Solvent System Ethyl Acetate Research Articles

Screening of Process Parameters for Aspirin Loaded PLGA Microsphere

Background amp Aim Aspirin a non-steroidal anti-inflammatory drug has the potential to stimulate various dental stem cells and promotes regeneration of dental tissues. Polylactic glycolic acid PLGA a biocompatible and biodegradable polymer has the potential to stimulate multiple processes such as proliferation osteogenic differentiation initiation of mechanical support and formation of three-dimensional niches for the growth of new tissue. Hence this study aimed to identify the most effective formulation and process variables for developing aspirin polylactic glycolic acid microspheres using the Placket-Burman method.Method In the screening study Placket-Burman screening was systematically employed to estimate all of the parameter effects and interactions. The microspheres were formulated using the double-emulsion solvent extraction method by varying various variables and performing statistical analysis. Eight independent variables were considered for screening with encapsulation efficiency as the dependent variable. These variables included the volume of the internal phase of primary emulsion solvent system ethyl acetate ethanol homogenization time stirring speed emulsification time flow rate poly vinyl alcohol concentration and PLGA concentration.Results Encapsulation efficiency varied between 50.11 and 88.98. The concentration of PLGA and Poly vinyl alcohol and the volume of the internal phase were found to have a significant influence on encapsulation efficiency in the synthesis of Aspirin PLGA microsphere.Conclusion Among the eight independent variables assessed for screening the concentrations of PLGA and polyvinyl alcohol and the volume of the internal phase were found to significantly impact the encapsulation efficiency in the formulation of aspirin PLGA microspheres.

Optimization of Color Fractionation of Monascus Pigment

Monascus fungi are unique among the various microorganisms that produce edible pigments, which consist of red, orange and yellow. These pigments are typically present in a mixture. Hence, pigment separation is needed. This study aims to optimize solvent systems that can effectively fractionate the pigments. The optimization was carried out using column chromatography and the data was analyzed using Central Composite Design (CCD). The interaction of the solvent systems ethyl acetate : formic acid : acetic acid : water was optimized. The optimal solvent system obtained was 98 : 14 : 12 : 28 (vol/vol.) for ethyl acetate : formic acid : acetic acid : water, respectively. The red and yellow pigment obtained were 2.42 mg and 0.93 mg, respectively. Based on the finding, 72.2% of the Monascus pigments was red while remaining was yellow.

Synthesis of Ketamine Derivatives by Mannich Reactions

Arylcyclohexylamine Ketamine (HCL) has played an important role in Veterinary and Human Medicine as safe and reliable anesthetic agent but asit produces dysphoria several new derivatives of Ketamine have been synthesized till date. In this Research we have Utilized Mannich reaction to Synthesize The ammonia, primary amine and secondary amine are treated with hydrochloric acid and then added to the formaldehyde. Three novel derivatives were synthesized namely2-(2-chlorophenyl)- 6-[(diethyl amino) methyl]-2-(methylamino) cyclohexanone by RXN:102 Mannich Reaction of Ketamine with Di-ethylamine , rxn 113 2-(2chlorophenyl-6-dinitrophenyl hydrazine-methyl2(methylamino) cyclohexanone by Reaction of Ketamine with Di-nitro Phenyl Hydrazine & Rxn: 601. Mannich Reaction of Ketamine with Piperazine to form (2-chlorophenyl)-2-(methylamino)-6-(piperazine -1-yl) methyl cyclohexanone. Other reactions were also undergone to prepare oxazolidine derivatives of ketamine namely Synthesis of ketamine dioxolane derivative:Ketal and Hemiaminal Formation , Rxn: 103 Reaction of Ketamine with Glycerin [6-(2-chlorophenyl)-6-(methylamino)-1, 4-dioxaspiro [4.5] dec-2-yl] methanol and Rxn: 801 Reaction of Ketamine with Ephedrine N ,3,4-trimethyl-2,6 diphenyl-1oxa-4azaspiro [4.5] decan-6-amine derivative was formed. Thin layer chromatography of the synthesized derivatives was performed by using a solvent system of ethyl acetate and chloroform (50:50) and the RF values were calculated. The solubility test proves that compounds obtained from ketamine are polar in nature. All derivatives show similarity in solubility of ketamine. Although the melting points were not exactly comparable to that of ketamine, the range was not more than 5-10°C. This relatively narrow range of melting points proved that these were pure compounds. NMR spectroscopy was performed on all newly synthesized derivatives of ketamine. The notable ones obtained from reactions i.e. Rxn 102, 113, 103, 601and 801. The results of spectroscopy demonstrate that the compounds obtained were completely new species however they were structurally related to ketamine. These derivatives synthesized and confirmed by NMR technology. The derivatives can potentially be formulated for therapeutical purpose. Despite some limitations which are being considered in current drug design, the derivatives have the potential to develop into chemically modified entities that can play a major role in clinical therapeutics. Additional research studies will potentially help to determine the advanced method for high and sophisticated yield of these derivatives. Moreover, synthesis of ketamine metabolites namely N-demethyl compound and N-demethyl-5,6-dehydro analogues is established. However, further studies and modifications of these compounds will open new ventures of drug design and development of clinical implications in health care system.

Phytochemical and In Vitro Cytotoxic Screening of Chloroform Extract of Ehretia microphylla Lamk

Ehretia microphylla of the Boraginaceae family has been extensively used as a folklore remedy for the treatment of a wide range of ailments such as cough, cancer, allergies, and gastrointestinal and venereal disorders. Extensive literature review reports have revealed these findings due to the presence of numerous phytomolecules. To validate traditional claims for cytotoxic activity of E. microphylla, the present study was undertaken. Dried leaves of the plant were powdered and defatted with petroleum ether followed by hot continuous extraction with chloroform. The chloroform extract was subjected to in vitro cytotoxic screening against a panel of human cancer cell lines such as HCT-116 (colon), MCF-7 (breast), PC-3 (prostate), A-549 (lung), HL-60 (leukemia) and MiaPaCa-2 (pancreatic) at 50 µM using SRB assay. The extract exhibited noteworthy cytotoxicity activity against breast and lung cancer. It exhibited 85.55% and 77.93% inhibition against MCF-7 and A-549 cancer cell lines, respectively. The mechanism behind cell death was determined using the DAPI staining method, which induces alteration in nuclear morphology in MCF-7 cell lines evidenced through DAPI staining. Phytochemical screening of E. microphylla extract showed the presence of saponins, steroids, lipids, tannins and triterpenoids. The chemoprofile of the chloroform extract of E. microphylla leaves was established using an n-hexane:ethyl acetate solvent system in a ratio of 6:4. The developed chromatogram showed five spots both in visible and UV light at 254 nm. The information provided in the present study will enable further studies on the isolation and characterization of bioactive compounds/fractions by following bioactivity-guided fractionation, and thus, the plant has the potential to reduce proliferation and may induce cell death via apoptosis in breast cancer cells.

Validated High-Performance Thin-Layer Chromatography-Mass Spectrometry Method and Stability Study of Linalool in the Volatile Oil of the Rhizomes of Homalomena aromatica Schott.

Homalomena aromatica is a herb of tremendous ethnomedicinal importance to various communities residing in northeast India. In this study, a high-performance thin-layer chromatography-based densitometric method was developed for identification, quantification and stability study of linalool. Mass spectrometry was hyphenated to HPTLC for streamlining the method. The stability of linalool was studied by analyzing the effect of acid, base, UV, sunlight, thermal stress and H2O2 on linalool. The chromatographic plates were developed to a height of 70mm in toluene:ethyl acetate solvent system at a ratio of 9.5:0.5 and visualized with p-anisaldehyde reagent. The developed method was found to be precise, accurate and reproducible according to International Conference on Harmonization guidelines, and compact bands of linalool were observed at Rf of 0.351 ± 0.001. The content of linalool in the volatile oil of H. aromatica was found to be 58% v/v. By application of the hyphenated MS technique, linalool was identified at m/z 137, (M + H)+. It was observed that acidic pH has the highest effect on linalool with a percentage degradation of 65. The developed method can be used in the analysis and quality control of herbal materials and volatile oils containing linalool and quality control of rhizomes of H. aromatica.

Synthesis, Spectroscopic Characterization, Crystal Structure and Theoretical Studies on New Organic Single Crystal of 1-(3,5-Difluorophenyl)-3-(2-Nitrophenyl)Urea

A new organic compound, 1-(3,5-difluorophenyl)-3-(2-nitrophenyl)urea was synthesized from 2-nitroaniline, 3,5-difluoroaniline and triphosgene in sequential two steps with 92% yield. The product was crystallized by the slow evaporation using THF and ethyl acetate solvent system to obtain its single crystal. The pure crystals were characterized with melting point, FT-IR, 1H NMR, 13C NMR and MS. The structure of the compound was brought to light by X-ray single-crystal structure determination. Density functional theory calculations were applied by using (DFT/B3LYP) method with the 6-311G(d,p) basis set level. The potential energy surface (PES) scanning was performed to determine the stability of the molecule. Frontier molecular orbitals of the compound were calculated. AIM charge and MEP analyzes were performed.

In-Vitro Anti-inflammatory Activity of Methanolic Extract of Convolvulus pluricaulis Choisy

Aim: In-Vitro Anti-inflammatory Activity of Methanolic Extract of Convolvulus pluricaulis Choisy.
 Material & Methods- The whole plant parts of Convolvulus pluricaulis Choisy were purchased from the local market. Whole plant materials were dried under shade and subjected to coarse powder for extraction process. Accurately weighed quantity of whole plant material was extracted using 95 % methanol by soxhlet apparatus for 72 h. Qualitative chemical tests of methanolic extracts were subjected to various chemical tests to detect various phytoconstituents. Solvent systems ethyl acetate: methanol: water (77:13:10) were found to be most satisfactory solvent system. After development of plates, they were air-dried and number of spots, color and Rf values were recorded. The % heamolysis was calculated by assuming the heamolysis produced by the control group as 100 %.
 Results: The preliminary phytochemical analysis revealed that different active constituent present in different extracts such as carbohydrates, proteins, amino acids, fat, oils, steroids, terpenoids, glycosides, alkaloids, tannins and other phenolics compounds. At a concentration of 500 µg/ml, the extract produced 71.59% protection of RBC haemolysis as compared with 72.73% produced by prednisolone. The methanolic extract of selected plant showed 39.70% inhibition. The Diclofenac sodium showed 55.88 % inhibition against denaturation of protein.
 Conclusion: In conclusion, it can be stated that the methanolic extract has beneficial effects in long lasting in membrane stabilizing method, inhibition of protein denaturation method and proteinase model.
 Keywords: In-Vitro, Anti-inflammatory Activity, Methanolic Extract, Convolvulus pluricaulis Choisy, Protein Denaturation Method

Quantitative Determination of Aloin in Aloe vera and Its Antioxidant Activity

Aloe vera is a popular medicinal plant used widely by the cosmetic, pharmaceutical and food industries. The Aloe vera gel, which is used mostly for its positive effects on human health,contains over 75 different bioactive compounds, including aloin. A sensitive and reliable densitometric High Performance Thin Layer Chromatography method has been developed for the quantification of aloin, an anthraquinone present in Aloe vera leaves. Chromatographic analysis was performed using methanol extract of leaves of Aloe vera using solvent system ethyl acetate: Methanol: water (100:13.5:10). Detection and quantification of aloin was done by densitometric scanning at 350nm. The results of linearity range and correlation coefficient show that there was a good correlation between peak area and corresponding concentration of aloin. The method developed here in can be implemented in the analysis and routine quality control of herbal materials. Antioxidant activity of Aloe vera was also done by using DPPH method and found that Aloe vera is also a natural antioxidant.

Separation of five flavonoids with similar polarity from Caragana korshinskii Kom. by preparative high speed counter‐current chromatography with recycling and heart cut mode

Five flavonoids with similar polarity were obtained from Caragana korshinskii Kom. by preparative high speed counter-current chromatography. Due to their similar polarity, it is difficult to select a suitable solvent system for one-step separation. Three fractions including one pure compound were only obtained with a relatively suitable solvent system of ethyl acetate: n-butanol:water (0.5% glacial acetic acid) (4:6:10, v:v:v). In order to improve the separation efficiency, recycling and heart cut modes were introduced. As a result, two flavonoids with separation factor value at 1.13 and the other two with separation factor value at 1.20 were successfully separated. Finally, five flavonoids with purity higher than 98% were obtained. Fortunately, a new compound named myricetin 3'-methyl ether 3-O-glucoside-7-O-rhamnoside was obtained. The results indicated that the recycling combined with heart-cut mode could be effective for the preparation of natural compounds with similar polarity.

Isolation and Characterization of Lupeol from the Stem of Tapinanthus globiferus (A Rich.) and its Antimicrobial Assay

Lupeol, a pentacyclic triterpenoid, was isolated from hexane and ethyl acetate solvent system. In antiquity, the stem and leaf infusion of Tapinanthus globiferus has been used ethno-medicinally as a remedy for stomach ache, diarrhea, dysentery, and wounds. Lupeol isolation from this species was carried out by column chromatography after concentrating the crude extract using a rotary evaporator, and the structure was determined by analysis of the isolate by IR, 13CNMR, 1HNMR, HSQC, and HMBC spectral analysis as well as comparison with reported data. This is the first isolation of lupeol from the stem of this species. 
 Keywords: Tapinanthus globiferus, Column chromatography, dysentery, Lupeol

Bioactivity of Couroupita guianensis Aubl. against filarial and dengue vectors and non-target fish

To evaluate the bioefficacy of crude extract and fractions from the leaves of Couroupita guianensis Aubl. against second and fourth instar larvae of Culex quinquefasciatus Say and Aedes aegypti L. The phytochemical parameters of hexane extract derived fraction were analyzed. Chromatographically enriched fractions from hexane extract were eluted by hexane: ethyl acetate solvent system through silica gel. Larval toxicity assay was conducted with water containing the crude extract from 125 to 1000 ppm and fractions of effective crude extract was tested from 25 to 200 ppm concentrations against C. quinquefasciatus and A. aegypti. Among the three solvent crude extracts hexane extract of C. guianensis showed promising larvicidal activity against the two vector mosquitoes followed by chloroform and ethyl acetate. Chromatographically eluted fraction 3 showed promising larvicidal activity followed by fractions 2 & 8 compared with the crude extract. It is concluded that C. guianensis extract has the potential to control Cx. quinquefasciatus and Ae. aegypti without harming nontarget fish.

Preparative Separation of Phenylethanoid and Secoiridoid Glycosides from Ligustri Lucidi Fructus by High-Speed Counter-Current Chromatography Coupled with Ultrahigh Pressure Extraction.

Three phenylethanoid glycosides, echinacoside (1), salidroside (3), and acteoside (6), and three secoiridoid glycosides, isonuezhenide (2), nuezhenoside G13 (4), and specnuezhenide (5), have been extracted and separated by a combined method of ultrahigh pressure extraction (UPE) and high-speed counter-current chromatography (HSCCC) from Ligustri Lucidi Fructus. For the UPE, the optimal extraction was developed with conditions including solvent of 90% ethanol, sample to solvent ratio of 1:20 g/mL, pressure of 200 MPa, and time of 2 min, which rendered the yields of compounds 4 and 5 were 15.0 and 78.0 mg/g, respectively. For the HSCCC separation, the strategy of changing flow rates between 1.0 and 2.0 mL/min allowed the acquisition for 2.7 mg of compound 1, 4.5 mg of compound 2, 6.8 mg of compound 3, 5.9 mg of compound 4, 11.2 mg of compound 5, and 2.2 mg of compound 6 in one separation run under the solvent system of ethyl acetate:n-butanol:water (2:1:3, v/v) from 200 mg of the UPE extract. The structures of these phenylethanoid and secoiridoid glycosides were elucidated by extensive spectroscopic methods.

Liquid-liquid extraction pretreatment samples method used for pharmacokinetic study of rhubarb in rats after oral administrated

Abstract Objective Pretreatment of biological samples is the most critical step in pharmacokinetic studies, especially pre-treatment of plasma samples. The pretreatment of biological samples in pharmacokinetic study of Traditional Chinese Medicines (TCM) is difficult due to the complexity of the ingredients. An organic solvent system ethyl acetate: acetone (10: 1) solution used for liquid-liquid extraction has been developed in this study and compared with the commonly used protein precipitation method. Methods Rats, Beagle dogs and humans plasma samples were adopted in this study in order to demonstrate the universality of the pretreatment method. Feasibility of this pretreatment method was also verified through its application to the pharmacokinetics of rhubarb in rats. Results According to the results of extraction recovery matrix effect, it was indicated that the liquid-liquid extraction methods with new organic solvent might be suitable for variety of structures of compounds and various types of plasma samples. The pharmacokinetic study result showed that the developed pretreatment method could successfully be used for simultaneous determination of three active compounds modin, emodin-8-O-β-D-glucopyranoside (EDG) and rhein in rat plasma with high sensitivity, accuracy, and recovery by liquid chromatography tandem mass spectrometry (LC-MS/MS). Conclusion: The pretreatment method of liquid-liquid extraction methods with new organic solvent could be successfully applied for multi-component pharmacokinetics of TCM.

Isolation, characterization and quantification of a bioactive compound from Cleome viscosa L. seeds by HPTLC-Densitometric method

AimThe current study presents the first report of isolation of bioactive compound (salicylic acid) from seeds of Cleome viscosa L. confirmed by its characterization by infrared (IR) spectroscopy, nuclear magnetic resonance (1H NMR and 13C NMR) spectroscopy and mass spectroscopy. The present work is the first report of HPTLC densitometric method, which has been developed and validated for quantification of a marker compound (salicylic acid) from ethyl acetate extract using the solvent system of ethyl acetate: acetonitrile (1.1:0.9, v/v). Material and methodsTLC glass plates pre-coated with silica gel F366 (stationary phase) were used. Densitometric analysis of salicylic acid was performed in reflectance mode at 366 nm. ResultThe developed compact spot for salicylic acid at Rf 0.51. ICH guidelines were used to validate this method in terms of precision, repeatability and accuracy. Linearity range for salicylic acid was 10–50 ng/spot and the content of salicylic acid was found to be 0.9 ± 0.04 μg/g of seed. The limit of detection (LOD) and limit of quanitification (LOQ) values for salicylic acid were 4 ng and 8 ng, respectively. Better resolution from other present constituents present in the extract were obtained by this developed simple, precise and accurate method. ConclusionThe study concluded the presence of Salicylic acid as active compound present in seeds of C viscosa. The present HPTLC-densitometric method may be used for routine quality control analysis for salicylic acid quantification from Cleome viscosa L.

Determination and Thermodynamic Analysis of the Solubility of Limonin in Eight Organic Solvents and Ethyl Acetate + 2-Propanol Binary Solvents from 283.2 to 323.2 K

In the present study, the solubility of limonin in eight organic solvents and a binary solvent system (ethyl acetate + 2-propanol) was determined by the high-performance liquid chromatography analysis method within the temperature range from 283.2 to 323.2 K. The results showed that the solubility of limonin increased with temperature in the eight pure solvents and the binary solvent mixtures. In the binary solvent system ethyl acetate + 2-propanol, the solubility decreased with the initial mole fraction of ethyl acetate. The experimental solubility of limonin was well correlated with temperature by the simplified thermodynamic equation, modified Apelblat equation, λh equation, and NRTL equation. Furthermore, the solubility of limonin in the binary solvent mixtures was correlated with both temperature and initial solvent composition of the binary solvent mixtures by the Jouyban–Acree model, van’t Hoff–Jouyban–Acree model, modified Apelblat–Jouyban–Acree model, Ma model, Sun model, and NRTL model. Finally,...

A Novel Validated Stability-indicating HPTLC Method to Quantitate Forskolin as a Bulk Drug and in a Nanosuspension

A novel, reproducible, precise, accurate and dependable high-performance thin-layer chromatographic technique was developed and authenticated as per International Conference on Harmonization guiding principle for quantitative evaluation of forskolin as a bulk drug and in a nanosuspension. Chromatographic plates 60 F-254 pre-coated with silica gel and a solvent system of ethyl acetate:hexane:formic acid (7:2.9:0.1 v/v) was used for the development of the plates. Densitometric scanning of the developed plates in the reflectance mode at 555 nm using a Camag TLC scanner after exposing the plates to anisaldehyde-sulphuric acid reagent, gave a compact spot for forskolin (Rf=0.46±0.012). In accordance to the ICH guidelines, the method was validated for linearity, precision (intra- and inter-day), limit of quantification and limit of detection, robustness and accuracy. Good linear relationship was observed for the calibration plots with linear range from 100-1000 ng/spot and a coefficient of correlation r2=0.994. Limit of detection and limit of quantification were found to be 10.83 and 36.12 ng/spot, respectively. The method appeared to be well suited for determining forskolin content in the developed nanoformulation. Forskolin was found to be susceptible to acid and alkali hydrolysis, oxidation, photochemical degradation with UV and sunlight. Statistical analysis confirmed that the process developed is precise, repeatable, accurate, and suitable for the analysis of forskolin in a formulation and could be used also for analysis of forskolin in plasma and other biological fluids. The ability of the developed method to separate the drug from its degradation products effectively confirmed that it could be a stability indicating method.

Chromatographic Fingerprinting of Sarasvata Churna-an Ayurvedic Polyherbal Formulation for Epilepsy

Sarasvata churna is an Ayurvedic formulation prescribed for Epilepsy and other brain related disorders. The objective of this study is to explore bioactive principles present in Sarasvata churna using GCMS and HPTLC techniques and establishing their correlation in Epilepsy treatment. The volatile oil obtained by hydro-distillation was used for the GCMS analysis and methanolic extracts were used for HPTLC studies. HPTLC fingerprints for the formulation and method for simultaneous qualitative analysis of biomarkers in the formulation was developed using HPTLC. The GCMS report shows the presence of α-Pinene, Propanal, 2-Caren-10-al, 3-Caren-10-al, Caryophyllene, Epiglobulol, Asarones, α-selinene, α-sesquiphellandrene, o-cymene, copaene etc. HPTLC fingerprints for methanolic extracts of Sarasvata churna was established with good separation and resolution in solvent system Toluene:Ethylacetate:Methanol (7.5:2.5:0.5v/v/v). The HPTLC method for simultaneous analysis of marker compounds in the formulation was developed in solvent system Ethylacetate:Glycial acetic acid:Toluene:Water (4:1:1:0.5v/v/v/v) with Rf 0.77 for Piperine, 0.07 for Bacosides and Rf 0.57 for Withnolides in the reference compounds and methanolic extracts of Sarasvata churna. Chromatographic patterns of reference compounds were easily detectable in the tracks of Sarasvata churna methanolic extracts. The chromatographic standards obtained from this study will help in chemical standardization of Sarasvata churna and enhance the use Sarasvata churna for management of Epilepsy.

Rapid separation of three proanthocyanidin dimers from Iris lactea Pall. var. Chinensis (Fisch.) Koidz by high‐speed counter‐current chromatography with continuous sample load and double‐pump balancing mode

The dried seeds of Iris lactea have been used in traditional Chinese medicine. Previous studies have been focused on irisquinones while other chemical components are rarely reported. To establish an efficient high-speed counter-current chromatography (HSCCC) separation method with continuous sample load (CSL) and double-pump balancing (DPB) mode to isolate proanthocyanidins from I. lactea. Firstly, an ethyl acetate extract of I. lactea was pre-fractionated by silica column chromatography for the enrichment of proanthocyanidins. Secondly, the enriched proanthocyanidins sample (EPS) was further fractionated by HSCCC with a two-phase solvent system ethyl acetate:n-butanol:water (9:1:10, v/v/v) using DPB mode. The flow rate of the two phases was 2.2 mL/min, the revolution speed was 900 rpm, the separation temperature was 30 °C and the detection wavelength was 280 nm. Finally, the structures of the three isolated proanthocyanidins were elucidated by spectroscopic methods and compared with published data. Under the optimized conditions, 600 mg of the EPS with six continuous injections (100 mg/time) was fractionated, yielding 57 mg of prodelphinidin B3, 198 mg of procyanidin B3, and 162 mg of procyanidin B1, at purities of 97.2%, 98.1% and 97.3%, respectively. The HSCCC separation method with CSL and DPB proved to be rapid, convenient and economical, constituting an efficient strategy for the isolation of proanthocyanidins.

2, 4-Di-tert-butylphenol, the bioactive compound produced by Streptomyces sp. KB1

Streptomyces sp. KB1(TISTR 2304), identified by 16S rDNA gene, was collected from the air sample at Ao-nang, Krabi province, Thailand and deposited in the GeneBank database (accession number KF939581.1). It could produce bioactive compounds and excrete into liquid culture medium within 4 days of incubation period at 30 °C, 200 rounds per minute in shaking incubator. Bioactive compounds in culture broth supernatant were extracted with 3-times of ethyl acetate (EA), evaporated by rotary evaporator at 45 °C under reduced pressure, purified by silica gel column chromatography and eluted by gradient solvent system of ethyl acetate: hexane at ratio of 1:9, 3:7, 4:6, 6:4 and 10:0, v/v. Eight pooled fractions were obtained, name PF1 - PF8, from basis of their TLC profile. Only PF4 displayed as purified active compound which showed anti-methicillin resistant Staphylococcus aureus (MRSA) activity. The PF4 was predicted as 2, 4-Di-tert-butylphenol (C14H22O) with molecular weight 206.2.

Development and validation of an HPTLC method for apigenin 7-O-glucoside in chamomile flowers and its application for fingerprint discrimination of chamomile-like materials

Brewed tea of chamomile flowers (Matricaria recutita L.) (Asteraceae) has been extensively consumed for centuries due to either its pleasant taste or medicinal purposes. On the other hand, the major problem is difficulty in distinguishing the genuine specimen when supplying chamomile through nature-picking. Consequently flowers of other Asteraceae members resembling to chamomile in appearance may frequently be practiced by lay people or marketed in spice shops or bazaars. Evidently detection of such adulterations plays a vital role in terms of public health to avoid risk of toxicity (i.e. pyrazolidin alkaloids) and ineffective treatments (lack or insufficient concentration of the active constituents). This work presents either development and validation of a high performance thin-layer chromatographic (HPTLC) method for apigenin 7-O-glucoside which is one of the active markers in chamomile flowers or its application for the fingerprint discrimination of chamomile-like materials i.e. Anthemis spp., Bellis spp., Chrysanthemum sp. and Tanacetum sp. gathered by local people assuming as chamomile. Separation was performed on the silica gel 60 NH2 F254s HPTLC plates using the developing solvent system of ethyl acetate–formic acid–acetic acid–water (30:1.5:1.5:3, v/v/v/v). The proposed HPTLC method may also be a leading guide for the quality assessment of chamomile tea products on the market.