HPTLC Fingerprint Analysis Research Articles

Aldose reductase inhibitory activity of alcoholic extract of Pedalium murex Linn fruit

ObjectiveTo investigate physico-chemical investigation and aldose reductase inhibitory activity in rat lens aldose reductase (AR) enzyme of the ethanolic extract of fruits of Pedalium murex (P. murex) Linn (Pedaleaceae). MethodsPhysico-chemical investigation of ethanolic extract of fruit of the P. murex (EPM) was carried out. EPM was screened for aldose reductase inhibitory activity using rat lens AR enzyme. HPTLC fingerprinting analysis of EPM was performed in chloroform: methanol (8:2) solvent system. ResultsTotal ash, acid insoluble ash, water soluble ash, alcohol and water soluble extractive value and loss on drying of P. murex were found to be 4.2%, 1.2%, 0.92%, 6.2%, 5.0% and 7.5% respectively. Eleven prominent spots were found to be present in the EPM in HPTLC fingerprint analysis. EPM showed significant level of aldose reductase inhibitory potential [IC50 value (57.20±3.68) μg/mL]. From the value of Vmax, Km and Ki, it was found that EPM exhibits non-compititive inhibition with AR enzyme. ConclusionThe present study reveals scientific basis of the P. murex, which can be used in the treatment of the cataract disorder.

Antidiabetic and Antioxidative activity of Ethyl acetate Fraction of Hydromethanolic Extract of Seed of Eugenia jambolana Linn Through In-Vivo and In-Vitro Study and its Chromatographic Purification

Introduction: Eugenia jambolana of Myrtaceae family is widely distributed and used as diabetic therapeutic traditional medicine in rural India. Antidiabetic potentiality of ethyl acetate fraction of hydromethanolic (40:60) extract of seed of E. jambolana was investigated following in-vivo models in experimental diabetic rat and antioxidative efficacy following in‑vitro models. Materials and Methods: Alteration in carbohydrate metabolism during hyperglycaemia was assessed by increased fasting blood glucose, glycated hemoglobin levels along with diminished body weight, level of serum insulin and glycogen contents in liver, skeletal and cardiac tissues of diabetic rat. In-vitro antioxidative potentiality was measured by inhibitory activity of lipid peroxidation, scavenging activity against hydrogen peroxide, nitric oxide, hydroxyl, ABTS and DPPH radicals along with preliminary phytochemical analysis. Bioactive phytoingredients were isolated from ethyl acetate fraction through column chromatography, HPTLC fingerprinting and RP-HPLC analysis. Results: Oral administration of 20 mg ethyl acetate fraction or 0.6 mg glibenclamide in 0.5 mL water/100 g body weight/rat for twice a day at fasting state to diabetic rats for 28 days significantly (p<0.05) resettled carbohydrate metabolomics towards the control levels. This fraction established its primary antioxidant attribute by scavenging hydroxyl, nitric oxide, hydrogen peroxide, ABTS and DPPH radicals along with the inhibition of lipid peroxidation with IC50 values 26.49,28.14,18.45,13.65,10.46, 28.88 μg/mL respectively. Phytochemical screening confirmed isolated compounds were chemically gallic acid in nature. Two separate spots of ethyl acetate fraction were recorded after scanning of HPTLC fingerprinting. RP-HPLC study also shows two completely resolved peaks. Its biosafety profile was established following guidelines. Conclusion: On the basis of experimental studies ethyl acetate fraction of E. jambolana proved its antihyperglycemic and antioxidant nature.

Occurrence of Bioactive compounds in Ananus comosus (L.): A quality Standardization by HPTLC

ObjectiveAnanus comosus (L.) is belonging to the family Bromeliaceae used to cure various diseases. The present study is focus to evaluate the HPTLC fingerprinting analysis and phytochemical analysis of Ananus comosus peel. MethodsHPTLC fingerprinting profiles was done by using Hamilton syringe and CAMAG LINOMAT 5 instrument. ResultsAmong all solvents, most of the secondary metabolites such as alkaloid, flavanoid, phenol, tannins, steroids etc were present in ethanolic extract of Ananus comosus peel and the yield of plant extract in different solvents were analyzed, from which ethanolic extract gave 2.40g/100g. The quantitative assay was also analyzed in flavanoid, alkaloid, phenols, carotenoid and lycopene. HPTLC fingerprinting profile confirms the presence of phenols, flavanoid and alkaloid. ConclusionFrom the above results ethanolic extract of Ananus comosus peel can be used as therapeutic agents to treat various disorders caused by free radical and chemical substances due to presence of its secondary metabolites.

HPTLC Fingerprint Analysis of Plant Staminal Cells Products

Utilization of natural products is radically changing. Changes were mainly due to the outcome in the market of a plethora of new food supplements, and in particular those generally named botanicals for their common plant origin. The validation of these novel products needs powerful analytical devices tailored for the study of herbal extracts in order to assess composition and face their natural complexity as a resource. The last item is important and crucial for the capacity and utility of the analytical results that means that each product should be analyzed with the right approach. Having in mind these arguments, we selected HPTLC as useful tool for the analysis of products based on plant staminal (stem) cells. Nowadays these products, generally named bud-derivatives, are waiting scientific validation to obtain their own place into food supplements regulation, after gained that in the market. Our analyses, based on HPTLC fingerprints, were able to show bud-derivatives complex compositions that resulted very similar, but also in part different, to those of the corresponding leaf hydro-alcoholic extract.

Study on Triterpenoic Acids Distribution in Ganoderma Mushrooms by Automatic Multiple Development High Performance Thin Layer Chromatographic Fingerprint Analysis

Ganoderma--"Lingzhi" in Chinese--is one of the superior Chinese tonic materia medicas in China, Japan, and Korea. Two species, Ganoderma lucidum (Red Lingzhi) and G. sinense (Purple Lingzhi), have been included in the Chinese Pharmacopoeia since its 2000 Edition. However, some other species of Ganoderma are also available in the market. For example, there are five species divided by color called "Penta-colors Lingzhi" that have been advocated as being the most invigorating among the Lingzhi species; but there is no scientific evidence for such a claim. Morphological identification can serve as an effective practice for differentiating the various species, but the inherent quality has to be delineated by chemical analysis. Among the diverse constituents in Lingzhi, triterpenoids are commonly recognized as the major active ingredients. An automatic triple development HPTLC fingerprint analysis was carried out for detecting the distribution consistency of the triterpenoic acids in various Lingzhi samples. The chromatographic conditions were optimized as follows: stationary phase, precoated HPTLC silica gel 60 plate; mobile phase, toluene-ethyl acetate-methanol-formic acid (15 + 15 + 1 + 0.1); and triple-development using automatic multiple development equipment. The chromatograms showed good resolution, and the color images provided more specific HPTLC fingerprints than have been previously published. It was observed that the abundance of triterpenoic acids and consistent fingerprint pattern in Red Lingzhi (fruiting body of G. lucidum) outweighs the other species of Lingzhi.

HPTLC Fingerprinting of different leaf extracts of Tylophora indica (Burm f.) Merill.

Tylophora indica is very popularly used for the treatment of asthma based on its traditional use for asthma. Tylophora is perennial climbing plant native to the plains, forests, and hills of southern and eastern India. A method has been developed for different extracts of Tylophora indica for HPTLC fingerprinting analysis for identification and quantification of marker compound. For chloroform extract-Chloroform(90): Methanol (5) : Ethyl acetate (5) v/v, Methanol Extract-Toluene(5): Chloroform(90), Ethyl acetate(5) v/v and for Petroleum ether extract-Hexane(40) : Ethyl acetate (60) v/v. The HPTLC fingerprinting profile developed for different extracts of Tylophora indica will help in proper identification and quantification of marker compound.

Comparative phytochemical investigation of the sources of ayurvedic drugPatha: A chromatographic fingerprinting analysis

Standardization of herbal drugs based on their chemical and biological activity profile is an important prerequisite for acquiring the herbal market. The main problem encountered in standardization of Ayurvedic drugs is proper identification of the source plant. The present study was aimed to establish identification characters, quality control parameters, chemical and biological parameters for roots of three plants Cissampelos pareira, Cyclea peltata and Stephania japonica (Fam. Menispermaceae) which are being used as source of Patha, in the market. All the three plant were subjected for evaluation of quality control parameters as per WHO guidelines and root extracts and total alkaloidal fractions were subjected for HPTLC and HPLC fingerprinting analysis using a marker compound Bebeerine isolated from roots of Cissampelos pareira. The parameters studied clearly indicated the significant differences among the three plant materials. The roots of Cissampelos pareira can be distinguished from other two plants by presence of high concentration of alkaloids especially the presence of high concentration of pharmacologically active alkaloid bebeerine, which was found to be present in very low concentration in Stephania japonica and absent in roots of Cyclea peltata. The roots of Cyclea peltata were found to contain high concentration of saponins and comparatively in low concentration in Cissampelos pareira where as it was found to be absent in roots of Stephania japonica.

Phytochemical analysis and antibacterial activity of Vitex agnus-castus

The leaves of Vitex agnus-castus was sequentially extracted in hexane, ethyl acetate, methanol and aqueous medium and studied for in vitro antibacterial property. The ethyl acetate extract was found to be most active against all the bacterial species tested except K. pneumoniae. The best MIC value (0.312 mg/ml) was seen against MRSA. Active ethyl acetate extract was further studied for HPTLC fingerprint and phytochemical analysis. HPTLC analysis confirmed segregation of eight individual compounds with individual Rf values and peak area percentage. The results of phytochemical screening of extract revealed the presence of terpenoids, steroids, flavonoids and carbohydrates. This analysis revealed the high antibacterial activity in active ethyl acetate from Vitex agnus- castus. Key words: Antibacterial activity, drug resistant, minimum inhibitory concentration, phytochemical analysis, vitex agnus-castus

Estimation of guggulsteronesEandZin solid dosage forms containingCommiphora mukul, Hook

The oleo-gum-resin known as ‘guggul’, obtained from the plant Commiphora mukul , Hook (family Burseraceae), is widely used, as a component of formulations containing other Ayurvedic drugs, for treatment of hypercholesterolemia, rheumatism, and arthritis. Chemically, guggul contains two active keto steroids, guggulsterones E and Z , which are reported to be responsible for its antihyperlipidemic activity. These cis and trans isomers have been isolated, purified by TLC and column chromatography, and characterized by determination of melting point and by acquisition of UV, IR, and NMR spectra. After confirmation of their identity, they were used as biomarkers in HPTLC fingerprinting analysis of five commercial Ayurvedic formulations, four tablets and a capsule, all of which contain guggul as a constituent. The analytical method was fully validated. Extrapolation was used to calculate probable amounts of guggul added to the formulations on the basis of E and Z isomer content.