IN VITRO CALLUS INDUCTION AND COMPARATIVE GC-MS ANALYSIS OF METHANOLIC EXTRACTS OF CALLUS AND LEAF SAMPLES OF AMPELOCISSUS LATIFOLIA (ROXB.) PLANCH

Objective: The aim of the present study was to develop a callus induction protocol and comparative study of therapeutic phytochemicals present in in vivo leaf and in vitro callus extracts through Gas Chromatography-Mass Spectrometry analysis.Methods: Murashige and Skoog media was used as culture media for callus induction. In vitro callus induction protocol was developed by studying the effects of various plant growth regulators like auxin, 2, 4-D (2,4-dichlorophenoxyacetic acid), NAA (naphthalic acetic acid), alone and in combination with cytokinin BAP (benzyl aminopurine), on leaf and stem explants. The GC-MS analysis of Ampelocissus latifolia was carried out on Shimadzu QP-2010 plus with thermal desorption system TD 20 to study the phytochemical profile.Results: In vitro callus induction protocol was developed for the plant and callusing was done from leaf and stem explants of Ampelocissus latifolia. The best result for callus induction was obtained using leaf explant, and callus production were maximum in Murashige and Skoog medium fortified with BAP (0.5 mg/l) and NAA (1.0 mg/l). Major compounds identified in the GC-MS analysis were Campesterol, Stigmasterol, Beta-Sitosterol, Docosanol, Dodecanoic acid, etc., in in vitro extract and Beta Sitosterol, Tocopherol, Squalene, Bergamot oil, Margarinic acid, Hexadecanoic acid, etc., in in vivo extract. The different active phytochemicals identified have been found to possess a wide range of biological activities, thus this analysis forms a basis for the biological characterization and importance of the compounds identified for human benefits.Conclusion: This is the first report on callus induction in Ampelocissus latifolia. From the results obtained through the in vitro callus induction and its comparative GCMS analysis with in vivo extract, it is revealed that Ampelocissus latifolia contains various bioactive compounds that are of importance for phytopharmaceutical uses. The GCMS analysis revealed that the amount of Beta-sitosterol and 5-Hydroxymethylfurfural (HMF) was very high in in vitro extract as compared to in vivo extract

HPTLC METHOD FOR ESTIMATION AND QUANTIFICATION OF β-SITOSTEROL FROM IN-VIVO AND IN-VITRO SAMPLES OF MERREMIA AEGYPTIA AND MERREMIA DISSECTA

Objective: A normal-phase high-performance thin-layer chromatography (HPTLC) method has been developed and validated for estimation and quantitation of beta-sitosterol from the methanolic fraction of different plant parts of two medicinally important plants viz. Merremia aegyptia and Merremia dissecta. These plants have been reported to possess antimicrobial, antioxidant, and anti-inflammatory activities. Methods: Chromatographic separation of beta-sitosterol from the methanolic extracts of plant parts of M. aegyptia and M. dissecta was performed on TLC aluminum plates pre-coated with silica gel 60F254 using a suitable mobile phase. The densitometric scanning was done after derivatization at ????-580 nm for ????-sitosterol. Result: Only M. dissecta leaf sample was reported to contain ????-sitosterol (4.6 ng/μl), whereas other samples such as seed, stem, and callus extracts of M. aegyptia and M. dissecta did not showed its presence. Conclusion: The developed HPTLC method is simple, rapid, and precise and can be used for routine analysis and quantification of ????-sitosterol and other useful plant bioactives that are phytopharmaceutically important

HPTLC method for isolation, identification and quantification of quercetin from in vivo and in vitro samples of Naringi Crenulata

Naringi crenulata(Roxb.) Nicolson, is a rare medicinal plant belonging to the family Rutaceae. It is a spinous tree and has great medicinal value. During the present study a rapid, simple, accurate and specific HPTLC method for quantitative estimation of quercetin present in the dried leaf powder and callus of N. crenulata has been developed .The method used in this work resulted in good peak shape and enabled good resolution of quercetin from N. crenulata samples. Quercetin was identified in in vivo (leaf) and in vitro (six weeks old callus) tissues. Presence of isolated quercetin was further confirmed by superimposable IR spectra of isolated and authentic samples of quercetin and NMR spectra of isolated quercetin. Variation in quercetin content in in vivo and in vitro samples in N. crenulata was observed. In vivo leaf had maximum amount of quercetin (0.13%) while minimum amount was found in in vitro callus (0.032%). High content of quercetin in leaf shows its potential of synthesizing quercetin. This study is also of practical importance because flavonoid quercetin is the most active of all flavonoids. Many medicinal plants owe their activity to their high quercetin content. Several studies revealed quercetin’s significant anti-inflammatory activity due to direct inhibition of initial processes of inflammation. It also has potent antitumor and antioxidant properties including the inhibition of cancer cell proliferation and migration.This study is of practical importance because compound quercetin was firstly reported to be isolated from callus of N. Crenulata

Extraction and application of natural dye preparations from the floral parts of <i style="mso-bidi-font-style:normal">Woodfordia fruticosa</i> (Linn.) Kurz

403-408India has a rich biodiversity and harbours a wealth of useful germplasm resources and there is no doubt that the plant kingdom is a treasure-house of diverse natural products. One such product from nature is the dye. Dyes are one of the most important uses of the plants. Recently, interest in the use of natural dyes has been growing rapidly due to the result of stringent environmental standards imposed by many countries in response to toxic and allergic reactions associated with synthetic dyes. Nature has gifted us more than 500 dye-yielding plant species. One such medicinally much used dye-yielding plant species, Woodfordia fruticosa (Linn.) Kurz is exploited particularly, in perfume, leather and textile industry and believed to be superior for woolen and silk fabrics. The present study deals with the extraction of natural dye from this species, commonly known as Fire-Flame Bush, and their application on textiles. Three different techniques/methods for extraction of dye from the collected flowers were evaluated to determine the best extraction method. Three different types of fabrics and three different types of yarns were used in the experiment to observe the strength of dye. Cotton Jute mix sample showed dark yellowish brown colour with Myrobalan, dark blackish brown colour with Ferrous Sulphate, Camel colour with Stannous Chloride and Yellowish brown with Potassium dichromate

In vitro propagation and quercetin quantification in callus cultures of Rasna (Pluchea lanceolata Oliver & Hiern.)

383-387A protocol for micropropagation of Pluchea lanceolata, an important medicinal herb was developed. Leaf explants obtained from field grown plants when tested for callus induction on Murashige and Skoog’s (MS) medium, supplemented with NAA in combination with BAP, produced the best callus. Maximum number of multiple shoots from the callus (26.6±0.67) was obtained on MS medium supplemented with BAP (1.0 mg/L) and Kn (1.0 mg/L). More or less uniform elongation of multiple shoots was obtained on MS medium with lower concentrations of cytokinins, i.e., BAP (0.25 mg/L) and Kn (0.5 mg/L). Further elongation and profuse rooting were achieved when the well-grown shoots were cultured on half strength MS medium supplemented with IBA (1.0 mg/L). The regenerated plantlets were hardened and established at 70% survival rate in pots. The bioactive secondary metabolite, quercetin, was isolated from callus tissues of different age groups and its identification and confirmation was carried out by the colour reaction, TLC behaviour, IR spectrum and HPLC techniques. Maximum quercetin content (0.23 mg/g dry wt of tissue) was obtained in 6-wk-old callus tissues

Isolation, Identification and quantitative analysis of Ellagic acid: a tannin compound from Helicteres isora

Plant derived secondary metabolites have widely attracted humans with great interest due to their immense medicinal and pharmacological properties. Ellagic acid, a natural phenolic compound found in many fruits exhibits both antimutagenic and anticarcinogenic activity. Qualitative analysis of the plant samples of Helicteres isora showed the presence of ellagic acid in in vivo (stem bark) and in vitro (callus) samples. Presence of isolated ellagic acid was confirmed by superimposable IR spectra of isolated and authentic samples of ellagic acid. The ellagic acid was further identified and confirmed by using different techniques such as TLC (Rf 0.41), and HPLC (Rt =5.546 min) studies