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UHPLC Determination of Besifloxacin Enantiomers on Immobilized Amylose Tris(3,5-dichlorophenylcarbamate) Chiral Stationary Phase

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Abstract

R-Besifloxacin hydrochloride is a novel chiral broad-spectrum fluoroquinolone developed for treatment of bacterial conjunctivitis and used in clinics due to its higher antibacterial activity compared to S-enantiomer. UHPLC separation of R- and S-enantiomers of Besifloxacin hydrochloride on a new Chiralpak IE-3 column consisting of an immobilized amylose tris(3,5-dichlorophenylcarbamate) chiral stationary phase on 3 µm silica was studied. The selection and effects of mobile phase solvents, additives, organic modifiers, and column temperature on retention, separation, and resolution were investigated and optimized. The separation was accomplished under normal-phase conditions using a mobile phase composed of dichloromethane:isopropyl alcohol:trifluoroacetic acid:diethylamine (90:10:0.3:0.05 v/v/v/v) under isocratic mode of elution at 15 °C. The interactions between the analytes, additives, and the stationary phase were discussed and the most probable separation mechanism was proposed. The developed method was validated according to ICH guidelines and found to be suitable for determining the enantiomeric purity of R-Besifloxacin in bulk drugs and formulations.

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References

  1. Appelbaum PC, Hunter PA (2000) Review: The fluoroquinolone antibacterials: past, present and future perspectives. Int J Antimicrob Agents 16:5–15

    Article  CAS  Google Scholar 

  2. Smith A, Pennefather PM, Kaye SB, Hart CA (2001) Fluoroquinolones: place in ocular therapy. Drugs 61:747–761

    Article  CAS  Google Scholar 

  3. Chang MH, Fung HB (2010) Besifloxacin: a topical fluoroquinolone for the treatment of bacterial conjunctivitis. Clin Ther 32:454–471

    Article  CAS  Google Scholar 

  4. Haas W, Pillar CM, Zurenko GE, Lee JC, Brunner LS, Morris TW (2009) Besifloxacin, a novel fluoroquinolone, has broad-spectrum in vitro activity against aerobic and anaerobic bacteria. Antimicrob Agents Chemother 53:3552–3560

    Article  CAS  Google Scholar 

  5. Haas W, Pillar CM, Hesje CK, Sanfilippo CM, Morris TW (2010) Bactericidal activity of besifloxacin against staphylococci, Streptococcus pneumoniae and Haemophilus influenza. J Antimicrob Chemother 65:1441–1447

    Article  CAS  Google Scholar 

  6. Haas W, Gearinger LS, Usner DW, Decory HH, Morris TW (2011) Integrated analysis of three bacterial conjunctivitis trials of besifloxacin ophthalmic suspension, 0.6%: etiology of bacterial conjunctivitis and antibacterial susceptibility profile. Clin Ophthalmol 5:1369–1379

    CAS  Google Scholar 

  7. http://www.accessdata.fda.gov/drugsatfda_docs/nda/2009/022308s000_MicroR.pdf (2009), pp 24–26. Accessed 25 Mar 2009

  8. Stephens TD, Bunde CJ, Fillmore BJ (2000) Mechanism of action in thalidomide teratogenesis. Biochem Pharmacol 59:1489–1499

    Article  CAS  Google Scholar 

  9. Andrea F, Michaela V, Janka Mydlova JL, Jan K (2006) Interconversion of oxazepam enantiomers during HPLC separation. Determination of thermodynamic parameters. J Liq Chromatogr Relat Technol 29:2889–2900

    Article  Google Scholar 

  10. ICH Topic Q 6 A Specifications: test procedures and acceptance criteria for new drug substances and new drug products: chemical substances. http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q6A/Step4/Q6Astep4.pdf. Accessed 6 Oct 1999

  11. Gu XF, Mao BY, Xia M, Yang Y, Zhang JL, Yang DS, Wu WX, Du YX, Di B, Su MX (2016) Rapid, sensitive and selective HPLC–MS/MS method for the quantification of topically applied besifloxacin in rabbit plasma and ocular tissues: application to a pharmacokinetic study. J Pharm Biomed 117:37–46

    Article  CAS  Google Scholar 

  12. Arnold DR, Granvil CP, Ward KW, Proksch JW (2008) Quantitative determination of besifloxacin, a novel fluoroquinolone antimicrobial agent, in human tears by liquid chromatography–tandem mass spectrometry. J Chromatogr B 867:105–110

    Article  CAS  Google Scholar 

  13. Costa MCN, Barden AT, Andrade JMM, Oppe TP, Schapoval EES (2014) Quantitative evaluation of besifloxacin ophthalmic suspension by HPLC, application to bioassay method and cytotoxicity studies. Talanta 119:367–374

    Article  CAS  Google Scholar 

  14. Wang Z, Wang S, Zhu F, Chen Z, Yu L, Zeng S (2012) Determination of enantiomeric impurity in besifloxacin hydrochloride by chiral high-performance liquid chromatography with precolumn derivatization. Chirality 24:526–531

    Article  CAS  Google Scholar 

  15. Mosiashvili L, Chankvetadze L, Farkas T, Chankvetadze B (2013) On the effect of basic and acidic additives on the separation of the enantiomers of some basic drugs with polysaccharide-based chiral selectors and polar organic mobile phases. J Chromatogr A 1317:167–174

    Article  CAS  Google Scholar 

  16. Gogaladze K, Chankvetadze L, Tsintsadze M, Farkas T, Chankvetadze B (2015) Effect of basic and acidic additives on the separation of some basic drug enantiomers on polysaccharide-based chiral columns with acetonitrile as mobile phase. Chirality 27:228–234

    Article  CAS  Google Scholar 

  17. ICH Q2 (R1) (2005) International conference on harmonization: validation of analytical procedures: test and methodology. http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf. Accessed Nov 2005

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Acknowledgements

The authors thank the Director, IICT, for encouragement and permission to communicate the results for publication. A. M. Swamy has received Senior Research (SRF) Fellowship from Council of Scientific and Industrial Research (CSIR) [Grant No. 31/14(2540/2013-EMR-I)] and N. N. Varma has received Senior Research (SRF) Fellowship from University Grants Commission (UGC) (Grant No. 2013/14-10454-94).

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Authors and Affiliations

  1. Discovery Laboratory, Analytical Chemistry Division, Indian Institute of Chemical Technology, Tarnaka, Hyderabad, Telangana, 500 607, India

    Nageswara Rao Ramisetti, Manikanta Swamy Arnipalli & Narendra Varma Nimmu

  2. Department of Chemistry, Government College for Men (A), Kadapa, Andhra Pradesh, 516004, India

    Ramachandra Bondigalla

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  1. Nageswara Rao Ramisetti
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  2. Manikanta Swamy Arnipalli
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  3. Narendra Varma Nimmu
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Correspondence to Nageswara Rao Ramisetti.

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Ramisetti, N.R., Arnipalli, M.S., Nimmu, N.V. et al. UHPLC Determination of Besifloxacin Enantiomers on Immobilized Amylose Tris(3,5-dichlorophenylcarbamate) Chiral Stationary Phase. Chromatographia 80, 1509–1515 (2017). https://doi.org/10.1007/s10337-017-3346-7

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