Analytical methods for determination of Amphotericin B in biological samples: a short review

  • Gurpreet Singh Department of Pharmaceutical Sciences Guru Nanak Dev University Amritsar (Punjab),143005
  • Tanurajvir Kaur
  • Anudeep Kaur
  • Ravinder Kaur
  • Ramandeep Kaur
Keywords: Amphotericin B, Biological samples, Plasma, Human CSF, Piroxicam, HPLC


Determination of Amphotericin B in biological samples is of great importance, not only in clinical and forensic toxicology, but also in drug testing and pharmacokinetic profile. Human biological samples including organs, tissues, biofluids such as blood and their derivatives, are increasingly important resources for biomedical research. They can help us to understand how a diagnosis can be carried out, categorized and treatment can be further imitated for a whole variety of medical condition and are particularly important when studying pharmacokinetic profile of drugs in animal models. This review paper is a compilation of last 60 years research articles from 1954 to 2013 that describe procedures for the detection of Amphotericin B, in enormous variety of biological samples.  The efficient analytical techniques assigned for accurately analyzing and quantifying drug in biological samples are HPLC and LC MS/MS (hyphenated techniques). Furthermore, this review provides all basic information regarding the sample preparation, type of biological sample and all detail concerning the determination through chromatography.

Author Biography

Gurpreet Singh, Department of Pharmaceutical Sciences Guru Nanak Dev University Amritsar (Punjab),143005
Assistant Professor [Pharmaceutics]Department of Pharmaceutical Sciences


1. Lederberg J. Antifungal agents. In: Schaechter M, editor. The Desk Encyclopedia of microbiology, 1st ed. UK: Academic Press; 2004.p. 47-67.
2. Hartsel S, Bolard J. Amphotericin B: new life for an old drug. Trends Pharmacol Sci. 1996;17:445-9.
3. Gallis HA, Drew RH, Pickard WW. Am-photericin B: 30 years of clinical expe-rience. Rev Infect Dis. 1990;12:308-29.
4. Hann IM, Prentice HG. Lipid-based am-photericin B: a review of the last 10 years of use. Int J Antimicrob Agents. 2001;17:161-9.
5. Hamill RJ. Amphotericin B formulations: a comparative review of efficacy and toxicity. Drugs. 2013;73:919-34.
6. Cifani C, Costantino S, Massi M, Berrino L. Commercially available lipid formulations of amphotericin B: are they bioequivalent and therapeutically equivalent? Acta Biomed. 2012;83:154-63.
7. Amphotericin B. In: Florey K, editor. Analytical Profiles of Drug Substances, vol 6. UK: Academic Press; 1977. p.1-42.
8. Amphotericin B. In: Moffat AC, editor. Clarkes's Isolation and Identification of Drugs, 4th ed. Pharmaceutical Press; 2011, p.897.
9. Antifungals. In: Sweetman SC, Martindale: The Complete Drug Reference, 36th ed. Pharmaceutical Press; 2009, p.523-7.
10. Amphotericin B. accessed on 1/3/2013
11. Tarbet JE, Sternberg TH. Microbiological estimation of antifungal blood levels fol-lowing administration of fungicides. Mycologia. 1954;46:263–75.
12. Halde C, Newcomer VD, Wright ET, Sternberg TH. An evaluation of amphotericin B in vitro and in vivo in mice against Coccidioides immitis and Candida albicans, and preliminary observations concerning the administration of amphotericin B to man. J Invest Dermatol. 1957;28:217-32.
13. Shadomy S, McCay JA, Schwartz SI. Bio-assay for hamycin and amphotericin B in serum and other biological fluids. Appl Microbiol. 1969;17:497-503.
14. Herd PA, Martin HF. A semi-quantitative spectrophotometric assay for amphotericin B in blood. Clin Biochem. 1974;7:359-65.
15. Millership JS, McCaffrey F, Tierney D. A novel application of ratio spectra derivative spectroscopy for the determination of amphotericin in highly icteric plasma. J Pharm Biomed Anal. 2008;48:408-13.
16. Ganière MC, Kergueris MF, Iooss P, Tho-mas L, Larousse C. Quantitation of amphotericin B in plasma by second-derivative spectrophotometry. J Pharm Biomed Anal. 1998;17:481-5.
17. Botsoglou NA, Fletouris DJ, Papageorgiou GE, Florou-Paner P, Mantis AJ. Rapid de-termination of amphotericin B in serum and urine by third-order derivative spectrophotometry. J Pharm Sci. 1996;85:402-6.
18. Pavan B, Dileep RJ, Nivesh KM, Bivash M, Laura AT, George CW. Rapid Quantitative Evaluation of Amphotericin B in Human Plasma, by Validated HPLC Method. Journal of Bioequivalence & Bioavailability. 2013;5:121-4.
19. Espada R, Josa JM, Valdespina S, Dea MA, Ballesteros MP, Alunda JM, Torrado JJ. HPLC assay for determination of amphotericin B in biological samples. Biomed Chromatogr. 2008;22:402-7.
20. Nilsson EI, Yoshikawa TT, Edwards JE, Schotz MC, Guze LB. Quantitation of amphotericin B with use of high-pressure liquid chromatography. J Infect Dis. 1977;135:414-22.
21. Koizumi T, Kubo K, Kaneki T, Hanaoka M, Hayano T, Miyahara T, et al. Pharmacokinetic evaluation of amphotericin B in lung tissue: lung lymph distribution after intravenous injection and airspace distribution after aerosolization and inhalation of amphotericin B. Antimicrob Agents Chemother. 1998;42:597-600.
22. Lemke A, Kiderlen AF, Petri B, Kayser O. Delivery of amphotericin B nanosuspen-sions to the brain and determination of activity against Balamuthia mandrillaris amebas. Nanomedicine. 2010;6:597-603.
23. Yang L, Xiao P, Shi C, You J, Dong X, Zhang Y. Comparative pharmacokinetics after administration of a novel formulation of amphotericin B nanoparticles and am-photericin B deoxycholate to rats and their antifungal activity in vitro. Asian J Pharm Sciences. 2007;2:122-9.
24. Bach PR. Quantitative extraction of amphotericin B from serum and its determination by high-pressure liquid chromatography. Antimicrob Agents Chemother. 1984;26:314-7.
25. Hosotsubo H, Hosotsubo K. Improved high-performance liquid chromatographic determination of amphotericin B in human serum and plasma. J Pharm Biomed Anal. 1989;7:975-9.
26. Liu H, Davoudi H, Last T. Determination of amphotericin B in cerebrospinal fluid by solid-phase extraction and liquid chromatography. J Pharm Biomed Anal. 1995;13:395-400.
27. Echevarría I, Barturen C, Renedo MJ, Di-os-Viéitez MC. High-performance liquid chromatographic determination of amphotericin B in plasma and tissue. Application to pharmacokinetic and tissue distribution studies in rats. Journal of Chromatography A. 1998; 819:171–6.
28. Zou SR, Li YQ, Li P, Zhou B, Wen YG. Determination of amphotericin B in human plasma by HPLC. Journal of Guangdong Pharmaceutical College. 2010;6:602-4.
29. Campanero MA, Zamarreno AM, Diaz M, .Dios-Vieitez MC, Azanza JR. Development and validation of an HPLC method for determination of amphotericin B in plasma and sputum involving solid phase extraction. Chromatographia. 1997;46:641-6.
30. Italia JL, Singh D, Ravi Kumar MN. High-performance liquid chromatographic analysis of amphotericin B in rat plasma using α-naphthol as an internal standard. Anal Chim Acta. 2009;634:110-4.
31. Egger P, Bellmann R, Wiedermann CJ. Determination of amphotericin B, liposomal amphotericin B, and amphotericin B colloidal dispersion in plasma by high-performance liquid chromatography. J Chromatogr B Biomed Sci Appl. 2001;760:307-13.
32. Uday SC, Tapan KP. Rapid and Sensitive High Performance Liquid Chromatography Method for the Determination of Amphotericin B in Rat Plasma. J Pharm Research. 2011;4:3194-7.
33. Goldblum D, Rohrer K, Frueh BE, Theurillat R, Thormann W, Zimmerli S. Ocular distribution of intravenously administered lipid formulations of amphotericin B in a rabbit model. Antimicrob Agents Chemother. 2002;46:3719-23.
34. Malone ME, Corrigan OI, Kavanagh PV, Gowing C, Donnelly M, D'Arcy DM. Pharmacokinetics of amphotericin B lipid complex in critically ill patients undergoing continuous venovenous haemodiafiltration. Int J Antimicrob Agents. 2013;42:335-42.
35. Alak A, Moy S, Bekersky I. A high-performance liquid chromatographic assay for the determination of amphotericin B serum concentrations after the administration of Ambisome, a liposomal amphotericin B formulation. Ther Drug Monit. 1996;18:604-9.
36. Bhatta RS, Chetan R, Chandasana H, Ku-mar D, Chhonker SY, Jain GK. LC–MS Method for Determination of Amphotericin B in Rabbit Tears and Its Application to Ocular Pharmacokinetic Study. Chromatographia. 2011;73:487-93.
37. Xiong X, Zhai S, Liu F. Determination of amphotericin B in human cerebrospinal fluid by. LC-MS-MS. Chromatographia. 2009;70:329-32.
38. Lee JW, Petersen ME, Lin P, Dressler D, Bekersky I. Quantitation of free and total amphotericin B in human biologic matrices by a liquid chromatography tandem mass spectrometric method. Ther Drug Monit 2001;23:268-76
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