Seven serial dilutions of this sample in hemolyzing
buffer (1 in 4 to approximately 1 in 20) were used to determine the range of readable area that produced a reliable Hb [A.
1) that the inclusion of a suitable hemolyzing
agent is a logical support to erythrocyte folate extraction.
Saponin was chosen as the hemolyzing
agent in the current studies not just because of its historical use for the hemolysis of erythrocytes, but because it was anticipated that the thick peptidoglycan outer cell wall of the gram-positive bacterium Lactobacillus rhamnosus (previously known as L.
Therefore, no other technologists saw that the wrong solution was being used as hemolyzing
reagent for 3 weeks.
Whole blood samples (20 [micro]L) were lysed by addition of 100 [micro]L of appropriate hemolyzing
Additives that did not have similar effects on ZPP/H included albumin and the hemolyzing
agents deoxycholate and saponin.
Therefore, we shall refer to these particular instruments as hemolyzing
For the latter, we rapidly prepared hemolysates by using hemolyzing
reagent straight from the refrigerator at 4-8 [degrees]C and immediately placed the prepared hemolysate in the Variant instrument with the sample tray at 8 [degrees]C.
According to our procedure there was no need for the glucose concentration to be known in advance; we stabilized the concentration by adding hemolyzing
agent at the same time that B-glucose concentration was being measured in the GP office.
Sample preparation involved dilution of 50 [micro]L of citrated whole blood to 1 mL with hemolyzing
reagent (10 mmol EDTA and 5 mmol KCN).
5 mL of hemolyzing
solution (acid potassium phthalate 25 mmol/L; KCN 8 mmol/L; Triton X-100 10 mL/L, pH 5.
Samples (Hb concentrations -2 g/L) were obtained by hemolyzing
20 [micro]L of blood in 1 mL of a buffer of potassium hydrogen phthalate (5 g/L) and potassium cyanide (5 g/L).