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Densitometric HPTLC has been used for determination of catechins in shoot and callus cultures of two species from the genus Phyllanthus, P. juglandifolius and P. grandifolius . The catechin composition of intact P. juglandifolius (leaves and stems) was also compared with that of biomass obtained by biotechnological methods. A mixture of the standards (−)-epicatechin, (+)-catechin, (−)-epigallocatechin, (−)-gallocatechin, (−)-catechin gallate, and (−)-epicatechin gallate was separated on C 18 plates, with methanol-water-formic acid 30:70:6 ( v/v ) as mobile phase, in an automated development chamber at 75% humidity. The catechins were detected at 440 nm after derivatization with diazotized sulfanilic acid. The total catechin content of the plant material was calculated on the basis of the amount of (+)-catechin. The method was validated for precision and repeatability. (−)-Epicatechin, (+)-catechin, (−)-epigallocatechin, and (−)-gallocatechin were shown to be present in all shoot and callus cultures of the species mentioned above and in the leaves of intact P. juglandifolius plants. Amounts of catechins were similar in all the samples — from 2.39 mg g −1 in the leaves of P. juglandifolius to 1.94–1.68 mg g −1 in the shoot cultures of both species.

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Summary

Gymnemic acid (GA), a well known anti-diabetic compound has been detected in methanol extracts of intact leaves and in vitro callus cultures derived from leaf explants of Gymnema sylvestre. Callus biomass was developed in MS medium with optimum plant growth regulators (OPGRs) of 2,4-D (1.5 mg L−1) + KN (0.5 mg L−1) under abiotic stress conditions at 45 days determined by growth curve analysis. GA detection and quantification were carried out using thin-layer chromatography (TLC), highperformance thin-layer chromatography (HPTLC), high-performance liquid chromatography (HPLC), and gravimetric techniques. GA detection peak area and their absorption spectra were evaluated through HPTLC and HPLC with the standard GA. Quantification of GA had showed the linearity, accuracy, robustness and precision by HPLC. GA content was significantly higher in gravimetric method than HPLC. All these methods were found to be simple, accurate, selective and rapid and could be successfully applied for the determination of GA. It could have potential as a pharmaceutical drug for Type 1 diabetes mellitus (IDDM) and obesity.

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, salicylic acid and dimethyl sulphoxide) and precursor (tryptamine) treatment in the in vitro plant cultures of R. serpentina was reported earlier [ 42 ]. Similarly there are some reports about the in vitro synthesis of reserpine using callus cultures

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