A red color with sodium amalgam and HCl acid. The flavone glycoside RS-2 was found to be soluble in water, ethanol and acetone and crystallized from methanol. RS-2 analyzed for molecular formula C29H34O13, m.p. 285–286° and M+ 590 (CIMS). The wavelengths of maximum absorption as observed with various shift reagents were at; λmax (MeOH) 270, 347 nm, λmax (NaOMe) 287, 395 nm, λmax (AlCl3) 278, 389, 405 nm, λmax (AlCl3 + HCl) 277, 389, 405 nm, and λmax (NaOMe) 272, 348 nm as depicted in Graph 2. The characteristic band observed in the IR spectrum of RS-2

and the structural assignments made with the help of available literature1, 2, 3 and 4 are described below: 3396.3 cm−1 (Hydrogen bonding intermolecular stretching), 2864.5 cm−1 (CH3 stretching of CH3), 1637.9 cm−1 (α,β-unsaturated C O), 1461.5 cm−1 (Aromatic ring system), 1219.0 cm−1 (C–O–C– stretching Osimertinib vibration), and 771 cm−1 (C–H out of plane bending) as portrayed in Graph 1. Significant band at Vmax (KBr) 3396.3 cm−1 as mentioned in Graph 1 in the IR spectrum of the glycoside (RS-2) indicated the presence of hydroxyl group(s) in it. The glycoside (RS-2) was acetylated with Ac2O/Pyridine to give an acetylated product having molecular formula, C41H46O19, m.p. 204–205° and M+ 842 (CIMS). The estimation of percentage of the

acetyl group (31.04%) in the acetylated derivative was given by Weisenberger method5 UMI-77 as described by Belcher and Godbert6 which showed that there were six acetylable hydroxyl groups in the glycoside (RS-2). The appearance of band in IR spectrum of the acetyl derivative at Vmax (KBr) 1725.4 cm−1 with disappearance of band at Vmax (KBr) 3396.3 cm−1 confirmed that the acetylation of all the hydroxyl groups present

in the glycoside RS-2 was complete. 7 and 8 The IR absorption spectrum of the flavone glycoside (RS-2) displayed important band at Vmax (KBr) 2925.9 cm−1 indicating the presence of methoxyl group(s) in it. The methoxyl group estimation (16.05%) was done by Zeisel’s method 9 which confirmed the presence of three methoxyl groups in RS-2. The 1H NMR spectrum Metalloexopeptidase of the flavonoidal glycoside (RS-2) showed three singlets at δ 4.0, δ 3.97 and δ 3.80 as depicted in Graph 3 each of these integrating for three protons, thereby suggesting the presence of three methoxyl groups in RS-2. Characteristic band at Vmax (KBr) 1461.5 cm−1 in the IR spectrum of glycoside RS-2 showed the presence of C C ring stretching. The structure of the glycoside (RS-2) was elucidated by its acid hydrolysis and identifying the components of hydrolyzate and the aglycone respectively. The glycoside (RS-2) on its acid hydrolysis with 7% alcoholic H2SO4 yielded an aglycone RS-2(A) as a solid residue and sugar moiety(ies) in the filtrate. They were separated by filtration and studied separately. The aglycone RS-2(A) was found to be homogenous on TLC examination (EtOAc–MeOH–H2O, 3:2:1). It crystallized from MeOH.