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  • Microextraction by packed sorbent (MEPS) to analyze catecholamines in innovative biological samples.

Microextraction by packed sorbent (MEPS) to analyze catecholamines in innovative biological samples.

Journal of pharmaceutical and biomedical analysis (2014-12-17)
Maria Addolorata Saracino, Laura Santarcangelo, Maria Augusta Raggi, Laura Mercolini
ABSTRACT

A new microextraction by packed sorbent (MEPS) procedure coupled to a high-performance liquid chromatographic method has been developed for quantitation of catecholamines (i.e. norepinephrine, epinephrine and dopamine) in innovative biological samples, namely dried plasma and urine spots. Analyses were carried out on a C18 reversed-phase column using a mobile phase composed of 2.5% methanol and 97.5% aqueous citrate buffer, containing octanesulfonic acid. Coulometric detection was used, setting the first analytical cell at -0.350 V and the second analytical cell at +0.400 V. Dried matrices were purified by means of a fast and feasible MEPS procedure, optimized on C18 sorbent and requiring only a small amount of biological sample. The availability of miniaturized procedures allowed satisfying specific requirements that ought to be considered during pre-treatment intended for catecholamine analysis. The extraction yield values were always higher than 85% and sensitivity was good, with a limit of quantitation of 100 pg mL(-1) for all the analytes. Satisfactory results were also obtained in terms of linearity, precision and accuracy. The method was successfully applied to dried plasma and urine spots derived from two socially diversified groups, namely psychiatric patients and poly-drug abusers, in comparison to healthy volunteers.

MATERIALS
Product Number
Brand
Product Description

Supelco
Sodium hydroxide concentrate, 0.1 M NaOH in water (0.1N), Eluent concentrate for IC
Sigma-Aldrich
Methanol, NMR reference standard
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Phosphate Standard for IC, TraceCERT®, 1000 mg/L phosphate in water (nominal concentration)
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Hydrogen chloride solution, 3 M in cyclopentyl methyl ether (CPME)
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Potassium chloride, tested according to Ph. Eur.
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Potassium chloride, BioUltra, for molecular biology, ≥99.5% (AT)
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Sodium chloride, tested according to Ph. Eur.
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Sodium chloride, BioUltra, for molecular biology, ≥99.5% (AT)
Supelco
Potassium chloride solution, conductance standard B acc. to ISO 7888, 0.01 M KCl
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Urea solution, BioUltra, ~8 M in H2O
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Potassium chloride solution, conductance standard C acc. to ISO 7888, 0.001 M KCl
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Ammonium chloride, BioUltra, for molecular biology, ≥99.5% (AT)
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Potassium chloride solution, BioUltra, for molecular biology, ~1 M in H2O
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Sodium chloride solution, BioUltra, for molecular biology, ~5 M in H2O
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Phosphoric acid solution, NMR reference standard, 85% in D2O (99.9 atom % D), NMR tube size 3 mm × 8 in.
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Phosphoric acid solution, NMR reference standard, 85% in D2O (99.9 atom % D), NMR tube size 5 mm × 8 in.
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Sodium hydroxide, BioUltra, for luminescence, ≥98.0% (T), pellets
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Sodium hydroxide solution, BioUltra, for molecular biology, 10 M in H2O
Supelco
Potassium chloride solution, conductance standard A acc. to ISO 7888, 0.1 M KCl
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Hydrogen chloride – ethanol solution, ~1.25 M HCl, for GC derivatization, LiChropur
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Potassium chloride solution, for Ag/AgCl electrodes, ~3 M KCl, saturated with silver chloride
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Ammonia solution, 0.4 M in THF
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Ammonia solution, 4 M in methanol
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Hydrochloric acid solution, ~6 M in H2O, for amino acid analysis
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ISA (ionic strength adjustment solution: 1 M KCl), 1 M KCl
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Sodium hydroxide solution, 49-51% in water, eluent for IC
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Sodium metabisulfite, tested according to Ph. Eur.
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Potassium chloride solution, BioUltra, ~3 M in H2O
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Sodium chloride, reference material for titrimetry, certified by BAM, >99.5%
Supelco
Hydrogen chloride – methanol solution, ~1.25 m HCl (T), for GC derivatization, LiChropur