Rapid validated high performance thin layer chromatography method for simultaneous estimation of mangiferin and scopoletin in Canscora decussata (South Indian Shankhpushpi) extract

AbstractMangiferin (polyphenolic xanthone) and scopoletin (phenolic coumarin) are well-studied biological markers present in Canscora decussata(Roxb.) Roem. & Schult., Gentianaceae. The objective set for the present studies is to establish and develop a new, simple, selective, sensitive, and precise high performance thin layer chromatography method for the simultaneous estimation of mangiferin and scopoletin in hydroalcoholic extract of C. decussata. The thin layer chromatographic separation of these biomarkers was carried out on aluminum plate pre-coated with silica gel 60F254, eluted with ethyl acetate:acetic acid:formic acid:water (10:0.5:0.5:1.5). The plate was then dried and densitometric scanning was performed at 254 nm using a Camag TLC scanner III. The system was found to give compact spots for mangiferin (RF 0.22) and scopoletin (RF 0.78). A good relationship of linear precision between the concentrations (100–600 ng/spot) and peak areas was obtained with correlation coefficient (r) of 0.9979 (mangiferin) and 0.9962 (scopoletin), respectively. The limits of detection and limit of quantification were determined to be 46 and 94 ng/spot for mangiferin and 31 and 78 ng/spot for scopoletin respectively. The percentage of recovery was found from 99.91 to 99.94% for mangiferin and 99.75 to 99.86% for scopoletin. Results obtained from recovery studies showed excellent reliability and reproducibility of the method. Present communication on validated high performance thin layer chromatography method may provide a new, selective, sensitive, and precise method to estimate mangiferin and scopoletin as phytomarkers in the hydroalcoholic extract of C. decussata used in Ayurvedic formulations.

Antioxidant markers based TLC-DPPH differentiation on four commercialized botanical sources of Shankhpushpi (A Medhya Rasayana): A preliminary assessment.

Shankhpushpi is a cognition boosting traditional ayurvedic brain supplement. Convolvulus pluricaulis (Convolvulaceae), Evolvulus alsinoides (Convolvulaceae), Clitoria ternatea (Papilionaceae), and Canscora decussata (Gentianaceae) are botanical claimants of Shankhpushpi. This investigation is to focus the identification of the compound based on biological marker differentiation of four botanical claimants of Shankhpushpi for their antioxidant evaluation on thin layer chromatography (TLC) by 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. A rapid TLC-DPPH method was developed to identify and differentiate four botanical claimants of Shankhpushpi in terms of presence of ß-carotene, rutin, scopoletin, chlorogenic acid, and mangiferin. C. pluricaulis shows presence of scopoletin; E. alsinoides shows presence of ß-carotene, scopoletin, and chlorogenic acid; C. ternatea shows presence of ß-carotene, scopoletin, and rutin; and C. decussata shows presence of ß-carotene, scopoletin, and mangiferin. The order, they followed, based on their antioxidant potential is ß-carotene < mangiferin < rutin < scopoletin < chlorogenic acid. Antioxidants are attributed for their beneficial role in age-related cognition decline. The proposed method provides an edge in terms of identification and quantification of antioxidant constituents in a multi-component system. This method may also provide application for identification of correct plant sources used in the name of Shankhpushpi in marketed ayurvedic formulation, food supplement, and extracts.

Protective role of standardized Feronia limonia stem bark methanolic extract against carbon tetrachloride induced hepatotoxicity.

OBJECTIVE: This study evaluates hepatoprotective potential of Feronia limonia stem bark (FSB) extracts and fractions using experimental models. MATERIALS AND METHODS: Activity levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) and cell viability were evaluated in HepG2 cells treated with carbon tetrachloride (CCl4) in presence or absence of FL extracts or fractions. Also, plasma markers of hepatic damage, hepatic antioxidants, lipid peroxidation and histopathological alterations were assessed in rats treated with CCl4 alone or in combination with 200 or 400 mg/kg bodyweight (BW) of FSB-7 or 25 mg/kg BW of silymarin. RESULTS: In vitro co-supplementation of FSB extracts or fractions recorded varying degree of hepatoprotective potentials. Also, pre-supplementation of FSB methanolic extract (FSB-7) followed by CCl4 treatment significantly prevented hepatic damage and depletion of cellular antioxidants. Also, CCl4+ FSB-7 group showed minimal distortion in the histoarchitecture of liver and results were comparable to that of CCl4+ silymarin treated rats. CONCLUSION: This inventory is the first scientific report on hepatoprotective potential of FSB methanolic extract.

Isolation, characterization and thin-layer chromatography method development of clerosterol palmityl ester: a chemical marker for standardization of leaves of Clerodendrum phlomidis.

OBJECTIVE: Clerosterol palmityl ester (CPE) is a unique clerosterol derivative isolated and characterized from the leaves of Clerodendrum phlomidis. Considering the uniqueness of this compound, the present study was planned to use CPE as a specific chemical marker and develop a new validated thin-layer chromatography (TLC) method for standardisation of C. phlomidis. METHODS: Separation and quantification of CPE were achieved by TLC using a mobile phase of petroleum ether (60 to 80 degrees centigrade) and ethyl acetate (95:5, volume ratio) (Rf 0.64) on precoated silica gel 60F(254) aluminium plates. Densitometric determination was carried out after derivatization with anisaldehyde sulphuric acid reagent in absorption mode at 527 nm. RESULTS: The calibration curve was linear in the concentration range of 100 to 500 ng/spot. The method was validated for precision, repeatability and accuracy. The proposed method was found to be simple, specific, precise, accurate, rapid and cost-effective. CONCLUSION: This TLC procedure may be used effectively for quantitative determination of CPE, identification of the plant and standardization of this plant or its derived products.

Isolation, characterization and thin-layer chromatography method development of clerosterol palmityl ester: a chemical marker for standardization of leaves of Clerodendrum phlomidis / 中西医结合学报

Clerosterol palmityl ester (CPE) is a unique clerosterol derivative isolated and characterized from the leaves of Clerodendrum phlomidis. Considering the uniqueness of this compound, the present study was planned to use CPE as a specific chemical marker and develop a new validated thin-layer chromatography (TLC) method for standardisation of C. phlomidis.

Standardization of Feronia limonia L. leaves by HPLC, HPTLC, physico-chemical and histological parameters / Estandarización de hojas de Feronia limonia L. por HPLC, HPTLC, parámetros fisicoquímicos e histológicos

Feronia limonia (Family Rutaceae, subfamily Aurantioideae), commonly known as kaitha or wood apple, is widely used as an ethnomedicine in India. Its leaves are prescribed for a wide variety of ailments like diarrhoea, urinary disorders, treatment of piles or haemorrhoids, acidity, ulcers, ringworm and other chronic skin infections. However, detailed scientific information is not available to identify the plant material and to ascertain its quality and purity. In the present communication, a qualitative fingerprinting of Feronia limonia (FL), extracts have been performed by HPTLC and HPLC methods, which provide qualitative insights into the bioactive constituents present in the extracts. Also, morphological anatomical and physico-chemical characters, along with phytochemical screening and fluorescence analysis of powdered crude drug were carried out for systemic identification and authentification of leaves. This study provides referential information for identification and characterization of FL leaf and its extracts.

Feronia limonia (Familia Rutaceae, subfamilia Aurantioideae), comúnmente conocida como kaitha o manzana de madera, y es extensamente usada como una ethnomedicina en India. Sus hojas son indicadas para una amplia variedad de dolencias como diarrea, desórdenes urinarios, el tratamiento de hemorroides, acidez, úlceras, tiña y otras infecciones crónicas de la piel. Sin embargo, la información científica detallada no está disponible para identificar el material de la especie y averiguar su calidad y pureza. En la presente comunicación, un análisis cualitativo de extractos de Feronia limonia (FL), se realizó por HPTLC y HPLC, lo que proporcionó información cualitativa de los componentes bioactivos presentes en los extractos. También, los caracteres anatómicos, fisico-químicos y morfológicos, junto con un examen fitoquímico y análisis de fluorescencia de la planta en polvo fueron realizados para la identificación sistémica y autentificación de las hojas. Este estudio proporciona la información de referencia para identificación y caracterización de las hojas de FL y sus extractos.

Comprehensive review of Clerodendrum phlomidis: a traditionally used bitter.

Clerodendrum phlomidis Linn. f., (syn. Clerodendrum multiflorum (Burm.f) O. Kuntze, Volkameria multiflorum Burm. f.) (Lamiaceae) is an important and well known medicinal plant extensively used in Ayurveda and Siddha system of medicine for treatment of various ailments. The popular therapies include on inflammation, diabetes, nervous disorder, asthma, rheumatism, digestive disorders, and urinary disorders as well as a bitter tonic. It was reported that pectolinaringenin, scutellarein, clerodin, clerodendrin, clerosterol, 24beta-ethylcholesta-5,22E,25-triene-3beta-ol, lup-20(29)-en-3-triacontanoate, 4,2',4'-trihydroxy-6'-methoxychalcone-4,4'alpha-D-diglucoside, 7-hydroxyflavone, 7-hydroxyflavanone-7-O-glucoside and alpha-L-rhamnopyranosyl-(1-2)alpha-D-glucopyranosyl-7-O-naringin-4'-O-alpha-D-glucopyranoside-5-methyl ether had been isolated from this plant. The alcoholic and aqueous extracts were reported active as analgesic, antidiarrhoeal, antiplasmodial, hypoglycemic, minor tranquilizers, anti-asthmatic, antifungal, nematicidal, anti-amnestic and anti-arthritic. There are coincidences between some of the traditional usages of this plant and experimentally observed effects of the extracts but very few biological studies available on bioactive fractions and/or pure compounds. This review is an attempt to compile the exhaustive literature on Clerodendrum phlomidis, to highlight, analyze and critically assess the pharmaceutical potential of this underestimated plant in a systematic way.

Comprehensive review of Clerodendrum phlomidis: a traditionally used bitter / 中西医结合学报

Clerodendrum phlomidis Linn. f., (syn. Clerodendrum multiflorum (Burm.f) O. Kuntze, Volkameria multiflorum Burm. f.) (Lamiaceae) is an important and well known medicinal plant extensively used in Ayurveda and Siddha system of medicine for treatment of various ailments. The popular therapies include on inflammation, diabetes, nervous disorder, asthma, rheumatism, digestive disorders, and urinary disorders as well as a bitter tonic. It was reported that pectolinaringenin, scutellarein, clerodin, clerodendrin, clerosterol, 24beta-ethylcholesta-5,22E,25-triene-3beta-ol, lup-20(29)-en-3-triacontanoate, 4,2',4'-trihydroxy-6'-methoxychalcone-4,4'alpha-D-diglucoside, 7-hydroxyflavone, 7-hydroxyflavanone-7-O-glucoside and alpha-L-rhamnopyranosyl-(1-2)alpha-D-glucopyranosyl-7-O-naringin-4'-O-alpha-D-glucopyranoside-5-methyl ether had been isolated from this plant. The alcoholic and aqueous extracts were reported active as analgesic, antidiarrhoeal, antiplasmodial, hypoglycemic, minor tranquilizers, anti-asthmatic, antifungal, nematicidal, anti-amnestic and anti-arthritic. There are coincidences between some of the traditional usages of this plant and experimentally observed effects of the extracts but very few biological studies available on bioactive fractions and/or pure compounds. This review is an attempt to compile the exhaustive literature on Clerodendrum phlomidis, to highlight, analyze and critically assess the pharmaceutical potential of this underestimated plant in a systematic way.

A novel triterpenoid isolated from the root bark of Ailanthus excelsa Roxb (Tree of Heaven), AECHL-1 as a potential anti-cancer agent.

BACKGROUND: We report here the isolation and characterization of a new compound Ailanthus excelsa chloroform extract-1 (AECHL-1) (C(29)H(36)O(10); molecular weight 543.8) from the root bark of Ailanthus excelsa Roxb. The compound possesses anti-cancer activity against a variety of cancer cell lines of different origin. PRINCIPAL FINDINGS: AECHL-1 treatment for 12 to 48 hr inhibited cell proliferation and induced death in B16F10, MDA-MB-231, MCF-7, and PC3 cells with minimum growth inhibition in normal HEK 293. The antitumor effect of AECHL-1 was comparable with that of the conventional antitumor drugs paclitaxel and cisplatin. AECHL-1-induced growth inhibition was associated with S/G(2)-M arrests in MDA-MB-231, MCF-7, and PC3 cells and a G(1) arrest in B16F10 cells. We observed microtubule disruption in MCF-7 cells treated with AECHL-1 in vitro. Compared with control, subcutaneous injection of AECHL-1 to the sites of tumor of mouse melanoma B16F10 implanted in C57BL/6 mice and human breast cancer MCF-7 cells in athymic nude mice resulted in significant decrease in tumor volume. In B16F10 tumors, AECHL-1 at 50 microg/mouse/day dose for 15 days resulted in increased expression of tumor suppressor proteins P53/p21, reduction in the expression of the oncogene c-Myc, and downregulation of cyclin D1 and cdk4. Additionally, AECHL-1 treatment resulted in the phosphorylation of p53 at serine 15 in B16F10 tumors, which seems to exhibit p53-dependent growth inhibitory responses. CONCLUSIONS: The present data demonstrate the activity of a triterpenoid AECHL-1 which possess a broad spectrum of activity against cancer cells. We propose here that AECHL-1 is a futuristic anti-cancer drug whose therapeutic potential needs to be widely explored for chemotherapy against cancer.