Identification Of Active Ingredients Research Articles

A Comparative GC-MS Analysis of Bioactive Compounds in the Different Fractions of Root Extract of Oroxylum indicum (L.) Vent.

The focus of this paper is on the analytical methodologies, which include the extraction, fractionization and identification of active ingredients of root extract of medicinal tree Oroxylum indicum (L.) Vent. The genus Oroxylum indicum (L.) Vent. (Family Bignoniaceae] is a small deciduous, soft wooded tree. Every part of tree produces a broad spectrum of valuable compounds mainly flavonoids and phenolic compounds. Root of O. indicum has long been used in different ayurvedic preparations like Dashmularisht and Chyavanprash. A comparative study of compounds present in the chloroform, ethyl acetate and n-butanol fractions of root extract through gas chromatography mass spectroscopy was performed. Maximum number of compounds (50) were isolated in chloroform fraction, whereas in ethyl acetate, n-butanol and in aqueous fractions the number of compounds isolated simultaneously were 35, 24 and 22. Two compounds (9, 12-Octadecadienoic acid and baicalein) were common in all fractions. Other important compounds isolated were myristyl myristate, hydroxy methyl furfural, ascorbic acid 6 -hexadecanoate, pseudopelletierine and squalene.

The therapeutic effect of Bletilla striata extracts on LPS-induced acute lung injury by regulation of inflammation and oxidation

Identification of active ingredients, and their structures, from Bletilla striata and investigation of the antioxidative and anti-inflammatory responses in vitro and in vivo.

Rapid screening and identification of active ingredients in licorice extract interacting with V3 loop region of HIV-1 gp120 using ACE and CE-MS

The binding of envelope protein gp120 to glycosphingolipids is very important during the human immunodeficiency virus entering into the host cell. This step occurs in the V3 loop region in particularly. The conserved core sequence of V3 loop in gp120 was named R15K. Anti-HIV drug targeting to R15K would avoid the drug-resistance caused by HIV-1 genetic diversity. Here, for the first time, affinity capillary electrophoresis (ACE) and capillary electrophoresis-mass spectrometry (CE-MS) were used for establishing a simple, rapid and effective method of screening the licorice extract for biological activity (anti-HIV), which avoided the complicated isolation and purification process. R15K, 3′-sialyllactose (the positive control), and d-galactose (the negative control) were used for the development and validation of ACE method. After the interaction between licorice extract and R15K was confirmed by ACE, the relative active ingredients were isolated by SPE and their structures were determined by CE-ESI-MS online. In this research, two mixtures from licorice extract were found to be active. Furthermore, glycyrrhizin and licorice saponin G2 were verified as the main ingredients that significantly interacted with R15K via CE-MS and LC–MS. The results of quantitative assays showed that the active mixture contained glycyrrhizin of 74.23% and licorice saponin G2 of 9.52%. Calculated by Scatchard analysis method, glycyrrhizin/R15K complex had the highest binding constant (1.69±0.08)×107L/mol among 27 compounds isolated from licorice extract. The anti-HIV activity of glycyrrhizin was further confirmed by bioactive experiment of cellular level. This strategy might provide a high throughput screening and identifying platform for seeking HIV-1 inhibitors in natural products.

Studies on free-radical scavenging activity and identification of active ingredients of different plant crude extracts of Mentha piperita collected from Sur, Sultanate of Oman

Studies on free-radical scavenging activity and identification of active ingredients of different plant crude extracts of Mentha piperita collected from Sur, Sultanate of Oman

Traditional Japanese oral mucositis medicine hangeshashinto to upregulate antimicrobial peptides in human salivary gland cells.

517 Background: The cause of chemotherapy-induced oral mucositis (COM) is thought to be direct injury and indirect stomatotoxic effects that result from the release of inflammatory mediators, loss of protective salivary constituents, and therapy-induced neutropenia. We previously found that gargling with hangeshashinto (HST), a traditional Japanese medicine, was effective for treatment of COM. Focusing on the pain and inflammation of COM, we reported at ASCO-GI 2013 and DDW 2013 that HST exhibited ameliorating effects in a hamster model of COM and multi-targeted effects on prostaglandin E2 (PGE2) production, followed by identification of active ingredients by PGE2 culture systems and LC-MS/MS. Our aim in this study was to address whether HST affects protective oral constituents. Methods: Human oral keratinocytes (HOK) and human salivary gland (HSG) cells were used for cell culture assays. Expression levels of mRNAs for antimicrobial peptides, extracellular matrixes, keratinocyte growth factor, amylase, COX-1/COX-2, and cNOS/iNOS in cells with or without HST (10–300 μg/mL) treatment were measured by RT-PCR. Results: HST increased gene expressions of some antimicrobial peptides (defensin β1, adrenomedullin, cathelicidin antimicrobial peptide), and amylase 1A in HSG cells. Further, HST dramatically inhibited COX-2 and iNOS mRNAs in IL-1β treated HOK cells, while it exerted no or little effect on expressions of any protective oral constituents in non-treated HOK cells. A random test to identify the ingredients that increase defensin β1 revealed that isoquinoline alkaloids like berberine were active. Conclusions: HST is expected to maintain oral homeostasis through two paths: increased production of antimicrobial peptides and decreased oral damage induced by excessive prostanoids and nitric oxide. HST thus functions as a multitarget agent, indicating that it may be beneficial in COM treatment.

Identification of active ingredients in Wuzhuyu decoction improving migraine in mice by spectral efficiency association.

Wuzhuyu decoction is a traditional Chinese medicine used for the effective treatment of migraines, termed 'Jueyin headache', in China. However, there have been few investigations to clarify the composition of Wuzhuyu decoction for the treatment of migraines. In the present study, 10 types of Wuzhuyu decoction were analyzed by chromatograms. 5-hydroxytryptamine (5-HT)-depletion mouse models of migraine were prepared by subcutaneous injection of reserpine and placement of autologous blood clots in the cerebral cortex. The levels of 5-HT, noradrenaline (NE), dopamine (DA), nitric oxide (NO) and nitric oxide synthase (NOS) in the brain tissues and sera of the mice were determined. The ingredients and pharmacodynamic indices of the Wuzhuyu decoctions were analyzed using spectral efficiency association by partial least squares regression. The levels of 5-HT, NE and DA in the mouse brain tissues were reduced to 337.785 ± 84.504, 171.173 ± 65.172 and 242.075 ± 158.621 mg/g brain tissue, respectively. The level of NO in the brain tissues increased to 0.425 ± 0.184 µmol/g protein and the activities of NOS in the brain tissues and sera increased to 0.719 ± 0.477 U/mg and 50.688 ± 8.132 U/ml, respectively. Regarding the ingredients of the Wuzhuyu decoction, those with significant regression coefficients were ginsenoside-Rg1, Re, Rb1, rutaevine (Rv), limonin (Li), evodiamine (Ev), rutaecarpine (Ru) and substance X (awaiting identification). Rg1, Re, Rb1, Rv, Li, Ev, Ru and X in the Wuzhuyu decoction were observed to yield the pharmacological effects, whereas Rb1, Rv and Ev were important in index improvement.

NMR spectroscopy and chemometrics as a tool for anti-TNFα activity screening in crude extracts of grapes and other berries

The identification of active ingredients in crude plant extracts offers great advantages. In this study, nuclear magnetic resonance and chemometrics were used for the screening of in vitro anti-TNFα activity in different berry types. Solid phase extraction was applied and the resulting water, methanol–water (1:1), and methanol fractions were tested for the activity. The methanol–water fraction contained most of the phenolics and showed significantly higher activity than the other two fractions. In the second phase of this study, grapes from ‘Trincadeira’, ‘Touriga Nacional’, and ‘Aragonês’, at four developmental stages were metabolically classified and tested for the TNFα inhibition. The initial stages of grape development, green and veraison, were found more active against TNFα production as compared to the later ripe and harvest stages. Among the cultivars, ‘Touriga Nacional’ was found to be the most potent inhibitor. Different multivariate data analyses algorithms based on projections to latent structures were applied to correlate the NMR and TNFα inhibition data. The variable importance in the projections plot showed that phenolics like quercetin, myricetin, (+)-catechin, (−)-epicatechin, caftarate, and coutarate, were positively correlated with high activity. This work demonstrates the great potential of NMR spectroscopy in combination with chemometrics for the screening of large set of crude extracts, to study the effects of different variables on the activity, and identifying active compounds in complex mixtures like plant extracts.Electronic supplementary materialThe online version of this article (doi:10.1007/s11306-012-0406-8) contains supplementary material, which is available to authorized users.

Defining the active ingredients of interactive computer play interventions for children with neuromotor impairments: A scoping review

Rehabilitation researchers who investigate complex interventions are challenged to describe the "active ingredients" of their interventions: the reason(s) why a treatment is expected to be effective. Interactive Computer Play (ICP) is an emerging complex intervention in rehabilitation practice and research. The purpose of this scoping review is to identify the active ingredients of ICP interventions that are designed to improve motor outcomes in children with neuromotor impairments. Eleven potential active ingredients were identified with the following foci: ICP system or game properties; intervention effects on the user; and therapist roles. However, few studies explicitly evaluate the impact of particular ingredients on outcomes. Identification of active ingredients in ICP interventions can inform trial design and clinical decision-making. Research and clinical practice will benefit from studies that utilize a framework such as motor learning theory to guide hypotheses and measurement of the active ingredients of complex interventions.

Multiple Chromatographic and Chemometric Methods for Quality Standardisation of Chinese Herbal Medicines

Abstract Quality standardisation of complementary medicine is fundamental for industry and practice as it strengthens their quality, safety and efficacy. Current herbal standardisations are often based on the quantitative analysis of a single compound, which may not reflect the total characteristic, bioactive and toxic nature of the herbs or products. Therefore, there is a need to establish internationally recognised methodology for quality standardisation of Chinese herbal medicines. The analytical methods reviewed in this article are pharmacognosy, TLC, HPLC, LCMS, CE and chemometrics. This article covers their developments and applications in quality standardisation. Recent advances show that a combination of these methods can create an overall chemical profile for each herb. This is supported by results reviewed in this article and obtained in our laboratories on medicinal herbs including Hypericum perforatum, Morinda officinalis and Centella asiatica . Significant variations in active ingredients have been observed between herbal samples and products. It is proposed that the identification of active ingredients, pharmacological activities and eventual clinical applications are required for a comprehensive quality standardisation system. The findings of this article indicate that the combination of various chromatographic and chemometric methods could advance the methodology of quality standardisation and enhance the overall confidence in herbal medicine for the health practitioner and the public.

Identification of active ingredients in dietary supplements using non-destructive mass spectrometry and liquid chromatography–mass spectrometry

A mid-forties woman purchased seven different dietary supplements from Thailand on the internet and subsequently died after taking these supplements. Since there were no ingredient labels on the supplements, we identified the active ingredients using direct analysis in real time–mass spectrometry (DART–MS), direct exposure probe–MS (DEP–MS), and liquid chromatography–MS (LC–MS). DART–MS gives exact molecular weights and DEP–MS shows the fragmentation of a molecule by electron ionization. Analyses using these two instruments are rapid and do not require extraction of the sample. The compounds predicted by DART–MS and DEP–MS were confirmed by LC–MS and the active ingredients of the seven dietary supplements were identified.

Ion mobility spectrometry for the rapid analysis of over-the-counter drugs and beverages

In the pharmaceutical industry, there are increasing requirements for analytical methods in quality assessment for the production of drugs. In this investigation, ion mobility spectrometry (IMS) was used for the rapid qualitative separation and identification of active ingredients in generic over-the-counter drugs and food additives in beverages. The active ingredients determined in drugs were acetaminophen, aspartame, bisacodyl, caffeine, dextromethorphan, diphenhydramine, famotidine, glucosamine, guaifenesin, loratadine, niacin, phenylephrine, pyridoxine, thiamin, and tetrahydrozoline. Aspartame and caffeine were determined in beverages. Fourteen over-the-counter drugs and beverages were analyzed. Analysis times below 10 s were obtained for IMS, and reduced mobilities were reported for the first time for 12 compounds. A quadrupole mass spectrometer coupled to a mobility spectrometer was used to assure a correct peak assignation. The combination of fast analysis, low cost, and inexpensive maintenance of IMS instruments makes IMS an attractive technique for the qualitative determination of the active ingredients in over-the-counter drugs and food additives in manufacture quality control and cleaning verification for the drug and food industries.

Geckos in traditional medicine: forensic implications

At least 14 species of geckos are utilized as ingredients in the pharmacopia of traditional medicine systems around the world. Chinese Traditional Medicine uses two types of geckos, large tokay geckos (Gekko gecko) and small geckos (Gekko spp., Hemidactylus spp.) to treat a diversity of ailments including asthma, tuberculosis, diabetes and cancer, and current research in China focuses on the identification of active ingredients in geckos and the verification of their efficacy. The actual number of species involved in the Chinese Traditional Medicine trade may be higher than currently realized due to the fraudulent sale of counterfeit geckos and accidental misidentification. The potential pool of species includes at least 37 species from throughout the broad area in which geckos and other wildlife are collected for use in China. Although powders and tablets pose difficulties for identification, the whole dried form in which geckos are often sold permits the identification of most species. A key is provided to the 12 species of geckos most likely to be encountered in the trade of Asian traditional medicine.

Botanical Therapeutics: Discovery, Development and Manufacture - Prospects and Constraints

Phytochemical and synthetic chemistry based pharmaceutical industry severed the historical connection between food, plants and medicines. Discovering new chemical entity (NCEs) based drugs through high throughput screening methods may reconnect plants and human health at a high level of technological sophistication. Botanical therapeutics with multi-component that comprise functional foods, nutraceuticals, dietary supplements and botanical drugs hold several advantages over conventional drugs that may earn them a more prominent place in the medicine of the next generation. Botanical therapeutics and deliver mixtures of multi functional molecules with synergistic and potentiating effects and pleiotropic targeting at a reasonable cost and with some regulatory constraints. They are well suited for long-term disease prevention in an era of genetic testing and increased life expectancy. They also provide additional vehicles for delivering health and wellness. Technologies that address the needs of discovery, development and manufacturing of multi-components botanical therapeutics are emerging, include computational and bioinformatics approaches, cell based gene expression and high-content screening systems. Phytochemical elicitation and unique plant cultivation / extraction methods designed to optimize the production of bioactive, standardized overall extract compositions and assure batch to batch product consistency are needed. Nevertheless, multi-components botanical therapeutics carry risks associated with potential interactions with conventional drugs and adverse reactions, which are difficult to detect and diagnose. Acceptance by medical community and pharmaceutical industry, safety, poor standardization and quality control, and difficulties in identification of active ingredients and determination of their complex mode of action are some of the problems. Solving these problems will accelerate the merger of grocery stores with pharmacies and agriculture with chemical manufacturing and provide physicians and patients with broader and more individualized choices of disease prevention and treatment. Multi-components botanical therapeutics will be useful for next generation.

<i>In vitro</i> antimicrobial activities of bark and leaf extracts of <i>Moringa stenopetala</i> against mastitis causing bacterial pathogens

Antimicrobial activities of methanolic and aqueous extracts of the bark and leaf of Moringa stenopetala (Moringaceae) were studied in vitro against major mastitis causing bacterial pathogens; Staphylococcus aureus, Streptococcus agalactiae and Streptococcus dysagalactiae. The test involved the use of agar well diffusion inhibition (AWDI) test and antimicrobial activity was determined by absence or presence of growth inhibition and measurement of inhibition zone diameter (mm) expressed as mean zone of inhibition ± standard error (MZI SE). Extract solutions were prepared at four concentrations (0.3, 0.7, 1.2 and 2.0 g/ml) using Tris buffer saline. Penicillin G (1.2 g/ml) was used as a standard drug for comparison with the test materials. The study showed significant response variation between the organisms to various concentrations of the test extracts. Growth inhibition of S. aurues was much pronounced at 0.3 g/ml of methanolic extract of the bark (MZI ± SE = 58.4 ± 2.4). Conversely, S. agalactiae and S. dysagalactiae exhibited significantly (p < 0.0005) higher degree of sensitivity to methanolic bark extract at 2.0 g/ml (54.0 ± 2.1 and 50.3 ± 0.7, respectively). Similarly, methanolic leaf extract showed a more significant (p < 0.0005) growth inhibition effect on S. aurues than on S. agalactiae and S. dysagalactiae. For aqueous extracts of bark and leaf, however, the variation in growth inhibition between test organisms was observed only at 2.0 g/ml. Bark and leaf extracts of M. stenopetala were demonstrated to have a potent microbial growth inhibition effects when tested in vitro against the major bacteria causing udder infection. Further in vitro studies, determination of optimal dose and identification of active ingredients were recommended. Keywords: antimicrobial activity, mastitis, Moringa stenopetala, moringaceae, bark and leaf extracts Ethiopian Pharmaceutical Journal Vol. 23 2005: 15-22

Kefir

Abstract Kefir is a fermented milk product that is well known and widely used in eastern Europe. Although the scientific literature is not extensive, there appears to be evidence about the beneficial effects of consuming kefir that go beyond oral tradition. Problems that must be overcome before kefir can receive official approval as being good for health include a clear definition of what kefir is, sound scientific studies in humans to demonstrate its efficacy and identification of active ingredients.