DEVELOPMENT AND VALIDATION OF RAPID REVERSE PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR SIMULTANEOUS ESTIMATION OF STIGMASTEROL AND β-SITOSTEROL IN EXTRACTS OF VARIOUS PARTS (LEAVES, STEMS, AND ROOTS) OF XANTHIUM STRUMARIUM LINN.

Objective: Xanthium strumarium is a cocklebur or burweed belonging to family Asteraceae and commonly found as a weed, is widely distributed in North America, Brazil, China, Malasia, and hotter part of India. The herb is traditionally used mostly in treating several ailments. The present study deals with development and validation of a reliable reverse phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of Stigmasterol and β-sitosterol in the various extracts of the plant.Methods: The proposed method utilizes a Qualisil Gold C18 column (250×4.6 mm), 5 µm particle size, under isocratic elution conditions with themixture of acetonitrile:ethanol:water (85:14:1 v/v/v) at 25° as a mobile phase. An effluent flow rate of 1 ml/minute and ultraviolet detection at202 nm was used for the analysis of Stigmasterol and β-sitosterol.Results: The described method was linear in the range of range of 100-500 µg/ml and 10-500 µg/ml for stigmasterol and β-sitosterol respectively, with excellent correlation coefficients. The precision, robustness, and ruggedness values were also within the prescribed limits (<2%). The recovery values were within the range, which indicates that the accuracy of the analysis was good and that the interference of the matrix with the recovery of phytosterols was low. The phytosterols were found to be stable in a stock solution for 24 hrs (percentage relative standard deviation was below 2%) and no interfering extra peaks were observed under controlled stress conditions.Conclusion: The proposed method is simple, specific, precise, accurate, and reproducible and thus can be used as appropriate method for routine analysis of X. strumarium phytosterols in quality control laboratories.Keywords: Xanthium strumarium, Reverse phase high-performance liquid chromatography method, Precision, Phytosterols, Quality contro

Phytochemical Screening and In-Vitro Evaluation of Antioxidant Activities of Various Extracts of Leaves and Stems of Kalanchoe crenata

Many diseases are associated with oxidative stress caused by free radicals. Current research is directed towards finding naturally occurring antioxidants of plant origin. Kalanchoe crenata Andr. (Crassulaceae), commonly known as "never die" or "Dog's liver," has been traditionally used for the treatment of ailments, such as, earache, smallpox, headache, inflammation, pain, asthma, palpitations, convulsion, and general debility. The aim of present research deals with phytochemical screening and in-vitro evaluation of antioxidant activities of the leaves & stems of K.crenata. Method: Successive extracts of leaves & stems was subjected for phytochemical screening. The preliminary screening reports the presence of saponins, phytosterols, flavanoids, phenols and alkaloids in the extracts. Various extracts of K.crenata leaves & stems was studied in-vitro for total antioxidant activity, for scavenging of nitric oxide, hydrogen peroxide, the antioxidant capacity by phosphomolybdenum, reducing power determination and determination of phenolic and flavonoid content in the extracts. 1,1-Diphenyl-2-picrylhydryzyl (DPPH) scavenging activity or the hydrogen donating capacity was quantified in presence of stable DPPH radical on the basis of Blois method. Nitric Oxide (NO) radical scavenging method was performed in the presence of nitric oxide generated from sodium nitroprusside using ascorbic acid as standard in both methods. The phenolic content was determined by using Folin-Ciocalteu reagent and flavonoid content was determined by aluminum chloride. Result: The radical scavenging activity was found to dose dependent. Thus extract has been established as an antioxidant. The reducing capacity serves as significant indicator of antioxidant activity. The reducing power was found to increase with the increasing concentration of extract. The 100mg plant powder yielded 0.34, 0.49, 0.72, 0.98%w/wand 0.15, 0.23, 0.39, 0.56%w/w phenolic content in the benzene, chloroform, acetone, ethanol extracts of leaves and stems respectively using gallic acid as standard. Plant contains about 0.19, 0.29, 0.48, 0.64%w/w and 0.11, 0.17, 0.32, 0.47 %w/w of flavonoid content in the benzene, chloroform, acetone, ethanol extracts of leaves and stems respectively using quercetin as standard. Conclusion: The present study provides evidence that different extracts of K.crenata leaves and stems is potential source of antioxidant activity. The extracts were found to contain phenolic compounds which could be responsible for the antioxidant properties. So K. crenata traditional use is justified in the present research work

Adaptation of the Wound Healing Questionnaire universal-reporter outcome measure for use in global surgery trials (TALON-1 study): mixed-methods study and Rasch analysis

BackgroundThe Bluebelle Wound Healing Questionnaire (WHQ) is a universal-reporter outcome measure developed in the UK for remote detection of surgical-site infection after abdominal surgery. This study aimed to explore cross-cultural equivalence, acceptability, and content validity of the WHQ for use across low- and middle-income countries, and to make recommendations for its adaptation.MethodsThis was a mixed-methods study within a trial (SWAT) embedded in an international randomized trial, conducted according to best practice guidelines, and co-produced with community and patient partners (TALON-1). Structured interviews and focus groups were used to gather data regarding cross-cultural, cross-contextual equivalence of the individual items and scale, and conduct a translatability assessment. Translation was completed into five languages in accordance with Mapi recommendations. Next, data from a prospective cohort (SWAT) were interpreted using Rasch analysis to explore scaling and measurement properties of the WHQ. Finally, qualitative and quantitative data were triangulated using a modified, exploratory, instrumental design model.ResultsIn the qualitative phase, 10 structured interviews and six focus groups took place with a total of 47 investigators across six countries. Themes related to comprehension, response mapping, retrieval, and judgement were identified with rich cross-cultural insights. In the quantitative phase, an exploratory Rasch model was fitted to data from 537 patients (369 excluding extremes). Owing to the number of extreme (floor) values, the overall level of power was low. The single WHQ scale satisfied tests of unidimensionality, indicating validity of the ordinal total WHQ score. There was significant overall model misfit of five items (5, 9, 14, 15, 16) and local dependency in 11 item pairs. The person separation index was estimated as 0.48 suggesting weak discrimination between classes, whereas Cronbach's α was high at 0.86. Triangulation of qualitative data with the Rasch analysis supported recommendations for cross-cultural adaptation of the WHQ items 1 (redness), 3 (clear fluid), 7 (deep wound opening), 10 (pain), 11 (fever), 15 (antibiotics), 16 (debridement), 18 (drainage), and 19 (reoperation). Changes to three item response categories (1, not at all; 2, a little; 3, a lot) were adopted for symptom items 1 to 10, and two categories (0, no; 1, yes) for item 11 (fever).ConclusionThis study made recommendations for cross-cultural adaptation of the WHQ for use in global surgical research and practice, using co-produced mixed-methods data from three continents. Translations are now available for implementation into remote wound assessment pathways

Proceedings Of The 23Rd Paediatric Rheumatology European Society Congress: Part Two

PubMe