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2.
Vet Res Forum ; 15(2): 97-104, 2024.
Article in English | MEDLINE | ID: mdl-38465318

ABSTRACT

Aminoglycoside antibiotics (AGs) can cause neuromuscular blockade and paralysis of skeletal muscles. To compare the paralytic effects of selected AGs on some motor behaviors in mice, 24 male mice were divided into four groups. Each group was given one of AGs (gentamicin, dihydro-streptomycin, apramycin and amikacin) at incremental doses that increased half-logarithmically compared to the therapeutic dose (16.00 mg kg-1). Motor behavioral tests included open field test, inclined plane, horizontal bars, static rods, parallel bars and rotarod. Finally, the data were analyzed using descriptive and analytical statistics. Gentamicin and dihydrostreptomycin at 32.00 times of the therapeutic dose produced complete paralysis of the limbs, respiratory arrest, and even death in some animals. However, apramycin and amikacin did not show significant effects on skeletal muscle and motor behaviors at 32.00 times of the therapeutic dose. After administration of apramycin at 100 times of the therapeutic dose, four out of six mice (66.67%) died from respiratory depression. Amikacin at this dose did not cause animal death, although it caused some changes in motor behaviors with a significant difference in comparison with control values. Gentamicin demonstrated significantly more potent effects on motor behaviors compared to the other AGs. Overall, the order of potency was gentamicin > dihydrostreptomycin > apramycin > amikacin. High doses of AGs could impair the skeletal muscle function and disrupt motor behaviors in mice. Furthermore, the paralytic potency of selected AGs on skeletal muscle was significantly different.

3.
Inflammopharmacology ; 31(4): 1913-1928, 2023 Aug.
Article in English | MEDLINE | ID: mdl-37119392

ABSTRACT

Minocycline is a semi-synthetic antimicrobial agent with claimed anti-inflammatory properties reported from different experimental models. This study was aimed to evaluate the anti-inflammatory effects of minocycline, compared to the actions of two common anti-inflammatory agents, on lipopolysaccharide (LPS)-induced paw oedema through some clinical, histopathological, haematological and molecular analyses. Forty-eight rats were divided into eight groups (n = 6). In control group (Ctrl), each animal was injected with normal saline into its sub-plantar region of hind paw. In groups 2-7, hind paw oedema was induced by injection of LPS. One hour before injections, groups 1 (Ctrl) and 2 (LPS) were treated orally with distilled water, 3 and 4 with methylprednisolone (Pred) and meloxicam (Melo) and 5-7 with minocycline in doses of 50, 150 and 450 mg/kg (M50, M150 and M450, respectively). The 8th group (MC) was given minocycline (150 mg/kg) orally and normal saline was injected into sub-plantar region. Paw swelling and body temperature were assessed at 0, 2, 4, 6 and 24 h post-injections. At 24 h, samples of blood and liver, kidney, spleen and hind paw tissues were taken for haematological and histopathological examinations. Some samples of the paw were also obtained for molecular analysis of some inflammatory-related cytokines at mRNA level. Paw swelling and body temperature increased in all LPS-injected groups 2 h post-injection. In LPS group, they remained significantly increased up to 24 h; however, these parameters decreased to normal in Pred, Melo and all minocycline groups. The histological findings showed mild-to-moderate signs of inflammation in tissue samples of groups 2-6, but not in group M450. Additionally, gene expression of pro-inflammatory cytokines (IL-1ß and IL-6) increased significantly in LPS group compared to other groups. In conclusion, this study supports the role of minocycline as an anti-inflammatory agent with effects comparable to those of meloxicam and methylprednisolone.


Subject(s)
Lipopolysaccharides , Minocycline , Rats , Animals , Minocycline/pharmacology , Lipopolysaccharides/pharmacology , Meloxicam/therapeutic use , Saline Solution/adverse effects , Anti-Inflammatory Agents/therapeutic use , Inflammation/chemically induced , Inflammation/drug therapy , Cytokines , Edema/chemically induced , Edema/drug therapy , Edema/metabolism , Methylprednisolone/adverse effects
4.
Anim Biotechnol ; 34(7): 2159-2165, 2023 Dec.
Article in English | MEDLINE | ID: mdl-35622407

ABSTRACT

To evaluate the immunomodulatory effect of minocycline, the present study was carried out on the gene expression of toll-like receptor type-4 (TLR4) and some pro-inflammatory (IL-1ß, IL-6) and anti-inflammatory cytokines (IL-10) associated with lipopolysaccharide (LPS) -induced inflammation in human peripheral blood mononuclear cells (PBMCs). The PBMCs were collected and then 5.4 × 106 PBMCs/mL were used in eight groups as follows: control group (only media), LPS group (only LPS), methylprednisolone (Pred) group (LPS plus Pred), meloxicam (Melo) group (LPS plus Melo), three minocycline groups [M1, M5 and M25] (LPS plus 1, 5, and 25 µg/mL minocycline, respectively) and minocycline control (MC) group (5 µg/mL minocycline). After incubation for 24 h, the PBMCs were subjected to quantitative PCR assays. Gene expression levels of TLR4 were not changed in any groups. The IL-1ß levels were increased in the LPS group but the increases were much more intense in the other groups except Pred group. Compared with control group, IL-6 levels increased significantly in Melo, M1 and M25 groups. Significant increases of IL-10 levels were also observed in Melo, M25 and MC groups. It can be concluded that minocycline had dual pro- and anti-inflammatory activities with potential clinical immunomodulatory effects.


Subject(s)
Cytokines , Lipopolysaccharides , Humans , Animals , Cytokines/metabolism , Lipopolysaccharides/pharmacology , Interleukin-10/genetics , Leukocytes, Mononuclear , Toll-Like Receptor 4/genetics , Toll-Like Receptor 4/metabolism , Minocycline/adverse effects , Minocycline/metabolism , Interleukin-6/genetics , Interleukin-6/metabolism , Interleukin-6/pharmacology , Inflammation/chemically induced , Inflammation/drug therapy , Inflammation/genetics , Anti-Inflammatory Agents/adverse effects , Anti-Inflammatory Agents/metabolism , Gene Expression
5.
J Adv Vet Anim Res ; 10(4): 620-629, 2023 Dec.
Article in English | MEDLINE | ID: mdl-38370882

ABSTRACT

This study conducted a literature review to investigate the immunomodulatory effect of levamisole in both humans and farm animals. The following procedure was followed for database searching: PubMed, Google Scholar, Web of Science, and Cochrane Library. All research works were updated to September 2022. The terms used in the literature search were included: ("levamisole" OR "immunity" OR "immune system") AND ("adjuvant" OR "fish" OR "poultry" OR "farm animal" OR "cattle" OR "sheep"). The current review enlightens the extensive potential of levamisole as an adjuvant immunotherapeutic agent and explains its divergent applications beyond its antiparasitic use as an adjuvant, dietary supplement, immunostimulant, antiviral, and anti-cancer drug in humans and farm animals. In the articles examined, various mechanisms have been proposed for levamisole immunoprotective effects, but hormonal alteration and stress hormone reduction are indicated as the main mechanisms in various animal species.

6.
Vet Res Forum ; 12(2): 253-257, 2021.
Article in English | MEDLINE | ID: mdl-34345395

ABSTRACT

Sustained release drug formulations are frequently developed to reduce dosage frequency and to improve outcomes of drug therapy. This study evaluates the pharmacokinetic (PK) parameters of a novel injectable danofloxacin (DANO) formulation in comparison with a conventional product in an animal model. A recently synthesized DANO formulation, prepared by incorporation of DANO-loaded mesoporous silica nanoparticles in liposomes and integration of liposomes in chitosan and ß-glycerophosphate solution (lipogel) along with the conventional DANO product were injected subcutaneously (SC) in rabbits. Blood samples were collected at specific time points and DANO concentrations in plasma samples were measured. The PK parameters including maximum concentration (Cmax), time to reach Cmax (Tmax), area under the concentration versus time curves (AUC), area under the first moment concentration-time curve (AUMC) and mean residence time (MRT) were studied by non-compartmental analyses. The values of MRT (156.00 ± 20.00 hr), AUC (15.30 ± 3.00 µg mL-1 per hr) and Tmax (4.70 ± 1.60 hr) for lipogel formulation were higher than those of the conventional product (8.50 ± 3.60 hr, 3.70 ± 2.00 µg mL-1 per hr and 0.80 ± 0.26 hr, respectively). However, Cmax values for lipogel formulation (0.41 ± 0.15 µg mL-1) were significantly lower than those of the conventional drug product (0.68 ± 0.09 µg mL-1). It was concluded that the novel DANO lipogel effectively slowed down the drug absorption and the incorporation of liposomes in hydrogel could be a useful approach to maintain the therapeutic drug level for a longer period; however, more studies are needed in this field.

7.
Toxicol Mech Methods ; 30(5): 317-323, 2020 Jun.
Article in English | MEDLINE | ID: mdl-32031029

ABSTRACT

Introduction: Nonsteroidal anti-inflammatory drugs (NSAIDs) intervene in the COX (cyclooxygenase) pathways which generate two important inflammation mediators, prostaglandins (PGs) and leukotriene (LTs). Contradictory claims regarding the effect of NSAIDs in asthmatic patients continues to be an issue. The present study investigated the effects of COX inhibitors on the responsiveness of the tracheal tract and on the levels of LTC4 and PGE2 in cells of the bronchoalveolar lavage fluid in an allergic guinea pig model.Materials and Methods: Adult male Dunkin-Hartley guinea pigs (250 - 300 g) were divided into seven groups of six animals each. Four COX inhibitors, aspirin (200 mg/kg and 20 mg/kg), indomethacin (10 mg/kg), ketoprofen (10 mg/kg), and celecoxib (25 mg/kg), were given orally on day 17 to allergy induced guinea pigs at 0, 12, and 24 h before ovalbumin challenge on day 18. PGF2 and LT4 were measured in the bronchoalveolar lavage fluid as well as inflammatory cell count and total protein. Tracheal responsiveness to acetylcholine (Ach) and histamine (His) also was evaluated.Results: An augment in the response of the trachea to Ach and His, as well as overt allergenic signs including short breath, wheezing and sneezing, was observed. The most significant increase in tracheal hyper-responsiveness was observed in the ketoprofen-treated group with similar but less pronounced changes observed in the indomethacin-treated group. Although some variables increased with the aspirin and celecoxib treatments, overall the tracheal sensitivity was reduced. Inflammatory cells including eosinophils and neutrophils corresponded to the changes observed for each treatment group.Conclusion: Ketoprofen and indomethacin increased the tracheal sensibility to Ach and His; therefore, their administration is not recommended in patients susceptible to allergy.


Subject(s)
Acetylcholine/pharmacology , Cyclooxygenase Inhibitors/adverse effects , Dinoprostone/analysis , Histamine/pharmacology , Hypersensitivity/immunology , Leukotriene C4/analysis , Trachea/drug effects , Animals , Bronchoalveolar Lavage Fluid/chemistry , Bronchoalveolar Lavage Fluid/immunology , Cyclooxygenase Inhibitors/therapeutic use , Disease Models, Animal , Guinea Pigs , Hypersensitivity/drug therapy , Male , Ovalbumin/immunology , Trachea/immunology
8.
Front Microbiol ; 10: 1310, 2019.
Article in English | MEDLINE | ID: mdl-31244816

ABSTRACT

The PK/PD cut-off (PK/PDCO) value of florfenicol for calf pathogens was determined for long acting formulations (MSD Nuflor® and a bioequivalent generic product). PK/PDCO is one of the three MICs considered by VetCAST, a sub-committee of the European Committee on Susceptibility Testing (EUCAST), to establish a Clinical Breakpoint for interpreting Antimicrobial Susceptibility Testing (AST). A population model was built by pooling three pharmacokinetic data sets, obtained from 50 richly sampled calves, receiving one of two formulations (the pioneer product and a generic formulation). A virtual population of 5,000 florfenicol disposition curves was generated by Monte Carlo Simulations (MCS) over the 96 h of the assumed duration of action of the formulations. From this population, the maximum predicted MIC, for which 90% of calves can achieve some a priori selected critical value for two PK/PD indices, AUC/MIC and T>MIC, was established. Numerical values were established for two bacterial species of the bovine respiratory disease (BRD) complex, Pasteurella multocida and Mannheimia haemolytica. It was concluded that the PK/PDCO of florfenicol for both AUC/MIC and T>MIC was 1 mg/L.

9.
Res Pharm Sci ; 14(1): 46-54, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30936932

ABSTRACT

Interaction of morphine and seizure is complex. Mouse brain hippocampal slices were used to estimate how acute and chronic morphine treatment alters the low-magnesium artificial cerebrospinal fluid (LM-ACSF)-induced seizure activity. Hippocampal slices were taken from the normal and morphine-treated mice. The normal mice received saline while the other group (morphine-treated mice) received morphine daily for 5 consecutive days. Saline/morphine administration was performed subcutaneously (s.c, 0.1 mL) at postnatal days 14-18. Hippocampal slices of all animals were perfused with LM-ACSF followed by different morphine concentrations (0, 10, 100, and 1000 µM) or naloxone (10 µM). Changes in the spike count were considered as indices for quantifying the seizure activity in the slices. In hippocampus of both groups perfused with 10 or 1000 µM morphine, epileptiform activity was suppressed while it was potentiated at 100 µM morphine. The excitatory effect of morphine at 100 µM was stronger in normal mice (acute exposure) than in dependent mice (chronic exposure). Naloxone suppressed the epileptiform activities in both groups. Suppressive effect of naloxone was more significant in morphine-treated mice than in normal mice. The seizure activity in morphine-dependent mice was more labile than that of normal mice. It can be concluded that morphine had a biphasic effect on LM-ACSF-induced epileptiform activities in both groups. The occurrence of seizure was comparable in acute and chronic exposure of morphine but strength of the effect was considerably robust in normal mice. The down regulation of opioid receptors in chronic exposure is likely to be responsible for these differences.

10.
Int J Dev Neurosci ; 60: 56-62, 2017 Aug.
Article in English | MEDLINE | ID: mdl-28455226

ABSTRACT

Hippocampal slices of mouse brain were used to estimate how selective agonist and antagonist of opioid receptors alter Low-Mg+2 artificial cerebrospinal fluid (LM-ACSF)-induced epileptiform activities in normal and morphine-dependent mice. Brain slices were obtained from control and morphine-dependent mice. The morphine-dependent group received morphine once a day for 5 consecutive days, and the control group received saline. All injections were administered subcutaneously (s.c) in a volume of 0.1mL on postnatal days 14-18. Brain slices were perfused with LM-ACSF along with selective agonist and antagonist of µ, κ and δ opioid receptors. Changes in spike count per unit of time were used as indices to quantify the effects of LM-ACSF exposure in the slices. In both groups, DAMGO (selective µ opioid receptor agonist) and DPDPE (selective δ opioid receptor agonist) suppressed while Dyn-A (selective κ opioid receptor agonist) potentiated the epileptiform activity. Meanwhile, BFN-A (selective µ opioid receptor antagonist) recovered epileptiform activity in normal brain slices but not in morphine-dependent ones. NTI (selective δ opioid receptor antagonist) and nor-BNI (selective κ opioid receptor antagonist) decreased epileptiform activity. It seems that the excitatory effect of morphine on epileptiform activity was mediated through kappa receptors and its inhibitory effect was mediated via the mu receptor and, to a lesser degree, through the delta receptor. The pattern of effect was similar in normal and morphine-dependent slices, but the intensity of the effect was significantly stronger in normal mice. Finding of this study might be considered for further research and attention in epilepsy treatment.


Subject(s)
Epilepsy/drug therapy , Epilepsy/physiopathology , Morphine Dependence/complications , Morphine Dependence/drug therapy , Morphine Dependence/physiopathology , Narcotic Antagonists/administration & dosage , Receptors, Opioid/agonists , Action Potentials/drug effects , Animals , Animals, Newborn , Cells, Cultured , Dose-Response Relationship, Drug , Epilepsy/etiology , Female , Hippocampus/drug effects , Hippocampus/physiopathology , Male , Mice , Treatment Outcome
11.
Iran J Pharm Res ; 15(4): 663-676, 2016.
Article in English | MEDLINE | ID: mdl-28261309

ABSTRACT

Tilmicosin (TLM) is an important antibiotic in veterinary medicine with low bioavailability and safety. This study aimed to formulate and evaluate physicochemical properties, storage stability after lyophilization, and antibacterial activity of three TLM-loaded lipid nanoparticles (TLM-LNPs) including solid lipid nanoparticles (SLNs), nanostructured lipid carriers (NLCs), and lipid-core nanocapsules (LNCs). Physicochemical parameters such as particle size-mean diameter, polydispersity index, zeta potential, drug encapsulation efficiency (EE), loading capacity, and morphology of the formulations were evaluated and the effects of various cryoprotectants during lyophilization and storage for 8 weeks were also studied. The profiles of TLM release and the antibacterial activities of these TLM-LNPs suspensions (against Escherichia coli and Staphylococcus aureus) were tested in comparison with their corresponding powders. TLM-LNPs suspensions were in nano-scale range with mean diameters of 186.3 ± 1.5, 149.6 ± 3.0, and 85.0 ± 1.0nm, and also EE, 69.1, 86.3, and 94.3% for TLM- SLNs, TLM-NLCs, and TLM- LNCs respectively. TLM-LNCs gave the best results with significantly low particle size and high EE (p<0.05). Mannitol was the most effective cryoprotectant for lyophilization and storage of TLM-LNPs. The drug release profiles were biphasic and the release times were longer at pH 7.4 where TLM-NLCs and TLM-LNCs powders showed longer release times. In microbiological tests, S. aureus was about 4 times more sensitive than E. coli to TLM-LNPs with minimum inhibitory concentration ranges of 0.5-1.0 and 2-4 µg/mL respectively, and TLM-LNCs exhibited the best antibacterial activities. In conclusion, TLM-LNP formulations especially TLM-LNCs and TLM-NLCs are promising carriers for TLM with better drug encapsulation capacity, release behavior, and antibacterial activity.

12.
Trop Anim Health Prod ; 43(4): 843-9, 2011 Apr.
Article in English | MEDLINE | ID: mdl-21197575

ABSTRACT

Artemisinin has been used for centuries to treat malaria, intestinal tract helminthosis, diarrhea, and used as an antipyretic and sedative agent, but the usage in veterinary medicine is a new field. Recently, it has been used successfully to control experimental poultry coccidiosis. The present study aimed to determine the effects of different doses of artemisinin in broiler chickens with chronic usage. Sixty birds divided into one control and four treatment groups that fed rations mixed with artemisinin at doses of 17, 34, 68, and 136 ppm for 36 days. During the experiment, birds showed no clinical signs except anemia. In microscopic examinations, heart, lung, and spleen had no lesion, but liver, kidney, and brain showed various lesions. Degenerative lesions like intracytoplasmic eosinophilic inclusions were seen in both kidney and liver but fatty change was seen only in liver. There was no relationship between severity of the liver lesions and drug dosage. Central chromatolysis, scattered neuronal necrosis, and mild spongy changes were observed in five regions of the brain that were chosen for sectioning (motor cortex, cerebellar nuclei, midbrain nuclei, and hindbrain nuclei at two separate levels). Severity of lesions in brain was dose-dependent, and cerebral cortex was the most vulnerable area. Haematologic tests showed lower values for hematocrit and red blood cell count dose-dependently. In conclusion, artemisinin is a promising drug for prevention and control of coccidiosis in broiler chickens and its side effects are not too much serious especially at therapeutic doses.


Subject(s)
Antiprotozoal Agents/adverse effects , Artemisinins/adverse effects , Chickens/parasitology , Coccidiosis/veterinary , Coccidiostats/adverse effects , Plant Extracts/therapeutic use , Poultry Diseases/drug therapy , Animals , Antiprotozoal Agents/administration & dosage , Antiprotozoal Agents/therapeutic use , Artemisinins/administration & dosage , Artemisinins/therapeutic use , Brain/pathology , Coccidia/drug effects , Coccidiosis/drug therapy , Coccidiostats/administration & dosage , Coccidiostats/therapeutic use , Dose-Response Relationship, Drug , Erythrocyte Count/veterinary , Iran , Kidney/pathology , Liver/pathology , Parasitic Diseases, Animal/drug therapy , Phytotherapy/veterinary , Poultry Diseases/parasitology
13.
Basic Clin Pharmacol Toxicol ; 105(1): 46-50, 2009 Jul.
Article in English | MEDLINE | ID: mdl-19302435

ABSTRACT

The present study was conducted to investigate the effects of eicosapentaenoic acid on glucocorticoid-induced bone changes in rats, and to compare them with those of alendronate. Thirty six male Wistar rats, 2.5 months of age, were divided into six groups (n = 6 each) and treated with 0.9% NaCl (control), methylprednisolone 7 mg/kg, once a week subcutaneously, methylprednisolone + alendronate 20 microg/kg, twice a week subcutaneously and methylprednisolone + 80 or 160 or 320 mg/kg eicosapentaenoic acid, per day orally, for 6 weeks. At the end of the experiment, serum and urinary parameters of bone metabolism determined and bone histomorphometric analyses performed on cancellous bone of femoral epiphysis and metaphysis and cortical bone of tibial diaphysis. There were no significant differences in serum and urinary parameters among groups. Decrease of epiphyseal and metaphyseal trabecular width, epiphyseal bone area/tissue area and increase of epiphyseal trabecular separation observed in the methylprednisolone group compared to control. Alendronate restored all of these parameters except metaphyseal trabecular width, which increased significantly by eicosapentaenoic acid at the doses of 80 and 160 mg/kg. Effects of alendronate and 160 mg/kg eicosapentaenoic acid on bone area/tissue area, alendronate and eicosapentaenoic acid at the doses of 80 and 160 mg/kg on trabecular separation and alendronate and eicosapentaenoic acid at doses of 160 and 320 mg/kg on epiphyseal trabecular width were statistically similar. Methylprednisolone did not significantly change cortical bone parameters including cortical width and marrow area/cortical area. Eicosapentaenoic acid, especially, at the dose of 160 mg/kg exerts beneficial effects on methylprednisolone-induced bone changes in rats; these effects are similar or sometimes even better than alendronate.


Subject(s)
Alendronate/pharmacology , Bone Density Conservation Agents/pharmacology , Bone Diseases, Metabolic/chemically induced , Bone and Bones/drug effects , Eicosapentaenoic Acid/pharmacology , Glucocorticoids/toxicity , Methylprednisolone/toxicity , Alendronate/administration & dosage , Animals , Biomarkers/blood , Biomarkers/urine , Body Weight , Bone Density Conservation Agents/administration & dosage , Bone Diseases, Metabolic/pathology , Bone Resorption/chemically induced , Bone and Bones/pathology , Dose-Response Relationship, Drug , Eicosapentaenoic Acid/administration & dosage , Femur/pathology , Injections, Subcutaneous , Male , Osteogenesis/drug effects , Random Allocation , Rats , Rats, Wistar , Tibia/pathology
14.
Phytochem Anal ; 18(4): 333-40, 2007.
Article in English | MEDLINE | ID: mdl-17623368

ABSTRACT

A simple and rapid microwave-assisted extraction (MAE) procedure combined with 1H-NMR spectrometry was developed and optimised for the extraction and quantitative determination of capsaicin in Capsicum frutescens. The influence of experimental variables, including irradiation power, extraction temperature and dynamic extraction time before reaching the selected extraction temperature, on the performance of the extraction procedure was systematically studied using a Box-Behnken experimental design followed by a conventional central composite design approach. Statistical treatment of the results together with results from some additional experiments suggested optimum extraction conditions as 120 degrees C and 150 W, for 15 min with acetone as extractant. The optimised MAE method provides extracts that can be analysed quantitatively using 1H-NMR without any preliminary clean-up or derivatisation steps. In the 1H-NMR spectrum of the crude extracts the doublet signal in the delta range 4.349-4.360 ppm was well separated from other resonances in deuterated chloroform. The quantity of the compound was calculated from the relative ratio of the integral value of the target peak to that of a known amount of dimethylformamide as internal standard. In comparison with traditional Soxhlet extraction, the proposed method is less labour-intensive and provides a drastic reduction of extraction time and solvent consumption. In addition, MAE showed higher extraction yield and selectivity, with comparable reproducibility and recovery, relative to both conventional Soxhlet and sonication methods.


Subject(s)
Capsaicin/analysis , Capsaicin/chemistry , Capsicum/chemistry , Microwaves , Capsaicin/isolation & purification , Magnetic Resonance Spectroscopy , Plant Extracts/chemistry
15.
J Sep Sci ; 28(4): 387-96, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15792254

ABSTRACT

A simple and rapid microwave-assisted extraction (MAE) procedure was developed and optimized for two common color pigments, alizarin and purpurin, in various samples of Rubiaceae plants. Several variables that can potentially affect the extraction efficiency, namely temperature, methanol concentration in the extractant mixture, time, and solvent volume were optimized by means of a central composite design approach. The results suggest that temperature and methanol concentration in the solvent mixture are statistically the most significant factors. The separation and quantitative determination of the pigments was carried out in less than 6 min by a developed high-performance liquid chromatographic method with UV detection at 250 nm. Under optimum operating conditions, MAE showed significantly higher recoveries than those obtained by the conventional extraction methods (ultrasonic and reflux extraction), ranging from 84 to 94%. In addition, a drastic reduction of the extraction time (20 min versus 6 h) and solvent consumption (20 versus 100 mL) was achieved with a reproducibility (RSDs < 10%) comparable with that provided by the reflux extraction as a reference method.


Subject(s)
Anthraquinones/chemistry , Rubiaceae/chemistry , Chromatography, High Pressure Liquid , Microwaves , Molecular Structure , Pigmentation , Plant Extracts/chemistry , Plant Roots/chemistry , Reproducibility of Results
16.
J Sep Sci ; 27(13): 1130-6, 2004 Sep.
Article in English | MEDLINE | ID: mdl-15495416

ABSTRACT

A simple and rapid microwave-assisted extraction (MAE) procedure was developed and optimized for the extraction of paclitaxel (Taxol) from the needles of yew trees Taxus baccata L. grown in Iranian habitats. The samples, immersed in a methanol-water mixture, were irradiated with microwaves in a closed-vessel system. The method was evaluated using a factorial design approach based on parameters such as extraction time, temperature, methanol concentration in water (v/v), and the ratio of grams of sample to 10 mL of solvent. Statistical treatment of the results revealed that the selected parameters were all significant except the extraction time. Optimum conditions would be 1.5 g samples in 10 mL solvent (90% methanol), an extraction temperature of 95 degrees C, and an extraction time of 7 min. The extracts has been analyzed by reverse-phase high-performance liquid chromatography with UV detection (LC/UV) at 227 nm for quantification. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used for confirmation. The main advantage of the proposed MAE method versus conventional solvent extraction (CSE) are the considerable reductions in time (7 min versus 16 h) and in solvent consumption (20 mL versus 150 mL). The MAE procedure yielded extracts that could be analyzed directly without any preliminary clean-up or solvent exchange steps. Both extraction methods show RSDs lower than 10% and lead to comparable recoveries of paclitaxel (87-92%).


Subject(s)
Microwaves , Paclitaxel/chemistry , Paclitaxel/isolation & purification , Taxus/chemistry , Chromatography, High Pressure Liquid , Mass Spectrometry , Methanol/chemistry , Molecular Conformation , Plant Leaves/chemistry , Plant Leaves/radiation effects , Reproducibility of Results , Sensitivity and Specificity , Water/chemistry
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