چاپ صفحه |  صفحه نخست سامانه |  نویسندگان |  اطلاعات تفضیلی |  دانلود مقاله | دانشگاه علوم پزشکی تبریز |
| نویسنده ثبت کننده مقاله | شهرام صادق وند |
| مرحله جاری مقاله | تایید نهایی |
| دانشکده/مرکز مربوطه | دانشکده پزشکی |
| کد مقاله | 72058 |
| عنوان فارسی مقاله | Simultaneous determination of valproic acid and its main metabolite in human plasma using a small scale dispersive liquid–liquid microextraction followed by gas chromatography–flame ionization detection detection |
| عنوان لاتین مقاله | Simultaneous determination of valproic acid and its main metabolite in human plasma using a small scale dispersive liquid–liquid microextraction followed by gas chromatography–flame ionization detection |
| نوع ارائه | پوستر |
| عنوان کنگره / همایش | نوزدهمین کنگره سالانه اعصاب کودکان و نوجوانان ایران |
| نوع کنگره / همایش | ملی |
| کشور محل برگزاری کنگره/ همایش | Iran (Islamic Republic) |
| شهر محل برگزاری کنگره/ همایش | قشم |
| سال انتشار/ ارائه شمسی | 1398 |
| سال انتشار/ارائه میلادی | 2019 |
| تاریخ شمسی شروع و خاتمه کنگره/همایش | 1398/11/30 الی 1398/12/02 |
| آدرس لینک مقاله/ همایش در شبکه اینترنت | http://congress.hums.ac.ir/canc19/fa/ |
| آدرس علمی (Affiliation) نویسنده متقاضی | pediatric Health Research Center, Tabriz University of Medical Sciences |
| نویسنده | نفر چندم مقاله |
|---|---|
| شهرام صادق وند | اول |
| محمد برزگر | دوم |
| بهروز فریدونی | سوم |
| مریم خوب نسب جعفری | چهارم |
| ابوالقاسم جویبان | پنجم |
| میر علی فرج زاده | چهارم |
| عنوان | متن |
|---|---|
| خلاصه مقاله | Sodium valproate powder (98%) was kindly donated by Rouz Darou Pharmaceutical Co. (Tehran, Iran). 3-eptanone (98%) was purchased from Sigma-Aldrich (Louis,USA). Trifluoroacetic acid, chloroform, 1,2-dichloroethane(1,2-DCE), 1,1,2,2-tetrachloroethane (1,1,2,2-TCE),dimethyl sulfoxide (DMSO), and carbon tetrachloride were purchased from Merck (Darmstadt, Germany). Acetonitrile,methanol, and acetone were from Sigma-Aldrich. Deionized water was obtained from Ghazi Pharmaceutical Company(Tabriz, Iran). A stock mixture solution of the studied analytes was prepared by dissolving appropriate amounts of the analytes in methanol at a concentration of 1000 mg L−1 of each analyte. Working standard solutions were prepared daily by spiking the drug-free plasma with the analytes.Another mixture standard solution (1000 mg L−1 of each analyte) in chloroform (extraction solvent) was prepared and directly injected into the separation system each day (three times) in order to evaluate the instrumental system quality and to calculate extraction recoveries (ERs) and enrichment factors (EFs) of the analytes. Real samples Drug-free (blank) human plasma was obtained from a healthy donor (Iranian Blood Transfusion Research Center, Tabriz, Iran) and stored in a refrigerator at −20 °C. In addition,ten blood samples were collected from the patients with seizure who had signed consent forms approved by the Ethics Committee of Tabriz University of Medical Sciences.These were collected in heparinized tubes and centrifuged at a rate of 8000 rpm for 5 min immediately after collection,and the plasmas were separated and stored at −20 °C until assay. Apparatus Analysis of the selected analytes was carried out using an Agilent 7890A gas chromatograph (Agilent Technologies, CA, USA) equipped with a split/splitless inlet operated at 300 °C in a splitless mode (sampling time 1 min and split ratio of 1:10) and an FID. Nitrogen (99.999%, Gulf Cryo,United Arabic Emirates) was used as the carrier gas (at a constant flow rate of 1 mL min−1) and make up gas (25 Ml min−1). Chromatographic separation was achieved on an HP-5 capillary column (30 m × 0.32 mm i.d. with a 0.25 μm stationary film thickness) (Hewlett–Packard, Santa Clara,USA). The oven temperature was programmed from 70 °C held (for 2 min) to 200 °C at a rate of 15 °C min− 1 and thenincreased to 300 °C at a rate of 20 °C min−1 for cleaning column. Chem Station software was used for data acquisition and processing. A 1-μL microsyringe (zero dead volume,Hamilton, Switzerland) was used for the injection of samples into GC. The FID temperature was maintained at 300 ◦C. Hydrogen gas was generated with a hydrogen generator(GLAIND–2200, Dani, Italy) for FID at a flow rate of 40 mL min−1. Air flow rate for FID was 400 mL min−1. A vortex from Labtron Company (Tehran, Iran) was used in sample preparation and extraction step. A Metrohm pH meter model 744 (Herisau, Switzerland) was used in pH measurements.Sigma centrifuge (Osterode, Germany) was used in proteins precipitation and extraction steps. Results Selection of extraction solvent The most important step in optimization of the proposed method is the selection of a suitable extraction solvent. In this method, the extraction solvent should have low solubility in water and higher density than water. Low solubility provides a distinct and well defined two-phase system with minimum loss of the analytes. An extraction solvent denser than water facilitates the removal of the extractant sedimented at the bottom of the microtube after centrifuging.Therefore, four extraction solvents including carbon tetrachloride, 1,1,2,2-TCE, 1,2-DCE, and chloroform were tested. A series of experiments was performed with different volumes of the mentioned extraction solvents to achieve a same sedimented phase volume in the proposed method.Solubility of the solvents in water is different. Therefore, to reach 15 ± 0.5 μL sedimented phase volume, it is necessary to add different volumes of the mentioned solvents. Thereby 19, 20, 22, and 25 μL of 1,1,2,2-TCE, chloroform, carbon tetrachloride, and 1,2-DCE, respectively, along with 60 μL acetonitrile as a disperser were used. As observed in Fig. 1,chloroform gives the highest peak areas for the analytes.Therefore, it was selected as the favorable extraction solvent. Selection of extraction solvent volume The extractant volume can influence EF and ER of the proposed method. To investigate the effect of extractant volume,a series of experiments was performed with different volumes of chloroform (15, 20, 25, and 30 μL). The results revealed that the analytical signals of VPA and 3-heptanone decreased proportionally by increasing chloroform volume,indicating that the ER was constant within the range of chloroform volumes used. Volumes less than 15 μL were not taken into account due to the difficulty in collecting the solvent after extraction. So, in order to obtain high EF, 20 μLof chloroform was selected as the optimum volume of the extractant. Selection of disperser solvent The most important criterion in the selection of a dispersive solvent is that it should form a cloudy state when it is injected along with the extractant into an aqueous phase.On the other hand, the dispersive solvent should be soluble in the aqueous phase and the extraction solvent. Hence,acetonitrile, methanol, acetone, and DMSO were tested as dispersive solvents and their effects on the extraction efficiency of the analytes were investigated. In investigation of this factor, a series of aqueous solutions was studied using a mixture containing 20 μL of chloroform and 60 μL of different dispersive solvents. The results in Fig. 2 indicate that the maximum analytical signals are achievable using acetonitrile as a dispersive solvent. Hence, acetonitrile was selected as the disperser solvent in this method. Conclusion A simple, efficient, and low-cost extraction method was proposed for the extraction of VPA and its metabolite (3-heptanone) from human plasma sample prior to their determination by GC-FID. The developed method was successfully employed in identification and quantitation of the analytes in the plasma samples of patients. The results showed that the proposed method may be useful for therapeutic drug screening of VPA in human plasma as well as in pharmacokinetic studies of this drug |
| کلمات کلیدی | Valproic acid • 3-Heptanone • Plasma |
| نام فایل | تاریخ درج فایل | اندازه فایل | دانلود |
|---|---|---|---|
| DR.pptx | 1399/02/13 | 1164031 | دانلود |
| scan.pdf | 1399/02/13 | 178246 | دانلود |
