HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC ANALYSIS OF DULOXETINE AND ITS METABOLITES IN RAT AND CHARACTERIZATION OF METABOLITES IN PLASMA, URINE, FECES AND BILE THROUGH RETRO-SYNTHESIS FOLLOWED BY NMR AND MS STUDY
Abstract
A simple and sensitive reverse phase high performance liquid chromatographic (RP-HPLC) method for determination of duloxetine and twelve of its metabolites, Sulfate conjugate of 4-hydroxy duloxetine (M1), N-desmethyl duloxetine (M2), Glucuronide conjugate of 4-hydroxy duloxetine (M3), Glucuronide conjugate of 6-hydroxy duloxetine (M4), Glucuronide conjugate of 4,6-dihydroxy duloxetine (M5), Glucuronide conjugate of 5-hydroxy-6-methoxy duloxetine (M6), 4-Hydroxy duloxetine (M7), 5-Hydroxy duloxetine (M8), 6-Hydroxy duloxetine (M9), Thienyl alcohol (M10), Glucuronide conjugate of 5-hydroxy duloxetine (M11), Glucuronide conjugate of dihydrodiol duloxetine (M12) in Wistar rat plasma, urine, feces and bile was developed. Analysis was carried out on a µ-Bondapak C18 column (250mm × 4.6mm, 5µm particle size) using methanol: phosphate buffer (pH 7.8, 50 mM) (7:3 v/v) as the mobile phase at a flow rate of 1ml/min. Detection was carried out at 221 nm with an UV detector. The above metabolites were characterized by retro-synthesis followed by 1H-NMR, 13C-NMR and M.S study for structure confirmation and finally injected separately into the HPLC system. All the twelve retention time matches with the metabolites present in the plasma, urine, feces and bile sample. This method has also been successfully applied in the pharmacokinetics study of duloxetine after orally administrating the duloxetine to Wistar rat.
Keywords:
Duloxetine, Metabolites, HPLC, Retro-synthesis, CharacterizationDOI
https://doi.org/10.25004/IJPSDR.2013.050207References
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18. Patel SK, Patel NJ, Prajapati AM, Patel DB, Patel SA. Stability-indicating RP-HPLC Method development and validation for Duloxetine Hydrochloride in Tablets. J AOAC Int. 2010; 93: 123-132.
19. Sinha VR, Anamika, Kumria R, Bhinge JR. Stress degradation studies on duloxetine hydrochloride and development of an RP-HPLC method for its determination in capsule formulation. J Chromatogr Sci. 2009; 47: 589-93.
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21. Chhalotiya UK, Bhatt KK, Shah DA, Baldania SL. Development and validation of a stability-indicating RP-HPLC method for duloxetine hydrochloride in its bulk and tablet dosage form. Sci Pharm. 2010; 78: 857-868.
22. Reddy PB.Validation and stability indicating reverse phase-high performance liquid chromatography for the determination of duloxetine in tablets. Int J Chem Tech Res. 2009; 1: 602-605.
23. Reddy PRM, Sreeramulu1 J, Naidu PY, Reddy AR. Stability indicating fast LC for the Simultaneous estimation of Intermediates and degradants of duloxetine hydrochloride. Chromatographia. 2010; 71: 95-100.
24. Laha TK, Mishra G, Sen S. A Validated Stability Indicating Reversed Phase High Performance Liquid Chromatographic Method of Duloxetine and Characterization of its Degradation Products through Retro-Synthesis. Pharm Anal Acta (Scientific Reports). 2012; 1: 1-6.
25. Adcock W, Alste J, Rizvi SQA, Aurangzeb M. Substituent effects in the naphthalene ring system by fluorine-19 NMR. J Am Chem Soc. 1976; 98: 1701-1711.
26. Adcock W, Dewar MJS. Substituent Effects VIII - Synthesis of Substituted α- and β-Fluoronaphthalenes. J Am Chem Soc. 1967; 89: 386-390.
27. Boswell GE, Licause JF. A Convenient Large-Scale Synthesis of 4-Fluoro-1-naphthaldehyde and Its Aromatic Nucleophilic Substitution Reactions. J Org Chem.1995; 60: 6592-6594.
28. Ishii H, Hanaoka T, Asaka, T, Harada Y, Ikeda N. Oxidation with Fremy's salt—VIII: Peri-effect of a group located at the C5 position of 1-naphthol and related compounds. Tetrahedron.1976; 32: 2693-2698.
29. Harvey RG, Pataki J, Lee H. Synthesis of the dihydrodiol and diol epoxide metabolites of chrysene and 5-methylchrysene. J Org Chem.1986; 51: 1407-1412.
30. Wheeler WJ and Kuo F. An asymmetric synthesis of duloxetine hydrochloride, a mixed uptake inhibitor of serotonin and norepinephrine and its C-14 labeled isotopomers. J Labelled Compd Radiopharm. 1995; 36: 213-223.
2. Mishra L. Duloxetine hydrochloride is a newer selective serotonin and norepinephrine reuptake inhibitor (SSNRI) used for major depressive disorders. Drugs today 2006; 1: 489.
3. Stephan AC, Luc AG, Francois RV, Peter RB, Frank PB, Melissa JJ, Christine H, William ZP. Duloxetine increases serotonin and norepinephrine availability in healthy subjects: A double-blind controlled study. J Neuropsychopharmacol. 2003; 28: 1685-1693.
4. National Institute for Health and Clinical Excellence, Urinary incontinence the management of urinary incontinence in women, NW, London, 2006.
5. National Institute for Health and Clinical Excellence, The pharmacological management of neuropathic pain in adults in non-specialist settings, High Holborn, London, 2010.
6. http://www.drugbank.ca/drugs/DB00476.
7. Wolfe F, Smythe HA, Yunus MB, Bennett RM, Bombardier C, Goldenberg DL, Tugwell P, Campbell SM, Abeles M, Clark P. Criteria for the Classification of Fibromyalgia. Arthritis Rheum.1990; 33: 160-72.
8. Douglas RD. Treating Patients for Comorbid Depression, Anxiety Disorders and Somatic Illnesses. J Am Osteopath Assoc. 2006; 106: S1-S8.
9. http://www.cancer.gov/dictionary/CdrID=589399.
10. Johnson JT, Oldham SW, Lantz RJ, Delong AF. High performance liquid chromatographic method for the determination of duloxetine and desmethyl duloxetine in human plasma. J Liq Chromatogr Rel Technol. 1996; 19: 1631-1641.
11. Soni P, Mariappan TT, Banerjee UC. High-performance liquid chromatographic method for the simultaneous estimation of the key intermediates of duloxetine. Talanta. 2005; 67: 975-978.
12. Brenna E, Frigoli S, Fronza G, Fuganti C, Malpezzi L. Isolation and characterization of a phenolic impurity in a commercial sample of duloxetine. J Pharm Biomed Anal. 2007; 43: 1573-1575.
13. Jansen PJ, Oren PL, Kemp CA, Maple SR, Baertschi SW. Characterization of impurities formed by interaction of duloxetine HCl with enteric polymers hydroxypropyl methylcellulose acetate succinate and hydroxypropyl methylcellulose phthalate. J Pharm Sci. 1998; 87: 81-85.
14. Kamila MM, Mondal N, Ghosh LK. A validated UV spectrophotometric method for determination of duloxetine hydrochloride. Pharmazie 2007; 62: 414-415.
15. Olsen BA, Argentine MD. HPLC method development for duloxetine hydrochloride using a combination of computer-based solvent strength optimization and solvent selectivity mixture design. J Liq Chromatogr Rel Technol. 1996; 19: 1993-2007.
16. Dhaneshwar SS, Deshpande P, Patil M, Vadnerkar G, Dhaneshwar SR. Development and validation of a HPTLC method for estimation of duloxetine hydrochloride in bulk drug and in tablet dosage form. Ind J Pharm Sci. 2008; 70: 233-236.
17. Musenga A, Amore M, Mandrioli R, Kenndler E, Martino L, Raggi MA. Determination of duloxetine in human plasma by capillary electrophoresis with laser-induced fluorescence detection. J Chromatogr B. 2009; 877: 1126-1132.
18. Patel SK, Patel NJ, Prajapati AM, Patel DB, Patel SA. Stability-indicating RP-HPLC Method development and validation for Duloxetine Hydrochloride in Tablets. J AOAC Int. 2010; 93: 123-132.
19. Sinha VR, Anamika, Kumria R, Bhinge JR. Stress degradation studies on duloxetine hydrochloride and development of an RP-HPLC method for its determination in capsule formulation. J Chromatogr Sci. 2009; 47: 589-93.
20. Raman NVVSS, Harikrishnaa KA, Prasada AVSS, Reddya KR, Ramakrishna K. Determination of duloxetine hydrochloride in the presence of process and degradation impurities by a validated stability-indicating RP-LC method. J Pharm Biomed Anal. 2010; 51: 994-997.
21. Chhalotiya UK, Bhatt KK, Shah DA, Baldania SL. Development and validation of a stability-indicating RP-HPLC method for duloxetine hydrochloride in its bulk and tablet dosage form. Sci Pharm. 2010; 78: 857-868.
22. Reddy PB.Validation and stability indicating reverse phase-high performance liquid chromatography for the determination of duloxetine in tablets. Int J Chem Tech Res. 2009; 1: 602-605.
23. Reddy PRM, Sreeramulu1 J, Naidu PY, Reddy AR. Stability indicating fast LC for the Simultaneous estimation of Intermediates and degradants of duloxetine hydrochloride. Chromatographia. 2010; 71: 95-100.
24. Laha TK, Mishra G, Sen S. A Validated Stability Indicating Reversed Phase High Performance Liquid Chromatographic Method of Duloxetine and Characterization of its Degradation Products through Retro-Synthesis. Pharm Anal Acta (Scientific Reports). 2012; 1: 1-6.
25. Adcock W, Alste J, Rizvi SQA, Aurangzeb M. Substituent effects in the naphthalene ring system by fluorine-19 NMR. J Am Chem Soc. 1976; 98: 1701-1711.
26. Adcock W, Dewar MJS. Substituent Effects VIII - Synthesis of Substituted α- and β-Fluoronaphthalenes. J Am Chem Soc. 1967; 89: 386-390.
27. Boswell GE, Licause JF. A Convenient Large-Scale Synthesis of 4-Fluoro-1-naphthaldehyde and Its Aromatic Nucleophilic Substitution Reactions. J Org Chem.1995; 60: 6592-6594.
28. Ishii H, Hanaoka T, Asaka, T, Harada Y, Ikeda N. Oxidation with Fremy's salt—VIII: Peri-effect of a group located at the C5 position of 1-naphthol and related compounds. Tetrahedron.1976; 32: 2693-2698.
29. Harvey RG, Pataki J, Lee H. Synthesis of the dihydrodiol and diol epoxide metabolites of chrysene and 5-methylchrysene. J Org Chem.1986; 51: 1407-1412.
30. Wheeler WJ and Kuo F. An asymmetric synthesis of duloxetine hydrochloride, a mixed uptake inhibitor of serotonin and norepinephrine and its C-14 labeled isotopomers. J Labelled Compd Radiopharm. 1995; 36: 213-223.
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01-04-2013
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“HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC ANALYSIS OF DULOXETINE AND ITS METABOLITES IN RAT AND CHARACTERIZATION OF METABOLITES IN PLASMA, URINE, FECES AND BILE THROUGH RETRO-SYNTHESIS FOLLOWED BY NMR AND MS STUDY”. International Journal of Pharmaceutical Sciences and Drug Research, vol. 5, no. 2, Apr. 2013, pp. 70-77, https://doi.org/10.25004/IJPSDR.2013.050207.
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How to Cite
“HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC ANALYSIS OF DULOXETINE AND ITS METABOLITES IN RAT AND CHARACTERIZATION OF METABOLITES IN PLASMA, URINE, FECES AND BILE THROUGH RETRO-SYNTHESIS FOLLOWED BY NMR AND MS STUDY”. International Journal of Pharmaceutical Sciences and Drug Research, vol. 5, no. 2, Apr. 2013, pp. 70-77, https://doi.org/10.25004/IJPSDR.2013.050207.
