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Biomedical subjects

F L Rodkey

Publications and source records attributed to F L Rodkey.

At least 19 recordsLinked to original sources

Spectrophotometric measurement of carboxyhemoglobin and methemoglobin in blood.

This paper describes separate spectrophotometric procedures for rapidly measuring carboxyhemoglobin (COHb) and methemoglobin (MetHb) in blood. Absorbance measurements are made in the Soret region at a blood dilution near 1000-fold. For COHb estimation, the diluent contains sodium hydrosulfite, providing the two-component system COHb--Hb for absorbance measurements at 420 and 432 nm. The NA2S2O4 effectively prevents dissociation of COHb by oxygen. For MetHb estimation, the diluent contains KCN and carbon monoxide, providing the two-component system COHb--CNMetHb. Absorbance measurements are made at 420 nm, before and after addiely analyzed for the total of all MetHb forms present. Each procedure requires about 3 muL of blood and can be applied to human or animal blood. Results obtained by the present methods are in satisfactory agreement with currently accepted procedures, which require much larger samples or more elaborate equipment.

Carboxyhemoglobin↗

Molar absorbance of cyanmethemoglobin from blood of different animals.

The molar absorbance of cyanmethemoglobin at 540 nm was measured in 35 species of mammals, birds, and reptiles. Values obtained were compared with those measured on human cyanmethemoglobin by the same procedure. There was no significant difference (P greater than or equal to 0.1) between the human and animal blood values observed in 29 samples. In six cases (4 mammals, 1 reptile, and 1 bird), the significant difference (P greater than or equal to 0.05) did not exceed 5% of the human value. The practice of applying the accepted absorbance of 1.10 x 10(4) M-1 cm-1 per mole of iron in human cyanmethemeglobin for the analysis of animal blood will not cause more than 5% uncertainty. Such results are satisfactory for use in clinical veterinary medicine except for those research projects concerned with animal hemoglobin function and structure.

Animals↗

Analytical precautions in measurement of blood cyanide.

Solutions of KCN in 0.1 mol/liter NaOH, stored for six months at 4 degrees C, were unchanged as determined by silver titration and color development with pyridine-pyrazalone reagent. At room temperature the cyanide concentration of such solutions decreased by 0.079% per day (half life = 880 days) and the extent of color development changed in direct proportion to the change in silver titer. Rubber stoppers adsorb (or dissolve) HCN when in contact with this gas. The cyanide tends to come out of the stopper long after the source of the gas is removed. Rubber stoppers and expired air contaminated by HCN must be carefully avoided during cyanide analysis. Expired air contains HCN, generally in larger amounts for persons who smoke.

Colorimetry↗

Determination of cyanide and nitroprusside in blood and plasma.

A procedure was refined for quantitative isolation of cyanide by gas transfer from acidified blood or plasma samples. The cyanide was trapped in dilute alkali and quantified as the pyridine/pyrazolone complex. The within-day coefficient of variation was 2%, which increased to about 2.5% for the day-to-day variation. Nitroprusside used as a hypotensive agent in clinical medicine provides a risk of cyanide toxicity when the rate of administration or the total amount of drug given is excessive. A procedure was developed for measuring nitroprusside in the plasma of man and animals. Nitroprusside in the sample is quantitatively converted to cyanide by incubation with cystein solution at slightly alkaline pH. Methemoglobin is added to combine with the cyanide formed and prevent its destruction. On acidification, the total amount of cyanide originally present as free cyanide or as nitroprusside is liberated as HCN, isolated by gas transfer into a sodium hydroxide trap, and quantified by spectrophotometry. Nitroprusside present in the sample is calculated from the increase in cyanide observed in the cysteine-treated sample compared to that obtained without cysteine treatment. The method has been used to estimate in vitro stability of nitroprusside in aqueous solution, blood, and plasma. Blood cyanide and plasma nitroprusside concentrations were measured when sodium nitroprusside was infused into a baboon. Over 90% of the nitroprusside in blood is present in the plasma, suggesting that the drug crosses the erythrocyte membrane slowly.

Analysis of Variance↗

Nitroprusside-induced cyanide poisoning: antidotal effect of hydroxocobalamin.

Sodium nitroprusside was investigated as a potential source of cyanide poisoning, Whole-blood cyanide determinations were performed on arterial samples from baboons receiving nitroprusside while anesthetized. There was a statistically significant increase in cyanide levels, as well as development of tachyphylaxis and severe metabolic acidosis. Hydroxocobalamin (vitamin B12a) infused simultaneously with nitroprusside significantly lessened the increase in cyanide levels and eliminated the development of metabolic acidosis. Nitroprusside can cause cyanide intoxication in the baboon, and hydroxocobalamin appears to be an effective antidote.

Acid-Base Equilibrium↗

A mechanism for the conversion of oxyhemoglobin to methemoglobin by nitrite.

Each mole of oxyhemoglobin iron converted to methemoglobin causes the oxidation of 1.5 mol of nitrite to nitrate and consumes 1 mol of protons. No oxygen is liberated. The overall reaction has two simultaneously occurring parts. In the beginning the rate-limiting reaction converting O2Hb to metHb is directly proportional to H+ and NO2- concentrations and is independent of metHb. The second portion accounts in major part for the stoichiometry and rate of the overall reaction. In this portion O2Hb tetramers and metHbNO2- are the reactants. Essentially no reaction takes place in the presence of CN-, which displaces nitrite from the metHbNO2-, nor in the presence of 0.5 mol/liter Nal, which converts the O2Hb to alphabeta-dimers. The autocatalytic nature of the overall reaction in the presence of excess nitrite is the result of metHb, which is formed in both parts of the reaction, associating with nitrite to increase the concentration of one reactant of the cyanide-sensitive part. The reaction rates at constant pH in excess nitrite are porportional to the product of the O2Hb concentration and the square of the metHb concentration. The rate increases up to about 66% conversion of O2Hb followed by a decrease as the O2Hb becomes limiting. The dissociation constant of metHbNO2- at 25 degrees C and pH = 6.4 was found to be 1.11+/-0.11 mmol/liter.

Chemical Phenomena↗