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Identification of Phospholipids in Soils and Sewage Sludges by High-Performance Liquid Chromatography¹

ABSTRACT

A method was developed for identification of phospholipids in soils and sewage sludges. This method involves extraction of the phospholipids from soils or sewage sludges with a chloroform-methanol mixture and analysis of an aliquot of the extract (10 µL) with a high-performance liquid chromatograph coupled with a normal-phase silica-gel analytical column. The eluent used was a mixture of acetonitrile-methanol-phosphoric acid (vol/vol/vol, 260:12:3). Absorbance at 205 nm was used as a means of detection. The following phospholipids were identified in soil and sewage sludge samples: phosphatidyl choline (PC), phosphatidyl ethanolamine (PE), phosphatidyl inositol (PI), and phosphatidyl serine (PS). The identities of the peaks were verified by standard additions technique and by two-dimensional thin-layer chromatography.

The concentration of lipid P in soils and sewage sludges ranged from 0.50 to 2.38 and from 38.2 to 330 mg kg^–1, respectively. The amounts of PE and PC present in six soils ranged from 5.4 to 16 µg P kg^–1 (avg = 10.1) and from 3.6 to 48 (avg = 20.9), respectively. The corresponding ranges for these phospholipids in six sewage sludge samples were 3.5 to 13 mg kg^–1 (avg = 6.1) and 0 to 6.8 (avg = 3.6). The other phospholipids were not detected in most of the soil and sewage sludge samples studied.

Key Words: thin-layer chromatography • phosphatidyl choline • phosphatidyl ethanolamine

¹ Journal Paper no. J-11404 of the Iowa Agric. Home Econ. Exp. Stn., Ames, IA 50011. Projects 2082, 2431, and 2274.

² Postdoctoral research associate and professor of soil chemistry, respectively. Dep. of Agron., Iowa State Univ., Ames, IA 50011.

Received for publication March 23, 1984.