HPTLC and TLC method development preliminary studies for damask rose

Rosa damascena Mill. (Rosaceae) has several pharmacological properties including anti-HIV, antibacterial, antioxidant, antitussive, hypnotic, antidiabetic, and relaxant effect on tracheal chains. Petals contain mainly essential oil and flavonoids [1].

The aim of this study is to develop TLC and HPTLC methods for quality control of marketed preparations. In this study, we identified the optimal solvent system and suitable reference molecules as biomarkers after several trials of extracts from Rosa damascena petals.

Extracts of R. damascena petals prepared by using the solvents in different polarity were tested with several mobile phases in thin layer chromatography. Subsequently, a simple, precise, accurate and rapid high performance thin layer chromatographic method was developed. Precoated silica gel 60 F254 was used as stationary phase. 9 mobile phase and 11 reference substance (kaempferol, kaempferol 3-O-β-D-glucopyranoside, kaempferol 3-O-β-D-galactopyranoside, kaempferol 3-O-β-D-gentiobioside, quercetin-3-O-rhamnopyranoside, quercetin-3-O-arabinopyranoside, quercetin 3-O-β-D-glucopyranoside, quercetin 3-O-β-D-galactopyranoside, rutin, RD1 (structure elucidation has not been carried out yet), gallic acid) were tested to find optimal mobil phase and marker substance.

According to the results, ethyl acetate: formic acid: acetic acid: water (100:11:11:10, v/v/v/v) was determined as the optimal solvent system and kaempferol 3-O-β-D-glucopyranoside, quercetin 3-O-β-D-glucopyranoside, quercetin 3-O-β-D-galactopyranoside as biomarkers. The detection of spots was carried out at 254, 366nm and vanilin/H₂SO₄ was used as revelator. The method validation in terms of linearity, accuracy, precision and specificity will be performed in further studies. The proposed method can be successfully used to determine the drug content of marketed preparations.

[1] Aslam M, Siddiqui A. Eur J Biomed Pharm Sci 2016; 3: 432 – 435.