International Journal of Horticulture and Floriculture

International Journal of Horticulture and Floriculture ISSN 2756-3790 Vol. 9 (1), pp. 001-004, January, 2021. © International Scholars Journals

Full Length Research Paper

Quantitative determination of DNA Methylation in tobacco leaves by HPLC

Feng Jia1, YunPeng Fu1, WeiQun Liu2*, Zheng Du1 and YiDan Zhao2

1College of Tobacco Science, Henan Agricultural University, Zhengzhou 450002, People’s Republic of China.

2College of Life Sciences, Henan Agricultural University, Zhengzhou, 450002, People’s Republic of China.

Accepted 20 October, 2020


A relatively straightforward high performance liquid chromatography (HPLC) method for determination of DNA methylation in flue-cured tobacco leaves was developed and validated. DNA methylation was measured as follows: DNA was hydrolyzed to nucleotide units in perchloric acid. The bases of cytosine and 5-methylcytosine were separated on a reversed-phase C18 column at 30°. The mobile phase used methanol: 5 mmol/L pentane sulfonate (10:90, v/v) and a flow rate of 0.7 mL/min. The absorbance of bases was measured at 273 nm. The results showed that the method was accurate, quick, and cost effective to determine DNA methylation of flue-cured tobacco.

Key words: Tobacco, DNA methylation, 5- Methylcytosine, high performance liquid chromatography (HPLC).