Abstract

Daidzein-natural isoflavones found in Pueraria tuberosa (Willd.) DC. (Fabaceae) which is a potent medicinal plant. It acts as signal carriers and responds to the pathogenic attacks and reduces pain during menopause, osteoporosis and also antidiabetic in nature. In the present work, in vitro callus development from tuber explants during both rainy and summer seasons was conducted. Further, a simple unambiguous and rapid high-performance thin-layer chromatography method was established for quantitative estimation of daidzein in callus and in vivo parts to assess the impact of seasonal change on valuable phytochemical accumulation. Maximum callusing (90%) was obtained on MS medium fortified with a combination of 6N-Benzylamino purine (2 mg/l) and 2,4-dichlorophenoxyacetic acid (2 mg/l) during the summer season from the tuber explant. During the HPTLC method validation, the linearity range obtained was 100–1000 ng/spot with a regression value (r) value of 0.99845. All the parts were found to contain a significant amount of daidzein. The maximum daidzein (2112.567±0.35 ng/g) content was obtained from a young tuber bark followed by callus (171.903±0.33 ng/g) during the summer season compared to rainy season parts and callus. Thus, it can be concluded that in vitro callus is an alternative source of daidzein without destroying the natural plant and the developed HPTLC method could be used for quality control analysis and recommended for daidzein quantification for different herbal formulation and drug preparation.

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