A new method for determining concentrations of Endod-S (Phytolacca dodecandra) in water during mollusciciding

A hemolytic method was developed for the quantitative determination of oleanane saponin (Endod-S) in Phytolacca dodecandra (Phytolaccaceae), a plant with potent molluscicidal properties. A chemical method based on thin-layer chromatography (TLC) was used as a control for comparison with a hemolysis test. Standard curves for calibration of both tests of Endod-S gave high correlation coefficients. The hemolysis method gave a correlation coefficient of 0.98 for saponin (Endod-S) levels ranging from 1.6 to 4.2 mg/l. The TLC method gave a correlation coefficient of 0.99 for saponin levels above 1.5 micrograms. Due to the action of saponins on erythrocytes, the hemolysis method could not be used at levels below 1.6 and above 4.2 mg/l. Similarly, sample size limited TLC quantitation of these saponins at levels below 1 microgram. Both the TLC and the hemolysis methods on identical Endod-S water samples gave comparable results at saponin levels between 2 and 6 mg/l. A snail bioassay showed that the LC50 and the LC90 concentrations of Endod-S to the snail Biomphalaria glabrata (albino) were 2.57 and 2.92 mg/l respectively. These values fell in the standard curve range for the hemolysis method thus indicating that the hemolysis method is of practical value for testing Endod-S concentrations that would actually be used in field applications for snail control. The hemolysis method is less expensive than TLC and can be easily adapted to field conditions.