year Abstract Summary Ref. No Etoricoxib Monograph IP 2010 Column: stainless steel column 30 cm x 3.9 mm, packed with phenyl silica gel Mobile phase : mixture of 26 volumes of acetonitrile and 74 volumes of a 0.272 per cent w/v solution of sodium acetate, Flow rate: 1.0 ml/min Wavelength: 256 nm 18 Pregabalin Monograph IP 2010 Column: stainless steel column 25 cm x 4.6 mm packed with octadecylsilane bonded to porous silica (5 μm) Mobile phase : mixture of62 volumes of a buffer solution prepared by diluting about 7.18 ml of triethylamine to 1000 ml water, adjusted to pH 3.0 with orthophosphoric acid and 38 volumes of acetonitrile, Flow rate: 2.0 ml/min Wavelength: 340 nm 19 21
- 22. Non - official Drugs Method Authors/year /article /journal Abstract Summary Ref. No Etoricoxib RP-HPLC Bhattacharya I Bhattacharya SP . Asian J. Research Chem 2009 297-299 Column: C18 column Mobile Phase: phosphate buffer ( pH-7.8) and methanol (90:10 v/v) Wavelength: 235 nm., Flow rate: 1 ml/min Retention times: 3.428 min Linearity range: Etoricoxib - 10-200 μg/ml 20 Etoricoxib HPLC Haque M , Shaema N. Am. J. Pharm Tech Res. 2012 275 – 283 Column: C18 column (250 mm × 4.6 mm. 5μm particle size) Mobile Phase: Ammonium Acetate Wavelength: 235 nm., Flow rate: 1 ml/min 21 22
- 23. Drugs Method s authors/ year /articals Abstract Summary Ref. No Etoricoxib RP- HPLC Venugopal S, Tripathi UM, Devanna N, E-Journal of Chemistry 2011 S119-S126 Column : Zorbax SB CN (250 × 4.6 mm , 5µm) Mobile phase : buffer : methanol 50:50 Flow rate : 0.8 ml/min Wavelength : 235 nm 22 Pregabalin RP- HPLC Kasawar GR, Faroogui MN . Indian Journal of Pharmaceutical Sciences. 2010 517-519 Column: Hypersil BDS(C8, 150×4.6 mm,5 μm) column Mobile Phase: phosphate buffer pH 6.9 and acetonitrile in the ratio of 95:05 Flow rate: 1 ml/min 23 Pregabalin RP- HPLC Prathima SSP. INDO AMERICAN JOURNAL OF PHARMACEUTICAL SCIENCES 2015 1038-1047 Column: AGILENT ZORBAX SB-ODS C18(250×4.6 mm)5 μm Mobile Phase: dipotassium hydrogen phosphate: methanol in the ratio of 60:40% v/v Wavelength: 247 nm. ,Flow rate: 1 ml/minRetention times: 2.00 min Linearity range: 50 μg/ml to 150 μg/ml LOQ: 0.53 𝜇g/ml LOD: 1.61 𝜇g/ml 24 23
- 24. Drugs Method Authors/year /article /journal Abstract Summary Ref. No Etoricoxib (ETR) and Thiocolchico side (THC) RP- HPLC Kumar US , Natraj D, Khan A INTERNATIONAL JOURNAL OF RESEARCH IN PHARMACY AND CHEMISTRY 2011 649-65 Column: C18 stainless steel column (InertSil ODS-3, 250 mm x 4.6 mm ID, particle size 5μm) Mobile Phase: phosphate buffer (PH6, adjusted with ortho phosphoric acid) and methanol (30:70 v/v) Wavelength: 255 nm. , Flow rate: 1.2 ml/min Retention times: Etoricoxib - 2.506 min Thiocolchicoside - 4.600 min Linearity range: Etoricoxib - 40-80 μg/ml Thiocolchicoside 2-6 μg/ml 25 Paracetamol (PCM) and Etoricoxib (ETO) HPLC Baheti KG Shaikh S International Journal of PharmTech Research vol -3 2011 1719-1727 Mobile Phase: phosphate buffer : acetonitrile (60:40v/v) Wavelength: 242 nm. Retention times: PCM – 1.51 min ETO – 4.31 min Linearity range: PCM - 5-30 μg/ml ETO 1-6 μg/ml 26 24
- 25. Drugs Method Authors/year /article /journal Abstract Summary Ref. No NSAIDs and their commonly prescribed combinatio n drugs RP-HPLC Gananadhamu S, Patel PN . Chromatography Research International 2013 1-13 Column: Kromasil C18 (250 × 4.6 mm, 5 𝜇m) Mobile Phase: phosphate buffer pH 3.25 and acetonitrile with gradient elution Wavelength: 230 nm. Flow rate: 1.1 ml/min LOQ: 0.64 to 3.24 𝜇g/Ml LOD: 0.04 to 0.97 𝜇g/mL 27 Etoricoxib and Paracetam ol RP-HPLC Rao KP, Damanu G . Journal of Advanced Studies in Agricultural, Biological and Environmental Sciences 2014 2394-2606 Column: Hypersil BDS C18 (250 x 4.6 mm i.d., 5μm particle size)Mobile Phase: 0.05 M sodium dihydrogen phosphate buffer (adjusted pH 3.2 with o-phosphoric acid): acetonitrile (35:65 v/v) Wavelength: 235 nm. Flow rate: 1 ml/min concentration range: Etoricoxib - 1000 to 3000μg/ml 40-80 μg/ml Paracetamol - 1200 to 3600μg/ml 28 25
- 26. Drugs Method Authors/ year/ artical Abstract Summary Ref. No Thiocolchic oside and Etoricoxib RP-HPLC Padmavati k, Rao MS. World Journal of Pharmaceutic al Sciences. 2016 76-79 Column: Hypersil BDS C18 (250 x 4.6 mm i.d., 5μm particle size)Mobile Phase: phosphate buffer(pH-3.4) and acetonitrile in the ratio of 35:65 v/v Wavelength: 260 nm., Flow rate: 1 ml/min Retention times: Etoricoxib – 6.92 min Thiocolchicoside – 2.83 min 29 Pregabalin And Methalcob alamine RP-HPLC Bhatt KK, Patelia EM, Mori A, Journal of Analytical & Bioanalytical Techniques 2018 1-4 Column : C18 ( 250× 4.6 mm) 5µm Mobile phase : Methanol : water 40:60 Wavelength : 218 nm Retension time : 6.4 PH :6.5 30 26
- 27. Drugs Method Authors / year / artical Abstract Summary Ref. No Epalrestat and Pregabalin RP- HPLC Parmeshweri SA ,Anunam G. International Journal of Pharmaceutica l Sciences and Research 2018 1844-1850 Column: C18 column 250 × 4.6 mm Mobile Phase: 0.1% ortho phosphoric acid buffer and acetonitrile ( ratio of 45: 55) Wavelength: 244 nm. , Flow rate: 1 ml/min Detector: PDA Linearity range: Epalrestat - 37.5 – 225 μg/ml Pregabalin - 18.75 - 112.5 μg/ml Retention times: Epalrestat – 2.407 min Pregabalin – 3.272 min 31 Nortriptyli ne and Pregabalin RP- HPLC Potkuri H, Rao SB. Journal of the Chilean Chemical Society.62 2017 62 Column: C18 column BDS (250mm x 4.6 mm, 5m) Mobile Phase: Perchloric acid (0.1%) and acetonitrile in the ratio of 55:45 Wavelength: 210 nm. , Flow rate: 1 ml/min concentration range: nortriptyline – 37.5 - 22.5 μg/ml Pregabalin - 37.5 – 22.5 μg/ml Retention times: nortriptyline – 2.407 min Pregabalin – 3.272 min 32 27
- 28. Drugs Method Authors /year/art. Abstract Summary Ref. No Epalrestat and Pregabalin RP-HPLC Goday S, Ragaman A., Asian journal of pharmaceuti cal and clinical reaserch, vol -112018 319-324 Precision : epalrestat - 0.2% Pregabalin – 0.3% LOQ : Epalrestat - 0.65 μg/ml Pregabalin – 0.25 μg/ml LOD : Epalrestat - 0.21 μg/ml Pregabalin – 0.08 μg/ml Linearity range: Epalrestat - 30–180 ppm Pregabalin - 15–90 ppm 33 Pregabalin and Celecoxib RP-HPLC Swapna G Merugu M, World Journal of Pharmaceuti cal Research, 2017 1354-1360 Column: Hypersil BDS(150 mm x 4.6 mm, 5m) Mobile Phase: potassium di hydrogen orthophosphate buffer of pH 6.5 and acetonitrile in the ratio of (70:30) Wavelength: 238 nm. Flow rate: 1 ml/min Linearity range: Celecoxib - 100 μg/ml -750 μg/ml Pregabalin-37.5μg/ml-281.25 μg/ml 34 28
- 29. Drugs Method Author/year / artical Abstract Summary Ref. No Pregabalin and Aeclofenac stability RP-HPLC Suchitta TJ Gurupadoyy a B M . Acta scientific Pharmaceuti cal sciences. 2018 20-26 Wavelength: Aeclofenac - 276 nm., Pregabalin - 406 nm concentration range: Aeclofenac – 10 - 50μg/ml μg/ml Pregabalin - 50 - 500μg/ml μg/ml LOQ: Aeclofenac - 2μg/ml , Pregabalin - 4μg/ml LOD: Aeclofenac – 0.50 μg/ml ,Pregabalin – 0.6 μg/ml 35 Epalrestat and Pregabalin stability- indicating RP-HPLC Prasad R , Madhuharik a MY. International journal of pharmacy 2017 157-164 Column: C18 (150x4.6mm, 5μ) column Mobile Phase: Ammonium acetate buffer (pH 10): ACN 70:30 % v/v , Flow rate: 1 ml/min LOQ : Epalrestat - 0.57 μg/ml Pregabalin – 1.51 μg/ml , LOD : Epalrestat - 0.19 μg/ml Pregabalin – 0.50 μg/ml Linearity range: Epalrestat - 37.5 – 225 μg/ml Pregabalin - 18.75 - 112.5 μg/ml Retention times: Epalrestat – 2.516 min Pregabalin – 3.132 min 36 29
- 30. Drugs Method Authors /year/ articals/ journal Abstract Summary Ref. No Etoricoxib and Drotaveri-ne RP- HPLC Syal PK, Sahoo M, Ingale KD, Ingale SS, Choudhari VP And Kuchekar BS, Scholars Research Library 2010 93-102 Column: kramosil C18 ( 250 × 4.6 mm, 5µm) Mobile phase : methanol: buffer 51:49 Flow rate : 0.9 ml/min Wavelength : 244 nm 37 Pregabalin And Nor- triptiline RP- HPLC Mewada NA, Patel BR, Patel JG, Vegad KL and Patel VS, International Journal for Pharmaceutical Research Scholars 2017 1-7 Column : BDS Hypersil C18 column (250×4.6 mm ,5 µm) Mobile Phase : buffer : methanol 70:30 Flow rate : 1 ml /min Wavelength : 210nm 38 30
- 31. Drugs Method Authors /year/ articals/ journal Abstract Summary Ref. No Epalrestat and Pregabalin RP-HPLC Sivagami I, Kavyalalitha S, INTERNATIONAL RESEARCH JOURNAL OF PHARMACY 2018 9 Column: C8 (250 x 4.6 mm, 5μm) Column. Mobile Phase: Potassium Dihydrogen Phosphate Buffer: Acetonitrile taken in the ratio 45:55 Flow rate: 1 ml/min, Wavelength: 274 nm LOQ : Epalrestat - 0.73 μg/ml Pregabalin – 0.07 μg/ml LOD : Epalrestat - 0.24 μg/ml Pregabalin – 0.02 μg/ml Retention times: Epalrestat – 2.373 min Pregabalin – 2.967 min 39 31
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