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  • Selective removal of phosphate for analysis of organic acids in complex samples.

Selective removal of phosphate for analysis of organic acids in complex samples.

Journal of chromatography. A (2015-03-01)
Sandeep Deshmukh, Andrej Frolov, Andrea Marcillo, Claudia Birkemeyer
ABSTRACT

Accurate quantitation of compounds in samples of biological origin is often hampered by matrix interferences one of which occurs in GC-MS analysis from the presence of highly abundant phosphate. Consequently, high concentrations of phosphate need to be removed before sample analysis. Within this context, we screened 17 anion exchange solid-phase extraction (SPE) materials for selective phosphate removal using different protocols to meet the challenge of simultaneous recovery of six common organic acids in aqueous samples prior to derivatization for GC-MS analysis. Up to 75% recovery was achieved for the most organic acids, only the low pKa tartaric and citric acids were badly recovered. Compared to the traditional approach of phosphate removal by precipitation, SPE had a broader compatibility with common detection methods and performed more selectively among the organic acids under investigation. Based on the results of this study, it is recommended that phosphate removal strategies during the analysis of biologically relevant small molecular weight organic acids consider the respective pKa of the anticipated analytes and the detection method of choice.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium phosphate dibasic, ReagentPlus®, ≥99.0%
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Sodium phosphate dibasic, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.0%
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Sodium phosphate dibasic, BioXtra, ≥99.0%
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Citric acid, 99%
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Sodium phosphate dibasic, for molecular biology, ≥98.5% (titration)
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Sodium phosphate dibasic, BioUltra, for molecular biology, ≥99.5% (T)
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Citric acid, BioUltra, anhydrous, ≥99.5% (T)
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Sodium fluoride, BioXtra, ≥99%
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Sodium fluoride, BioReagent, suitable for insect cell culture, ≥99%
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Sodium phosphate dibasic, purum p.a., anhydrous, ≥98.0% (T)
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Acetic anhydride, ACS reagent, ≥98.0%
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Acetic anhydride, ReagentPlus®, ≥99%
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Citric acid, ACS reagent, ≥99.5%
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Sodium fluoride, 99.99% trace metals basis
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Sodium phosphate dibasic, 99.95% trace metals basis
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Sodium fluoride, anhydrous, powder, 99.99% trace metals basis
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Acetic anhydride, ≥99%
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Sodium phosphate dibasic, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.0%, free-flowing, Redi-Dri
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Citric acid, anhydrous, free-flowing, Redi-Dri, ACS reagent, ≥99.5%
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Formic acid, ACS reagent, ≥88%
Supelco
Sodium Fluoride, Pharmaceutical Secondary Standard; Certified Reference Material
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Sodium phosphate dibasic, puriss. p.a., ACS reagent, anhydrous, ≥99.0% (T)
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Formic acid, ACS reagent, ≥96%
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Formic acid, puriss., meets analytical specifications of DAC, FCC, 98.0-100%
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Citric acid, meets analytical specification of Ph. Eur., BP, USP, E330, anhydrous, 99.5-100.5% (based on anhydrous substance)
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Sodium fluoride, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99%
Sigma-Aldrich
Sodium phosphate dibasic, ACS reagent, ≥99.0%
Sigma-Aldrich
Sodium fluoride, ACS reagent, ≥99%
Sigma-Aldrich
Formic acid, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥98%
Sigma-Aldrich
Acetic anhydride, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99% (GC)