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High-performance thin-layer chromatographic methods in the evaluation of the antioxidant and anti-hyperglycemic activity of Myrmecodia platytyrea as a promising opportunity in diabetes treatment

Agatonovic-Kustrin S, Morton DW, Adam A, Mizaton HH, Zakaria H

J Chromatogr A, 2017 Dec 29;1530:192-196.
PMID: 29132827 DOI: 10.1016/j.chroma.2017.11.012

The steady increase of diabetes is becoming a major burden on health care systems. As diabetic complications arise from oxidative stress, an antioxidant therapy along with anti-diabetic drugs is recommended. Myrmecodia or ant plant is highly valued as a traditional medicine in West Papua. It is used as an alternative treatment for diabetes, as the substances produced by ants can reduce blood sugar levels. The aim of this study was to develop and establish high-performance thin-layer chromatographic (HPTLC)-bioautographic methods to measure the antioxidant and hypoglycemic effects in different extracts from Myrmecodia platytyrea and to compare them with sterol content. Antioxidant activity in methanol, ethanol, dichloromethane (DCM) and ethyl acetate (EA) extracts were measured with a direct HPTLC-2,2-diphenyl-1-picrylhydrazyl free radical (DPPH) assay, while hypoglycemic effects were assessed using a newly developed α-amylase inhibitory activity assay. Stigmasterol is observed, after derivatization with anisaldehyde, as purple colored zones under visible light at hRF values of 0.66. The highest antioxidant activity was observed in the ethanol extract which is rich in polyphenols and flavonoids, while the DCM extract did not show antioxidant activity, but had significant α-amylase inhibitory activity. The highest α-amylase inhibitory activity was observed in the EA and DCM extracts and was related to their stigmasterol content.

Matched MeSH terms: Chromatography, Thin Layer/methods*
Multivariate analysis of PRISMA optimized TLC image for predicting antioxidant activity and identification of contributing compounds from Pereskia bleo

Sharif KM, Rahman MM, Azmir J, Khatib A, Sabina E, Shamsudin SH, et al.

Biomed Chromatogr, 2015 Dec;29(12):1826-33.
PMID: 26033701 DOI: 10.1002/bmc.3503

Multivariate analysis of thin-layer chromatography (TLC) images was modeled to predict antioxidant activity of Pereskia bleo leaves and to identify the contributing compounds of the activity. TLC was developed in optimized mobile phase using the 'PRISMA' optimization method and the image was then converted to wavelet signals and imported for multivariate analysis. An orthogonal partial least square (OPLS) model was developed consisting of a wavelet-converted TLC image and 2,2-diphynyl-picrylhydrazyl free radical scavenging activity of 24 different preparations of P. bleo as the x- and y-variables, respectively. The quality of the constructed OPLS model (1 + 1 + 0) with one predictive and one orthogonal component was evaluated by internal and external validity tests. The validated model was then used to identify the contributing spot from the TLC plate that was then analyzed by GC-MS after trimethylsilyl derivatization. Glycerol and amine compounds were mainly found to contribute to the antioxidant activity of the sample. An alternative method to predict the antioxidant activity of a new sample of P. bleo leaves has been developed.

Matched MeSH terms: Chromatography, Thin Layer/methods*
Rapid evaluation and comparison of natural products and antioxidant activity in calendula, feverfew, and German chamomile extracts

Agatonovic-Kustrin S, Babazadeh Ortakand D, Morton DW, Yusof AP

J Chromatogr A, 2015 Mar 13;1385:103-10.
PMID: 25666499 DOI: 10.1016/j.chroma.2015.01.067

The present study describes a simple high performance thin layer chromatographic (HPTLC) method for the simultaneous quantification of apigenin, chamazulene, bisabolol and the use of DPPH free radical as a post-chromatographic derivatization agent to compare the free radical scavenging activities of these components in leaf and flower head extracts from feverfew, German chamomile and marigold from the Asteraceae family. Feverfew (Tanacetum parthenium) leaves have been traditionally used in the treatment of migraine with parthenolide being the main bioactive compound. However, due to similar flowers, feverfew is sometimes mistaken for the German chamomile (Matricaria recutita). Bisabolol and chamazulene are the main components in chamomile essential oil. Marigold (Calendula officinalis) was included in the study for comparison, as it belongs to the same family. Parthenolide was found to be present in all leaf extracts but was not detected in calendula flower extract. Chamazulene and bisabolol were found to be present in higher concentrations in chamomile and Calendula flowers. Apigenin was detected and quantified only in chamomile extracts (highest concentration in flower head extracts). Antioxidant activity in sample extracts was compared by superimposing the chromatograms obtained after post-chromatographic derivatization with DPPH and post-chromatographic derivatization with anisaldehyde. It was found that extracts from chamomile flower heads and leaves have the most prominent antioxidant activity, with bisabolol and chamazulene being the most effective antioxidants.

Matched MeSH terms: Chromatography, Thin Layer
Safety assessment of standardised methanol extract of Cinnamomum burmannii

Ahmad M, Lim CP, Akowuah GA, Ismail NN, Hashim MA, Hor SY, et al.

Phytomedicine, 2013 Sep 15;20(12):1124-30.
PMID: 23827665 DOI: 10.1016/j.phymed.2013.05.005

The present study aims to evaluate the safety of methanol extract of Cinnamomum burmannii (MECB) by acute 14-day (single dose) and sub-chronic 28-day (repeated doses) oral administration to Sprague-Dawley rats. Our results showed that no toxicity was found in either acute or sub-chronic toxicity studies. MECB (containing 0.07% and 0.20% (w/w) of coumarin and trans-cinnamaldehyde, respectively), which was given orally at doses of 500, 1000 and 2000 mg/kg caused neither visible signs of toxicity nor mortality. No significant differences were observed in general condition, growth, organ weight, hematological parameters, biochemical values, or the gross and microscopic appearance of the organs from the treatment groups as compared to the control group. In conclusion, MECB did not cause any mortality nor did it cause any abnormalities in the necropsy and histopathology findings of treated rats. The LD50 for the MECB was found to be more than 2000 mg/kg. No adverse effects were observed in the treated rats at all the doses tested. The no-observed-adverse-effect level (NOAEL) for the 28-day study was determined to be 2000 mg/kg body weight/day.

Matched MeSH terms: Chromatography, Thin Layer
Fulltext Antihyperglycemic effect of orthosiphon stamineus benth leaves extract and its bioassay-guided fractions

Mohamed EA, Mohamed AJ, Asmawi MZ, Sadikun A, Ebrika OS, Yam MF

Molecules, 2011 May 04;16(5):3787-801.
PMID: 21544041 DOI: 10.3390/molecules16053787

Preliminary investigations were carried out to evaluate the antidiabetic effects of the leaves of O. stamineus extracted serially with solvents of increasing polarity (petroleum ether, chloroform, methanol and water); bioassay-guided purification of plant extracts using the subcutaneous glucose tolerance test (SbGTT) was also carried out. Only the chloroform extract, given at 1 g/kg body weight (b.w.), significantly reduced (P < 0.05) the blood glucose level of rats loaded subcutaneously with 150 mg/kg (b.w.) glucose. The active chloroform extract of O. stamineus was separated into five fractions using a dry flash column chromatography method. Out of the five fractions tested, only chloroform fraction 2 (Cƒ2), at the dose of 1 g/kg (b.w.) significantly inhibited (P < 0.05) blood glucose levels in SbGTT. Active Cƒ2 was split into two sub-fractions Cƒ2-A and Cƒ2-B, using a dry flash column chromatography method. The activities Cƒ2-A and Cƒ2-B were investigated using SbGTT, and the active sub-fraction was then further studied for anti-diabetic effects in a streptozotocin-induced diabetic rat model. The results clearly indicate that Cƒ2-B fraction exhibited a blood glucose lowering effect in fasted treated normal rats after glucose-loading of 150 mg/kg (b.w.). In the acute streptozotocin-induced diabetic rat model, Cƒ2-B did not exhibit a hypoglycemic effect on blood glucose levels up to 7 hours after treatment. Thus, it appears that Cƒ2-B functions similarly to metformin, which has no hypoglycemic effect but demonstrates an antihyperglycemic effect only in normogycemic models. The effect of Cƒ2-B may have no direct stimulatory effects on insulin secretion or on blood glucose levels in diabetic animal models. Verification of the active compound(s) within the active fraction (Cƒ2-B) indicated the presence of terpenoids and, flavonoids, including sinensitin.

Matched MeSH terms: Chromatography, Thin Layer
Fulltext Usage of traditional medicines among elderly and the prevalence of prednisolone contamination

Ismail Z, Mohamed R, Mohd Hassan MH, Wan Su K

Malays J Med Sci, 2005 Jul;12(2):50-5.
PMID: 22605958

The elderly consume many medications including traditional medicines. In 1986, it was found that 29% of elderly took traditional medicines although in 1996, the National Health Morbidity survey reported a 2.3% prevalence. However, studies from other countries showed much higher percentages. The Ministry of Health in Malaysia is concerned that some of these preparations maybe contaminated with steroids, antihistamines, hormones and other poisons. The aims of the study were to determine a). the health seeking behaviour of elderly Malays living in rural areas, b). the utilization of both modern and traditional medicines and c). the steroid content of the traditional medicines used. Methodology included interviews using structured questionnaires of elderly Malays living in rural areas of Kelantan, aged above 60 years. Samples of traditional medications collected were sent to the Pharmacology Department, School of Medical Sciences, Universiti Sains Malaysia, for steroid content analysis using Thin Layer Chromatography. A total of 599 elderly respondents were interviewed comprising 62.4% females and 37.6% males. The 60-69 years cohort group made up 48.7%, followed by 70-79 years at 36.1% and the remainder 15.2% were more than 80 years. There were 82% of elderly taking medicines. The trends of utilization of modern and traditional medicine in the last two weeks among elderly were 59.3% and 40.9% respectively. The utilization of traditional medicine by rural elderly Malays was therefore much higher than that reported in the previous study and nearly similar to that of France and Australian studies. There were 102 samples of traditional medications collected and analysed for steroid content. Results showed that 27.5% were positive for prednisolone, 34.3% positive for unknown steroids (a total of 61.8%) and 38.2% were negative for both steroids. The present study therefore once again confirmed the high usage of traditional medicines where some of which are contaminated with steroids.

Matched MeSH terms: Chromatography, Thin Layer
Fulltext Bioguided Fractionation of Local Plants against Matrix Metalloproteinase9 and Its Cytotoxicity against Breast Cancer Cell Models: In Silico and In Vitro Study

Hariono M, Rollando R, Karamoy J, Hariyono P, Atmono M, Djohan M, et al.

Molecules, 2020 Oct 14;25(20).
PMID: 33066411 DOI: 10.3390/molecules25204691

Matrix metalloproteinase9 (MMP9) is known to be highly expressed during metastatic cancer where most known potential inhibitors failed in the clinical trials. This study aims to select local plants in our state, as anti-breast cancer agent with hemopexin-like domain of MMP9 (PEX9) as the selective protein target. In silico screening for PEX9 inhibitors was performed from our in house-natural compound database to identify the plants. The selected plants were extracted using methanol and then a step-by-step in vitro screening against MMP9 was performed from its crude extract, partitions until fractions using FRET-based assay. The partitions were obtained by performing liquid-liquid extraction on the methanol extract using n-hexane, ethylacetate, n-butanol, and water representing nonpolar to polar solvents. The fractions were made from the selected partition, which demonstrated the best inhibition percentage toward MMP9, using column chromatography. Of the 200 compounds screened, 20 compounds that scored the binding affinity -11.2 to -8.1 kcal/mol toward PEX9 were selected as top hits. The binding of these hits were thoroughly investigated and linked to the plants which they were reported to be isolated from. Six of the eight crude extracts demonstrated inhibition toward MMP9 with the IC50 24 to 823 µg/mL. The partitions (1 mg/mL) of Ageratum conyzoides aerial parts and Ixora coccinea leaves showed inhibition 94% and 96%, whereas their fractions showed IC50 43 and 116 µg/mL, respectively toward MMP9. Using MTT assay, the crude extract of Ageratum exhibited IC50 22 and 229 µg/mL against 4T1 and T47D cell proliferations, respectively with a high safety index concluding its potential anti-breast cancer from herbal.

Matched MeSH terms: Chromatography, Thin Layer
Fulltext The effect of environmental conditions on biofilm formation of Burkholderia pseudomallei clinical isolates

Ramli NS, Eng Guan C, Nathan S, Vadivelu J

PLoS One, 2012;7(9):e44104.
PMID: 22970167 DOI: 10.1371/journal.pone.0044104

Burkholderia pseudomallei, a Gram-negative saprophytic bacterium, is the causative agent of the potentially fatal melioidosis disease in humans. In this study, environmental parameters including temperature, nutrient content, pH and the presence of glucose were shown to play a role in in vitro biofilm formation by 28 B. pseudomallei clinical isolates, including four isolates with large colony variants (LCVs) and small colony variants (SCVs) morphotypes. Enhanced biofilm formation was observed when the isolates were tested in LB medium, at 30 °C, at pH 7.2, and in the presence of as little as 2 mM glucose respectively. It was also shown that all SVCs displayed significantly greater capacity to form biofilms than the corresponding LCVs when cultured in LB at 37 °C. In addition, octanoyl-homoserine lactone (C(8)-HSL), a quorum sensing molecule, was identified by mass spectrometry analysis in bacterial isolates referred to as LCV CTH, LCV VIT, SCV TOM, SCV CTH, 1 and 3, and the presence of other AHL's with higher masses; decanoyl-homoserine lactone (C(10)-HSL) and dodecanoyl-homoserine lactone (C(12)-HSL) were also found in all tested strain in this study. Last but not least, we had successfully acquired two Bacillus sp. soil isolates, termed KW and SA respectively, which possessed strong AHLs degradation activity. Biofilm formation of B. pseudomallei isolates was significantly decreased after treated with culture supernatants of KW and SA strains, demonstrating that AHLs may play a role in B. pseudomallei biofilm formation.

Matched MeSH terms: Chromatography, Thin Layer
Fulltext Induction of cytotoxicity by Bruguiera gymnorrhiza in human breast carcinoma (MCF-7) cell line via activation of the intrinsic pathway

Chaudhry GE, Rahman NH, Sevakumaran V, Ahmad A, Mohamad H, Zafar MN, et al.

J Adv Pharm Technol Res, 2020 10 10;11(4):233-237.
PMID: 33425710 DOI: 10.4103/japtr.JAPTR_81_20

Breast cancer is among the frequently occurring cancer worldwide. The foremost underline aim of this study was to determine the growth inhibitory effect along with mechanistic study of a Bruguiera gymnorrhiza extract on MCF-7. The cytotoxicity activity was determined by using the MTS assay. Butanol extract exhibited the maximum cytotoxicity activity against the MCF-7 cells with IC50 of 3.39 μg/mL, followed by diethyl ether and methanol extract (IC50 at 16.22 μg/mL and 37.15 μg/mL, respectively) at 72 h. The DeadEndTM Colorimetric Apoptosis Detection System confirmed the induction of apoptosis (via DNA fragmentation) in MCF-7 cells. Both butanol and diethyl ether extracts of B. gymnorrhiza significantly increase the caspase-3 level. However, the diethyl ether extract induced higher caspase-9 levels compared to caspase-8, suggesting that the intrinsic pathway was the major route in the process of apoptosis. Thin-layer chromatography profiling demonstrated the presence of phenolic, terpene, and alkaloid compounds in crude methanol, diethyl ether, and butanol extracts. The phytochemicals present in the extracts of B. gymnorrhiza might have the potential to be a future therapeutic agent against breast cancer.

Matched MeSH terms: Chromatography, Thin Layer
Male sex pheromonal components derived from methyl eugenol in the hemolymph of the fruit fly Bactrocera papayae

Hee AK, Tan KH

J Chem Ecol, 2004 Nov;30(11):2127-38.
PMID: 15672660 DOI: 10.1023/B:JOEC.0000048778.02561.70

Pharmacophagy of methyl eugenol (ME)--a highly potent male attractant, by Bactrocera papayae results in the hydroxylation of ME to sex pheromonal components, 2-ally-4,5-dimethoxyphenol (DMP) and (E)-coniferyl alcohol (CF). These compounds, which are also male attractants, are then sequestered and stored in the rectal gland prior to their release during courtship at dusk. Chemical analyses of the digestive tract (excluding the crop and rectal gland) showed the absence of the sex pheromonal components and their precursor, ME. However, B. papayae males were attracted to and fed on the ME-fed male hemolymph extracts but not on hemolymph extracts of ME-deprived males. After thin layer chromatography in a hexane:ethyl acetate solvent system, flies were attracted to and fed on the original point on the TLC plate where the hemolymph extract had been spotted, suggesting that the pheromone components were bound in polar complexes. Chemical analyses of the ME-fed male hemolymph and crop extracts revealed the presence of the sex pheromonal components. The presence of the ME-derived pheromonal components and the absence of ME in the hemolymph suggest that the hemolymph is involved in the transportation of sex pheromonal components from the crop to the rectal gland.

Matched MeSH terms: Chromatography, Thin Layer
Assay of 3-hydroxy-3-methylglutaryl CoA reductase activity using anionic-exchange column chromatography

Ong KK, Khor HT, Tan DT

Anal Biochem, 1991 Aug 01;196(2):211-4.
PMID: 1776669

A rapid, easy, and sensitive method is described in this paper for the assay of 3-hydroxy-3-methylglutaryl CoA (HMG CoA) reductase, a key enzyme in cholesterol biosynthesis. [14C]HMG CoA was used as the substrate and the product formed, i.e., [14C]mevalonate, was allowed to be converted to its lactone form (mevalonolactone) in the presence of HCl. The reaction mixture was applied to a column containing an anionic exchanger. The column was made up of QAE-Sephadex (A25, formate form) packed to a height of 4 cm in Pasteur pipets. Under these conditions, mevalonolactone was not retained by the column and was eluted with ammonium formate solution while HMG CoA, being negatively charged, was retained by the gel and eluted by HCl above 0.05 M. Determination of the amount of radioactivity in mevalonolactone was then used to quantitate the activity of HMG CoA reductase. This assay has been successfully used for determining the activity of this enzyme in a microsomal fraction prepared from the liver of the rat.

Matched MeSH terms: Chromatography, Thin Layer/methods
Heterogeneity of mycolactones produced by clinical isolates of Mycobacterium ulcerans: implications for virulence

Mve-Obiang A, Lee RE, Portaels F, Small PL

Infect Immun, 2003 Feb;71(2):774-83.
PMID: 12540557

Mycobacterium ulcerans is the causative agent of Buruli ulcer, a severe necrotizing skin disease endemic in tropical countries. Clinical evidence suggests that M. ulcerans isolates from Asia, Mexico, and Australia may be less virulent than isolates from Africa. In vivo studies suggest that mycolactone, a polyketide-derived macrolide toxin, plays a major role in the tissue destruction and immune suppression which occur in cases of Buruli ulcer. Mycolactones were extracted from 34 isolates of M. ulcerans representing strains from Africa, Malaysia, Asia, Australia, and Mexico. Thin-layer chromatography, mass spectroscopic analysis, and cytopathic assays of partially purified mycolactones from these isolates revealed that M. ulcerans produces a heterogeneous mixture of mycolactone variants. Mycolactone A/B, the most biologically active mycolactone species, was identified by mass spectroscopy as [M(+)Na](+) at m/z 765.5 in all cytotoxic isolates except for those from Mexico. Mycolactone C [M+Na](+) at m/z 726.3 was the dominant mycolactone species in eight Australian isolates, and mycolactone D [M+Na](+) m/z 781.2 was characteristic of two Asian strains. Mycolactone species are conserved within specific geographic areas, suggesting that there may be a correlation between mycolactone profile and virulence. In addition, the core lactone, [M+Na](+) m/z 447.4, was identified as a minor species, supporting the hypothesis that mycolactones are synthesized by two polyketide synthases. A cytopathic assay of the core lactone showed that this molecule is sufficient for cytotoxicity, although it is much less potent than the complete mycolactone.

Matched MeSH terms: Chromatography, Thin Layer
Ergosterol from the soilborne fungus Ganoderma boninense

Toh Choon RL, Sariah M, Siti Mariam MN

J Basic Microbiol, 2012 Oct;52(5):608-12.
PMID: 22143962 DOI: 10.1002/jobm.201100308

Ergosterol is the main component of the fungal membrane and is not found in plants or other microbial cells. Therefore, it can be a useful biomarker for the quantification of fungal biomass. We are now reporting the first isolation and characterisation of ergosterol from the mycelium of G. boninense. The ergosterol structure was detected by Thin Liquid Chromatography (TLC) and Ultra Performance Liquid Chromatography (UPLC) and confirmed with Gas Chromatography coupled with Mass Spectrometry (GCMS) and Nuclear Magnetic Resonance (NMR) analysis.

Matched MeSH terms: Chromatography, Thin Layer
Fulltext Antibacterial and Antispore Activities of Isolated Compounds from Piper cubeba L

Alqadeeri F, Rukayadi Y, Abbas F, Shaari K

Molecules, 2019 Aug 26;24(17).
PMID: 31454974 DOI: 10.3390/molecules24173095

Piper cubeba L. is the berry of a shrub that is indigenous to Java, Southern Borneo, Sumatra, and other islands in the Indian Ocean. The plant is usually used in folk traditional medicine and is an important ingredient in cooking. The purpose of this study was to isolate and purify the bioactive compounds from P. cubeba L. fractions. In addition, the isolated compounds were tested for their antibacterial and antispore activities against vegetative cells and spores of Bacilluscereus ATCC33019, B. subtilis ATCC6633, B.pumilus ATCC14884, and B.megaterium ATCC14581. The phytochemical investigation of the DCM fraction yielded two known compounds: β-asarone (1), and asaronaldehyde (2) were successfully isolated and identified from the methanol extract and its fractions of P. cubeba L. Results showed that exposing the vegetative cells of Bacillus sp. to isolated compounds resulted in an inhibition zone with a large diameter ranging between 7.21 to 9.61 mm. The range of the minimum inhibitory concentration (MIC) was between 63.0 to 125.0 µg/mL and had minimum bactericidal concentration (MBC) at 250.0 to 500.0 µg/mL against Bacillus sp. Isolated compounds at a concentration of 0.05% inactivated more than 3-Log10 (90.99%) of the spores of Bacillus sp. after an incubation period of four hours, and all the spores were killed at a concentration of 0.1%. The structures were recognizably elucidated based on 1D and 2D-NMR analyses (1H, 13C, COSY, HSQC, and HMBC) and mass spectrometry data. Compounds 1, and 2 were isolated for the first time from this plant. In conclusion, the two compounds show a promising potential of antibacterial and sporicidal activities against Bacillus sp. and thus can be developed as an anti-Bacillus agent.

Matched MeSH terms: Chromatography, Thin Layer
Fulltext Reinventing nano drug delivery systems for hydrophilic active ingredients in Ayurvedic lipid based formulations containing poly herbal decoction

Duraipandi S, Selvakumar V

J Ayurveda Integr Med, 2019 01 08;11(3):224-227.
PMID: 30635246 DOI: 10.1016/j.jaim.2018.01.008

BACKGROUND: Anu Tailam, an Ayurvedic medicated oil where 'anu' meant for atom and 'tailam' meant for oil and virtually meant for 'oil of subtle or atomic size particles'. Since the major active ingredients in this formulation are incorporated from the polyherbal decoction, it is expected to contain predominantly water soluble ingredients.
OBJECTIVES: It is hypothesized that these polar active botanical ingredients are present in the formulation should be either suspended in the form of submicron particles or entrapped in the submicron vesicular structures since the formulation did not show any precipitation or phase separation instead showed a monophasic oily liquid with very little moisture.
MATERIALS AND METHODS: In the present investigation, the micro architecture of the anu tailam is studied via column chromatography and high performance thin layer chromatography to prove the contents are polar hydrophilic compounds followed by optical microscopy, photon correlation Spectroscopy (PCS) and environmental scanning electron microscope (ESEM) to study the particle/vesicle size of the formulation.
RESULTS: In this study, it was proved that the formulation contained only polar ingredients and can be extracted in polar solvents like methanol and ethanol. It was also found that the formulation taken for study contained nano particles of the active botanical ingredients embedded in a network of vesicular structures of the lipid base.
CONCLUSION: The selected Ayurvedic formulation 'anutailam' found to contain novel nano drug delivery system to deliver water soluble ingredients across barriers.

Matched MeSH terms: Chromatography, Thin Layer
Fulltext Bioassay-directed isolation of active compounds with antiyeast activity from a Cassia fistula seed extract

Jothy SL, Zakaria Z, Chen Y, Lau YL, Latha LY, Shin LN, et al.

Molecules, 2011 Sep 05;16(9):7583-92.
PMID: 21894090 DOI: 10.3390/molecules16097583

BACKGROUND AND OBJECTIVE: Cassia fistula L belongs to the family Leguminosae, and it is one of the most popular herbal products in tropical countries. C. fistula seeds have been used as a herbal medicine and have pharmacological activity which includes anti-bacterial, anti-fungal, and antioxidant properties. The goal of this study was to identify compounds from C. fistula seeds which are responsible for anti-Candida albicans activity using bioassay-directed isolation.
RESULTS: The preliminary phytochemical screening of the plant seed revealed the presence of anthraquinones, flavonoids, saponins, tannins and terpenoids. The isolation of active compounds was carried out in four steps: multiple extractions, fractionation using column chromatography and purification using preparative thin-layer chromatography (TLC) and liquid chromatography/mass spectrometry (LC/MS). The structure of separated compounds was determined on the basis of mass spectrometry data. One compound was identified is roseanone.
CONCLUSIONS: The MS analysis on the active fraction from seed extract of C. fistula confirmed the presence of roseanone with antiyeast activity.

Matched MeSH terms: Chromatography, Thin Layer
Occurrence of opium alkaloids in commercial herbal stomach remedy preparation

Gan EK, Sam TW

Med J Malaysia, 1976 Sep;31(1):33-5.
PMID: 1023010
Matched MeSH terms: Chromatography, Thin Layer
Optimization of enzymatic synthesis of palm-based kojic acid ester using response surface methodology

Ashari SE, Mohamad R, Ariff A, Basri M, Salleh AB

J Oleo Sci, 2009;58(10):503-10.
PMID: 19745577

Kojic acid monooleate is a fatty acid derivative of kojic acid which can be widely used as a skin whitening agent in a cosmetic applications. In avoiding any possible harmful effects from chemically synthesized product, the enzymatic synthesis appears to be the best way to satisfy the consumer demand nowadays. The ability of immobilized lipase from Rhizomucor meihei (lipozyme RMIM) to catalyze the direct esterification of kojic acid and oleic acid was investigated. Response Surface Methodology (RSM) and 5-level-4-factor central composite rotatable were employed to evaluate the effects of synthesis parameters such as enzyme amount (0.1-0.4 g), temperature (30-60 degrees C), substrate molar ratio (1-4 mmol, kojic acid:oleic acid) and reaction time (24-48 h) on percentage molar conversion to kojic acid monooleate. Analysis of the product using TLC, GC and FTIR showed the presence of kojic acid monooleate. The optimal conditions for the enzymatic reaction were obtained after analysis with backward elimination using 0.17 g of enzyme and 4 mmol of substrate at 52.50 degrees C for 42 h. Under these conditions the esterification percentage was 37.21%. The results demonstrated that response surface methodology can be applied effectively to optimize the lipase-catalysed synthesis of kojic acid monooleate. The optimum conditions can be used to scale up the process.

Matched MeSH terms: Chromatography, Thin Layer
Secretory expression and characterization of a highly Ca2+-activated thermostable L2 lipase

Sabri S, Rahman RN, Leow TC, Basri M, Salleh AB

Protein Expr Purif, 2009 Dec;68(2):161-6.
PMID: 19679187 DOI: 10.1016/j.pep.2009.08.002

Thermostable lipases are important biocatalysts, showing many interesting properties with industrial applications. Previously, a thermophilic Bacillus sp. strain L2 that produces a thermostable lipase was isolated. In this study, the gene encoding for mature thermostable L2 lipase was cloned into a Pichia pastoris expression vector. Under the control of the methanol-inducible alcohol oxidase (AOX) promoter, the recombinant L2 lipase was secreted into the culture medium driven by the Saccharomyces cerevisiae alpha-factor signal sequence. After optimization the maximum recombinant lipase activity achieved in shake flasks was 125 U/ml. The recombinant 44.5 kDa L2 lipase was purified 1.8-fold using affinity chromatography with 63.2% yield and a specific activity of 458.1 U/mg. Its activity was maximal at 70 degrees C and pH 8.0. Lipase activity increased 5-fold in the presence of Ca2+. L2 lipase showed a preference for medium to long chain triacylglycerols (C(10)-C(16)), corn oil, olive oil, soybean oil, and palm oil. Stabilization at high temperature and alkaline pH as well as its broad substrate specificity offer great potential for application in various industries that require high temperature operations.

Matched MeSH terms: Chromatography, Thin Layer
The Production of 9-methoxycanthin-6-one from Callus Cultures of (Eurycoma longifolia Jack) Tongkat Ali

Maziah M, Rosli N

Methods Mol Biol, 2009;547:359-69.
PMID: 19521859 DOI: 10.1007/978-1-60327-287-2_29

Plant cell culture technology is potentially useful in producing high-valued secondary metabolites. Eurycoma longifolia root extracts are consumed as a health tonic but more popularly used as an aphrodisiac. Studies on the aphrodisiac properties and the possible compounds involved have been widely studied. There are many potentially useful compounds reported from the root extracts of E. longifolia. However, studies on the in vitro production of useful compounds from this plant have not been reported. This chapter will describe methods of callus induction and extraction of 9-methoxycanthin-6-one from E. longifolia Jack explants with emphasis on the tap and fibrous roots. This compound, known to have anti-tumour activity, is present in intact plant parts and in callus tissues of different explants.

Matched MeSH terms: Chromatography, Thin Layer

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