Abstract
The emergence and prevalence of multi-drug-resistant bacterial strains increase the potential for outbreaks of incurable infections. The discovery of novel antibiotics and pharmacological preparations requires the identification of novel bioactive small molecules. A specific, sensitive, and reliable quantification method using high-performance liquid chromatography (HPLC) with UV detection was developed for the determination of total persipeptides (A and B), which are cyclic pentapeptides found in the fermentation broth of Streptomyces zagrosensis UTMC 1154 that exhibit bioactivity against methicillin-resistant Staphylococcus aureus (MRSA). A simple liquid–liquid extraction (LLE) method using butanol was employed to extract persipeptides from the fermentation broth prior to HPLC analysis. The chromatographic separation of persipeptides and the internal standard, virginiamycin, was achieved with a gradient of acetonitrile and water on a C18 reversed-phase analytical column in a 25-min analytical run utilizing a flow rate of 0.8 mL min−1 and detection at 210 nm. The whole assay was validated, and the method presented a linear response range with a regression coefficient of determination R 2 of 0.9996 for the quantification of persipeptides in the concentration range of 3.9–250.0 µg mL−1, as well as extraction recoveries ranging from 54.78 ± 9.83 % to 56.45 ± 16.33 %. The bias and the precision of the proposed method were <10 %. The detection and quantification limits for the persipeptides were 27 and 83 µg L−1, respectively.
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Wise R, Blaser M, Carrs O, Cassell G, Fishman N, Guidos R, Levy S, Powers J, Norrby R, Tillotson G (2011) The urgent need for new antibacterial agents. J Antimicrob Chemother 66:1939–1940
Walsh C (2003) Antibiotics: actions, origins, resistance. American Society for Microbiology (ASM), Washington, DC
Fischbach MA, Walsh CT (2009) Antibiotics for emerging pathogens. Science 325:1089–1093
Hancock Robert EW, Chapple DS (1999) Peptide antibiotics. Antimicrob Agents Chemother 43:1317–1323
Perlman D, Bodanszey M (1971) Biosynthesis of peptide antibiotics. Annu Rev Biochem 40:449–464
Mohammadipanah F, Matasyoh J, Hamedi J, Klenk H, Laatsch H (2012) Persipeptides A and B, two cyclic peptides from Streptomyces sp. UTMC 1154. Bioorg Med Chem 20:335–339
Mohammadipanah F, Hamedi J, Spröer C, Rohde M, del Carmen Montero-Calasanz M, Klenk HP (2014) Streptomyces zagrosensis sp. Nov., isolated from soil. Int J Syst Evol Microbiol 64:3434–3440
Laatsch H (2011) A data base for rapid dereplication and structure determination of microbial natural products. Wiley-VCH, Weinheim
Paim CS, Führ F, Barth AB, Gonçalves CE, Nardi N, Steppe M, Schapoval EE (2011) Gemifloxacin mesylate (GFM) stability evaluation applying a validated bioassay method and in vitro cytotoxic study. Talanta 83:1774–1779
Berridge N, Barrett J (1952) A rapid method for the turbidimetric assay of antibiotics. J Gen Microbiol 6:14–20
Salem H (2004) Selective spectrophotometric determination of phenolic β-lactam antibiotics in pure forms and in their pharmaceutical formulations. Anal Chim Acta 515:333–341
Frutos P, Torrado S, Perez-Lorenzo M, Frutos G (2000) Validated quantitative colorimetric assay for gentamicin. J Pharm Biomed Anal 21:1149–1159
Ašperger D, Mutavdžić D, Babić S, Horvat AJ, Kaštelan-Macan M (2006) Solid-phase extraction and TLC quantification of enrofloxacin, oxytetracycline, and trimethoprim in wastewater. J Planar Chromat 19:129–134
Sulc M, Fadrhoncova I, Jelinkova M, Chudomelova M, Felsberg J, Olsovska J (2011) Determination of sibiromycin and its natural derivatives using new analytical and structural approaches. J Chromatogr A 1218:83–91
Dong MW (2013) A three-pronged template approach for rapid HPLC method development. LC-GC N Am 31:456
Gilroy JJ, Dolan JW (2004) Gradient performance checks. LC-GC Eur 17:566–572
Schwalbe R, Steele-Moore L, Goodwin AC (2007) Antimicrobial susceptibily testing protocols. CRC, New York
US FDA (1994) Reviewer guidance: validation of chromatographic methods. Center for Drug Evaluation Research (US FDA), Washington, DC
ICH (2008) IHT guideline: validation of analytical procedures: text and methodology, Q2 (R1), current step 4 version, parent guidelines on methodology dated November 6 1996, incorporated in November 2005. International Conference on Harmonisation (ICH), Geneva. http://www.ich.org
Dong MW (2006) Modern HPLC for practicing scientists. Wiley, Hoboken
Dejaegher B, Heyden YV (2007) Ruggedness and robustness testing. J Chromatogr A 1158:138–157
Snyder LR, Kirkland JJ, Dolan JW (2010) Basic concepts and the control of separation. In: Introduction to modern liquid chromatography. Wiley, Hoboken, pp 19–86
USP (2006) Chromatography: system suitability. The United States Pharmacopeia, 30th edn. The United States Pharmacopeia Convention (USP), Rockville, p 254
US FDA (2001) Guidance for industry: bioanalytical method validation. Center for Veterinary Medicine (US FDA), Rockville
Islam M, Toledo R, Hamdy M (1999) Stability of virginiamycin and penicillin during alcohol fermentation. Biomass Bioenerg 17:369–376
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Mohammadipanah, F., Kazemi Shariat Panahi, H., Imanparast, F. et al. Development of a Reversed-Phase Liquid Chromatographic Assay for the Quantification of Total Persipeptides in Fermentation Broth. Chromatographia 79, 1325–1332 (2016). https://doi.org/10.1007/s10337-016-3140-y
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DOI: https://doi.org/10.1007/s10337-016-3140-y