Naslov
Determination of veterinary pharmaceuticals by HPLC-FLD chromatography

Autori
Ašperger. Danijela ; Tišler, Vesna ; Kaštelan-Macan, Marija ; Babić, Sandra

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
XXII. hrvatski skup kemičara i kemijskih inženjera : knjiga sažetaka / Tomašić, Vesna ; Maduna Valkaj, Karolina - Zagreb : Hrvatsko društvo kemijskih inženjera i tehnologa (HDKI), 2011, 103-103

ISBN
978-953-6894-42-0

Skup
Hrvatski skup kemičara i kemijskih inženjera (22 ; 2013)

Mjesto i datum
Zagreb, Hrvatska, 13.02.2011. - 16.02.2011

Vrsta sudjelovanja
Poster

Vrsta recenzije
Domaća recenzija

Ključne riječi
fluorokinolonski antibiotici; enrofloksacin; norfloksacin; tekućinska kromatografija visoke djelotvornosti (HPLC); fluorescentni detektor (FLD)
(fluoroquionolone antibiotics; enrofloxacin; norfloxacin; high-performance liquid chromatography (HPLC); fluorescence detector (FLD))

Sažetak
Everyday use of antibiotics causes necessity of developing new analytical methods for their determination in environment. These methods have to be accessible, simple, fast, and specific for the investigated analyte, but they should also enable determination of very low analyte concentrations. The aim of this work was to develop high performance liquid chromatography with fluorescence detector (HPLC-FLD) method for separation, detection and determination of fluoroquionolone antibiotics (enrofloxacin [ENRO] and norfloxacin [NOR]) at very low concentration levels. Therefore, optimal excitation (λex) and emission (λem) wavelengths of those antibiotics were determined. Experimental results showed that the highest signal responses for enrofloxacin and norfloxacin were obtained at λex / λem = 227 nm/445 nm. The LC analyses were performed using a Varian ProStar 500 HPLC-FLD system. The column temperature was set to 30 oC and injection volume was 10 μL. Chromatographic column LiChrosphere 100 CN, 125mm×4.0 mm, particle size 5 μm was used to separate compounds in the mixture. The mobile phase used in the chromatographic separation consisted of a binary mixture of solvents A (0.01 M oxalic acid) and B (acetonitrile). Simultaneous mobile phase gradient program and flow rate gradient program was used for separation of investigated antibiotics. The investigated pharmaceuticals were eluted within 20 min. Hence, HPLC-FLD method was validated by determining linearity, limit of detection (LOD), limit of quantification (LOQ) and repeatability of the method. The linearity was evaluated for both pharmaceutical in the mixture using eight concentrations in the range of 6.25 μg/L – 10 mg/L depending on the pharmaceutical. Coefficients of correlation were higher then 0.9985 thus confirming the linearity of the method. Limits of detection (ENRO 3.00 μg/L, NOR 0.05 mg/L) and quantification (ENRO 6.00 μg/L, NOR 0.10 mg/L) were experimentally estimated from the injection of standard solution serially diluted until the signal-to-noise ratio for any single analyte reached a value of ten for LOQ and three for LOD. Intra-day repeatability was determined by analysing five replicates of water samples using same equipment and same analytical procedure. Intra-day repeatability was expressed as relative standard deviation (R.S.D.) (R.S.D. for ENRO is 0.54% and for NOR 0.77%).

Izvorni jezik
Engleski

Znanstvena područja
Kemija

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