GC-MS Analysis of Bioactive Compounds and Hosttoxicity Studies of Azolla caroliniana Symbiotic with the Cyanobacterium Anabaena azollae

Abstract:

Objective: The present research work was conducted to do the gas chromatography and mass spectrometry (GC-MS) analysis of the water fern, Azolla caroliniana symbiotic with the cyanobacterium Anabaena azollae, along with in vitro host toxicity testing to check its toxicity level. Materials and methods: The GC-MS analysis of the methanol extract of A. caroliniana was carried out using a GC-MS instrument and in vitro host toxicity testing of A. caroliniana extract was carried out with cultured lymphocytes from human umbilical cord blood. Results: It was evident that only one phytochemical, 3-o-methyld- glucose with retention time 16.581 min and a peak area of 91.89% as the major phytochemical was present in methanolic extract of the fern, while the rest 7 of the total 8 chemicals namely (with peak area values), ‘2-butanone, 3-methoxy-3-methyl’ (0.865%), ‘2,2-dimethyl propionic acid, cyclopentyl ester’ (0.670%), ‘butane, 1-bromo- 2-methyl’ (0.398%), ‘2-hexen-1-ol,2-ethyl’ (0.269%), ‘5-hydroxy-2,2-dimethyl hexan- 3-one’ (0.212%), ‘phthalic acid, ethyl pentyl ester’ (0.21%), ‘pentanoic acid, 2-methyl’ (0.182%), were present in minor quantities. Host toxicity testing was done with in vitro cultured lymphocytes from human umbilical cord blood. The LC₂₅ values were 870.96 and 691.83 mg/L with the trypan blue and acridine orange/ethidium bromide staining, respectively by methanolic extract of A. caroliniana in cellular toxicity experiments, against the minimum inhibitory concentration values of 300 mg/L extract. During the assessment of nuclear toxicity, no comet was found in the cells grown with 0 to 1000 mg/L of the extract. Conclusion: The extract had neither cellular nor nuclear toxicity to cultured human lymphocytes.

Key words: Azolla Caroliniana, Fern, Gc-Ms Analysis, 3-O-Methyl-D-Glucose, Host Toxicity Testing, In Vitro Cultured Lymphocytes.