Development of a validated high-performance thin-layer chromatography method for quantification of lupeol from different parts of Bauhinia acuminata.

The goal of the study was to determine how much lupeol was present in different Bauhinia acuminata samples. Lupeol was analyzed using a unique (verified) high-performance thin-layer chromatography (HPTLC) method. Separation of various compounds was achieved on silica gel (66 F 254) plates with toluene and methanol (99:1) as the mobile phase, followed by derivatization with Liebermann-Burchard reagent (r2 = 0.996). The proposed approach demonstrated a satisfactory linear relationship in the concentration range of 4-8 µg/mL. The limit of detection and limit of quantification were 63.84 ng/spot and 208.57 ng/spot, respectively. The proposed method was a simple and precise procedure that provided good resolution with other constituents of extracts. The validated HPTLC method described was simple, dependable, and practical and could be useful for standardizing and evaluating the quality of herbal material and formulations containing various Bauhinia species. The leaf, stem, unsaponified matter leaf, and stem petroleum ether extract had good amounts of lupeol (25.75, 38.25, 1.65, and 1.75, respectively). This research will be helpful for the quantitative determination of lupeol from herbal formulations as well as useful for further studies in this area.

Neuroprotective effect of Cubebin: A dibenzylbutyrolactone lignan on scopolamine-induced amnesia in mice.

BACKGROUND & OBJECTIVES: Acetylcholinesterase (AChE) inhibitors represent a major class of drugs which provide symptomatic relief and improvement in cognitive function in Alzheimer's disease (AD). In this study, cubebin, a dibenzylbutyrolactone lignan, was isolated from Piper cubeba and investigated for its AChE inhibitory activity in an attempt to explore its potential for memory-enhancing activities in mice. METHODS: Molecular docking of cubebin was carried out followed by in vitro AChE activity. Mice were treated with cubebin (25 & 50 mg/kg; i.p.), for three days and memory impairment was induced by scopolamine (3 mg/kg; i.p.). Memory function was evaluated by Morris water maze (MWM) test. Biochemical parameters of oxidative stress and cholinergic function were estimated in brain. RESULTS: Molecular docking study revealed that cubebin was well bound within the binding site of the AChE enzyme showing interactions such as π-π stacking and hydrogen bonding with residues present therein. Cubebin inhibited AChE enzyme in an in vitro assay with IC50value of 992 µM. Scopolamine administration caused a significant impairment of learning and memory in mice, as indicated by a marked decrease in MWM performance. Scopolamine administration also produced a significant enhancement of brain AChE activity and oxidative stress in mice brain. Pre-treatment of cubebin (25 and 50 mg/kg; i.p.) significantly prevented scopolamine-induced learning and memory deficits along with attenuation of scopolamine-induced rise in brain AChE activity and oxidative stress level. INTERPRETATION & CONCLUSIONS: Cubebin showed promising protective activity in scopolamine-induced spatial memory impairment in mice. This could be attributed to its brain AChE inhibition and antioxidant activity.

Assessment of conventional and novel extraction techniques on extraction efficiency of five anthraquinones from Rheum emodi.

Rheum emodi is principally known to consist 1,8-dihydroxyanthraquinones (DHAQs) that find immense use in the chemical, pharmaceutical, cosmetic industries and in herbal medication and food sector. The aim of this study was to compare non-conventional and classical methods for extraction of anthraquinones from R. emodi. Optimisation of the extraction parameters for various methods was done and their extraction efficiency was evaluated. In preliminary screening experiments, choice of solvent and solid : solvent ratio was optimised. Comparison of extraction efficiency for classical methods like maceration, heat-reflux, soxhletion and non-conventional methods like ultra-sonication and sublimation was done for five DHAQs - aloe emodin, rhein, emodin, chrysophanol and physcion using HPLC-UV and fluorescence detection in native and acid hydrolysed samples. It was observed that ethanol was the best solvent for extraction of anthraquinones with a solid : solvent ratio of 1:20. A prior acid hydrolysis led to significant increase in anthraquinone extraction. Among the extraction methods heat reflux for 45 min was the most prominent extraction method with highest recovery of the DHAQs. In ultrasonic assisted extraction, an increase in the anthraquinone extraction was seen till 45 min after which the concentration declined. A novel, solvent-free, green and selective method of extraction by sublimation was found to be effective for extraction of anthraquinones.

Simultaneous quantification of furanocoumarins from Aegle marmelos fruit pulp extract.

Aegle marmelos Correa (Rutaceae) is a prominent species in the Sub-Himalayan region, also found in central and south India, and has been widely used as remedy for diarrhea and dysentery. Unripe fruits show antiviral activity and prove to be a potent hypoglycemic agent. In this study, a high-performance liquid chromatography (HPLC) method for the simultaneous quantification of major furocoumarin components marmelosin, psoralen and bergapten in the extract from fruit pulp of A. marmelos has been developed. Components were found in the range of 5-6.5, >0.4-0.75 and >0.25 mg/g of dried fruit weight, respectively. The method was validated for linearity, precision, accuracy, robustness, limit of detection and limit of quantification. Linearity was determined over the range of 5-40 µg/mL (r > 0.965). Intra- and interday precision showed a relative standard deviation of <2.5%. The accuracy of the method was determined by a spike recovery study, and the average recoveries were 99.46, 101.04 and 100.8% for marmelosin, psoralen and bergapten, respectively. The proposed HPLC method was found to be simple, precise and specific and can be used simultaneously for the routine quality control of raw materials of A. marmelos fruit extracts and their products, and also other products containing these markers.

Fate of ß-asarone in Ayurvedic Sodhana process of Vacha.

BACKGROUND: Calamus (Acorus calamus Linn., Araceae) rhizome synonymously called sweet flag or Vacha is an aromatic herb indigenous to Central Asia and Eastern Europe. It has been used by the Ayurvedic practitioners since time immemorial for diseases ranging from weakness of memory to being used as an anthelminthic. Reports of its use have been found in books like Charak Samhita, Sushruta Samhita, etc., The major constituent of the oil of Vacha is a phenyl propanoid called ß-asarone, which is reported to show carcinogenic properties. Due to the toxic effects of ß-asarone, sodhana prakriya (detoxification process) has been prescribed for Vacha before its inclusion in the Ayurvedic medicines. Shodhanaprakriya (S. prakriya) of Vacha has been mentioned in the Ayurvedic texts. OBJECTIVES: This study was undertaken with an aim to find out the mechanism involved in the S. prakriya of Vacha and also to suggest an alternate method for the conventional one. MATERIALS AND METHODS: The conventional method was studied in the laboratory and equivalent alternate methods were designed based on the mechanism involved. Vacha samples were subjected to the conventional method as well as the alternate methods and the content of ß-asarone in the different samples was monitored using Gas Chromatography technique. RESULTS: Various alternate methods have been devised based on the mechanism involved in the S. prakriya which have given results comparable with those of the conventional method. CONCLUSION: The scientific mechanism involved in the S. prakriya of Vacha has been established and alternate methods have been proposed.