Colorimetric phospholipid determination with erythrosin B.
Binding of phospholipid vesicles to erythrosin B results in a shift of the absorption maximum of the dye from 528 to 549 nm. This effect was employed to develop a simple, rapid, and sensitive quantification method for unilaminar phospholipid vesicles. At room temperature the color development of the phospholipid-dye complex at 549 nm is essentially complete in 5 min and only a slight decrease is observed in the following hours. The pH optimum of 4.5 for the assay is related to the tight binding at this pH (Kd = 3.6 micrograms/ml). A maximal binding of one erythrosin to seven phospholipid molecules is found. The sensitivity of the assay is high for zwitterionic phospholipids (e.g., 0.023 delta A549 x ml/micrograms for dioleoyl phosphatidyl choline) and lower for anionic phospholipids. The assay depends on the size of the phospholipid liposomes, indicating the importance of the phospholipid surface area for binding.
Andree
H A
HA
Mount Sinai School of Medicine, Department of Biochemistry, New York, New York 10029.
Soedjak
H S
HS
D003124:Colorimetry / Q000379:methods*; D004923:Erythrosine / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D008081:Liposomes; D008823:Micelles; D010743:Phospholipids / Q000032:analysis*; D012680:Sensitivity and Specificity; D013053:Spectrophotometry; D017186:Titrimetry
D008081:Liposomes; D008823:Micelles; D010743:Phospholipids; D004923:Erythrosine