Performance Thin Layer Chromatography Fingerprint Research Articles

High performance thin layer chromatography fingerprint on cyano-bonded stationary phase of selected Stachys and Betonica species and their antioxidant activity with chemometric calculations

The methanolic extracts of selected Stachys and Betonica species were analyzed by high performance thin layer chromatography (HPTLC) method using cyano bonded stationary phase and mobile phase consisted of methanol and water 45:55 (v/v) with Natural product reagent derivatization. The HPTLC method was validated (linearity, precision, accuracy, robustness, LOD, LOQ) and the validation results confirm the usefulness of the method used. The obtained images of fingerprint chromatograms presented as numerical data (using the TLC analyzer program) were next processed using Excel and SpecAlign softwares. The phytochemical similarity between analyzed plant extracts was evaluated using the similarity index (Pearson correlation coefficient) and the results were presented as hierarchical cluster analysis dendrograms and as graph for principal component analysis (PCA). The total phenolic content (TPC) by Folin-Ciocalteu method and the antioxidant activity with Ferric Ion Reducing Antioxidant Potential (FRAP) technique were determined for studied extracts using gallic acid as standards in both methods.

Herbal Emulgel Containing Azadirachta indica (Neem) and Nigella Sativa L. (Black Cumin) Oils in Wound Management: Preclinical Investigations

Abstract Context: In wound management, traditional use of plants has received attention by the scientific community, as traditional medicines are a valuable source of affordable, comprehensive medical care. Azadirachta indica A. Juss (Neem) and Nigella sativa Linn (Black cumin) are well known versatile medicinal plants with wide spectrum of biological activity. Methods: This study was undertaken to evaluate the wound healing activity of neem oil (NO) and black cumin oil (BCO) in rats. The oils were standardized using standard procedures. Gas chromatography mass spectrometry analysis and high performance thin layer chromatography fingerprinting of oils were carried out to identify major phytoconstituents present in the oils. The oils were suitably incorporated in carbopol based emulgel formulation, evaluated for physicochemical parameters and stability. Results: The wound healing potential assessed by the measurement of wound contraction and histopathology studies in excision and burn wound models, demonstrated comparable efficacy with standard nano crystalline silver gel (0.002% w/w). This may be attributed to the presence of phytoconstituents such as nimbidin and thymoquinone present in selected oils. Polyunsaturated fatty acid profile of BCO may be responsible for stimulating the release of vascular endothelial growth factor, which plays a significant role in accelerating wound healing process. Conclusion: Topical carbopol 934 base emulgel containing natural oils such as BCO and NO have been successfully prepared for wound management. However, further studies to establish the mechanistic aspects of wound healing needs to be undertaken.

High Performance Thin Layer Chromatography Fingerprinting Analysis of Piper betle L. Leaves

Introduction: Many modern medicines used today based on plants and plant products. Piper betle is generally known as the betle vine, it is an important medicinal and recreational plant. High performance thin layer chromatography (HPTLC) is an advanced powerful analytical method with more separation power, high performance and superior reproducibility than classic thin layer chromatography (TLC). A chromatographic fingerprint of a plant extract is a chromatographic pattern of some common chemical constituents of pharmacologically active and/or chemical characteristics. Chromatographic fingerprints are useful in authentication and identification of plant.
 Objectives: Objectives of present research was to establish HPTLC fingerprinting of methanolic extract of Piper betle L. leaves.
 Materials and Methods: Methanolic extract of Piper betle leaves was prepared using soxhlet apparatus. HPTLC studies were performed using a CAMAG HPTLC system equipped with automatic TLC sampler-4 (ATS 4), TLC scanner 4, and vision CATS 3.0 software.
 Results: The study revealed the presence of alkaloids with Rf value 0.65, flavonoids with Rf values 0.19, 0.29, 0.72, 0.95., and phenolic compound with Rf value 0.7.
 Conclusion: The HPTLC fingerprinting profile developed for the methanolic extract of Piper betle L. leaves will help in proper identification of the plant.Piper betle

Chemical Profiling of Tylophora indica (Burm. F.) Merrill. Shoot Cultures Established Through Leaf Explant

Tylophora indica (Burm. F.) Merrill. is an important medicinal plant of Asclepiadaceae family. Leaf explants were utilized for establishment of shoot cultures in Murashige and Skoog’s medium fortified with sucrose (3%) and different cytokinins individually as well as in combinations. Optimum 28.90 ± 0.41 shoots regenerated in medium supplemented with 6-benzyladenine (BA, 4 µM) and kinetin (Kn, 8 µM), when leaves of second node were placed adaxially with upright position for 8 weeks. Further transferring it to same medium for another 4 weeks formed 63.80 ± 1.45 shoots. The shoots were successfully rooted using earlier published media for the same plant and healthy rooted shoots were hardened and acclimatized to field. High performance thin layer chromatography fingerprint revealed that the in vitro shoots were able to synthesize metabolites at par with in vivo shoots.

High performance thin layer chromatography fingerprint and antioxidant activities of Cyclea barbata in Thailand

High performance thin layer chromatography fingerprint and antioxidant activities of Cyclea barbata in Thailand

Tamarindus indica Fruit: Pharmacognostical Standardization, Detection of Contaminant, and In Vitro Antioxidant Activity.

Objective:The main objective of the current study was to perform pharmacognostical, physiochemical analysis and in vitro antioxidant activity of Tamarindus indica Linn. (Family: Fabaceae) fruit.Materials and Methods:The present study deals with pharmacognostical characters as identification parameters of the fruit, which were subjected to macro- and microscopical studies. Physiochemical analysis was performed as per World Health Organization–recommended parameters. Toxicological parameters such as heavy metals, aflatoxins, and pesticide residue and presence of microbial count were analyzed. This study also deals with the thin layer chromatography/high performance thin layer chromatography fingerprinting and antioxidant activity.Results:The microscopy study revealed the presence of epidermis, vascular bundles, parenchyma cells, mucilage fiber, starch grain, and nonlignified fibers. Physiochemical parameters such as loss on drying, moisture content, ash value, and extractive value were also determined. Heavy metal, aflatoxin, total microbial load, and pesticide residues were found to be variable but under the permissible limits.Conclusion:It is critical to analyze these parameters in each crude herbal drug before further processing to ensure their safety and efficacy for better approval at the international podium. This study revealed precise identification for the meticulous crude drug which will be valuable in detection and manage to adulterations of the raw material.

High performance thin layer chromatography fingerprint analysis of guava (Psidium guajava) leaves

High-performance thin layer chromatography (HPTLC) fingerprint analysis is commonly used for quality control of medicinal plants in term of identification and authentication. In this study, we have been developed HPTLC fingerprint analysis for identification of guava (Psidium guajava) leaves raw material. A mixture of chloroform, acetone, and formic acid in the ratio 10:2:1 was used as the optimum mobile phase in HPTLC silica plate and with 13 bands were detected. As reference marker we chose gallic acid (Rf = 0.21) and catechin (Rf = 0.11). The two compound were detected as pale black bands at 366 nm after derivatization with sulfuric acid 10% v/v (in methanol) reagent. Validation of the method was met within validation criteria, so the developed method could be used for quality control of guava leaves.

GC-MS Analysis and HPTLC Fingerprinting Profile of Hydroalcoholic Extract of Polygonum barbatum Linn. Leaves

An important medicinal plant, Polygonum barbatum was undertaken for gas chromatography-mass spectrometry analysis and high performance thin layer chromatography finger print profile to investigate the chemical constituent present in the hydroalcoholic extract of Polygonum barbatum leaves. Gas chromatography-mass spectrometry analysis shown that twenty two compounds were identified and named from the extract. Among these constituents, terpenoids are the major constituents present in this extract. Other major constituents were present in the extracts were carbohydrate and fatty acids. High performance thin layer chromatography fingerprint analysis of hydroalcoholic extract of P. barbatum was carried out by using ethyl acetate-hexane (7:3 v/v) as a mobile phase. From the high performance thin layer chromatography analysis, peak numbers 4, 5 and 6 shown highest % of peak area of 29.79, 21.42 and 10.67, respectively at 366 nm.

Standardization of Stamens of Mesua ferrea Linn.

India has a rich culture of medicinal herbs and spices, which includes Ayurvedic, Unani, Siddha and other traditional medicines but only very few have been studied chemically and pharmacognostically evaluated for their potential medicinal value. According to the World Health Organization, most populations still rely on traditional medicines for their psychological and physical health requirements. Nagkesar is a very famous ayurvedic herb used in treating fever, vomiting, urinary tract disorders, migraine and also used to preparation of several compound formulation viz. Candanabalalaksadi Taila, Kumaryasava, Nagakesaradi Curna, Kankasava, Eladi curna, Jatiphaladi curna and Karpuradi churna etc. The present paper provides a detailed account of the scientific evaluation of Mesua ferrea stamens. The study includes macro and microscopic characters, powder microscopic characteristics, High Performance Thin Layer Chromatography fingerprint profile, preliminary phytochemical screening and physicochemical parameters. The information generated by this particulars study provides relevant pharmacognostical and physicochemical data needed for proper identification and authentication of Mesua ferrea stamens.

High Performance Thin Layer Chromatography Fingerprint Profile of Rhizome Extracts of Five Important Curcuma Species

The genus Curcuma (Family: Zingiberaceae) has 80 different species. Some of the species have been used world-wide as functional food and traditional medicines. The bright coloured pigments in the rhizome of different Curcuma sp. come mainly from fat soluble polyphenolic compounds known as curcuminoids. It comprises of three different pigments: curcumin, demethoxycurcumin and bisdemethoxycurcumin whose concentration varies from the species to species. A variety of spectrophotometric methods have been used for the quantification of the curcuminoids. However, quantification of the individual curcuminoids is not possible with spectro-photometric methods. Very little information is available on the determination of the individual constituents of curcuminoids using high performance thin layer chromatography. This method has advantages in terms of accuracy, flexibility, parallel analysis of a large number of samples and different modes of development. The present study was undertaken to provide information on the physico-chemical characteristics of individual curcuminoid pigments of five important Curcuma sp. to facilitate their proper extraction, separation and identification of individual phyto-constituents and their further utilization in the crop improvement programme.

Alyssum homolocarpum seeds: phytochemical analysis and effects of the seed oil on neural stem cell proliferation and differentiation.

Pharmacognostic evaluation of medicinal plants may assess their current applications and possibly results in finding new active components. In this study, ash and extractive values and high performance thin layer chromatography fingerprints of Alyssum homolocarpum (Brassicaceae) seed extracts were investigated to elucidate its composition. Differential scanning calorimetry and gas chromatography-mass spectrometry analysis were employed to determine the components of A. homolocarpum seed oil (AHO). Neurosphere assay, in vitro differentiation and immunofluorescence analysis were performed to evaluate the effects of oral administration of AHO (0.5 or 1g/kg/day for 14days) on proliferation and differentiation of neural stem cells (NSCs) in adult male BALB/c mice. Total, acid-insoluble and water-soluble ash values were determined as 45.83±5.85, 6.67±2.89 and 28.33±2.89mg/g, respectively. The extractive values were 4.90, 0.43 and 0.56% (w/w) for n-hexane, dichloromethane and ethanolic extracts, respectively. Interestingly, AHO was mainly composed of α-linolenic acid (89.71%), β-sitosterol (3.3mg/g) and campesterol (0.86mg/g). Administration of AHO at 1g/kg/day significantly increased proliferation of NSCs, as evidenced by an increase in mean neurosphere-forming frequency per brain (872.7±15.17) and neurosphere diameter (101±2.48µm) compared to the control group (424.3±59.29 and 78.63±1.7µm, respectively; P<0.05). AHO treatment did not affect in vitro differentiation of the harvested NSCs. Our data show that A. homolocarpum seed oil is a rich source of α-linolenic acid and β-sitosterol with potential therapeutic application to enhance NSC proliferation and recruitment in neurological diseases.

Quantification of total phenol, flavonoid content and pharmacognostical evaluation including HPTLC fingerprinting for the standardization of Piper nigrum Linn fruits

ABSTRACT Objective: To carry out the physicochemical and phytochemical standardization with high performance thin layer chromatography fingerprinting of Piper nigrum L. (P. nigrum) fruits in order to ascertain the standard pharmacognostical parameters of this king of spices. Methods: Many standardization parameters like extractive values, total ash value, water soluble ash value and acid insoluble ash, moisture content, loss on drying and pH values of P. nigrum L. fruits were analyzed. The method of Harborne was adopted for the preliminary phytochemicals screening. Analysis of total phenolic and flavonoid contents, pesticides residues, aflatoxin and heavy metals were also performed. CAMAG-high performance thin layer chromatography system was used for fingerprinting of methanolic extract of P. nigrum L. fruits. Results: The results of phytochemicals testing indicated the presence of carbohydrates, phenolic compounds, flavonoids, alkaloids, proteins, saponins, lipids, sterols and tannins in various solvent extracts. Total phenolic and flavonoid contents in methanolic extract were found to be 1.728 1 mg/g and 1.087 μg/g, respectively. Heavy metals concentrations were found to be within standard limits. Aflatoxins and pesticides residues were absent. Conclusions: The outcome of this study might prove beneficial in herbal industries for identification, purification and standardization of P. nigrum L. fruits.

High performance thin layer chromatography fingerprinting, phytochemical and physico-chemical studies of anti-diabetic herbal extracts.

Introduction:Herbal medicines have gained increasing popularity in the last few decades, and this global resurgence of herbal medicines increases their commercial value. However, this increasing demand has resulted in a decline in their quality, primarily due to a lack of adequate regulations pertaining to herbal medicines.Aim:To develop an optimized methodology for the standardization of herbal raw materials.Materials and Methods:The present study has been designed to examine each of the five herbal anti-diabetic drugs, Gymnema sylvester R. Br., Pterocarpus marsupium Roxburgh., Enicostema littorale Blume., Syzygium cumini (L.) Skeels. and Emblica officinalis Gaertn. The in-house extracts and marketed extracts were evaluated using physicochemical parameters, preliminary phytochemical screening, quantification of polyphenols (Folin–Ciocalteu colorimetric method) and high performance thin layer chromatography (HPTLC) fingerprint profiling with reference to marker compounds in plant extracts.Results:All the plants mainly contain polyphenolic compounds and are quantified in the range of 3.6–21.72% w/w. E. officinalis contain the highest and E. littorale contain the lowest content of polyphenol among plant extracts analyzed. HPTLC fingerprinting showed that the in-house extracts were of better quality than marketed extracts.Conclusion:The results obtained from the study could be utilized for setting limits for the reference phytoconstituents (biomarker) for the quality control and quality assurance of these anti-diabetic drugs.

Antioxidant potential of Rumex vesicarius L.: in vitro approach

To assess in-vitro antioxidant activity of different fraction and perform high performance thin layer chromatography fingerprint analysis of most active fraction of Rumex vesicarius L. (R. vesicarius). In the present study, acetone, ethyl acetate, n-butanol, and methanol extracts of R. vesicarius were evaluated for radical scavenging activity by studying the inhibition of the level of lipid peroxidation induced by Fe(++)/ascorbate, DNA sugar damage, scavenging of hydrogen peroxide, diphenylphosphine DPPH radical scavenging activity, total phenolic content, total flavonoids content and total proanthocyanidin. High performance thin layer chromatography finger print profiling of R. vesicarius L. was also done. Lipid peroxidation induced by the iron/ascorbate system, hydrogen peroxide, diphenylphosphine and DNA sugar damage were inhibited by the addition of different extract of R. vesicarius. Among them, methanolic extract showed maximum efficacy. The methanolic extract showed the highest total phenolic, total flavonoids and total proanthocyanidin contents. The results suggest that the extracts can be a vital source of phytochemical antioxidants.