Separation Of Active Constituents Research Articles

Concept of a six-fold multiplex planar bioassay to distinguish endocrine agonist, antagonist, cytotoxic and false-positive responses

To analyze a complex sample for endocrine activity, different tests must be performed to clarify androgen/estrogen agonism, antagonism, cytotoxicity, anti-cytotoxicity, and corresponding false-positive reactions. This means a large amount of work. Therefore, a six-fold planar multiplex bioassay concept was developed to evaluate up to the mentioned six endpoints or mechanisms simultaneously in the same sample analysis. Separation of active constituents from interfering matrix via high-performance thin-layer chromatography and effect differentiation via four vertical stripes (of agonists and end-products of the respective enzyme–substrate reaction) applied along each separated sample track were key to success. First, duplex endocrine bioassay versions were established. For the androgen/anti-androgen bioassay applied via piezoelectric spraying, the mean limit of biological detection of bisphenol A was 14 ng/band and its mean half maximal inhibitory concentration IC50 was 116 ng/band. Applied to trace analysis of six migrate samples from food packaging materials, 19 compound zones with agonistic or antagonistic estrogen/androgen activities were detected, with up to seven active compound zones within one migrate. For the first time, the S9 metabolism of endocrine effective compounds was studied on the same surface and revealed partial deactivation. Coupled to high-resolution mass spectrometry, molecular formulas were tentatively assigned to compounds, known to be present in packaging materials or endocrine active or previously unknown. Finally, the detection of cytotoxicity/anti-cytotoxicity and false-positives was integrated into the duplex androgen/anti-androgen bioassay. The resulting six-fold multiplex planar bioassay was evaluated with positive control standards and successfully applied to one migrate sample. The streamlined stripe concept for multiplex planar bioassays made it possible to assign different mechanisms to individual active compounds in a complex sample. The concept is generic and can be transferred to other assays.

Molecular Docking and in vitro Enzyme Assay of Bioactive Compound Isolated from Rhus tripartite Collected from Hail Region of Saudi Arabia as Potential Anti-Diabetic Agent

Abstract: Aim: In the present investigation, we have studied the inhibitory potential of a bioactive compound isolated from Rhus tripartite on Glucokinase (GK), Dipeptidyl Peptidase-IV (DPP-IV), alpha-glucosidase, and alpha-amylase enzymes. Materials and Methods: The plant leaves were subjected to Soxhlet extraction followed by qualitative phytochemical screening and the separation of active constituents by applying column chromatography. The obtained fraction was subjected to spectral analysis to identify the compound. Molecular docking followed by in vitro enzyme assays were used to study the inhibitory effect of the isolated compound. Results: The plant leaves were used for the extraction, and the identified compound was S-benzo[d]oxazol-2-yl 2-(piperazine-1-yl)ethanethioate, confirmed by spectral analysis. From in-silico ADMET analysis, the isolated compound displayed most drug-likeness features, and in molecular docking studies, it has developed many crucial hydrogen bonding and hydrophobic interactions with enzymes (PDB IDs: 1V4S, 2P8S, 3BAX, and 3WY2). An in vitro enzyme assay validated the virtual screening results of isolated compounds. Isolated compound at 250 μgm/ mL displayed 96.29±2.56, 89.23±2.1, 72.72±0.75, and 69.76±0.85% activity against GK, DPP-IV, alpha-amylase, and alpha-glucosidase enzymes, respectively. Conclusion: Our study concluded that DM could be treated using an isolated compound after further in-vivo and in-vitro studies. Keywords: DPP-IV, in-vitro, Rhus tripartite, Isolation, Characterization.

Preclinical Evidence of Curcuma longa and Its Noncurcuminoid Constituents against Hepatobiliary Diseases: A Review.

Hepatobiliary disease currently serves as an important public health issue due to the fact that it is one of the major causes of death among economically active individuals and can easily progress to chronic diseases. Despite the development of vaccines and numerous drugs, a definite treatment remains lacking owing to different stages of the disease itself, its intricate pathogenesis, an effect uncertainty for long-term use, resistance, and side effects. Curcuma longa (C. longa), which belongs to the family Zingiberaceae and the genus Curcuma, has long been used not only as spice for curry or dye but also as a constituent of herbal formula for the treatment of different diseases due to its bioactive activities. Recently, many studies on the experimental results of C. longa have been published relative to hepatobiliary diseases such as fatty liver, hepatitis, cirrhosis, and tumors. Therefore, in this review, we aimed to summarize the pharmacological effects and underlying molecular mechanisms of C. longa and its four compounds, β-elemene, germacrone, ar-turmerone, and bisacurone, against hepatobiliary diseases. C. longa exhibited antioxidant, hepatoprotective, antisteatotic, anti-inflammatory, antifibrotic, antitumor, and cholagogic effects by regulating apoptosis, CYP2E1, Nrf, lipid metabolism-related factors, TGF-β, NF-κB, CYP7A1, and so on. In particular, β-elemene could be an attractive compound owing to its remarkable hepatoprotective, anti-inflammatory, antifibrotic, and antitumor activities. Altogether, the present review provides a preclinical basis for the efficacy of C. longa as an effective therapeutic agent for the prevention and treatment of hepatobiliary diseases, despite the need for further studies to establish the extraction conditions and separation of active constituents with high bioavailability, and warrants further evaluation in clinical trials.

Separation of active constituents in some medicinal plants by thin layer chromatography

In the context of rising unemployment of college-level workforce while the need for business exists, the development of a set of competency criteria is needed to help students have a better preparation in order to meet the needs of employers in the future. This research aimed to meet the requirements. The research used data from two surveys, including 30 employers and 300 students from 5 universities. The analysis methods were the descriptive statistics and the T-test, conducted on the Stata 12 software. As a result, the study selected 15 most important competencies in 30 initial aggregated ones. Analyzing the set of competency criteria, the results showed that there was a significant difference in perceptions about the importance of criteria between students and employers. While employers considered the skill and attitude criteria to be more important, the students punctuated the criteria of knowledge. In addition, the findings showed that students had not yet met what the business needed in all 15 criteria at the current time.

Phytochemical and Pharmacological Study of Valepotriates in Valeriana officinalis L. F.Valerianeceae Cultivated in Iraq

This study concerned with phytochemical investigation and methods of extraction and separation of active constituents from Valeriana officinalis plant cultivated in Iraq. Due to the large number of active constituents in Valeriana officinalis, it was necessary to make analytical study of its constituents to determine the chemical nature of these constituents and then determine the main classes (terpenes and iridoids) using chemical reagents specific for each class. Different organic solvents like ethanol (70%) used in soxhlet apparatus and hexane, ethyl acetate and methanol were used separately to extract the main active constituents by maceration. Through comparison between these solvents using thin layer chromatography (TLC), it has been found that hexane was best suited to extract most of the active constituents from the plant by maceration method. Analytical study showed that terpens was separated and purified using preparative TLC and preparative HPLC. Identification of isolated component (valtrate) from roots of plant was obtained using HPLC and PHPLC also depending on some material-specific constants such as infrared spectroscopy (FTIR). Results of analysis showed that Valeriana officinalis cultivated in Iraq is a good source of many constituents with different important pharmacological activities. The pharmacological study showed that the total organic plant extract has sleep induction effect when injected I.P. to experimental mice.
 Key wards: Valeriana officinalis, Valepotriate, Insomnia.

Simultaneous extraction and separation of flavonoids and alkaloids from Crotalaria sessiliflora L. by microwave-assisted cloud-point extraction

Abstract A novel method of simultaneous extraction and separation of flavonoids and alkaloids from Crotalaria sessiliflora L., was developed by microwave-assisted cloud-point extraction (MACPE) using Triton X-100−NaCl−HCl system as extractant. The optimum conditions including Triton X-100 concentration, NaCl concentration, HCl concentration, solvent-to-material ratio, extraction temperature and time were investigated by orthogonal test on the single factor experiment. The extraction yields of vitexin, isovitexin and monocrotaline reached respectively up to 601.4, 386.7 and 105.5 μg/g under the optimum conditions. Compared with heating reflux extraction (HRE) and ultrasonic-assisted extraction (UAE), MACPE exhibited a significantly higher extraction yields and took a shorter time (only 10 min). The MACPE mechanism was discussed based on phase’s shift and extraction property. As a result, flavonoids and alkaloids can be transferred respectively to bottom phase and top phase after simultaneous extraction. The proposed method integrated MAE with cloud-point extraction (CPE) in one-step procedure took both advantages. It provided a simple and efficient alternative to extraction and separation of active constituents from the herb materials.

Application of molecularly imprinted solid phase extraction in the separation and determination of active constituents from natural compounds

The active constituents of natural medicine resources are complicated and in low content, which are difficult to be extracted and separated by general methods. Molecularly imprinted polymers are functional porous materials with molecular-specific recognition sites to a particular target molecule. The molecularly imprinted solid phase extraction has been used as a sample preparation technique for the separation of active constituents from natural medicine resources. The target molecules in a mixture of chemical species can be recognized selectively. In this review, the applications of the molecularly imprinted solid phase extraction for the determination and separation of active constituents (e. g. flavonoids, polyphenols, alkaloids, organic acids, phenylpropanoids, terpenoids, etc. ) from natural compounds in recent years are summarized.

HPTLC-Fingerprinting Profile of Calotropis procera Leaves

The present investigation was designed to assess the phytochemical profile of Calotropisprocera (family: Asclapiadacea). It is one of the important medicinal plants that is being used since years totreat many ailments. A new simple high performance thin layer chromatography - densitometry method wasdeveloped for the separation of active constituents in Calotropis procera extracts. The extracts in differentsolvents of the powdered leaves of Calotropis procera were examined for the qualitative and quantitativedistribution of the active principles like alkaloids, glycosides, saponins, proteins, terpenoids, sterols, flavonoids.