Identification and quantification of the steroidal sapogenin diosgenin by HPTLC in rhizomes with roots of Dioscorea caucasica Lipsky and Dioscorea nipponika Makino

Introduction. The original article presents the results of an experimental study conducted using the method of high-performance thin-layer chromatography (HPTLC) on plant samples – rhizomes with roots of Dioscorea caucasica Lipsky and Dioscorea nipponika Makino. These species of Dioscorea are used in various fields of medicine as fungicides, antimicrobials, and anti-sclerotic agents. The HPTLC method is used at the screening stage of plant samples for preliminary identification and densitometric quantitative determination of diosgenin in plant extracts.

Aim. Development of a HPTLC methodology for quantitative assessment of diosgenin content after acid hydrolysis of extracts in air-dried raw materials – rhizomes with roots of Dioscorea caucasica Lipsky and Dioscorea nipponika Makino.

Materials and methods. The extracts were obtained by preliminary degreasing and depigmentation of air-dried rhizomes with roots of the studied Dioscorea species using high-purity dichloromethane, followed by double extraction. The first extraction was carried out in a 50 % aqueous solution of isopropanol with ultrasonication, followed by acid hydrolysis of O-glycosidic bonds and evaporation. The second extraction was carried out by redissolving the dry residue in methanol; it was purified from suspended particles by filtration through syringe filters with a perforation diameter of 20 μm. HPTLC was performed on a «CAMAG» (Switzerland) apparatus using HPTLC Aluminum sheets Silica gel 60 F254 20 × 20 cm, which were cut to a size of 20 × 10 cm.

Results and discussion. After performing scanning densitometry at 366 and 542 nm, it was established that the chromatography of methanol extracts in a toluene-chloroform-acetone solvent system (2 : 8 : 2 v/v) allows for satisfactory separation and subsequent densitometric quantitative determination of diosgenin. The tracks of plant extracts from the rhizomes and roots of Dioscorea caucasica Lipsky and Dioscorea nipponika Makino were compared with a standard sample of diosgenin.

Conclusion. The study found that the content of the steroid sapogenin diosgenin in air-dried Dioscorea nipponica Makino raw material (286,4–296,3 μg/g) slightly exceeds its content in Dioscorea caucasica Lipsky raw material (257–277,1 μg/g). The quantitative values obtained, calculated separately for peak height and area, show good convergence, confirming the correctness of the method. The developed method of high-performance thin-layer chromatography with densitometric detection of diosgenin can be recommended for routine quantitative analysis of this compound in plant raw materials.

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