HPLC determination of colistin and aminoglycoside antibiotics in feeds by post-column derivatization and fluorescence detection

Morovján, Gy.; Csokan, P; Németh-Konda, L.

doi:10.1007/bf02467512

Cited by 48 publications

(24 citation statements)

References 7 publications

(5 reference statements)

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“…In order to increase the UV absorption of the kanamycin A molecule, chemical derivatization is often applied. The commonly used derivatizing agents are 3,5-dinitrobenzoyl chloride [10], phenyl isocyanate [11], o-phthalaldehyde [12], and 2,4,6-trinitrobenzenesulfonic acid [13]. There are many functional groups in the kanamycin A molecule.…”

Section: Introductionmentioning

confidence: 99%

Analysis of kanamycin A in human plasma and in oral dosage form by derivatization with 1-naphthyl isothiocyanate and high-performance liquid chromatography

2006

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A simple and sensitive HPLC method has been developed for trace determination of kanamycin A by derivatization. Plasma proteins are precipitated by acetonitrile and chemical derivatization is performed on the supernatant containing kanamycin A with 1-naphthyl isothiocyanate in pyridine at 70 degrees C. After the derivatization reaction, a methylamine/acetonitrile solution was added to the reaction mixture to eliminate the excess of derivatizing agent and shorten the analysis time. The resulting derivative was separated using a Lichrocart Purospher STAR RP-18e column and water/methanol (33:67, v/v) as a mobile phase (detection at 230 nm). Optimization conditions for the derivatization of kanamycin A were investigated by HPLC. The linear range for the quantitation of kanamycin A in spiked plasma was over 1.2-40 microg/mL; the detection limit (signal to noise ratio = 3; injection volume, 10 microL) was about 0.3 microg/mL. The relative standard deviation was less than 2.9% for intra-day assay (n = 6) and inter-day assay (n = 6) and relative recoveries were found to be greater than 98%. Preliminary application of the method for monitoring kanamycin A in humans upon intramuscular injection of the injection product demonstrated the usefulness of the assay for clinical studies. The proposed method can also be used to analyze the compound in pharmaceutical formulations.

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Section: Introductionmentioning

confidence: 99%

Analysis of kanamycin A in human plasma and in oral dosage form by derivatization with 1-naphthyl isothiocyanate and high-performance liquid chromatography

2006

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“…A fluorescence detector operated at excitation wavelength 355 nm and emission wavelength 415 nm yielded a linear range from 0.2 to 250 mg/L for colistin and from 0.4 to 320 mg/L for amikacin, kanamycin, gentamycin and neomycin. Recoveries obtained were: 87.5% for colistin, 89.1%, for amikacin, 91.1% for kanamycin, 84.6% for gentamycin and 92.4% for neomycin [49].…”

Section: Analytical Strategiesmentioning

confidence: 99%

Chromatographic analysis of banned antibacterial growth promoters in animal feed

Samanidou¹,

Evaggelopoulou

2008

J of Separation Science

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The issue of antimicrobial use in animals used as food is of global concern. Antimicrobials are used in animal agriculture to improve health and welfare of animals, meat quality, the economic efficiency of growth and production and public health by decreasing shedding of zoonotic pathogens. However, large quantities are often used without professional supervision. The growth-promotant (now reclassified as zootechnical feed additives) effect of low levels of antibiotics in animal feeds was first described in the late 1940s. Already in 1969 the Swann Committee recommended that use of antibiotics as a supplement in animal feedstuff should be restricted to those with little or no application as therapeutic agents for humans and animals, which would not impair the efficacy of therapeutic antibiotics through the development of resistant strains of organisms. Antimicrobials like avoparcin, ardacin, zinc bacitracin, virginiamycin, tylosin, spriramycin, carbadox and olaquindox were withdrawn within the period 1997-1999. Four others (monensin sodium, salinomycin sodium, avilamycin and flavophospholipol) were still permitted for use as growth promoters in animal feed to animals marketed in the European Union (EU). Since January 2006, they have been banned as well. This review focuses on the analytical methods developed to be an effective tool for monitoring compliance with the ban.

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“…Hence, it cannot be quantified with high sensitivity by conventional techniques without derivatization with UV-absorbing or fluorescent reagents. Ortho-phthaldehyde (OPA) was used as a derivatizing agent for the determination of colistin by HPLC using pre-column derivatization [3] with UV detection at 220 nm, and by post-column derivatization [9] with fluorescence detection at an emission wavelength of 415 nm. On-line and off-line pre-column HPLC derivatization methods were also used with OPA and fluorimetric detection and was performed at 340 and 440 nm for excitation and emission wavelengths, respectively [10,11].…”

Section: Introductionmentioning

confidence: 99%

Mixed Micellar Electrokinetic Chromatographic Analysis of Colistin, Polypeptide Antibiotic, Using Laser-Induced Fluorescence Detection

Ahmed¹,

Elbarbry²,

Clark³

2012

AJAC

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The main goal of this work was to quantify the detection of colistin at low levels in urine samples through the practical application of mixed surfactant micellar electrokinetic chromatography-laser-induced fluorescence (MEKC-LIF) analysis method using its advantage of sensitivity and to examine direct injection of biological samples. Colistin (polymyxin E) has neither strong UV chromophore nor fluorophore. So, its assay for metabolism, pharmacokinetics studies for bioavailability and bioequivalence are difficult because of poor detectability. Therefore an enhanced UV or fluorescence detection by chemical derivatization is required. MEKC-LIF method was proposed for colistin with a 488/520 nm argon-ion laser using a pre-CE derivatization with fluorescein isothiocyanate (FITC). Borate buffer was used as background buffer (BGB). The different parameters affecting the proposed derivatization reaction including concentration of the derivatizing reagent, reaction time and temperature were studied and optimized. The derivative was stable for up to 3 days. Different micelles (TX-100 and SDS) were examined as BGB additives separately but negative-charged mixed micelles (SDS/TX-100) were shown to be the best additive to BGB for the analysis of colistin particularly in human urine as they enhance both selectivity and sensitivity of the proposed method. BGB was used with pH 9.5, 10 kV, 8 s inj time, capillary length 75 cm × 75 µm ID (66 cm effective length), detection was LIF Ex 488 nm; Em 520 nm. The method was applied to colistin analysis in human urine and the recovery was >98% (n = 5). LOD and LOQ in urine after pre-column derivatization using FITC were 100 and 250 ng/ml, respectively. Urine samples were analysed by direct injection without sample pre-treatment. The mechanism of enhancement of fluorescence of the derivative by surfactant was proposed.

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HPLC determination of colistin and aminoglycoside antibiotics in feeds by post-column derivatization and fluorescence detection

Cited by 48 publications

References 7 publications

Analysis of kanamycin A in human plasma and in oral dosage form by derivatization with 1-naphthyl isothiocyanate and high-performance liquid chromatography

Analysis of kanamycin A in human plasma and in oral dosage form by derivatization with 1-naphthyl isothiocyanate and high-performance liquid chromatography

Chromatographic analysis of banned antibacterial growth promoters in animal feed

Mixed Micellar Electrokinetic Chromatographic Analysis of Colistin, Polypeptide Antibiotic, Using Laser-Induced Fluorescence Detection

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