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  • HPLC-UV method validation for the identification and quantification of bioactive amines in commercial eggs.

HPLC-UV method validation for the identification and quantification of bioactive amines in commercial eggs.

Talanta (2015-05-25)
Tadeu Chaves de Figueiredo, Débora Cristina Sampaio de Assis, Liliane Denize Miranda Menezes, Guilherme Resende da Silva, Isabela Pereira Lanza, Luiz Guilherme Dias Heneine, Silvana de Vasconcelos Cançado
ABSTRACT

A quantitative and confirmatory high-performance liquid chromatography with ultraviolet detection (HPLC-UV) method for the determination of bioactive amines in the albumen and yolk of commercial eggs was developed, optimized and validated by analyte extraction with trichloroacetic acid and pre-column derivatization with dansyl chloride. Phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine standards were used to evaluate the following performance parameters: limit of detection (LoD), limit of quantification (LoQ), selectivity, linearity, precision, recovery and ruggedness. The LoD of the method was defined from 0.2 to 0.3 mg kg(-1) for the yolk matrix and from 0.2 to 0.4 mg kg(-1) for the albumen matrix; the LoQ was from 0.7 to 1.0 mg kg(-1) for the yolk matrix and from 0.7 to 1.1 mg kg(-1) for the albumen matrix. The validated method exhibited excellent selectivity and separation of all amines with coefficients of determination higher than 0.99. The obtained recovery values were from 90.5% to 108.3%, and the relative standard deviation (RSD) was lower than 10% under repeatability conditions for the studied analytes. The performance parameters show the validated method to be adequate for the determination of bioactive amines in egg albumen and yolk.

MATERIALS
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Product Description

Sigma-Aldrich
Spermine tetrahydrochloride, BioUltra, for molecular biology, ≥99.5% (AT)
Pricing and availability is not currently available.
Sigma-Aldrich
Spermine tetrahydrochloride, ≥99.0% (AT), powder or crystals
Pricing and availability is not currently available.
Sigma-Aldrich
Sodium trichloroacetate, 97%
Pricing and availability is not currently available.
Sigma-Aldrich
L-Proline, BioUltra, ≥99.5% (NT)
Pricing and availability is not currently available.
Sigma-Aldrich
Trichloroacetic acid, BioUltra, ≥99.5% (T)
Pricing and availability is not currently available.
Sigma-Aldrich
Trichloroacetic acid, ACS reagent, for the determination of Fe in blood according to Heilmeyer, ≥99.5%
Pricing and availability is not currently available.
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Dansyl chloride, suitable for fluorescence, BioReagent, ≥99.0% (HPLC)
Pricing and availability is not currently available.
Sigma-Aldrich
L-Proline, 99%, FCC, FG
Pricing and availability is not currently available.
Sigma-Aldrich
Acetone, ≥99%, meets FCC analytical specifications
Pricing and availability is not currently available.
Sigma-Aldrich
Acetonitrile solution, contains 0.1 % (v/v) trifluoroacetic acid, suitable for HPLC
Pricing and availability is not currently available.
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Acetone, natural, ≥97%
Pricing and availability is not currently available.
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Acetonitrile solution, contains 0.1 % (v/v) formic acid, suitable for HPLC
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Spermine tetrahydrochloride, powder or crystals
Pricing and availability is not currently available.
Sigma-Aldrich
Sodium bicarbonate, BioXtra, 99.5-100.5%
Pricing and availability is not currently available.
Sigma-Aldrich
Dansyl chloride, BioReagent, suitable for amino acid labeling, powder and chunks, ≥99% (HPLC)
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Trichloroacetic acid, suitable for electrophoresis, suitable for fixing solution (for IEF and PAGE gels), ≥99%
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Sodium bicarbonate, powder, BioReagent, for molecular biology, suitable for cell culture, suitable for insect cell culture
Pricing and availability is not currently available.
Sigma-Aldrich
L-Proline, from non-animal source, meets EP, USP testing specifications, suitable for cell culture
Pricing and availability is not currently available.
Sigma-Aldrich
L-Proline, ReagentPlus®, ≥99% (HPLC)
Pricing and availability is not currently available.
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Trichloroacetic acid, BioXtra, ≥99.0%
Pricing and availability is not currently available.
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Putrescine dihydrochloride, ≥98% (TLC)
Pricing and availability is not currently available.
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Putrescine dihydrochloride, powder, BioReagent, suitable for cell culture
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Trichloroacetic acid, ≥99.0% (titration)
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Putrescine dihydrochloride, γ-irradiated, lyophilized powder, BioXtra, suitable for cell culture
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Sigma-Aldrich
Hydrochloric acid, 36.5-38.0%, BioReagent, for molecular biology
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SAFC
L-Proline
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Supelco
Hydrochloric acid solution, volumetric, 0.1 M HCl (0.1N), endotoxin free
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Hydrochloric acid solution, 1.0 N, BioReagent, suitable for cell culture
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Sigma-Aldrich
Hydrogen chloride solution, 3 M in cyclopentyl methyl ether (CPME)
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Sigma-Aldrich
Spermine tetrahydrochloride, BioReagent, for molecular biology
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