Thin-layer Chromatographic Technique Research Articles

Identification of Plasma Fatty Acids in Four Lipid Classes to Understand Energy Metabolism at Different Levels of Ketonemia in Dairy Cows Using Thin Layer Chromatography and Gas Chromatographic Techniques (TLC-GC).

Simple SummaryDairy cows with a high milk production require an excessive amount of energy in the first phase of lactation. Different metabolic processes are involved to reach the high energy requirement, in particular the mobilization of the adipose tissue which could lead to the development of metabolic diseases, such as subclinical ketosis. Early detection of excessive lipid mobilization could improve the health status of animals and reduce economic losses in farms. The aim of this research is to identify the plasma fatty acids in four lipid classes in order to understand energy metabolism at different levels of ketonemia in dairy cows using Thin Layer Chromatography and Gas Chromatographic techniques (TLC-GC). The research establishes the concentrations of fatty acids, belonging to four plasma lipid classes, in animals with different degrees of hyperketonemia.Excessive mobilization of adipose tissue in high milk producing dairy cows predisposes to metabolic diseases. The aim of this research was to identify the plasma fatty acids in four lipid classes as biomarkers for the diagnosis of hyperketonemia in bovines using thin layer chromatography and gas chromatographic techniques (TLC-GC). Sixty multiparous Holstein–Friesian dairy cows were enrolled in the study. Blood samples from the coccygeal vein were collected and β-hydroxybutyrate (BHB) was evaluated. All animals were divided into three groups on the basis of ketonemia: BHB < 0.50 mmol/L, 0.50 < BHB < 1.0 mmol/L, and BHB > 1.0 mmol/L. Plasma fatty acid concentrations were evaluated in four lipid classes: Free Fatty Acids (FFA), Triglycerides (TG), Cholesterol Esters (CE) And Phospholipids (PL). The concentration of fatty acids was analyzed using TLC-GC. The results showed the following significance in the lipid classes: 19 fatty acids were significant (p < 0.053) in FFA, nine fatty acids were significant (p < 0.050) in TG, eight fatty acids were significant (p < 0.050) in CE and three fatty acids were significant (p < 0.049) in PL. Eleven parameters were considered as predictive fatty acids related to animals in hyperketonemia. The FFA increased simultaneously with blood BHB levels, although the identified predictive fatty acids related to the TG and CE lipid classes decreased, meanwhile the BHB values increased. In the PL lipid class, no fatty acids were predictive.

In Vitro Cholesterol Esterase Inhibitory Activity of Some Purified Phenolic Acids From Agaricus bisporus: An Investigation of Cardioprotective Properties

Background: Mushrooms with significant traditional medicine properties are rich sources of vitamins, proteins, fibres, minerals, and polyphenolic compounds. Objectives: <i>Agaricus bisporus</i> (edible white button mushroom) extracts and their purified polyphenolic content were examined in terms of cholesterol esterase (CEase) inhibitory activity. Methods: Organic solvent-based extraction methodologies were adopted to prepared the extracts. Two dimensional preparative thin layer chromatography (2D PTLC) technique and direct MS/ESI⁺ method were also used to purify and elucidate the polyphenolic structures. Standard spectrophotometric analyses were employed to determine the flavonoid and phenolic acid contents as well as CEase inhibitory activity in both extract and purified polyphenols. Results: The findings supported the strong cholesterol esterase inhibitory activity of aqueous and methanolic extracts (separating funnel: 97.9 ± 1.1% and 97 ± 0.8%), and acetone extract (shake flask: 97.6 ± 0.2%). The investigation also documented the remarkable quantity of flavonoid (32.61 ± 4.1 mg/g tissue) and phenolic acid (12.38 ± 1.32 mg/g tissue). A fraction of strong blue colour phenolic acid was purified using 2D PTLC technique, and the direct MS/ESI⁺ method adopted for the purified eluate also revealed the presence of p-hydroxy benzoic acid (m/z 136), ferulic acid (m/z 194), cinnamic acid (m/z 146), and protocatechuic acid (m/z 154). The purified phenolic acids represent a moderate cholesterol esterase inhibitory activity (24.7 ± 1.7%; IC₅₀ = 2.61 µg). Conclusions: The study revealed the remarkable CEase inhibitory property, implying the cardioprotective attribute of <i>A. bisporus</i>.

Avaliação da atividade antibacteriana de plantas medicinais de uso popular: Alternanthera brasiliana (penicilina), Plantago major (tansagem), Arctostaphylos uva-ursi (uva-ursi) e Phyllanthus niruri (quebra-pedra)

Studies of substances obtained from vegetables have been acquiring new perspectives, due to problems associated with the treatment of several infections, mainly about the resistance to antimicrobials. OBJECTIVES: To evaluate the antibacterial activity of medicinal plants against different species of bacteria, as well as to analyze their chromatographic profiles. MATERIALS AND METHODS: Hydroalcoholic extract and dichloromethane, ethyl acetate, n-butanol, and aqueous fractions were obtained from Alternanthera brasiliana, Plantago major, Arctostaphylos uva-ursi, and Phyllanthus niruri, which were tested by the broth microdilution method using strains of Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum, Escherichia coli, Staphylococcus aureus, Staphylococcus saprophyticus, Proteus mirabilis, and Klebsiella pneumoniae. Thus, the minimum inhibitory concentration (MIC) of those samples was determined. The chromatographic profile of the extracts and fractions was determined by thin layer chromatography technique. RESULTS: The ethyl acetate fraction of A. uva-ursi presented MIC of 125 µg/mL against M. genitalium and 31.25 µg/mL against S. aureus, the latter has been an excellent antibacterial activity. For the same fraction, P. niruri showed moderate antibacterial activity, with MIC of 250 µg/mL against M. genitalium and S. aureus. The species A. brasiliana and P. major showed little or no activity against the tested strains. CONCLUSION: It was not possible to determine a close link between popular medicine, in vitro antibacterial activity, and the phytochemical profile of plants, since the antibacterial effects of the studied plant species have varied compared to different bacteria with and without cell walls.

Thin-layer chromatographic fingerprint of selected Paulownia species with chemometrics and antioxidant activity

Sixty-seven methanolic extracts from leaves of different genetic Paulownia species were analyzed using thin-layer chromatographic (TLC) technique to obtain their chromatographic fingerprint profiles. Analysis was performed using the TLC silica gel 60 F254 plates and mobile phase consisting of ethyl acetate–formic acid–acetic acid–water (12:1,5:1,5:4; v/v/v/v) with Naturstoff reagent derivatization. Next, the images of fingerprint chromatograms were saved as numerical data using the TLC analyzer program and processed using Excel and SpecAlign software. Finally, the data were exported to Excel and presented as a correlation matrix. The next stage of work was to compare the chemical similarity between studied samples using the similarity (Pearson correlation coefficient) and distance (Euclidean) indices and the results were presented as dendrograms (cluster analysis, CA) or graphs (principal component analysis, PCA). All studied extracts were also analyzed using the DPPH–TLC test to confirm their antioxidant activity.

Comparative study of HPLC-DAD and HPTLC for the simultaneous determination of a new multitarget antidiabetic ternary mixture in combined tablets

Qternmet XR® (FDA approval, May 2019) is a multitarget anti-diabetic drug combination composed of metformin (MET), saxagliptin (SAX) and dapagliflozin (DAP). To our present knowledge, no analytical reports were found in the scientific databases for the simultaneous quantification of MET, SAX and DAP in their ternary combined tablets, moreover, no articles have attempted the simultaneous estimation of the cited drugs in any matrix using high-performance liquid chromatography with diode-array detection (HPLC–DAD) or high-performance thin-layer chromatography (HPTLC) technique. The current work represents a comparative study on two developed and validated chromatographic methods for the simultaneous determination of the ternary mixture (MET, SAX and DAP) in pure form and in combined tablet dosage form. The first method is reversed-phase HPLC using Agilent C18 column (4.6 × 250 mm, 5 μm p.s.) with a mobile phase consisting of acetonitrile and acidic aqueous phase pH 3 with a photodiode array detection at 230 nm. The second method is HPTLC in which drug solutions were applied to Merck HPTLC silica gel plates developed with a mixture of chloroform:methanol:water:acetic acid (7.4:2.6:0.5:0.01, v/v) and scanned at 224 nm. Both methods were fully validated following the ICH guidelines in terms of linearity, accuracy, precision, selectivity and robustness.

The use of thin-layer chromatography in the assessment of the quality of lutein-containing dietary supplements

Bivariant multiple development thin-layer chromatography technique (BMD–TLC) along with high-performance liquid chromatography–diode array detection–electrospray ionization–mass spectrometry (HPLC–DAD–ESI–MS) analysis was used in determination of lutein or lutein mixed with zeaxanthin in eight dietary supplements. The developed two-step TLC separation procedure combined purification, compaction of samples and separation of the analyzed compounds what significantly shortened and simplified samples preparation. Qualitative analysis was based on co-chromatography with reference substances and HPLC–DAD–ESI–MS analysis. It was revealed that three of eight dietary supplements did not contain lutein. In turn, quantitative analysis with the use of developed TLC conditions along with densitometry showed that the amount of lutein or its mixture with zeaxanthin in the others differed from that claimed by producers.

Physical-Chemical Characterization and Thin Layer Chromatographic Comparison of a Local Propolis with the Mangifera indica Resin

Mango (Mangifera indica) is an abundant tree of Cuban flora that is known for many healing properties and a deposit of bees as a source of raw materials for the propolis production. This work aimed to determine if the resin of mango felled bark was used by honeybees in the elaboration of a specific type of propolis. This could be possible by qualitatively comparison of its samples with the mango resin and the quality of this propolis through its physicochemical characteristics. To do this, propolis was collected and hydroalcoholic extracts were prepared. From this, botanical origin was determined by the technique of thin layer chromatography. Humidity, oxidation index, wax, resin and mechanical mixtures were evaluated from raw material. Results demonstrated that the samples of propolis collected presented a chromatographic profile similar to mango resin. In the samples of propolis, it was determined that quercetin and mangiferin are present in their qualitative composition of secondary metabolites. On the other hand, the mango propolis satisfies the requirements established by international regulations for good quality propolis in terms of humidity, oxidation index and mechanical mixtures.

Descubrimiento de alcaloides en la piel de la rana Pristimantis gaigei, indicios de mimetismo Mülleriano en dos poblaciones de Panamá

La rana Pristimantis gaigei ha sido utilizada como modelo de mimetismo batesiano, adoptando el aposematismo (coloración llamativa) de su modelo, la rana venenosa Phyllobates lugubris. Los estudios sobre la composición química de su piel son escasos. Considerando que se han reportado alcaloides en otras especies de ranas consideradas miméticas y sin toxinas por mucho tiempo, queremos corroborar la presencia o ausencia de algún tipo de alcaloide en el extracto de piel de dos poblaciones de P. gaigei (PJIC001-Parque Nacional Altos de Campana y BJIC002-Parque Internacional La Amistad-Atlántico) en Panamá. Para la caracterización fitoquímica de los alcaloides se utilizó las pruebas de Dragendorff, vapores de yodo y radiación de luz ultravioleta (365nm). Igualmente se empleó la técnica de cromatografía de capa fina (CCF), con dos fases móviles diferentes (cloroformo/metanol 9:1 y acetonitrilo/agua 9:1) y poder identificar el perfil químico de los compuestos. Tanto las muestras PJI001 Y BJI002, dieron positivas para la prueba de Dragendorff. Con capa fina se logró separar y caracterizar la presencia de más de un compuesto de alcaloide. Existió diferencia en la composición y comportamiento químico entre los dos extractos, donde la muestra dePJIC001 presenta compuestos entre una polaridad media y muy polares, con una composición química más abundante, para la muestra BJIC002, presentó compuestos con alta polaridad y baja polaridad.Estos resultados demuestran la presencia de diferentes compuestos de alcaloide en la piel de la P. gaigei, siendo un indicio de un mimetismo mülleriano, logrando adquirir un posible modelo químico para su defensa.

Gas and high performance thin layer chromatographic based-determination method for quantification of thymol in semisolid traditional dosage forms

The control and standardization process of herbal products is a critical point in preparation of those medicaments. In Traditional Persian Medicine (TPM) literature, out of all the different pharmaceutical dosage forms, Jawarish is a semisolid gastrointestinal dosage form with positive related effects. Jawarish-e-Khuzi, including Zataria multiflora, Lepidium sativum, Trachyspermum ammi, Terminalia chebula, ferrous sulfate, and also honey is one of the popular mentioned traditional oral formulations. However, there have been no noticeable and proven control and standardization for this formulation. In this study, Jawarish-e-Khuzi was prepared based on one of the pharmaceutical textbooks of Traditional Persian Medicine (TPM). Using gas chromatography/mass spectroscopy (GC/MS), the volatile composition of this formulation was analyzed. Subsequently, Gas chromatography/flame ionization detector (GC/FID) and High-Performance Thin-Layer Chromatography (HPTLC) techniques were employed to determine the main component. The GC/MS results showed thymol as the main constituent. In the content determination process via GC/FID, thymol was proved to be 0.02% of the whole preparation. The outcome of HPTLC method also corresponded with that of GC/FID. Based on the method validation parameters, both GC/FID and HPTLC methods are useful for the volatile content determination of semisolid dosage forms.

New planar assay for streamlined detection and quantification of β-glucuronidase inhibitors applied to botanical extracts

The inhibition of the β-glucuronidase released from gut bacteria is associated with specific health-related benefits. Though a number of β-glucuronidase inhibition assays are currently in use, none of them can directly measure the relevant activity of each single constituent in a complex mixture, without prior separation and tedious isolation of the pure compounds. Thus, the hyphenation of the high performance thin layer chromatography (HPTLC) technique with a β-glucuronidase inhibition assay was investigated and successfully demonstrated for the first time. A colorimetric as well as fluorometric detection of the inhibitors was achieved using 5-bromo-4-chloro-3-indolyl-β-D-glucuronide as a substrate. Hence, β-glucuronidase inhibitors were detected as bright zones against an indigo blue or fluorescent background. The established method was optimized and validated employing the well-known inhibitor d-saccharic acid 1,4-lactone monohydrate. As proof of concept, the suitability of the new workflow was verified through analysis of two botanical extracts, Primula boveana and silymarin flavonolignans from Silybum marianum fruits. The found inhibitors were identified by spectroscopic methods; one of them, 3ʹ-O-(β-galactopyranosyl)-flavone, is here described as a newly isolated natural compound. The new hyphenation HPTLC-UV/Vis/FLD-β-glucuronidase inhibition assay-HRMS covers four orthogonal dimensions, i.e. separation, spectral detection, biochemical activity and structural characterization, in a highly targeted time- and material-saving workflow for analysis of complex or costly mixtures.

Potential antidiabetic and antioxidant activities of a heliangolide sesquiterpene lactone isolated from Helianthus annuus L. leaves.

Heliangolide is a naturally occurring sesquiterpene lactone and its derivatives are biologically active compounds present in most medicinal plants. This study evaluated the antioxidant and antidiabetic properties of a heliangolide sesquiterpene lactone isolated from Helianthus annuus L. leaves. The heliangolide sesquiterpene lactone was isolated through a combination of solvent-solvent partitioning, column chromatography, thin layer chromatography and high-performance liquid chromatography techniques. The antioxidant activity of the compound was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide radical scavenging assays while the antidiabetic effects were investigated in alloxan-induced diabetic rats. The heliangolide derivative at the concentration of 954.2 µmol L-1 showed 23.7 % DPPH and 26 % nitric oxide radical inhibitions compared with 96.6 and 50.9 %, resp., displayed by the controls (2,271.2 µmol L-1). It also reduced the fasting blood glucose (FBG) levels in a time-dependent manner. The highest activity was recorded within 6 h post-treatment at 0.2 mmol kg-1 bm. The heliangolide derivative exhibited significant (p < 0.05) antioxidant and antidiabetic properties and provides a basis for further development of constituents of Helianthus annuus leaves for the management of such diseases.

Hydrotropy: A Boon in Thin layer Chromatography A Review

Around a century ago, in 1916, the term ‘hydrotropy’ was coined by the scientist Carl A. Neuberg to concentrate on anionic organic salts which significantly enhanced the aqueous solubility of weakly soluble solutes1. The thin layer chromatography (TLC) of drugs are usually performed with the help of solvents like butyl amine, toluene, ether, chloroform, light petroleum, ethyl acetate, ethanol, hexane, xylene and phenol. The majority of which are much expensive and toxic1-3. Now a days, aqueous solubility of drugs is major problem in pharmaceutical field. Hydrotropic agents are employed as mobile phase for estimation of poorly aqueous soluble drugs by TLC technique. The current review takes the readers throughout a brief summary, geometrical Features of hydrotropic agents, and their different advances toward Chromatographic techniques. Moreover, this review would provide an insight of the future perspectives concerned with the TLC and hydrotropism. In future, hydrotropic solutions shall prove a boon in TLC and high performance thin layer chromatography (HPTLC) analysis of a vast number of drugs thereby limiting the use of organic solvents to a great extent.

Synthesis of anti-corrosion polymers from thiadiazole

One of the biggest problems facing many industries particularly oil, is the problem of corrosion, where the metal parts under the influence of the vital factors are eroded during use and storage, therefore, to lift the metal’s ability and to protect it against corrosion, corrosion inhibitors are used. For the first time in this research, polymers which contain sulfur - heterocyclic ring with a thiadiazole base were made. Anti- corrosion polymers were made on two stages, in the first stage, thiadiazole was made from hydrazine hydrate reaction to carbon disulfide, afterwards the first product was reacted with an excess of hydrazine. In the second stage, polymers were prepared by the reaction of the monomer produced from benzaldehyde reaction with [dichloro methane, dichloro ethane or vanillin] and the reactions progress were monitored using thin layer chromatography technique. Prepared compounds were characterized using FT-IR, 1HNMR spectra, TLC, DSC, m.p measurements and weight loss measurement. More than one type of polymers prepared with different concentrations (0.5, 1.0%) was tested as a weight loss inhibitor and the results were very good.

Evaluation of &amp;lt;i&amp;gt;In-Vitro&amp;lt;/i&amp;gt; Anti-Tuberculosis Activity of &amp;lt;i&amp;gt;Tetrapleura tetraptera&amp;lt;/i&amp;gt; Crude and Fractions on Multidrug Resistant &amp;lt;i&amp;gt;Mycobacterium tuberculosis&amp;lt;/i&amp;gt;

The management, control and elimination of tuberculosis (TB) have been difficult with the advent of HIV and cases of multidrug resistant (MDR-TB) tuberculosis. The cases of multidrug resistance to rifampicin and isoniazid pose greater challenges on first line and second line drugs to eliminate TB. The study is aimed at establishing anti-tuberculosis activity of Tetrapleura tetraptera against Mycobacterium tuberculosis and MDR-TB and the phytochemical present. The leaves of Tetrapleura tetraptera were collected, weighed, dried and pulverized to powder. The pulverized leaves of Tetrapleura tetraptera were subjected to 70% methanol extraction and screened for phytochemical. The crude extract was further purified into fractions using silica gel and thin layer chromatography techniques. M. tuberculosis and MDR-TB were obtained from positive acid fast bacilli sputa of TB patients and confirmed using GeneXpert to differentiate genotypic drug susceptible M. tuberculosis and MDR-TB. The sputa were digested using sodium hydroxide-cysteine technique and cultured in Middlebrook 7H9. The crude extract and fractions were screened for anti-tuberculosis activity using tetrazolium microtitre plate assay. The results showed that Tetrapleura tetraptera crude had activities against M. tuberculosis at 7.4 ± 0 mg/ml and 27.5 ± 0 mg/ml for MDR-TB. One of the fractions inhibited the growth of M. tuberculosis at 0.24 ± 0 mg/ml and MDR-TB at 0.89 ± 0 mg/ml. The phytochemical screened includes tannins, alkaloids, saponins, flavonoids, phenols and resins. T. tetraptra possesses anti-tuberculosis potential at low concentration on MDR-TB and can be a lead compound in drug development for the treatment of tuberculosis and multidrug resistant tuberculosis.

Characterization of the Lipid Components in &amp;lt;i&amp;gt;Desmodesmus&amp;lt;/i&amp;gt; and &amp;lt;i&amp;gt;Scenedesmus&amp;lt;/i&amp;gt; Strains: Lipid Content, Lipid Classes and Fatty Acid Profile

The lipid composition in terms of the amount of neutral lipids, free fatty acids and polar lipid content is of great importance to make full use possible of this fraction and to define the suitability of its application, either as a raw material for fuel production, nutraceutical purposes or feed. In addition to the fatty components present in the lipid extract, other components may be included, such as: carotenoids, pigments and sterols. The microalgae studied in this work, Scenedesmus sp. and Desmodesmus sp., were subjected to the same growth conditions and evaluated for lipid content, quantification and diversity of lipid components as well as its fatty acid profile. For lipid determination two extraction methods were compared: the J: Schmid-Bondzynski-Ratzlaff and Bligh & Dyer method. For Desmodesmus sp. 5.43% ± 0.41% and 9.18% ± 0.33% of lipids were obtained on an ash-free dry weight basis and for Scenedesmus sp. 12.46% ± 0.38% and 8.16% ± 0.42% of lipids were obtained on an ash-free dry weight basis using for both methods J: Schmid-Bondzynski-Ratzlaff and Bligh & Dyer, respectively. For the identification of the main lipid components present in the extracts, the Thin layer chromatography (TLC) technique was used. This made it possible, using a simple and inexpensive method, to identify the compounds extracted by different extraction methods, that is, it was possible to verify the selectivity of the different extraction methods. In addition, it has been shown that using these methods, widely described in the literature as methods of extracting lipids in practice, extracts a wide diversity of compounds. The major lipid class for both microalgae was fatty acids with amounts between 23.62% - 38.02%. The triglycerides percentage in biomasses without chemical treatment did not exceed 18.26%. In the lipid extract obtained with Bligh & Dyer, the microalgae Desmodesmus sp. presented 55.26% of unsaponifiable material, higher than the amount present using the same extraction method for Scenedesmus sp. 49.06%. Among the main unsaponifiables identified are hydrocarbons (carotenes) and sterols esters. The acid treatment of biomass, method J: Schmid-Bondzynski-Ratzlaff, showed selectivity of 72.84% and 76.66% for obtaining fatty material from the microalgae Desmodesmus sp. and Scenedesmus sp., respectively. The results showed that depending on the method used for extraction, the lipid fraction will be different in relation to the percentage of fatty components.

Quantification of Lipid Content and Identification of the Main Lipid Classes Present in Microalgae Extracts Scenedesmus sp. for Obtaining Fatty Compounds for Biofuel Production

Microalgae biomass has been reported in the literature as one of the most promising sources for obtaining different products of industrial interest such as lipids, fatty acids, carotenoids, proteins and fibers. The lipid fraction of microalgae comprises neutral lipids, free fatty acids and polar lipids. It is of great importance to estimate the composition of the lipid fraction to define the potential for use, either as a raw material for the production of biofuels or for use for nutraceuticals and/or food purposes. The microalgae Scenedesmus sp. cultivated in a photobioreactor, the sky open raceway type, was evaluated for lipid content, identification and quantification of lipid components obtained from different extracts. In the quantification of the lipid content, extraction methods were proposed without chemical treatment (use of solvents only) such as chloroform:methanol (2:1 v/v)—Bligh & Dyer, Ethanol, Ethyl acetate:Hexane (1:1 v/v) and others with chemical treatment such as J-Schmid-Bondzynski-Ratzlaff (acid) and saponification (basic). For the identification of the main lipid components present in the extracts, the Thin layer chromatography (TLC) technique was used. This made it possible, using a simple and inexpensive method, to identify the compounds extracted by different extraction methods, that is, it was possible to verify the selectivity of the different extraction methods. In addition, it has been shown that using these methods, widely described in the literature as methods of extracting lipids in practice, extracts a wide diversity of compounds. The levels of lipids obtained via solvent extraction were up to 50% higher than those obtained with chemical treatment. In lipid extracts, obtained via solvent extraction, the presence of polar compounds, glycerides, carotenoids, pigments and sterols was identified, with up to 53% being composed of an unsaponifiable fraction, thus, presenting low selectivity for extracting fatty components. The acidic and basic treatments applied to the biomass of Scenedesmus sp. showed greater selectivity for obtaining fat components of 71.47% and 94.99%, respectively. The results showed that depending on the solvent/method used to quantify the lipids, the selectivity for obtaining the grease fraction, fundamental for conversion into biofuels, varies and the total lipid content may be overestimated.

Simple Thin-Layer Chromatographic and UV-Spectrophotometric Analysis of Promethazine and its N-Demethylation Metabolites from Biological Fluids

Several Ultra-Violet spectrophotometric analytical method has been made available in the analysis of phenothiazine group of drugs but Thin-Layer Chromatographic techniques are not fully utilized. Hence, we were able to develop a succinct, simple and cost-effective TLC and UV-spectrophotometric quantification method for the analysis promethazine and metabolites from biological fluids, validating previous studies. The proposed method was found to be precise, accurate and phenotypic determination and categorization were successfully estimated among the test samples. From the chromatogram intensities, 33.3% of the study population were classified as poor metabolizers, 40% were intermediate metabolizers, and 26.67% extensive metabolizers. Little or no elimination of N-desmethylpromethazine was observed for subjects with poor metabolism in correlation with the severity. Therefore, the phenotypic knowledge will help in the clinical choices as an individual from the same family may likely metabolize a giving drug in the same manner due to their genetic similarities. Keywords : Promethazine, Metabolites, Thin-Layer Chromatography, cytochrome P450.

Evaluación del efecto acaricida de Momordica charantia, Megaskepasma erythrochlamys y Gliricidia sepium sobre Rhipicephalus microplus

Objetivo. Se evaluó la actividad acaricida de Momordica charantia (Mc), Megaskepasma erythrochlamys (Me) y Gliricidia sepium (Gs) sobre Rhipicephalus microplus (Rm). Materiales y métodos. Se realizó la marcha fitoquímica preliminar de hojas del extracto metanólico de Mc (EMc), del extracto etanólico de Me (EMe) y del extracto acetónico de Gs (EGs) a través de la técnica de colorimetría y cromatografía en capa delgada (CCD). La actividad acaricida se realizó a través de pruebas in-vitro utilizando la prueba de inmersión de larvas (LIT) y la prueba de inmersión de adultos (AIT). Para las pruebas in-situ se usaron bovinos en pastoreo infestados naturalmente con garrapatas, utilizando las CL50 obtenidas en las pruebas in-vitro AIT; posteriormente las teleoginas se llevaron a incubación para evaluar su capacidad reproductiva. Resultados. Se determinó la presencia de varios grupos de metabolitos secundarios de interés acaricida. Se demostró el efecto acaricida de los extractos de las plantas sobre teleoginas; aunque sólo EGs mostró actividad larvicida. Los extractos a 160 mg/mL afectaron el ciclo de vida de Rm inhibiendo la ovoposición en un 46.9%, 66.1% y 84.03% (p&lt;0.05) para EGs, EMc y EMe, respectivamente. Por otro lado, en las pruebas in situ se observó diferencia significativa (p&lt;0.05) entre el tratamiento de EMc y EMe respecto a los grupos controles. Conclusiones. Los resultados obtenidos son prometedores para fortalecer la posibilidad de vinculación de los extractos de estas plantas dentro de planes integrados de control de garrapatas en sistemas de producción de bovinos.

Effect of Moisture Content on Aflatoxin B1 production in Wheat Flour Samples Collected from Lahore, Pakistan

The present research was conducted to study the effects of moisture contents on the level of aflatoxin B1 (AFB1) in wheat flour samples collected from different areas of Lahore. Total 30 samples were collected from different locations and shops in Lahore- Pakistan, making sure that they have different levels of moisture contents. Thin layer chromatography technique was (TLC) selected to check the AFB1 contamination levels in wheat flour samples. The AFB1 was detected in 10 samples (33.33%), out of which 7 samples (23.33%) had AFB1 levels within permissible range, i.e. 2 μg/kg, while 3 samples (10%) contained AFB1 levels i.e. 2.67 μg/kg, 2.93 μg/kg, 3.01 μg/kg above permissible range. From the study it was concluded that the contaminated samples had greater moisture contents than that of non contaminated samples. Sample with 35% moisture content has 3.01 μg/kg AFB1 levels. The food stuff can be protected from AFB1 production by minimizing the moisture contents, the lesser the moisture content, the lesser will be the aflatoxin levels in food samples.

Secondary Metabolites in Arctium lappa L.: Variation Among Plant Parts and Phenological Stages

Arctium lappa L. (Asteraceae) occupies a prominent position in the ethnotherapeutics practices. The herb is known to contain a number of bioactive metabolites, which have immense therapeutic value. The objective of our present investigation was to estimate the variation of metabolites in different parts and phonological stages of A. lappa. This investigation has been done using a high-performance thin-layer chromatography technique. The present analysis reveals that the root (vegetative stages) holds the significantly high content of triterpenoids (betulinic acid, oleanolic acid, and lupeol), in comparison to the reproductive stage.