Thin Layer Chromatography Fingerprinting Research Articles

ISOLATION AND CHARACTERIZATION OF β-SITOSTEROL FROM ELAEAGNUS ANGUSTIFOLIA CULTIVATED IN IRAQ

Objective: The aim of our study was to investigate chemical constituents of leaves and seeds of Elaeagnus angustifolia since no phytochemical investigation had been done previously in Iraq.Methods: Phytochemical screening of the extracts obtained from the leaves of E. angustifolia indicated the presence of flavonoids (type of flavonols), saponins, alkaloids, steroids, and terpenoids. Thin-layer chromatography fingerprinting and the spraying reagent (concentrated H2SO4 and vanillin in ethanol) were used to identify the hexane extract containing phytosterols.Results: The different chromatographic and spectroscopic results revealed the presence of β‐sitosterol isolated from E. angustifolia. Conclusion: The isolation and purification afforded white crystalline powder which was subjected to chemical and spectral identification by infrared and 1H-nuclear magnetic resonance. The compound was identified as β‐sitosterol.

Adjustment of the formula of local surface water quality assessment

Separation and identification of the active pharmacological compounds from some medicinal plants (Citrus aurantifolia, Rhodomyrtus tomentosa, Camellia sinensis) by Thin Layer Chromatography (TLC) showed the yields of crude extracts obtained from Citrus aurantifolia, Rhodomyrtus tomentosa and Camellia sinensis leaves at 1.5%, 5.62% and 10.4% respectively. Toluene: ethyl acetate (93:7) (v:v) solvent was suitable for the separation of active compounds in crude extract of Citrus aurantifolia, Rhodomyrtus tomentosa leaves while chloroform: ethylacetate: formic acid (5:4:1) (v:v:v) solvent was suitable for the extraction of active compounds in crude extract from Camellia sinensis leaves. Furthermore, the results showed that the number of compounds in extract from Citrus aurantifolia were likely affected by the vacuum evaporator effects. The TLC fingerprints of all three medicinal plants had the same visual ability when the fingerprint detected by UV (λ = 254 nm) and the reagent containing 0.1 g vanillin in 28 mL of methanol: 1 mL of sulfuric acid. In brief, the three active compounds including citral (Citrus aurantifolia), rhdomyrtone (Rhodomyrtus tomentosa) and catechin hydrate (Camellia sinensis) contained in the extract of 3 medicinal herbs had the limited detection (LOD) at 195 ng/spot, 321.5 ng/spot and 625 ng/spot, respectively.

HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY ANALYSIS AND FREE RADICAL SCAVENGING POTENTIAL OF SOUTH INDIAN ORTHODOX BLACK TEA

Objective: The objective of this study was to evaluate the free radical scavenging potential and high-performance thin-layer chromatography (HPTLC) fingerprinting of the ethanolic extract of south Indian orthodox black tea (OBT).Methods: Phytochemical analysis was carried out using standard methods, and free radical scavenging activity of the extract was determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH), nitric oxide (NO), superoxide anion (SOD), and hydroxyl radical scavenging capacities. The ethanolic extract of OBT was loaded in the pre-coated HPTLC plates (silica gel 60 F 254) E-MERCK KGaA. HPTLC was carried out with toluene: ethyl acetate: diethylamine (7:2:1), chloroform: methanol:formic acid (8.5:1.0:0.5), and butanol: isopropyl alcohol (1:1) as mobile phase for alkaloids, flavonoids, and terpenoids, respectively.Results: HPTLC results confirmed that the extract contained several potential active components such as flavonoids, alkaloids, and terpenoids as the slides revealed multicolored bands of varying intensities. Extract of OBT reliably showed the total phenolics 132.27 mg/g, flavonoids 72.52 mg/g, and alkaloids 66.01 mg/g of dry matter. The IC50 value of OBT for DPPH was found to be 372.22 μg/ml, SOD 311.93 μg/ml, NO 362.17 μg/ml, hydroxyl radical 342.14 μg/ml, and reducing power 178.54 μg/ml.Conclusion: The HPTLC fingerprinting profile developed for ethanolic extract will help in proper identification and quantification of marker compounds. The ethanolic extract of OBT was found to possess a wide range of phytochemicals with excellent antioxidant properties. This information may help to choose the best beverage to be consumed in the future.

Bioprofiling for the quality control of Egyptian propolis using an integrated NIR-HPTLC-image analysis strategy

Quality control of propolis being a complex mixture of compounds that are very difficult to analyze and standardize is certainly challenging. Shown on the example of 35 Egyptian propolis samples, a strategy for an improved quality control was demonstrated in which efficacy-directed fingerprint analysis of high-performance thin-layer chromatography (HPTLC) fingerprints were evaluated in combination with Near Infra-Red spectrometry (NIRS) to realize precise discrimination and high quality assessment of chemical and effective consistency of propolis samples that are collected from various geographical locations in Egypt. The fingerprints obtained after derivatization and fluorescence detection (FLD) at λ 366 nm were analyzed by using multivariate data analysis and data were used to identify nine marker ingredients of the different propolis samples. These markers were then quantified by a new validated HPTLC method and an attempt to classify the samples by using a targeted approach was implemented. All investigated propolis samples were initially divided into two types in the untargeted and targeted HPTLC image multivariate analysis. The antimicrobial activities of propolis samples were then evaluated and the results showed that there was significant variation in the biological activities of the different samples. The quantitative data of marker compounds as well as the samples bioactivity results were then subjected to partial least squares regression (PLS-R) analysis which revealed that chrysin, galangin‑5‑O‑methylether, pinostrobin, and islapinin were the main bioactive markers. Based on the efficacy-associated marker ingredients, pretreated spectral patterns of NIRS were selected as a complementary evaluation technique directed to the comprehensive efficacy-directed discrimination of the different Egyptian propolis samples which could successfully classify the propolis samples tested into three different types. Hierarchical cluster analysis and orthogonal projection to latent structures discriminant analysis (OPLS-DA) were applied to the quantitative results from the efficacy-associated markers only. From this systematic method, the 35 samples were subsequently divided into three types; orange, green and blue, displaying a more intricate division than OPLS-DA which is based only on the untargeted HPTLC analysis or the full NIR spectral data. This study initiated the research on the Egyptian propolis and confirmed the existence of three different types of Egyptian propolis, the blue, green and orange types. The study also emphasizes how it is critical to implement the fingerprint- efficacy relationship analysis in finding out the main health-relevant biomarkers for the quality assessment especially for samples displaying delicate differences in their chemical composition and bioactivity.

Bioactivity guided fractionation of potent antiacne plant extract against Propionibacterium acnes

Propionibacterium acnes (P. acnes) is a commensal anaerobic, Gram positive bacterium that belongs to the normal microflora. P. acnes play an important role in the pathogenesis of various skin infections and diseases. However, the available agents are associated with number of side effects and P. acnes show the genetic base of antibacterial resistance against erythromycin and clindamycin. Therefore, alternative natural approaches for the treatment are needed for an hour for alleviation of infection caused by P. acnes. To find out the most effective anti P. acnes extract amongst the selected four plants commonly used for anti-acne potential and to obtain the characterized fraction of the extract with highest anti P. acnes potential, in vitro antimicrobial activity was evaluated using disk diffusion method and the zone of inhibition was compared using Hi-media zone measuring ruler. Afterwards, minimum inhibitory concentration (MIC) was determined for the potent fraction and the results were compared with Clindamycin. The active fraction was characterized chemically and its thin-layer chromatography (TLC) finger printing profile was prepared in order to standardize it. The methanolic extracts of Rubia cordifolia showed maximum (14.0 and 24.0 mm) anti P. acnes activity. The petro-ether fraction showed highest potential compared to other fractions of the extract. Sub-fraction D of petro-ether fraction with maximum activity was further purified. But, the results were similar to the sub-fraction D. The MIC was found to be 3 µg/ml of sub-fraction. R. cordifolia extracts have significant anti P. acnes activity in comparison to Tephrosia purpurea, Viola tricolor and Serenoa repens. The fractionation of R. cordifolia extract enhanced the activity but only up to certain extent. The TLC finger printing profile and chemical identification tests can be used for the identification/standardization purpose of this active fraction. Key words: Rubia cordifolia, Tephrosia purpurea, Viola tricolor, Serenoa repens, antibacterial activity.

DETERMINATION OF PHYSICOCHEMICAL PROPERTIES, ANTIOXIDANT CONSTITUENTS BY HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY FINGERPRINTING, AND ANTIOXIDANT ACTIVITY OF CUCURBITA MAXIMA SEEDS

Objective: The present study is designed to evaluate preliminary phytochemical constituents, physiochemical properties evaluation, free radical scavenging activity, and high-performance thin-layer chromatography (HPTLC) fingerprinting analysis of Cucurbita maxima seed oil.Methods: Total phenol content, total flavonoid content, total tannin content, 1, 1-diphenyl, 2-picryl-hydrazyl (DPPH) radical scavenging method, and HPTLC fingerprinting analysis for compounds responsible for antioxidant activity were used.Result: Phytochemical screening shows the presence of constituents such as alkaloids, glycosides, carbohydrates, fixed oil and fats, phytosterols, saponin, phenolic compounds, tannins, proteins, amino acids, gums, and mucilage. Physicochemical studies show that the oil contains an acid value, saponification value, iodine value of 9.53 mg KOH/g, 183 mg KOH/g, and 116.51 mg I2/100 g, respectively. Total phenol content was 3.9498±3.2 mg gallic acid equivalent/g, total flavonoid content was 56.115±4.6 mg rutin equivalent/g, and total tannin content was 19.4±0.40 mg tannic acid equivalent/g. The radical scavenging activity by DPPH Model shows that the methanolic extract has maximum antioxidant activity with IC50 value of 238.31. HPTLC fingerprinting also proofs the presence of phenol and flavonoid compounds in the seed oil.Conclusion: The results in the paper show that the C. maxima seed oil is an natural antioxidant and can be used for the treatment of various disorders.

Thin-layer chromatography fingerprint and chemometric analysis of selected Bryophyta species with their cytotoxic activity

Twenty land mosses were extracted by double maceration and ultrasonic extraction techniques using the mixture of 80% ethanol and water. The obtained extracts were analyzed using thin-layer chromatography (TLC) with silica gel (the mobile phase was consisted of ethyl acetate–formic acid–acetic acid–water, 14.0:1.5:1.5:2.0, v/v) and RP-18 (the composition of eluent methanol–water–formic acid, 7.0:2.5:0.5, v/v) chromatographic plates. After developing and drying, the plates were sprayed using the Naturstoff reagent, and after drying, the chromatograms were photographed and the obtained images were processed using the ImageJ program. Principal component analysis was performed to confirm the chemical similarity between the studied moss extracts. The cytotoxic activity of the ethanolic extracts of Bryophyta species was studied using the cell lines CCRF/CEM and CEM. For most of the moss samples, the cytotoxic activity was confirmed.

HPTLC Fingerprinting Analysis of Tannin Profile on Canthium coromandelicum and Flueggea leucopyrus willd.

The study of chemical composition of medicinal plants or drugs derived from plants is said to be Phytochemistry. Tannins are polyphenolic compounds; they are identified as better antiseptic due to the presence of phenolic group. The main objective of this study was to progress the High Performance Thin Layer Chromatography (HPTLC) fingerprinting of tannin profile on methanolic extracts of leaves and stem of Canthium coromandelicum and Flueggea leucopyrus willd aerial parts. The HPTLC fingerprinting analysis was carried for tannin using CAMAG LINOMAT 5 instrument which revealed that the methanolic extracts of Canthium coromandelicum leaves, stem and aerial parts Flueggea leucopyrus showed the presence of 10, 8 and 7 peaks with Rf values in the range of 0.04 to 0.97 respectively. Thus, the HPTLC methods developed in this study against the methanolic extract of selected plants provide useful and simple tools for the quality assessment and identification of phytochemical marker compounds. By isolating and identifying the biomarker compounds, new drugs can be synthesised to treat various ailments.

Antidiabetic, toxicological, and metabolomic profiling of aqueous extract of Cichorium intybus seeds

Background: Cichorium intybus has a wide range of therapeutic applications in Indian traditional systems of medicine, especially in metabolic disorders. Objective: To evaluate the toxicity profile and to investigate the antidiabetic, antihyperlipidemic, and antioxidative efficacy of C. intybus seeds in Wistar rats. Materials and Methods: The aqueous extract of seeds was prepared by decoction, and its quality control analysis was carried out by thin-layer chromatography and ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) fingerprinting. Wistar rats were fed with high-fat diet for 5 weeks followed by a single dose of streptozotocin intraperitoneally to induce diabetes. The protective group of rats was given aqueous extract during and after the induction of type 2 diabetes mellitus. Further, repeated dose 28-day (subacute) and repeated dose 90-day (chronic) toxicity studies were conducted as per the OECD guidelines. Results: A total of 18 metabolites have been tentatively identified by UPLC-MS profiling in aqueous extract of C. intybus seeds. No significant changes in mortality and biochemical parameters have been observed during toxicity studies. Moreover, administration of the extract to a protective group of diabetic rats attenuated serum glucose and triglyceride levels by 52.7% and 65.3%, respectively, supported by similar results for parameters related to insulin resistance and oxidative stress. The beneficial effect of extract has also been confirmed through in silico screening. Conclusion: C. intybus can be used as a natural dietary supplement for the prevention and management of diabetes and can be explored to develop a potent phytopharmaceutical for diabetes. Abbreviations used: AECIS: Aqueous extract of Cichorium intybus seeds; HOMA-IR: Homeostatic model assessment of insulin resistance; OGTT: Oral glucose tolerance test; TLC: Thin-layer chromatography; UPLC-MS: Ultra-performance liquid chromatography-mass spectrometry.

Comparative Evaluation of Thin-layer Chromatography/ High-performance Thin-layer Chromatography Fingerprint Profiles of Successive Extracts in Development of Standardization Protocol for Ayurvedic, Siddha or Unani Formulations

Aim: Trikatu churna is an important and unique polyherbal formulation used for various therapeutic indications. The World Health Organization (WHO) has emphasized the need to ensure the quality of medicinal plant products and herbal formulations by using any modern controlled techniques. High-performance thin-layer chromatography (HPTLC) is a frequently used technique to develop the chromatographic fingerprints. Therefore, the present study was carried out for quality evaluation of Trikatu churna through the development of standardization protocol with comparative HPTLC fingerprint profiles of successive extracts besides routine analysis. Materials and methods: Successive extracts of various solvents of polarity in increasing order through soxhlet apparatus for Trikatu churna and its authenticated raw materials were prepared; comparative chromatographic fingerprint profiles developed along with the evaluation of qualitative and quantitative tests parameters have been carried out by using the Ultraviolet (UV)–visible spectrophotometer, flame photometer, and atomic absorption spectrophotometer. Results: The following test parameters physicochemical parameters, evaluation of qualitative, quantitative phytochemicals such as total tannins, sugars and phenols, micronutrients such as sodium (Na), potassium (K); safety parameters such as microbial load, heavy metals and aflatoxins; comparative TLC and HPTLC fingerprint profiles of successive extracts of ingredients in increasing the polarity order of the solvents (n–hexane to alcohol) with respect to that of consecutive extracts of the final polyherbal formulation have been carried out for the quality evaluation. Conclusion: Trikatu churna was evaluated for its identity and purity by the systematic analysis of the above parameters for raw botanical ingredients and formulation in the course of development of standardization protocol for quality control.

Central nervous system activity of phenol-rich fraction of Piper sylvaticum roots

Obective: Piper sylvaticum Roxb. is an important folk medicine in Indian Traditional System of Medicine widely used by different tribes in many countries. In the present study, the anticonvulsant activity of extract/fractions of Piper sylvaticum (PS) roots was investigated. Materials and Methods: 70% ethanolic extract of PS roots was successively extracted using hexane and ethyl acetate to prepare various fractions. Total phenol content was found at maximum 324.65 mg/gallic acid equivalent/g in ethyl acetate fraction (EAF) (phenol-rich fraction [PRF]). High-performance thin-layer chromatography fingerprinting profiling of PS roots was performed. The anticonvulsant properties of the EAF of roots of PS were examined by maximal electroshock method as compared to standard phenytoin (25 mg/kg body weight). Result: It was found that EAF shows potent anticonvulsant activity at different dose levels against maximum electroshock seizure-induced convulsions in Swiss albino mice. Conclusion: From the observation, it can be concluded that the current study has expressed that the phenol-rich EAF of the ethanolic extract of the roots of PS has shown the dose-dependent anticonvulsant effect in mice. The anticonvulsant potential may be due to the presence of phenolic compounds in PRF. The outcomes suggested a high potential for application of EAF of PS root as an anticonvulsant agent. Abbreviations Used: HPTLC: High-performance thin-layer chromatography, OECD: Organization for Economic Co-operation and Development, GAE: Gallic acid equivalent, PRF: Phenol-rich fraction.

English

Cassia sieberiana DC. (Leguminosae - Caesalpinioideae), commonly known as drumstick and ‘aridantooro’ in Yoruba, is a perennial tree native to Africa. It is used in ethno-medicine to manage arthritis and rheumatism. Pharmacological activities such as myorelaxant, antispasmodic, anti-inflammatory, and antimicrobial have been reported in literature. Pharmacognostic investigation including microscopy, chemomicroscopy, physicochemical analysis and phytochemical investigations including thin layer chromatographic finger printing were conducted on fresh and powdered leaf and root samples of this plant. The macro and microscopic studies revealed the leaves to be simple, petiolated, glabrous and pinnately veinnated. The lower epidermal surface is characterized by abundant anomocytic stomata, polygonal epidermal cells and numerous uniseriate, unicellular trichomes. Quantitative leaf analysis revealed the following: stomatal number (163.8), stomatal index (19.04), palisade ratio (17.01), vein islet number (56.45) and vein termination number (61.45). Chemomicroscopic characters present include lignins, tannins, mucilage, starch, oils and calcium oxalate crystals. The physicochemical parameters evaluated are moisture content of 6.3%, total ash of 4.2%, acid-insoluble ash of 3.4%, sulphated ash of 11.0%, water-soluble ash of 0.8%, alcohol-soluble extractive of 21.3%, and water-soluble extractive of 16.7%. Chromatographic fingerprints of ethanol 70% extracts show major spots at Rf = 0.18 daylight (brown), UV366 (deep brown), spray reagent at 100°C (brown); Rf = 0.57 daylight (brown), UV366 (deep brown), spray reagent at 100°C (brown); Rf = 0.89 daylight (green), UV366 (red), spray reagent at 100°C (brown). The pharmacognostic evaluation of the leaves of C. sieberiana is reported here for the first time. The results of this research provide information which can be included in official monograph of the plant for its proper identification and quality control. Key words: Cassia sieberiana, pharmacognostic studies, physicochemical studies, chemomicroscopy.

Quality assessment of Herba Leonuri based on the analysis of multiple components using normal- and reversed-phase chromatographic methods.

A novel, improved, and comprehensive method for quality evaluation and discrimination of Herba Leonuri has been developed and validated based on normal- and reversed-phase chromatographic methods. To identify Herba Leonuri, normal- and reversed-phase high-performance thin-layer chromatography fingerprints were obtained by comparing the colors and Rf values of the bands, and reversed-phase high-performance liquid chromatography fingerprints were obtained by using an Agilent Poroshell 120 SB-C18 within 28min. By similarity analysis and hierarchical clustering analysis, we show that there are similar chromatographic patterns in Herba Leonuri samples, but significant differences in counterfeits and variants. To quantify the bio-active components of Herba Leonuri, reversed-phase high-performance liquid chromatography was performed to analyze syringate, leonurine, quercetin-3-O-robiniaglycoside, hyperoside, rutin, isoquercitrin, wogonin, and genkwanin simultaneously by single standard to determine multi-components method with rutin as internal standard. Meanwhile, normal-phase high-performance liquid chromatography was performed by using an Agilent ZORBAX HILIC Plus within 6min to determine trigonelline and stachydrine using trigonelline as internal standard. Innovatively, among these compounds, bio-active components of quercetin-3-O-robiniaglycoside and trigonelline were first determined in Herba Leonuri. In general, the method integrating multi-chromatographic analyses offered an efficient way for the standardization and identification of Herba Leonuri.

Thin-layer chromatography—fingerprint, antioxidant activity, and gas chromatography—mass spectrometry profiling of several Origanum L. species

Essential oils obtained from Origanum species are characterized by high diversity in composition. In this work, thin-layer chromatography (TLC)—fingerprinting of Origanum species was performed for the first time and enabled the discrimination of three chemotypes (carvacrol, caryophyllene oxide, and terpineol/sabinyl). Gas chromatography—mass spectrometry and statistical analyses confirmed the presence of the mentioned chemotypes and deepened the characterization of the studied essential oils. TLC—bioautography showed that thymol and carvacrol are the main antioxidant compounds in Origanum sp. essential oils (EOs).

IMPACT OF FERMENTING AGENTS ON MARKETED PATENT PROPRIETARY FORMULATION ASHWAGANDHA KALPA

Objective: The present work is aimed to formulate Ashwagandha kalpa and evaluate the impact of fermenting agent. Methods: A. kalpa is a marketed patent proprietary Ayurvedic product prepared by adopting the Ayurvedic principles of madya sandhana and it belongs to the class of formulations known as Sandhana kalpana . It is given to the patients suffering from hypertension, insomnia, paralysis, loss of concentration, etc. In the preparation of madya sandhana , the sandhana dravya (fermenting agents) play a key role. The present study was taken up to see the effect of three different sandhana dravya (fermenting agents), i.e., Dhataki pushpa (DAK) , Madhuka (MAK), and yeast (YAK) on the formulation characterization of A. kalpa . Results: The prepared samples were tested for their physicochemical parameters, i.e., total solid, pH, specific gravity, sugar content, alcohol content, refractive index, phytochemical screening, quantitative assay for secondary metabolites, and high-performance thin-layer chromatography (HPTLC) fingerprinting. The findings of phytochemical evaluation, quantitative assay, and HPTLC fingerprint show marked variation among three different prepared samples of A. kalpa. The preliminary phytochemical estimation for the detection of secondary metabolites was done. The study revealed the presence of highest percentage of alkaloids in YAK, tannins in DAK and saponins in YAK. Total solid content was found to the highest in MAK. Physicochemical parameters such as pH, total solid content, refractive index, alcohol content, and specific gravity also showed marked variation. Alcohol content for the DAK and YAK was found same. Conclusion: s andhana dravya (fermenting agent) causes variations in different physicochemical and phytochemical parameters in the formulation and development of A. kalpa.

IMPACT OF FERMENTING AGENTS ON MARKETED PATENT PROPRIETARY FORMULATION ASHWAGANDHA KALPA

Objective: The present work is aimed to formulate Ashwagandha kalpa and evaluate the impact of fermenting agent.Methods: A. kalpa is a marketed patent proprietary Ayurvedic product prepared by adopting the Ayurvedic principles of madya sandhana and it belongs to the class of formulations known as Sandhana kalpana. It is given to the patients suffering from hypertension, insomnia, paralysis, loss of concentration, etc. In the preparation of madya sandhana, the sandhana dravya (fermenting agents) play a key role. The present study was taken up to see the effect of three different sandhana dravya (fermenting agents), i.e., Dhataki pushpa (DAK), Madhuka (MAK), and yeast (YAK) on the formulation characterization of A. kalpa.Results: The prepared samples were tested for their physicochemical parameters, i.e., total solid, pH, specific gravity, sugar content, alcohol content, refractive index, phytochemical screening, quantitative assay for secondary metabolites, and high-performance thin-layer chromatography (HPTLC) fingerprinting. The findings of phytochemical evaluation, quantitative assay, and HPTLC fingerprint show marked variation among three different prepared samples of A. kalpa. The preliminary phytochemical estimation for the detection of secondary metabolites was done. The study revealed the presence of highest percentage of alkaloids in YAK, tannins in DAK and saponins in YAK. Total solid content was found to the highest in MAK. Physicochemical parameters such as pH, total solid content, refractive index, alcohol content, and specific gravity also showed marked variation. Alcohol content for the DAK and YAK was found same.Conclusion: sandhana dravya (fermenting agent) causes variations in different physicochemical and phytochemical parameters in the formulation and development of A. kalpa.

PHARMACOGNOSTIC SCREENING STUDIES OF JUSTICIA GENDARUSSA BURM. LEAVES FOUND IN DEHRADUN DISTRICT OF UTTARAKHAND

Objective: Justicia gendarussa Burm. (family Acanthaceae) commonly known as nilinirgundi, is found in Southern India possesses multifarious biological activities due to large range of phytoconstituents. The present study is designed to evaluate the various pharmacognostic parameters of the leaves of J. gendarussa, found in Dehradun district of Uttarakhand for its authentication.Methods: Fresh leaves were taken for the morphological and microscopical (histology and powder) evaluation. Physicochemical parameters (ash values, extractives values, florescence analysis, microbial contamination, and loss on drying) were also performed. Phytochemical screening and thin-layer chromatographic fingerprinting of extracts were also performed to check the presence of various phytoconstituents.Results: The microscopy of the leaves evinced the presence of anisocytic stomata, cuboidal calcium oxalate crystals, cystoliths, multicellular covering trichomes, starch grains and oil globules. The quantitative estimation of total ash, acid insoluble, and water soluble ash values were 13.8%, 1.2%, and 4.5% w/w, respectively. The alcohol soluble and water soluble extractives were estimated as 11.45% and 15.67% w/w, respectively. Foreign organic matter and loss on drying values obtained were 0.23% and 11.2% w/w. Phytochemical screening of petroleum ether, chloroform, methanol and aqueous extracts ascertained the presence of alkaloids, phenolic compounds, saponins, tannins, carbohydrates, flavonoids, glycosides, steroids and triterpenoids. The thin-layer chromatography (TLC) profiling of different extracts revealed the presence of potential compounds which can be further isolated with the help of high-performance liquid chromatography or high-performance TLC.Conclusion: The results of this study provide suitable standards for the authentication of this plant. In the present study, there are certain variations observed from the evaluations done on the same species by other research groups. The probable reason suggested for such disparity is due to the environmental and geographical differences in the locations of the plant collected.

High-performance thin-layer chromatographic fingerprint method for the detection of sennosides in Cassia senna L. and Cassia angustifolia Vahl

A Ph. Eur. herbal drug monograph requires for identification testing besides macroscopic (A) and microscopic (B) evaluation a thin-layer chromatography (TLC) fingerprint, prescribed as identification testing in subsection C. The main active constituents of Cassia senna L. and Cassia angustifolia Vahl are the dianthrone glucosides (mainly sennoside A and B). Therefore, this study presents a high-performance thin-layer chromatography (HPTLC) method which uses the sennosides as characteristic constituents. The presented method results in a fast and simple fingerprint with characteristic yellow fluorescence for the sennosides due to derivatization with a potassium hydroxide solution. The use of a senna herbal drug dry extract as system suitability test (SST) allows for a cost-effective test control. The resolution of the sennosides zones is dependent on the extractives concentration. An overload of the analytes leads to tailing (u-shaped form) and broadening as well as change in the retardation factor (RF). T...

Clinical Evaluation of a Polyherbal Nutritional Supplement in Dyslipidemic Volunteers

ABSTRACTTen important plant parts routinely used in South Indian ethnic food preparation as spices and condiments were investigated for their potential antidyslipidemic properties. The aim of the study was to characterize the biochemical properties of the polyherbal formulation (nutritional supplement) and evaluate its use to control dyslipidemia in patients. Phytochemical evaluation, in vitro α-amylase inhibitory assay, and high performance thin layer chromatography (HPTLC) fingerprinting were carried out with alcoholic extracts of all 10 individual plants and with the nutritional supplement. Investigation in human volunteers was conducted to evaluate the effect on dyslipidemia as measured by serum lipid biomarkers. Sixty-five volunteers were recruited for this study. Biomarker values at baseline and at 6th visit (end of review, 8/9 months) were compared to assess the usefulness of the nutritional supplement in the normalization of lipid biomarkers. In the qualitative analysis of metabolites, the results revealed the presence of various bioactive primary and secondary metabolites that might be responsible for their medicinal attributes. In human volunteers, after supplement intake along with standard therapy, we observed significant decrease in serum cholesterol, triglyceride, low-density lipoprotein (LDL), and very low-density lipoprotein (VLDL) levels. High-density lipoprotein (HDL) level did not change in test patient volunteers. Reductions in hemoglobin A1C (HBA1C) and postprandial blood sugar levels were observed; the difference was not statistically significant. We believe that the polyherbal formulation of 10 medicinal plants has potent antidyslipidemic activity. Our results contribute for the first time toward documentation of augmented dyslipidemia control by use of the formulation.

Thin layer chromatography fingerprint, antioxidant, and antibacterial activities of rhizomes, stems, and leaves of Curcuma aeruginosa Roxb.

Fingerprints of 5 temu hitam (Curcuma aeruginosa Roxb.) accessions (Malang, Cirebon, Kuningan 1, Bogor, and Liwa) were determined by thin-layer chromatography (TLC) and compared to fingerprints of turmeric (Curcuma longa L), temu putih (Curcuma zedoaria (Christm.) Roscoe), and temu lawak (Curcuma zanthorriza Roxb.). Maceration method with ethanol as the solvent was used for extraction. The eluent used for fingerprint by TLC was chloroform:dichloromethane (9:1v/v). Five accessions of temu hitam show similar fingerprint patterns, but different in band thickness. Temu hitam rhizomes have bands of curcuminoid (Rf 0.22, 0.10, 0.03), and characteristic bands of Rf 0.42, 0.27, and 0.77, which can be distinguished from turmeric and temu lawak and Rf 0.13, which is different from temu putih. Leaves and stems of temu hitam can be distinguished from temu putih, turmeric, and temu lawak at Rf 0.60. Rhizomes of all plants reveal strong antibacterial activity against Staphylococcus aureus and antioxidant activity on DPPH radicals than its corresponding stems and leaves. Antibacterial and antioxidant activities were determined by microdilution and TLC-bioautography. Antibacterial activity of rhizomes of Cirebon and Kuningan 1 accessions are higher than that of other accessions (MIC = 250 μg/mL; MBC = 500 μg/mL, but lower as compared to that of temu lawak (MIC = 62.5 μg/mL, MBC = 250 μg/mL) and tetracycline (MIC = MBC = 15.63 μg/mL). Rhizome of Liwa accession exhibits the highest antioxidant activity (IC50 = 124.88 μg/mL) amongst all accessions, but lower than that of temu lawak (IC50 = 18.45 μg/mL), turmeric (IC50 = 18.82 μg/mL), and temu putih (IC50 = 94.35 μg/mL).