Formononetin (FMN) is a methoxylated isoflavone which is the major constituent in red clover and in commercially available extracts of this plant. In this study, we investigated the parallel ...artificial membrane permeability assay (PAMPA) permeability, protein binding, blood uptake characteristics, pharmacokinetics and metabolism of FMN. The permeability study samples were analyzed by HPLC–PDA method; whereas the pharmacokinetic study, protein binding and whole blood partitioning samples were analyzed by LC–MS/MS method. The PAMPA permeability of FMN was found to be high at pH 4.0 and 7.0. Plasma protein binding of FMN was found to be 93.61±0.44% and 96.14±0.15% at the tested concentration of 50 and 150ng/mL, respectively. FMN reached equilibrium fast between red blood cells (RBCs) and plasma, and the partition coefficients between RBCs and plasma (KRBC/PL) were independent of the initial rat blood concentrations of FMN. The bioavailability of unchanged/free FMN was found to be poor, i.e. approximately 3%. FMN was found to have a high clearance (5.13L/h/kg) and a large apparent volume of distribution (14.16L/kg). Circulating conjugates (glucuronides/sulfates) of FMN and daidzein (DZN) were quantified using enzymatic hydrolysis of plasma samples. The levels of isoflavone glucuronides/sulfates were found to be much greater than that of the corresponding aglycones.
Gugulipid, an ethyl acetate extract of the resin of plant
Commiphora whighitii is an established hypolipidemic agent in clinical practice. The major constituent of gugulipid is guggulsterone 4, 17 ...(20)-pregnadiene-3, 16-dione. It has been observed recently that patients receiving lipid-lowering drugs like statins have a reduced risk of dementia. Therefore, the present study was planned to explore the potential of gugulipid as cognitive enhancer. Gugulipid (12.5, 25 and 50 mg/kg, p.o.) showed dose dependent improvement in scopolamine-induced deficits in passive avoidance test. The maximal effective dose of gugulipid i.e. 50 mg/kg, p.o. was used for further studies on streptozotocin (STZ) model of dementia in mice. Gugulipid was investigated for its effect on learning and memory, parameters of oxidative stress (GSH and MDA) and acetylcholinesterase (AChE) activity in the STZ (ic)-treated mice. Intracerebral (ic) injections of STZ (0.5 mg/kg) on 1st and 3rd day caused significant deficit in memory in passive avoidance and Morris water maze test after the 14th day of first dose. In passive avoidance, transfer latency time (TLT) was not increased on retention trials in STZ (ic) group while gugulipid treatment resulted in significant increase in TLT on retention trials in STZ (ic)-treated mice. In Morris water maze test the latency time to reach platform in STZ (ic)-treated mice was significantly higher than control and vehicle (artificial CSF). Pre-treatment of gugulipid (50 mg/kg, p.o.) daily for 14 days started with the first dose of STZ (ic), significantly prevented STZ (ic)-induced memory deficit. Post-treatment i.e. after 14 days of first dose of STZ (ic) of gugulipid (50 mg/kg, p.o.) significantly decreased the latency time indicating anti-dementia activity. Effect of gugulipid and STZ in visible platform test was similar to those seen with hidden platform. Gugulipid and STZ-treated mice did not cause significant change in locomotor activity. Furthermore, STZ (ic) resulted into increase in AChE activity, low level of GSH and high concentration of MDA in brain on 21st day as compared to control. Gugulipid treatment caused significant decrease in AChE activity, low level of MDA and high concentration of GSH in brain following STZ (ic) as compared to vehicle administration in STZ (ic)-treated mice. The study demonstrated that gugulipid has significant protective affect against streptozotocin-induced memory deficits model of dementia that can be attributed to anti-oxidant and anti-AChE activity of gugulipid. These observations suggest gugulipid as a potential anti-dementia drug (CDRI, Lucknow has obtained US patent No. 6896901 for use of gugulipid as cognitive enhancer).
A new simple, rapid, sensitive and accurate quantitative detection method using liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) for the measurement of formononetin (FMN) and ...daidzein (DZN) levels in rat plasma is described. Analytes were separated on a Supelco Discovery C18 (4.6
×
50
mm, 5.0
μm) column with acetonitrile: methanol (50:50, v/v) and 0.1% acetic acid in the ratio of 90:10 (v/v) as a mobile phase. The method was proved to be accurate and precise at linearity range of 5–100
ng/mL with a correlation coefficient (
r) of ≥0.996. The intra- and inter-day assay precision ranged from 1.66–6.82% and 1.87–6.75%, respectively; and intra- and inter-day assay accuracy was between 89.98–107.56% and 90.54–105.63%, respectively for both the analytes. The lowest quantitation limit for FMN and DZN was 5.0
ng/mL in 0.1
mL of rat plasma. Practical utility of this new LC–MS/MS method was demonstrated in a pharmacokinetic study in rats following intravenous administration of FMN.
Herein, the current review deals with the distinctive properties of carbon-based molecularly imprinted polymers (MIPs) and their applications towards the diversified biological analysis that includes ...micro to macromolecules. Apart from several advantages like specific recognition site, selectivity, and stability, MIPs also have some disadvantages such as loss of conductivity, electro-catalytic activity and expensiveness, which restrict their application in different fields. To improve their analytical presentation and overcome their disadvantages, researches shifted towards carbon-based MIPs using carbon nanotubes, carbon dots, carbon nanoparticles, carbon electrodes, and graphene as a substrate. These carbon-based nanomaterials have high selectivity and specificity towards molecular identification and have been used in diverse applications including environmental, biological and food sample analysis. This review covers the development of carbon-based MIPs and its biological applications.
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•Development of carbon-based MIPs and its biological applications.•Described different CBNM based MIPs for selective analysis of target molecules.•Application of CBNM based MIPs using different methodology.
Malaria is the leading parasitic disease in emerging countries. Therapeutic drug monitoring of antimalarial drugs is becoming increasingly important due to their spreading resistance. Measuring ...systemic antimalarial drug concentrations is also vital for safety and PK evaluations during clinical development. The dried blood spot (DBS) technique is a convenient alternative sample-collection method to venipuncture, especially in resource -limited areas where the clinical studies of antimalarials are usually carried out. Various bioanalytical methods for antimalarial drug estimation utilizing DBS sampling have been reported. This review discusses the applicability and relevance of DBS in quantitative assessment of antimalarial drugs, the advantages and drawbacks of DBS, and the difficulties encountered during its implementation.
► To the best of our knowledge, there is no simultaneous analytical method reported for at least two model drugs from each permeability class. ► A simple and reliable RP-HPLC method has been ...developed and validated for simultaneous determination of nine permeability model drugs. ► Applicability of the cassette dosing principle and its practice to intestinal perfusion studies for the first time.
A simple, sensitive and specific reversed phase high performance liquid chromatographic (RP-HPLC) method for simultaneous determination of atenolol, paracetamol, hydrochlorothiazide, caffeine, cephalexin, metoprolol, propranolol, ketoprofen along with phenol red (a non-absorbable compound) in samples obtained from intestinal in situ single-pass perfusion studies, was developed and validated. Chromatography was carried out on RP18 column with mobile phase comprising of 10mM phosphate buffer (pH 2.5) and methanol in gradient mode. The calibration curves were linear for all nine permeability model compounds (r2>0.999) across the concentration range of 1.25–40μg/ml. The coefficient of variation for intra and inter-day assay precision was between 0.04 and 3.08% and the accuracy was between 98.39 and 109.45%. Stability studies were carried out at different storage conditions and all the analytes were found to be stable. The method was successfully applied for analysing the permeability samples obtained from in situ single pass perfusion studies. The effective permeability (Peff) values obtained upon cassette administration were in close proximity to the permeability values obtained upon single administration of model compounds. In conclusion, the developed RP-HPLC method can be used for high throughput cassette validation of rat in situ perfusion model for intestinal permeability assessment.
► Study was conducted in leukocytes and macrophages by using in vivo, ex vivo and in vitro test systems. ► Lipopolysaccharide (LPS) causes significant DNA damage in leukocytes and macrophages ...irrespective of test system. ► Piracetam offered significant protection against LPS induced DNA damage in both leukocytes and macrophages.
Piracetam is a nootropic drug that protects neurons in neuropathological and age-related diseases and the activation and modulation of peripheral blood cells in patients with neuropathological conditions is well known. Therefore, in the present study, in vivo, ex vivo, and in vitro tests were conducted to investigate the effect of piracetam on leukocytes and macrophages. Lipopolysaccharide (LPS) causes oxidative DNA damage; thus, in the present study, LPS was used as a tool to induce DNA damage. In vivo experiments were conducted on Sprague Dawley rats, and piracetam (600
mg/kg, oral) was provided for five consecutive days. On the fifth day, a single injection of LPS (10
mg/kg, i.p.) was administered. Three hours after LPS injection, blood leukocytes and peritoneal macrophages were collected and processed, and a variety of different assays were conducted. Ex vivo treatments were performed on isolated rat blood leukocytes, and in vitro experiments were conducted on rat macrophage cell line J774A.1. Cell viability and the level of reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and DNA damage were estimated in untreated (control) and piracetam-, LPS- and LPS
+
piracetam-treated leukocytes and macrophages. In vivo experiments revealed that rats pretreated with piracetam were significantly protected against LPS-induced increases in ROS levels and DNA damage. Ex vivo isolated leukocytes and J774A.1 cells treated with LPS exhibited augmented ROS levels and DNA damage, which were attenuated with piracetam treatment. Thus, the present study revealed the salutary effect of piracetam against LPS-induced oxidative stress and DNA damage in leukocytes and macrophages.
This study describes a versatile, robust and fast sample pre-concentration novel method based on chemical vapour deposition grown iron nanoparticles dispersed hierarchical carbon fiber forest ...(Fe-ACF/CNF) for the determination of multi-pesticide residue in water samples. This method was developed by the implementation of Fe-ACF/CNF to magnetic solid-phase extraction method (MSPE) for the adsorption of twenty-nine pesticides of various classes using gas chromatography equipped with an electron capture detector. Fe-ACF/CNF was grown via tip growth mechanism and Fe-nanoparticles are moved to the tip of CNF. The presence of Fe-nanoparticles is responsible for the magnetic property of proposed adsorbents. The Fe-ACF/CNF is competent enough to extract twenty-nine pesticides of different physico-chemical characteristics from water samples. All the predominant parameters including the amount of sorbent desorption time, temperature, sonication effect, regeneration, and reusability of Fe-ACF/CNF were thoroughly investigated. Acceptable linearity was obtained in the range of 20–500 μg/L with a correlation coefficient value ≥ 0.990 for all pesticides. The accuracy of the developed method was evaluated and the obtained recovery of the spiked samples was within 70–120% (standard deviation ≤ 15%) and reusability up to the 4th cycle. The limit of detection and quantification values was in the range of 1.44–5.15 and 4.76–17.0 μg/L, respectively. The obtained results are also cross verified with real water samples from the Gomti river (Lucknow, India) and shown the excellent extraction efficiency of Fe-ACF/CNF.
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•Twenty-nine pesticides were extracted by CVD grown iron-doped sorbents (Fe-ACF/CNF).•Fe-ACF/CNF showed good magnetic property and high surface area for the adsorption.•Optimized amount and time for adsorption were found to be 2 mg and 10 min respectively.•LOD and LOQ were found to be 1.44–5.15 and 4.76–17.0 ng/mL respectively.•Recoveries are found within range (~70–~120%) for water samples with RSD ≤15%.
Tamoxifen is the agent of choice for the treatment of estrogen receptor-positive breast cancer. Tamoxifen is a substrate of P-glycoprotein (P-gp) and microsomal cytochrome P450 (CYP) 3A, and ...biochanin A (BCA) is an inhibitor of P-gp and CYP3A. Hence, it could be expected that BCA would affect the pharmacokinetics of tamoxifen. In the present study we have developed and validated a simple, sensitive and specific LC–ESI-MS/MS method for the simultaneous quantification of tamoxifen and its metabolite 4-hydroxytamoxifen with 100
μL rat plasma using centchroman as an internal standard (IS). Tamoxifen, 4-hydroxytamoxifen and IS were separated on a Supelco Discovery C18 (4.6
mm
×
50
mm, 5.0
μm) column under isocratic condition using 0.01
M ammonium acetate (pH 4.5):acetonitrile (10:90, v/v) as a mobile phase. The mobile phase was delivered at a flow rate of 0.8
mL/min. The method was proved to be accurate and precise at linearity range of 0.78–200
ng/mL with a correlation coefficient (
r) of ≥0.996. The intra- and inter-day assay precision ranged from 1.89 to 8.54% and 3.97 to 10.26%, respectively; and intra- and inter-day assay accuracy was between 87.63 and 109.06% and 96 and 103.89%, respectively for both the analytes. The method was successfully applied to study the effect of oral co-administration of BCA (an isoflavone) on the pharmacokinetics of tamoxifen and 4-hydroxytamoxifen in female rats. The coadministration of BCA caused no significant changes in the pharmacokinetics of tamoxifen and 4-hydroxytamoxifen. However, the peak plasma concentration (
C
max) of 4-hydroxytamoxifen in BCA pretreated rats was significantly (
P
<
0.05) lower than those from control group.
Bisphenol A (BPA) is a well-recognized endocrine disruptor, and considering its adverse effects its use in infant bottles has been banned in many countries. Growing concern on the use of BPA has led ...to its replacement with its analogues in numerous applications. Present is the first report determining the occurrence of seven bisphenols (BPs: BPA, BPAF, BPC, BPE, BPFL, BPS, and BPZ) in Indian infant formula. A reliable and efficient UPLC-MS/MS method for their simultaneous determination was developed and validated in powdered infant formula (n = 68). The limit of quantification of the method was 0.19 ng/g for BPA, BPAF, BPE, BPS and BPZ and 0.78 ng/g for BPC and BPFL. The highest concentration was detected for BPA (mean = 5.46 ng/g) followed by BPZ and BPS. BPAF, BPFL, BPC and BPE were detected in none of the samples. The estimated daily intake (EDI) of total BPs in infants (0–12 months old infants) was determined to be 54.33-213.36 ng/kg b.w./day. BPA mainly contributed to the total intake (EDI = 92.76 ng/kg b.w./day). The dietary exposure to total BPs evaluated in the present study was approximately 1 order of magnitude lower than the reference value of BPA set by EFSA (4 μg/kg b.w./day) and, thus, may not pose considerable risks to infants.
