SIMULTANEOUS ESTIMATION OF CLOPIDOGREL AND ATORVASTATIN IN HUMAN PLASMA USING BIO-ANALYTICAL RP-ULTRA FAST LIQUID CHROMATOGRAPHIC METHOD
Keywords:
Bioanalytical, Clopidogrel, Atorvastatin, RP-UFLC, USFDAAbstract
This manuscript depicts a simple, accurate, sensitive, precise and robust Ultra fast liquid chromatographic (UFLC) method developed and validated for the simultaneous determination Clopidogrel and Atorvastatin in human plasma as per USFDA draft guidelines.
In the current study, the analysis was performed on phenomenex C8 (250 × 4.6 mm, 5μm) column using phosphate buffer (pH-2.5) and acetonitrile (50: 50 v/v) as mobile phase at flow rate of 1.3 mL/min. In the current developed method, Clopidogrel and Atorvastatin eluted at a retention time of 4.06 and 10.31 min respectively. The proposed method is having linearity in the concentration range from 10 to 50μg/mL of Clopidogrel and Atorvastatin. The current method was validated with respect to linearity; precision, lowest limit of detection (LLOD), accuracy and recovery according to the USFDA guidelines.
The system consisted of a pump (Shimadzu, prominence, UFLC), with 20 µl sample injector, along with a PDA detector at a wavelength of 243 nm and 220 nm for Atorvastatin and Clopidogrel respectively. Data was compiled using Shimadzu LC Solution software.
A good linear relationship over the concentration range of 10-50µg/ml was shown. Validation of the method was carried out as per the USFDA draft guidelines. The method developed was found to be precise, accurate, specific, linear and selective.
Statistical analysis shows that the method is selective and reproducible for the estimation of Clopidogrel and Atorvastatin in combined dosage forms.
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References
Panda SS. Ion-pairing RP-HPLC method for simultaneous determination of aspirin and clopidogrel bisulphate in tablet and capsule dosage form. Int J Pharm Tech Res 2010;2(1):269-73.
Patel RB, Shankar MB, Patel MR, Bhatt KK. Simultaneous estimation of acetylsalicylic acid and clopidogrel bisulfate in pure powder and tablet formulations by high-performance column liquid chromatography and high-performance thin-layer chromatography. J AOAC Int 2008;91(4):750-5.
Anandakumar T, Ayyappan V, Raghu Raman, Vetrichelvan T, Sankar ASK, Nagavalli D. RP-HPLC analysis of aspirin and clopidogrel bisulphate in combination. Indian J Pharm Sci 2011;69:597-9.
Londhe SV, Mulgund SV, Deshmukh RS, Jain KS. Simultaneous HPTLC analysis of aspirin, atorvastatin calcium and clopidogrel bisulphate in the bulk drug and in capsules. Acta Chromatogr 2010;22(2):297-305.
Durga Rao D, Kalyanaraman LS, Sait S, Venkata Rao PA. A validated stability-indicating normal phase LC method for clopidogrel bisulfate and its impurities in bulk drug and pharmaceutical dosage form. J Pharm Biomed Anal 2010;52(1):160-5.
Fayed AS, Weshahy SA, Shehata MA, Hassan NY, Pauwels J, Hoogmartens J, et al. Separation and determination of clopidogrel and its impurities by capillary electrophoresis. J Pharm Biomed 2009;49(2):193-200.
Kalaichelvi R, Fatima Rose M, Vadivel K, Jayachandran E. Simple extractivecolorimetric determination of Atorvastatin sodium by aciddye complexation method in solid dosage form. Int J Chem Res 2010;1(1):6-8.
Kakde RB, Gedam SN, Chaudhary NK, Barsagade AG, Kale DL, Kasture AV. Three-wavelength spectrophotometric method for simultaneous estimation of Atorvastatin and domperidone in pharmaceutical preparations, Int J Pharm Tech Res 2009;1(2):386-9.
Pimpodkar NV, Nalawade RS, Kuchekar BS, Mahajan NS, Jadhav RL. New spectrophotometric method for the estimation of Atorvastatin in bulk and pharmaceutical formulation. Int J Chem Sci 2011;6(2):993-9.
Challa BR, Boddu SH, Awen BZ, Chandu BR, Bannoth CK, Khagga M, et al. Development and validation of a sensitive bioanalytical method for the quantitative estimation of Atorvastatin in human plasma samples by LC-MS/MS: application to bioequivalence study. J Chromatogr B 2010;878(19):1499-505.
Prasanna Reddy B, Kiran Kumar Reddy N. Development and validation of rp-hplc for the Atorvastatin sodium sesquihydrate in pharmaceutical dosage forms and human plasma. Int J Chem Tech Res 2011;1(2):95-198.
Rajnish Kumar, Pinderjit Singh, Harinder Singh. Development of UV Spectrophotometric method for estimation of Atorvastatin in pharmaceutical dosage forms. Int J Pharm Res Dev 2011;3(2):113-7.
Prasanna Kumar Reddy B, Ramanjaneya Reddy Y, Ramachandra D. Determination of atorvastatin sodium and lansoprazole in individual tablet dosage forms by RP-HPLC using single mobile phase. E-J Chem 2011;6(2):489-94.
Gupta KR, Chawla RB, Wadodka SG. Spectrophotometric methods for simultaneous estimation of Atorvastatin and itopride hydrochloride in capsules Orbital the Electro. J Chem 2010;2(2):181-8.
Sivakumar T, Manavalan R, Valliappan K. Development and validation of a RP-HPLC method for simultaneous determination of domperidone and Atorvastatin in pharmaceutical dosage forms. Acta Chromatogr 2007;18:130-42.
FDA Guidance for Industry, Bioanalytical Method Validation, Biopharmaceutics, September; 2013.
United States Pharmacopoeia: USP; 2009. p. 31.
British Pharmacopoeia; 2009.
Panda SS. Ion-pairing RP-HPLC method for simultaneous determination of aspirin and clopidogrel bisulphate in tablet and capsule dosage form. Int J Pharm Tech Res 2010;2(1):269-73.
Patel RB, Shankar MB, Patel MR, Bhatt KK. Simultaneous estimation of acetylsalicylic acid and clopidogrel bisulfate in pure powder and tablet formulations by high-performance column liquid chromatography and high-performance thin-layer chromatography. J AOAC Int 2008;91(4):750-5.
Anandakumar T, Ayyappan V, Raghu Raman, Vetrichelvan T, Sankar ASK, Nagavalli D. RP-HPLC analysis of aspirin and clopidogrel bisulphate in combination. Indian J Pharm Sci 2011;69:597-9.
Londhe SV, Mulgund SV, Deshmukh RS, Jain KS. Simultaneous HPTLC analysis of aspirin, atorvastatin calcium and clopidogrel bisulphate in the bulk drug and in capsules. Acta Chromatogr 2010;22(2):297-305.
Durga Rao D, Kalyanaraman LS, Sait S, Venkata Rao PA. A validated stability-indicating normal phase LC method for clopidogrel bisulfate and its impurities in bulk drug and pharmaceutical dosage form. J Pharm Biomed Anal 2010;52(1):160-5.
Fayed AS, Weshahy SA, Shehata MA, Hassan NY, Pauwels J, Hoogmartens J, et al. Separation and determination of clopidogrel and its impurities by capillary electrophoresis. J Pharm Biomed 2009;49(2):193-200.
Kalaichelvi R, Fatima Rose M, Vadivel K, Jayachandran E. Simple extractivecolorimetric determination of Atorvastatin sodium by aciddye complexation method in solid dosage form. Int J Chem Res 2010;1(1):6-8.
Kakde RB, Gedam SN, Chaudhary NK, Barsagade AG, Kale DL, Kasture AV. Three-wavelength spectrophotometric method for simultaneous estimation of Atorvastatin and domperidone in pharmaceutical preparations, Int J Pharm Tech Res 2009;1(2):386-9.
Pimpodkar NV, Nalawade RS, Kuchekar BS, Mahajan NS, Jadhav RL. New spectrophotometric method for the estimation of Atorvastatin in bulk and pharmaceutical formulation. Int J Chem Sci 2011;6(2):993-9.
Challa BR, Boddu SH, Awen BZ, Chandu BR, Bannoth CK, Khagga M, et al. Development and validation of a sensitive bioanalytical method for the quantitative estimation of Atorvastatin in human plasma samples by LC-MS/MS: application to bioequivalence study. J Chromatogr B 2010;878(19):1499-505.
Prasanna Reddy B, Kiran Kumar Reddy N. Development and validation of rp-hplc for the Atorvastatin sodium sesquihydrate in pharmaceutical dosage forms and human plasma. Int J Chem Tech Res 2011;1(2):95-198.
Rajnish Kumar, Pinderjit Singh, Harinder Singh. Development of UV Spectrophotometric method for estimation of Atorvastatin in pharmaceutical dosage forms. Int J Pharm Res Dev 2011;3(2):113-7.
Prasanna Kumar Reddy B, Ramanjaneya Reddy Y, Ramachandra D. Determination of atorvastatin sodium and lansoprazole in individual tablet dosage forms by RP-HPLC using single mobile phase. E-J Chem 2011;6(2):489-94.
Gupta KR, Chawla RB, Wadodka SG. Spectrophotometric methods for simultaneous estimation of Atorvastatin and itopride hydrochloride in capsules Orbital the Electro. J Chem 2010;2(2):181-8.
Sivakumar T, Manavalan R, Valliappan K. Development and validation of a RP-HPLC method for simultaneous determination of domperidone and Atorvastatin in pharmaceutical dosage forms. Acta Chromatogr 2007;18:130-42.
FDA Guidance for Industry, Bioanalytical Method Validation, Biopharmaceutics, September; 2013.
United States Pharmacopoeia: USP; 2009. p. 31.
British Pharmacopoeia; 2009.
Panda SS. Ion-pairing RP-HPLC method for simultaneous determination of aspirin and clopidogrel bisulphate in tablet and capsule dosage form. Int J Pharm Tech Res 2010;2(1):269-73.
Patel RB, Shankar MB, Patel MR, Bhatt KK. Simultaneous estimation of acetylsalicylic acid and clopidogrel bisulfate in pure powder and tablet formulations by high-performance column liquid chromatography and high-performance thin-layer chromatography. J AOAC Int 2008;91(4):750-5.
Anandakumar T, Ayyappan V, Raghu Raman, Vetrichelvan T, Sankar ASK, Nagavalli D. RP-HPLC analysis of aspirin and clopidogrel bisulphate in combination. Indian J Pharm Sci 2011;69:597-9.
Londhe SV, Mulgund SV, Deshmukh RS, Jain KS. Simultaneous HPTLC analysis of aspirin, atorvastatin calcium and clopidogrel bisulphate in the bulk drug and in capsules. Acta Chromatogr 2010;22(2):297-305.
Durga Rao D, Kalyanaraman LS, Sait S, Venkata Rao PA. A validated stability-indicating normal phase LC method for clopidogrel bisulfate and its impurities in bulk drug and pharmaceutical dosage form. J Pharm Biomed Anal 2010;52(1):160-5.
Fayed AS, Weshahy SA, Shehata MA, Hassan NY, Pauwels J, Hoogmartens J, et al. Separation and determination of clopidogrel and its impurities by capillary electrophoresis. J Pharm Biomed 2009;49(2):193-200.
Kalaichelvi R, Fatima Rose M, Vadivel K, Jayachandran E. Simple extractivecolorimetric determination of Atorvastatin sodium by aciddye complexation method in solid dosage form. Int J Chem Res 2010;1(1):6-8.
Kakde RB, Gedam SN, Chaudhary NK, Barsagade AG, Kale DL, Kasture AV. Three-wavelength spectrophotometric method for simultaneous estimation of Atorvastatin and domperidone in pharmaceutical preparations, Int J Pharm Tech Res 2009;1(2):386-9.
Pimpodkar NV, Nalawade RS, Kuchekar BS, Mahajan NS, Jadhav RL. New spectrophotometric method for the estimation of Atorvastatin in bulk and pharmaceutical formulation. Int J Chem Sci 2011;6(2):993-9.
Challa BR, Boddu SH, Awen BZ, Chandu BR, Bannoth CK, Khagga M, et al. Development and validation of a sensitive bioanalytical method for the quantitative estimation of Atorvastatin in human plasma samples by LC-MS/MS: application to bioequivalence study. J Chromatogr B 2010;878(19):1499-505.
Prasanna Reddy B, Kiran Kumar Reddy N. Development and validation of rp-hplc for the Atorvastatin sodium sesquihydrate in pharmaceutical dosage forms and human plasma. Int J Chem Tech Res 2011;1(2):95-198.
Rajnish Kumar, Pinderjit Singh, Harinder Singh. Development of UV Spectrophotometric method for estimation of Atorvastatin in pharmaceutical dosage forms. Int J Pharm Res Dev 2011;3(2):113-7.
Prasanna Kumar Reddy B, Ramanjaneya Reddy Y, Ramachandra D. Determination of atorvastatin sodium and lansoprazole in individual tablet dosage forms by RP-HPLC using single mobile phase. E-J Chem 2011;6(2):489-94.
Gupta KR, Chawla RB, Wadodka SG. Spectrophotometric methods for simultaneous estimation of Atorvastatin and itopride hydrochloride in capsules Orbital the Electro. J Chem 2010;2(2):181-8.
Sivakumar T, Manavalan R, Valliappan K. Development and validation of a RP-HPLC method for simultaneous determination of domperidone and Atorvastatin in pharmaceutical dosage forms. Acta Chromatogr 2007;18:130-42.
FDA Guidance for Industry, Bioanalytical Method Validation, Biopharmaceutics, September; 2013.
United States Pharmacopoeia: USP; 2009. p. 31.
British Pharmacopoeia; 2009.