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Biomedical subjects

Jeff Rohrer

Publications and source records attributed to Jeff Rohrer.

3 recordsLinked to original sources

Determination of residual trifluoroacetate in protein purification buffers and peptide preparations by ion chromatography.

Trifluoroacetate (TFA) is commonly used in a variety of pharmaceutical applications. Because of its toxic nature, it is important to reliably measure the effective removal of TFA. We developed an ion chromatography (IC) method to determine the concentration of residual TFA in samples found in the pharmaceutical industry. A high-capacity anion-exchange column was used to separate trace trifluoroacetate from an excess of chloride, phosphate, and other anions without the need for sample preparation. TFA was detected by suppressed conductivity. A method with four KOH eluent step changes was optimized and reproducibly executed using automated generation of the KOH eluent. We used this method to determine TFA in the following samples: a phosphate-buffered saline (PBS), an acetate-buffered saline containing protein, and a commercial peptide. The method detection limits for TFA in these samples were all less than 90 ng/ml.

Buffers↗

Monitoring trace anion contamination in disk drive components.

Ion chromatography was used to determine trace anionic contamination on the surface of hard disk drive components. These contaminants can have a detrimental effect on device reliability and yield. Disk drive components were soaked in deionized water and these extracts were analyzed for anions. The anions fluoride, acetate, formate, acrylate, methacrylate, chloride, nitrite, bromide, nitrate, benzoate, sulfate, oxalate, phthalate and phosphate were separated on a high-performance anion-exchange column and determined at concentrations less than 1 microg/l with suppressed conductivity detection. The extract solutions were analyzed either by injecting 1 ml or by preconcentrating 5 ml. We evaluated the performance of both methods.

Anions↗

Determination of nitrite, nitrate, and glucose-6-phosphate in muscle tissues and cured meat by IC/MS.

The endogenous nitrate concentration in fresh meat and the residual nitrate and nitrite contents after curing are related to food quality and safety. Most ion chromatography (IC) methods suffer from interferences, especially in fresh meat samples, in which the endogenous nitrate content is low, and in cured meat products, in which other nitrogenous compounds can interfere with the separation of inorganic anions. One of the major classes of interfering compounds in fresh meat are sugar phosphates, which originate from glycolysis during the conversion of muscle glycogen to lactic acid. Nitrate can be separated from interfering compounds with a high-capacity anion-exchange column that was manufactured for use with hydroxide eluents (i.e., hydroxide-selective). This column has a different selectivity than traditional IC columns that use carbonate eluents and facilitates the determination of nitrate in both fresh and cured meats. Nitrate was detected by both suppressed conductivity measurement and mass spectrometry (MS). The identifications of nitrate and glucose-6-phosphate were confirmed by MS detection. The described IC/MS method is robust, sensitive to nitrate concentrations as low as 0.10 mg/kg, and can determine sugar phosphates that are useful for monitoring meat freshness. We successfully used this method to determine nitrate in nearly 100 muscle tissues and cured meat samples.

Chemistry Techniques, Analytical↗