Changes in the composition of erythrocyte membrane during storage of blood in di-(2-ethyl hexyl) phthalate [DEHP] plasticized poly vinyl chloride (PVC) blood storage bags.

Very little information is available on the changes in the erythrocyte membrane composition during storage of blood at 4 degrees C, particularly with respect to the glycosaminoglycans and glycoproteins. In view of this, a detailed study was carried out on the changes in the membrane proteins, glycosaminoglycans (GAG), carbohydrate components of glycoproteins, cholesterol, phospholipids and vitamin E in blood stored in glass bottles and a di-(2-ethyl hexyl) phthalate (DEHP) plasticized PVC bag (Penpol blood bag). Blood was collected in CPDA solution in glass bottles and in Penpol blood bags and kept at 4 +/- 1 degrees C. Analysis was made immediately after blood collection and after 28 and 42 days. Significant increase in the total protein in the erythrocyte membrane was observed during storage of whole blood in glass bottles and Penpol blood bag at 4 degrees C. This increase was progressively more with increase in storage time. Significant changes were also observed in GAG, carbohydrate components of glycoproteins, cholesterol, phospholipids and vitamin E in the erythrocyte membrane under these conditions. The protein:GAG ratio, protein:carbohydrate ratio, cholesterol:phospholipid ratio as well as protein:lipid ratio showed significant increase in the membrane. The extent of these changes was lower in the Penpol bag, indicating the stabilizing effect of DEHP on the erythrocyte membrane.

Synthesis of NAD+ in erythrocytes incubated with nicotinic acid and the effect of di-(2-ethyl hexyl) phthalate (DEHP).

Synthesis of NAD+ from nicotinic acid by erythrocytes incubated in SAGM phosphate solution and effect of di-[2-ethyl hexyl] phthalate, a plasticizer commonly used in PVC blood/component storage bags, on this synthesis was studied. Erythrocytes are able to synthesise NAD+ in SAGM (sodium chloride, adenine, glucose, mannitol) phosphate solution and this synthesis was more in the presence of added nicotinic acid (optimum concentration 1 mM). The level of NAD+ decreased when the incubation period was increased from 24 to 48 hr. Glutamine had a deleterious effect on this synthesis, possibly due to the decrease in pH. Di-[2-ethyl hexyl] phthalate had an inhibitory effect on NAD+ synthesis when the cells were incubated in SAGM phosphate solution, either alone or in the presence of added nicotinic acid. There was significant decrease in the release of potassium and haemoglobin from the cells in the presence of nicotinic acid, indicating increased red cell stability.

Effect of DEHP [di-(2-ethyl hexyl) phthalate] on lipid peroxidation in liver in rats and in primary cultures of rat hepatocytes.

The effect of DEHP [di-(2-ethly hexyl) phthalate] on lipid peroxidation in the liver in rats and in primary cultures of rat hepatocytes incubated with it was studied. The doses of DEHP used in this study corresponded to the low levels of this substance leaching into blood stored in DEHP plasticised PVC bags. Increased activity of superoxide dismutase (SOD) and catalase, increased concentration of malondialdehyde (MDA) and conjugated dienes and decrease in the concentration of glutathione and vitamin E have been observed in the liver of rats administered DEHP. Primary cultures of rat hepatocytes incubated with DEHP also showed increase in the activity of these enzymes, increase in the concentration of MDA and decrease in vitamin E. These results indicate that DEHP promotes lipid peroxidation. Incorporation of vitamin E along with DEHP into the culture medium containing hepatocytes counteracted these effects.