Efficient and Sensitive Method for Quantitative Analysis of Alkaloids in Hardinggrass (Phalaris aquatica L.)

• Published in: Journal of agricultural and food chemistry Vol. 54; no. 25; pp. 9287 - 9291
• Main Authors: Zhou, Lili, Hopkins, Andrew A, Huhman, David V, Sumner, Lloyd W
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 13-12-2006
• Subjects: alkaloids; Alkaloids - analysis; Biological and medical sciences; chemical constituents of plants; Chromatography, High Pressure Liquid - instrumentation; Chromatography, High Pressure Liquid - methods; Food industries; forage grasses; Fundamental and applied biological sciences. Psychology; high-performance thin layer chromatography; Mass Spectrometry; Phalaris - chemistry; Phalaris aquatica; Plant Extracts - chemistry; poisonous plants; quantitative analysis; Solvents; thin layer chromatography; High performance; Monocotyledones; high-performance thin-layer chromatography (HPTLC); alkaloids; hardinggrass; Thin layer chromatography; Alkaloid; Analysis method; Gramineae; Phalaris; Angiospermae; thin-layer chromatography (TLC); Spermatophyta; Phalaris aquatica; Quantitative analysis
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf061819k

An efficient high-performance thin-layer chromatography (HPTLC) method for the analysis of alkaloids in hardinggrass (Phalaris aquatica L.) was developed. The method employed HPTLC glass plates precoated with silica gel 60F-254 as the stationary phase. The solvent system consisted of ethyl acetate/chloroform/7 N NH4OH in methanol (8:2:1, v/v/v). Using unidimensional double-development, bands were well separated for 10 alkaloid standards as well as alkaloids observed in hardinggrass plant extracts. Identities of compounds observed using HPTLC were validated by high-performance liquid chromatography−mass spectrometry (HPLC-MS). Software was used to quantify individual alkaloids in plant samples based on HPTLC retention factors and intensities relative to standards of known concentration. Correlation coefficients of 0.99 were obtained between estimated and actual concentrations for four standards (methyltyramine, hordenine, gramine, and 5-methoxydimethyltryptamine), with linearity in the range of 120−3840 ng/spot. The HPTLC method is repeatable and specific for β-carboline, tryptamine, gramine, and tyramine type alkaloids in mixed standard and plant extracts. Initial results indicate substantial variation in alkaloid composition among and within hardinggrass populations. Keywords: Phalaris aquatica; thin-layer chromatography (TLC); high-performance thin-layer chromatography (HPTLC); alkaloids; quantitative analysis; hardinggrass