Determination of the nucleic acid constituents in mushroomsby liquid chromatography

Aim: The remarkable reservoir of nucleotides in mushroom has led to a recent interest in the qualitative and quantitative determination of their distribution. In the present research, it was intended to analyse some nucleosides and nucleobases in edible mushroom species collected from Tekirdağ region by HPLC system. Material and Method: The analyzed mushroom species are Truffle (Tuber borchii), Truffle (Tuber aestivum), Black Trumpet (Craterellus fallax), Lamb Belly (Morchella esculenta), Porcini (Boletus edulis), Sığır Dili (Hydnum repandum), Chanterelle (Cantharellus cibarius) and Kanlıca (Lactarius salmonicolor) mushrooms, respectively. After the mushrooms were lyophilized, they were stored at -20°C. The nucleotides in the mushrooms were analyzed by the reverse phase liquid chromatography method. Separation of compounds was accomplished by gradient elution on a SWEA C18 column (4.6 mm x 250 mm, 5µm). A mixture of MeOH: 50 mM sodium phosphate buffer (1:99, v/v) at pH 3.7 as solvent A; MeOH: 50 mM sodium phosphate buffer (20:80) at pH 6.2 was used as solvent B. The internal standard is 3-methyl xanthine. The method has been validated. Two different extraction methods were used to extract the lyophilized mushroom samples, namely, sonication-assisted liquid extraction and pressurized liquid extraction. Deionized water was used as the extraction solvent. Parameters such as temperature and time have been optimized. Results: The 14 nucleotides were simultaneously separated at a wavelength of 254 nm in 25 minutes. Sonication-assisted liquid extraction method has been found to be more effective. This is because the pressurized liquid extraction system is a closed system and matrix-solvent interaction is insufficient. Among the edible mushroom species studied, Lamb's Belly was found to have the highest nucleotide content. Conclusion: Adenosine, guanosine and uridine were found to be the most abundant compounds in all mushrooms studied. Keywords: HPLC-DAD, nucleobase, nucleoside, edible mushrooms, sound wave assisted liquid extraction