جستجوی مقالات مرتبط با کلیدواژه "human plasma" در نشریات گروه "پزشکی"
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Background and Aims
To prepare human plasma for therapeutic purposes and serum with a low risk of contamination for cell culture use, a technique is needed to reduce blood borne pathogens. Silver nanoparticles (AgNPs) are a suitable option due to their low-risk of microbial resistance and strong antimicrobial activity.
Materials and MethodsAs representative microorganisms, we tested Herpes simplex type 1 (HSV-1), Escherichia coli (ATCC 25922), and Staphylococcus aureus (ATCC 1413). The cytotoxicity of biosynthesized AgNPs was assessed on the Vero cell line. The Maximum Non-Toxic Dose (MNTD) of AgNPs did not affect the virus. In order to address the high toxicity of AgNPs and utilize the highest colloidal concentration of AgNPs (300 µg/ml) that did not precipitate within the 24-hour examination period, we developed a removal process. AgNPs were removed after being exposed to viruses and bacteria. This removal step was done by addition of magnetic iron oxide nanoparticles (iron oxide NPs). Then, treated virus and bacteria were exposed to neodymium magnet, and were used for cell inoculation.
ResultsBoth AgNPs and iron oxide NPs reduced the titer of virus by at least 3.5 log when used together. Additionally, they inactivated 106 CFU/ml of bacteria in human plasma. While each nanoparticle exhibited antiviral activity individually, but also their effect was enhanced when used simultaneously.
ConclusionHuman plasma pathogens were reduced using AgNPs and/or iron oxide NPs. The plasma could be treated with these nanoparticles, and subsequently the magnetic nanoparticles that adsorbed silver nanoparticles were separated from the plasma using a neodymium magnet, as one of the techniques for pathogen reduction.
Keywords: Pathogen reduction, Human plasma, Herpes simplex type 1, Silver nanoparticles, Iron oxide nanoparticles -
A fast and simple method for extraction of carvedilol in human plasma samples based on salting out assisted liquid-liquid extraction (SALLE) is described. The method involving extraction of carvedilol with water-miscible organic solvent acetonitrile when solvent phase separation occurs using NaCl as a salt. The extracted phase was analyzed by high- performance liquid chromatography with ultra violet detection at 240 nm. The procedure has been optimized with respect to type and amount of salt, volume of sample, extraction solvent and the pH of solution. In the optimal condition the linear calibration range was 5-500 μg L-1 and the correlation coefficient was 0.9965. The limit of detection and limit of quantification were 1.0 µg L-1 and 3.3 μg L-1, respectively and a relative standard deviation of 3.5 % for five replicates were obtained. In spiking experiments on real samples, the average recoveries found by the present method were between 96.0% and 105.0%.Keywords: Salting out assisted liquid, liquid chromatography, Carvedilol, HPLC, Human Plasma
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BackgroundMalondialdehyde (MDA) is a commonly used biomarker of oxidative stress in clinical studies and has been measured in many pathological conditions during last decades. Different analytical methods have been reported for determination of MDA in biological samples in which MDA was adducted with thiobarbituric acid (TBA) to produce more sensitive chromophore and also convert it to a fluorescent compound. In spite of the routine applications of this derivatization and subsequent analysis of MDA in biomedical studies, its reliability, repeatability and reproducibility is questionable. The aim of this work is to investigate the effects of some factors on the repeatability of MDA determinations in standard solutions and also in plasma samples using spectroscopic method.MethodsMDA-TBA adduct is prepared in standard solutions and the effects of pH, temperature, reaction time, open, closed and reflux systems and the ratio of MDA and TBA is investigated by measuring the absorbance of the solution at 532 nm. These effects are also investigated in human plasma samples.ResultsThe best results are obtained at pH 2.5, temperature of 70 °C, reaction time of 150 minutes, reflux system and ratio of 2.ConclusionUsing the optimized conditions are resulted in better repeatability.Keywords: Malondialdehyde, Thiobarbituric acid, Derivatization, Reproducibility, Human plasma
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Background And AimsA selective and sensitive high performance liquid chromatography-electrospray ionization mass spectrometry method has been established for determination of methotrexate in human plasma.MethodsMethotrexate was extracted from plasma with acetonitrile. The mobile phase consisted of acetonitrile-water-formic acid 74: 25: 1(v/v). Twenty µl of sample was chromatographically analyzed using a repacked ZORBAX-XDB-ODS C18 column (2.1mm×30mm, 3.5 micron). The mode of mass spectrometry was selected-ion monitoring (SIM).ResultsStandard curve was linear (r= 0.998) over the concentration range of 0.1100.0 ng/ml and showed a suitable accuracy and precision. The limit of detection (LOD) was 0.05ng/ml.ConclusionThe method is quick, easy, very steady and precise for the partition, assignment and evaluation of methotrexate in human plasma.Keywords: Methotrexate, LC, MS, Human plasma
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Cefixime is essential member of orally energetic third generation cephalosporin and has tremendous activity aligned with many pathogens. The virtual bioavailability of a newly industrial dispersible tablet as compared with a recognized identified formulation. A simple and available reversed-phase HPLC method with UV detection has been urbanized and validate for cefixime evaluate in human plasma using a C18 analytical column and a mobile phase of tetrabutylammonium hydroxide (pH 6.5)-acetonitrile (3:1 v/v). The detection wavelength was 280 nm. To method observed major linear response-concentration association all through the cefixime concentration range of 15-100 ng/ml, with the average accuracy within-run and between-run values of 97.29% and 99.27%. The average drug recovery from plasma was 98.2% throughout the linear concentration range. The limits of detection (LOD) and quantitation (LOQ) of the method were 5 and 15 ng/ml, respectively. The method is quick, easy, very steady and precise for the partition, assignment, evaluation of cefixime in human plasma.Keywords: Cefixime, human plasma, reversed-phase HPLC
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A simple and sensitive high performance liquid chromatography-electrospray ionization mass spectrometry method has been evaluated for the assignment of clonidine hydrochloride in human plasma. The mobile phase composed of acetonitrilewater 60:40 (v/v), and 0.2% formic acid 20 µL of sample was chromatographically analyzed using a repacked ZORBAX-XDB-ODS C18 column (2.1 mm×30 mm, 3.5 micron). Detection of analytes was achieved by tandem mass spectrometry with electrospray ionization (ESI) interface in positive ion mode was operated under the multiple-reaction monitoring mode (m/z 230.0 → 213). Sample pretreatment involved in a one-step protein precipitation (PPT) with methanol and percholoric acid (HClO4) of 0.15 mL plasma. Standard curve was linear (r = 0.998) over the concentration range of 0.01-10.0 ng/ml and showed suitable accuracy and precision. The limit of quantification (LOQ) was 0.01 ng/ml. The method is rapid, simple, very steady and precise for the separation, assignment, evaluation of clonidine healthy in human plasma.Keywords: Clonidine hydrochloride, LC-MS, Human plasma
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A quick and thoughtful liquid chromatography–tandem mass spectrometry (LC-MS) method has been established and authorized for the estimation of amlodipine and atorvastatin in human plasma.MethodsLC-MS with electrospray ionization (ESI) interface in positive ion mode was functioned under the multiple-reaction monitoring (MRM) mode was used for detection of analytes. Ethyl acetate was secondhand for extraction of analytes from plasma by simple liquid–liquid extraction technique. The re-formed samples with a C18 column by pumping acetonitrile-ammonium acetate buffer (10 mM, pH = 3.0), 70:30 (v/v) at a flow rate of 0.15 mL/min were chromatographed. The standard curves were established to be linear in the range of 0.2–20 ng/mL for atorvastatin and 0.1–10 ng/mL for amlodipine with mean correlation coefficient of ≥0.999 for each analyte.ResultsThe lower limit of quantification for amlodipine and atorvastatin were demonstrated to be 0.1 ng/ml and 0.2 ng/ml respectively. The mean (SD) Cmax and Tmax values of amlodipine later supervision of the test and reference were: 6.58 (0.22) versus 6.64 (0.37) ng/mL, 6.12(0.86) versus 6.13 (0.73) hours respectively. The mean (SD) Cmax and Tmax values of atorvastatin later government of the test and reference, were 61.66 (3.05) versus 62.16 (0.76) ng/mL, 4.21(0.86) versus 4.22 (0.73) hours respectively.ConclusionThe results proposed the test formulation of amlodipine and atorvastatin is bioequivalence with reference formulation and the established evaluate method was successfully realistic to a pharmacokinetic and bioavailability trainings in 20 human male volunteers following oral administration of amlodipine and atorvastatin.Keywords: Amlodipine, Atorvastatin, LC, MS, Pharmacokinetics, Human plasma, Bioequivalence
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A rapid and sensitive liquid chromatography–tandem mass spectrometry (LC-MS) method for the estimation of enalapril and enalaprilat in human plasma. Detection of analytes was achieved by tandem mass spectrometry with electrospray ionization (ESI) interface in positive ion mode was operated under the multiple-reaction monitoring mode. Sample pretreatment involved in a one-step protein precipitation (PPT) with percholoric acid (HClO4) of 0.15ml plasma. The reconstituted samples were chromatographed on C18 column by pumping methanol: water: acid formic74:24:2 (v/v)at a flow rate of 0.2 mL/min.Each plasma samplewaschromatographedwithin1.25min.The standard curves were found to be linear in the range of 0.1–20ng/mL for enalapril and enalaprilat with mean correlation coefficient of ≥0.999 for each analyte. The intra-day and inter-day precision and accuracy results were well within the acceptable limits.The limit of quantification(LOQ) was 0.1ng/ml for enalapril andenalaprilat. The mean (SD) Cmax, Tmax, AUC0–tand AUC0–∞ values of enalaprilversusenalaprilatafter administration of the 10 mg enalapril, respectively, were in this manner: 141.33(3.51) versus73.33 (5.03) ng/mL, 1.15(1.45) versus 4.12 (1.74) hours, 142.57 (34.34) versus 425.94(13.09) ng/mL/h, and 150.74 (16.69) versus 455.80 (65.11) ng/mL/h. The mean (SD) t1/2 was 2.72 (2.01) hours for the enalapril and 6.34 (2.13) hours for the enalaprilat. The developed assay method was successfully applied to a pharmacokinetic study in human male volunteers.
Keywords: Enalapril, enalaprilate, LC-MS, Human plasma, Pharmacokinetic, Bioequivalence Study -
Objective(s)Dispersive liquid-liquid microextraction coupled with gas chromatography (GC)-flame ionization detector was developed for the determination of valproic acid (VPA) in human plasma.Materials And MethodsUsing a syringe, a mixture of suitable extraction solvent (40 µl chloroform) and disperser (1 ml acetone) was quickly added to 10 ml of diluted plasma sample containing VPA (pH, 1.0; concentration of NaCl, 4% (w/v)), resulting in a cloudy solution. After centrifugation (6000 rpm for 6 min), an aliquot (1 µl) of the sedimented organic phase was removed using a 1-µl GC microsyringe and injected into the GC system for analysis. One variable at a time optimization method was used to study various parameters affecting the extraction efficiency of target analyte. Then, the developed method was fully validated for its accuracy, precision, recovery, stability, and robustness.ResultsUnder the optimum extraction conditions, good linearity range was obtained for the calibration graph, with correlation coefficient higher than 0.998. Limit of detection and lower limit of quantitation were 3.2 and 6 μg/ml, respectively. The relative standard deviations of intra and inter-day analysis of examined compound were less than 11.5%. The relative recoveries were found in the range of 97 to 107.5%. Finally, the validated method was successfully applied to the analysis of VPA in patient sample.ConclusionThe presented method has acceptable levels of precision, accuracy and relative recovery and could be used for therapeutic drug monitoring of VPA in human plasma.Keywords: Dispersive liquid, liquid, microextraction, Gas chromatography, ame, ionization detector, Human plasma, Valproic acid
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A rapid, sensitive and reproducible HPLC method was developed and validatedfor the analysis of amifostine (AMF) and/or its metabolite, WR-1065 inhuman plasma. The method involves the alkylation of free sulfydryl group with iodoacetic acid followed by derivatization of the drug and its metabolite witho-phthaldialdehyde (OPA) andUVdetection at 340 nm. The derivatized AMF and WR-1065 were eluted inless than 11 min, and in the case of the metabolite with no interferences from the endogenous plasma peaks. Cystein was used as the internal standard. Analysis was carried out on a Eurosphere Performance (RP-18e, 100 × 4.6 mm) analytical column. The mobile phase was a mixture of methanol and phosphate buffer 0.03 M pH = 2.7 at a ratio of 40: 60v/v, respectively, with a flow rate of 1.5 mLmin-1.Limit of detection was 0.5µgmL-1.The method involved a simple extraction procedure for AMF and/or its metabolite and analytical recovery was 90 ± 0.9%.The calibration curve was linear over the concentration range of 1-200 µgmL-1. The coefficients of variationfor intra-day and inter-day assayswere less than 10%.Keywords: amifostine, WR, 1065, human plasma, HPLC, OPA derivatization
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In previous studies, mebudipine, a dihydropyridine calcium channel blocker, showed a considerable potential to be used in cardiovascular diseases. The aim of the current study was to develop a valid method using reversed-phase high performance liquid chromatography coupled with electrospray ionization mass spectrometry to assay mebudipine in the human plasma. Separation was achieved on a Zorbax Eclipse® C18 analytical column using a mobile phase consisted of methanol/water (90:10, v/v). The flow rate was 0.6 mL/min and carbamazepine was used as an internal standard (IS). This method involved the use of [M +Na]+ ions of mebudipine and IS at m/z 411 and 259, respectively with the selected ion monitoring (SIM) mode. There were no interfering peaks from endogenous components in blank plasma chromatograms. Standard curves were linear (r2>0.99) between 5 to 100 ng/mL. The mean extraction efficiency was about 84% and the limit of quantification for mebudipine was 5 ng/mL in plasma. The coefficient of variation and error at all of the intra-day and inter-day assessments were less than 11%. The results indicated that this method is a fast, accurate, sensitive, selective and reliable method for the determination of mebudipine in the human plasma. The assay method has been successfully used to estimate plasma concentration of mebudipine after the oral administration of 2.5 mg tablet in healthy adults.Keywords: Mebudipine, Liquid chromatography, mass spectrometry, human plasma
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زمینه و هدفپلاسمای انسانی منشا پروتئین ها و آنزیم های متفاوتی می باشد که دارای اثرات دارویی متفاوتی می باشند. از این داروها می توان فاکتورهای انعقادی هشت و نه، ایمونوگلوبولین ها، و آلبومین را نام برد. آلبومین دارای مولکولی زنجیره ای با 585 آمینو اسید می باشد. آلبومین دارای 17 پل دی سولفیدی می باشد که ایجاد یک پروتئین کروی پایدار می نماید. وزن مولکولی آلبومین حدود 66500 دالتون می باشد که حدود 80 درصد فشار انکوتیک خون را سبب می شود. هدف از این مطالعه، تهیه ی آلبومین از پلاسمای انسانی با استفاده از اتانول و دمای پایین بوده است.روش بررسیدر این مطالعه پلاسمای مورد استفاده از نوع FFP و اتانول مصرفی از نوع تجاری با خلوص 94 تا 96 درصد بود. مواد شیمیایی مصرفی شامل سدیم هیدروکساید، سدیم کلراید، اکتانوئیک اسید Analytical Grade و از شرکت مرک بود. در این مطالعه جداسازی آلبومین از پلاسمای انسانی با استفاده از اتانول در دمای پایین صورت پذیرفت. با تنظیم پارامترهایی از قبیل pH، درجه حرارت، و درصد غلظت الکل در شرایط مختلف اجزای مختلف خونی از پلاسما جدا گردید که شامل قسمت های یک تا پنج بود. هر کدام از این اجزا می تواند منبع خوبی جهت جداسازی پروتئین ها و آنزیم ها در مراحل بعدی باشد. جزء اول منبع خوبی برای فیبرینوژن و فاکتور هشت انعقادی، جزء دوم مناسب برای تهیه ی ایمونوگلوبولین ها و در نهایت جزء پنجم منبع بسیار مناسب جهت تهیه ی آلبومین بود.
یافته هادر این مطالعه آلبومین با دو غلظت پنج و بیست درصد تهیه گردید و مقایسه ی کیفیت در دو غلظت حکایت از خلوص بیش از 95 درصد داشت. در تهیه ی آلبومین با غلظت 5 درصد راندمان 71 درصد و در تهیه ی آلبومین با غلظت 20 درصد راندمان 5/75 درصد به دست آمد.نتیجه گیریآلبومین تهیه شده خلوص بیش از 95 درصد را نشان داد و میزان پلیمر آن کمتر از 5 درصد بود. در تهیه ی آلبومین 5 درصد می توان از غلظت پایین تر پایدار کننده ی سدیم کاپریلات استفاده نمود و همچنین در فرمولاسیون نشان داده شده که فرآیند تهیه را می توان با pH حدود7/4 به جای 5/6 شروع نمود، زیرا امکان آلودگی کاهش می یابد.
کلید واژگان: خالص سازی, پلاسمای انسانی, پروتئین, آلبومینBackground And ObjectiveHuman plasma is a suitable source for different proteins and enzymes with a variety of therapeutic effects. Examples include coagulation factors VIII and IX, immunoglobulin, and albumin, which consists of a 585 amino-acid chain, weighing 66.5 kD. Albumin molecule also has 17 di-sulfide bridges which form a stable spherical protein structure, which it cause of 80% of oncotic blood pressure. The aim of this study was to separate albumin from the human plasma by using ethanol at low temperature.Materials And MethodsFresh frozen plasma was used as the starting material. In this study, albumin was separated from human plasma by using ethanol (commercial grade with 94-96% purity) in low temperature. By adjustment of different parameters such as pH, temperature, and concentration of alcohol, different factions of plasma can be separated which are from fraction I to fraction V. Fraction I is a good source for fibrinogen and coagulation factor VIII preparation, fraction II is suitable for preparation of immunoglobulin, and fraction V is a good source for albumin preparation.ResultsAlbumin was prepared with 5 and 20 percent concentrations. Quality comparison of the two concentrations showed a purity of more than 95%. The average yields of the 5 % and 20 % albumin preparations were 71% and 75.5%, respectively.ConclusionOur data show that the produced albumin has a purity of higher than 95% and contains less than 5% polymer. In preparation of 5 % albumin, it is possible to use lower concentrations of sodium caprylate as stabilizer. It has also been shown in the formulation that the preparation process can be started at the acidic pH of 4.7, instead of 6.5, in order to avoid bacterial contamination.Keywords: Purification, Human plasma, Protein, Albumin -
A simple, reproducible and rapid gas chromatographic method for precise determination of valproic acid (VPA) in human plasma has been developed. Total time for sample preparation and GC analysis is less than 45 min. After plasma protein precipitation, VPA was extracted into chloroform with suitable recovery. By using Stabilwax®-DA capillary GC column, a symmetrical gas chromatographic peak was obtained without the need for derivatization. The calibration curve was proved to be linear (r2 = 0.998) in a wide concentration range (0.45-100 μg/ml). Inter-day and intra-day accuracy and precision of this method was investigated during the method validation and the method has good precision and accuracy. This method is highly reproducible with a limit of detection 150 ng/ml of VPA in human plasma and could be used in TDM and pharmacokinetic studies.Keywords: Determination, Gas chromatography, Human plasma, Valproic acid
نکته
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