Colorimetry and constant-potential coulometry determinations of transferrin-bound iron, total iron-binding capacity, and total iron in serum containing iron-dextran, with use of sodium dithionite and alumina columns.

After the parenteral administration of iron-dextran (imferon), the increased total iron concentrations in serum can be determined by atomic absorption spectroscopy and by colorimetric methods involving sodium dithionite, which reductively dissociates iron from the dextran complex. We report that constant-potential coulometry detects only about 55-70% of dextran-bound iron before dithionite reduction and variable amounts after reaction with the reducing agent. In addition, we have developed a procedure for determining transferrin-bound iron, total iron-binding capacity (TIBC), total iron, and dextran-bound iron with the Kodak Ektachem colorimetric system. In determining total serum iron, the sample is first mixed with sodium dithionite, which rapidly dissociates all dextran-bound iron, but does not remove iron from either transferrin or hemoglobin. After the mixture is applied to an Ektachem slide, transferrin-bound iron is released at pH 4 and is detected together with the iron previously bound to dextran. TIBC is determined by mixing serum with ferric citrate in moderate excess and filtering through a small alumina (Al2O3) column, which binds excess free iron and iron-dextran; the iron in the column eluate represents the TIBC. Transferrin-bound iron is determined by applying diluted serum without added ferric citrate to an alumina column and measuring the iron in the column eluate. Dextran-bound iron is equivalent to the difference between total and transferrin-bound iron. Using this method, we found that transferrin iron-binding sites are saturated in vitro by excess iron-dextran less efficiently than by ferric citrate.

Definitive liquid-chromatographic demonstration that N-ethylglycine is the metabolite of lidocaine that interferes in the Kodak sarcosine oxidase-coupled method for creatinine.

Patients receiving lidocaine may show false increases of serum creatinine as assayed by the single-slide method on the Kodak Ektachem 700. Bissell et al. (Clin Chem 1987;33:951) suggested that this interference was due to oxidation of N-ethylglycine (NEG), a previously uncharacterized metabolite of lidocaine, by the sarcosine oxidase preparation used in the Ektachem creatinine slide. To investigate this possibility, we synthesized NEG, added it to drug-free human serum, and analyzed the NEG-supplemented sera for creatinine with the Ektachem 700. We found the following linear relationships between creatinine bias (y, mg/L) and NEG concentration (x, mg/L) for first (I), third (III), and fourth (IV) generation slides: I: y = 1.70x - 0.8 mg/L (n = 13, r = 1.0) III: y = 0.39x - 0.3 mg/L (n = 3, r = 1.0) IV: y = 0.79x - 1.8 mg/L (n = 13, r = 1.0) Using HPLC, we directly demonstrated the presence of NEG in sera of patients receiving lidocaine and quantified NEG concentrations in sera from four of these patients. The increasing artifactual bias in creatinine with increasing NEG concentration unequivocally confirmed that NEG is responsible for the lidocaine-associated interference in the Kodak Ektachem single-slide creatinine method.

Effects of prolonged ingestion of glucose or ethanol on fatty acid synthesis by mitochondria and cell sap of rat liver and adipose tissue.

Effects of prolonged ingestion of glucose and ethanol on the rate of fatty acid synthesis by liver and adipose tissue have been investigated in male rats. Ethanol significantly enhanced the rate of fatty acid synthesis from malonyl-2-(14)C CoA in liver cell sap; glucose feeding enhanced the rate of fatty acid synthesis from both malonyl-2-(14)C and acetyl-1-(14)C CoA. Neither dietary supplement modified the types of fatty acid synthesized in this enzyme system. Palmitic acid was the principal product synthesized from a mixture of malonyl and acetyl CoA, whereas myristic and palmitic acids were the predominant products formed from acetyl CoA alone. Neither glucose nor ethanol affected fatty acid synthesis by adipose tissue cell sap. Mitochondria derived from liver and adipose tissue of control, glucose-fed, and ethanol-fed animals all incorporated acetyl-1-(14)C CoA into lipid at about the same rate, but did not utilize malonyl CoA for lipid synthesis to any significant degree. The label appeared in fatty acids, one-half of which were contained in phospholipid. Both unsaturated and saturated fatty acids synthesized by mitochondria contained isotope, most of which was present in the carboxyl groups. Ethanol and glucose feeding stimulated the labeling of monoenoic fatty acids in liver mitochondria, but only glucose did so for adipose tissue. These findings agree with results previously obtained when lipogenesis was measured with acetate-(14)C in vivo.