QUANTIFICATION OF GLYCYRRHIZIN BIOMARKER IN GLYCYRRHIZA GLABRA RHIZOME AND BABY HERBAL FORMULATIONS BY VALIDATED RP-HPTLC METHODS

Background: A simple and sensitive thin-layer chromatographic method has been established for quantification of glycyrrhizin in Glycyrrhiza glabra rhizome and baby herbal formulations by validated Reverse Phase HPTLC method. Materials and Methods: RP-HPTLC Method was carried out using glass coated with RP-18 silica gel 60 F254S HPTLC plates using methanol-water (7: 3 v/v) as mobile phase. Results: The developed plate was scanned and quantified densitometrically at 256 nm. Glycyrrhizin peaks from Glycyrrhiza glabra rhizome and baby herbal formulations were identified by comparing their single spot at Rf = 0.63 ± 0.01. Linear regression analysis revealed a good linear relationship between peak area and amount of glycyrrhizin in the range of 2000-7000 ng/band. Conclusion: The method was validated, in accordance with ICH guidelines for precision, accuracy, and robustness. The proposed method will be useful to enumerate the therapeutic dose of glycyrrhizin in herbal formulations as well as in bulk drug

Absolute configuration of (1S,2S)-3-methyl-2-phenyl-2,3-dihydro­thia­zolo[2,3-b]quinazolin-5-one

The absolute structure of the molecule in the crystal of the title compound, C17H14N2OS, was determined by the refinement of the Flack parameter to 0.0 (2) based on 1011 Friedel pairs. The quinazoline ring is essentially planar, with a maximum deviation of 0.037 (2) Å. The thia­zole ring is distorted from planarity [maximum deviation = 0.168 (2) Å] and adopts a slightly twisted envelope conformation, with the C atom as the flap atom. The central thia­zole ring makes dihedral angles of 7.01 (8) and 76.80 (10)° with the quinazoline and phenyl rings, respectively. The corresponding angle between the quinazoline and phenyl rings is 3.74 (9)°. In the crystal, there are no classical hydrogen bonds but stabilization is provided by weak C—H⋯π inter­actions, involving the centroids of the phenyl rings

COMPARATIVE PROFILING OF BIOMARKER PSORALEN IN ANTIOXIDANT ACTIVE EXTRACTS OF DIFFERENT SPECIES OF GENUS FICUS BY VALIDATED HPTLC METHOD

Background: A simple but sensitive HPTLC method was developed for the comparative evaluation of psoralen in antioxidant active extracts of leaves of five different species of genus Ficus (Ficus carica, Ficus nitida, Ficus ingens, Ficus palmata and Ficus vasta). Materials and Methods: HPTLC studies were carried out using CAMAG HPTLC system on Glass-backed silica gel 60F254 HPTLC pre-coated plates using selected mobile phase toluene: methanol (9:1). The antioxidant activity was carried out, using DPPH free radical method. Results: Among all the five species of genus Ficus, F. palmata and F. carica exhibited comparatively good antioxidant activity in DPPH assay. The developed HPTLC method was found to give a compact spot for psoralen (Rf = 0.55±0.001) at 305 nm. The regression equation and r2 for psoralen was found to be Y= 4.516X+35.894 and 0.998. The quantification result revealed the presence of psoralen in only two species, F. carica (0.24%, w/w) and F. palmata (1.88%, w/w) which supported their supremacy for anti-oxidant potential over other species. The statistical analysis proved that the developed method was reproducible and selective. Conclusion: The developed method can be used as an important tool to assure the therapeutic dose of active ingredients in herbal formulations as well as for standardization and quality control of bulk drugs and in-process formulations. This method can also be employed for the further study of degradation kinetics and determination of psoralen in plasma and other biological fluids

Utilization of gel electrophoreses for the quantitative estimation of digestive enzyme papain

SDS-PAGE densitometric method for analysis of papain in pharmaceutical formulations was developed and validated for the first time. Standard and samples were mixed with SDS sample buffer and denatured at 95 °C for 5 min and the gel was run at 20 mA and 200 V for 30–40 min in SDS-PAGE buffer. Gels were stained in Coomassie blue solution and distained by 5% methanol and 10% acetic acid. Destained gels were imaged and analyzed using the ChemiDoc™ XRS+ System. Bands of papain appeared at Rf value 0.78 ± 0.03 corresponding to molecular weight 23406 Da between proteins with molecular weight 31,000 and 21,500 Da of the broad range protein standard. The generated calibration curve was used for quantitative estimation of papain in pharmaceutical formulations. The developed method was validated for precision, accuracy, specificity and robustness as described by the ICH guidelines. The proposed method gives an alternative approach for enzymes and protein analysis

New microbial source of the antifungal allylamine “Terbinafine”

The isolated active compound “F12” from the culture media of the Streptomyces sp. KH-F12 was identified using different spectroscopic techniques. Both 1D- and 2D-NMR as well as HRESIMS were utilized to characterize the structure of the isolated compound. ‘F12” was found to be the known systemic antifungal drug terbinafine marketed under the name “Lamisil”. Full analysis of the COSY, HSQC and HMBC enables the full assignment of proton and carbon atoms. Terbinafine is a synthetic allylamine and is reported here for the first time from natural source

In Vivo Hepatoprotective and Nephroprotective Activity of Acylated Iridoid Glycosides from Scrophularia hepericifolia

Phytochemical investigation of the chloroform fraction obtained from Scrophularia hypericifolia aerial parts led to the isolation of nine acylated iridoid glycosides. The new compounds were identified as 6-O-α-L(2″-acetyl, 3″,4″-di-O-trans-cinnamoyl) rhamnopyranosyl-6′-acetyl catalpol (6′-acetyl hypericifolin A) (1), 6-O-α-L(2″, 4″-diacetyl, 3″-O-trans-cinnamoyl) rhamnopyranosyl-6′-acetyl catalpol (6′-acetyl hypericifolin B) (2), 6-O-α-L(2″-acetyl, 3″,4″-di-O-trans-cinnamoyl) rhamnopyranosyl catalpol (hypericifolin A) (3) and 6-O-α-L(2″, 4″-diacetyl, 3″-O-trans-cinnamoyl) rhamnopyranosyl catalpol (hypericifolin B) (4). Previously reported compounds were identified as laterioside (5), 8-O-acetylharpagide (6), 6-O-α-L(4′-O-trans-cinnamoyl) rhamnopyranosyl catalpol (7), lagotisoside D (8) and harpagoside (9). Identification achieved via analyses of physical and spectral data including 1D, 2D NMR and High Resolution Electrospray Ionization Mass spectroscopy (HRESIMS). Compounds 2–4 and 6 were subjected to biological evaluation against paracetamol-induced toxicity. The biochemical parameters aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and gamma glutamyl transpeptidase (GGT) as well as total bilirubin were used to access the liver condition. Measurement of serum levels of urea, creatinine, sodium and potassium cations were indicators for kidney condition. Liver and kidney samples were subjected to histopathological study. The best protection was found in the group treated with 3 followed by 4 and 6, while 2 was almost inactive

Fumigant Toxicity and Feeding Deterrent Activity of Essential Oils from <i>Lavandula dentata</i>, <i>Juniperus procera</i>, and <i>Mentha longifolia</i> against the Land Snail <i>Monacha obstructa</i>

Land mollusks are one of the most destructive agricultural pests worldwide, the management of which depends on synthetic molluscicides. However, many of these molluscicides are harmful to nontarget organisms. Hence, there is a need to develop alternative ecofriendly molluscicides that are less impactful toward nontarget organisms. So, an investigation into the fumigant toxicity and feeding deterrent effect of essential oils (EOs) from Lavandula dentata L. (Lamiaceae), Juniperus procera Hochst. (Cupressaceae), and Mentha longifolia (L.) Huds. (Lamiaceae) against the land snail Monacha obstructa (Pfeiffer, 1842) (Hygromiidae) was performed. L. dentata EO exhibited the highest fumigant toxicity with LC50 values of 8.68 μL/L air and 7.24 μL/L air after 24 h and 48 h exposure periods, respectively. Its main components were camphor, 1,8-cineole, fenchone, and β-myrecene. The fumigant toxicity of J. procera EO was lower than that of L. dentata, with LC50 values of 25.63 μL/L air and 20.11 μL/L air after 24 h and 48 h exposure periods, respectively. The major constituents of J. procera EO were α-pinene, p-cymene, and β-ocimene. The analysis of M. longifolia EO showed that pulegone, and menthol were the major constituents. However, it displayed no fumigant toxicity up to 50 μL/L air. The three EOs exhibited a strong feeding deterrent effect at sublethal concentrations. The EOs extracted from L. dentata, J. procera and M. longifolia are promising ecofriendly botanical molluscicides against the land snail M. obstructa