Development and validation of HPLC method for the determination of tobramycin in urine samples post-inhalation using pre-column derivatisation with fluorescein isothiocyanate

Mashat, M.; Chrystyn, Henry; Clark, Brian J.; Assi, K.H.

doi:10.1016/j.jchromb.2008.05.012

Cited by 38 publications

(28 citation statements)

References 26 publications

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“…The tobramycin concentration was determined by using validated HPLC method following pre-column derivatization with fluorescein isothiocyanate (FITC) [53]. The …”

Section: Tobramycin Assaymentioning

confidence: 99%

Assessment of recent nebulizer delivery systems using urinary pharmacokinetics method and aerodynamic characteristics of TOBI ® nebulized dose following inhalation

Mashat¹,

Clark

Assi

et al. 2017

Journal of Drug Delivery Science and Technology

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“…The tobramycin concentration was determined by using validated HPLC method following pre-column derivatization with fluorescein isothiocyanate (FITC) [53]. The …”

Section: Tobramycin Assaymentioning

confidence: 99%

Assessment of recent nebulizer delivery systems using urinary pharmacokinetics method and aerodynamic characteristics of TOBI ® nebulized dose following inhalation

Mashat¹,

Clark

Assi

et al. 2017

Journal of Drug Delivery Science and Technology

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“…The derivatization procedure was reported by our group [12]. However it was optimized for the CE analysis.…”

Section: Fitc Derivatization Proceduresmentioning

confidence: 99%

“…The main drawback of using OPA as derivatizing agent is that its derivatives are not very stable. However, fluoresceinisothiocyanate (FITC) derivatives were practicaly stable for days [12]. Dansyl chloride was used for the derivatization of colistin A and separated by HPLC and the fluorescence detection at 344 nm and 518 nm for excitation and emission wavelengths, respectively [13].…”

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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“…Gentamicins C1, C1a and C2 were determined as their 2,4-dinitrophenyl derivatives in plasma and urine using a C18 RP column and as mobile phase 680ml/l acetonitrile-320ml/l Tris buffer (8.3mmol/l titrated to pH 7.0) (Isoherranen and Sobak, 2000). A HPLC method for the analysis of tobramycin in urine samples used pre-column derivatization of tobramycin with fluorescein isothyociante followed by fluorescence detection; the chromatographic separation was carried out on a Phenomenex Luna C18 column and the limit of quantitation of the method in urine was 250 ng/mL (Mashat et al, 2008). Another derivatization agent used for the RP-HPLC analysis of aminoglycoside antibiotics was phenylisocyanate in the presence of triethylamine.…”

Section: Chromatography Of Aminoglycosidesmentioning

confidence: 99%

HPLC-MS/MS of Highly Polar Compounds

Silvestro¹,

Tarcomnicu²,

Savu³

2012

Tandem Mass Spectrometry - Applications and Principles

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Development and validation of HPLC method for the determination of tobramycin in urine samples post-inhalation using pre-column derivatisation with fluorescein isothiocyanate

Cited by 38 publications

References 26 publications

Assessment of recent nebulizer delivery systems using urinary pharmacokinetics method and aerodynamic characteristics of TOBI ® nebulized dose following inhalation

Assessment of recent nebulizer delivery systems using urinary pharmacokinetics method and aerodynamic characteristics of TOBI ® nebulized dose following inhalation

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

HPLC-MS/MS of Highly Polar Compounds

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