شیمى کاربردى روز

شیمى کاربردى روز

توسعه و بهینه سازی مرحله به مرحله روش کروماتوگرافی مایع با عملکرد بالا با تکنیک آماده سازی نمونه رقیق و تزریق مستقیم برای اندازه‌گیری بنیدیپین هیدروکلرید در نمونه‌های دارویی و بیولوژیکی

نوع مقاله : مقاله علمی پژوهشی

نویسنده
دانشکده علوم پایه، گروه شیمی، دانشگاه یاسوج، یاسوج، ایران
چکیده
در این کار پژوهشی از روش کروماتوگرافی مایع با کارایی بالا با آشکارساز مرئی- فرابنفش (HPLC-UV-Vis) برای اندازهگیری داروی بنیدیپین، در نمونههای دارویی، اورین انسان و نمونه‌ی سرم استفاده شده است. روش پیشنهادی پس از بهینه‌سازی شرایط مختلف کروماتوگرافی و سایر پارامترهای آزمایشی، بر اساس دستورالعمل‌های کنفرانس بین‌المللی هماهنگ‌سازی اعتبارسنجی شد.
نتایج مطلوب با ستون ACE 5µm C18 (150 میلی متر × 60/4 میلی متر؛ 00/5 میکرومتر) در 45 درجه سانتی گراد به دست آمد. فاز متحرک شامل استونیتریل: بافر (40 میلی مولار استات آمونیوم) با غلظت 75: 25 درصد حجمی-حجمی (V/V%)تنظیم شده در pH برابر 75/6 و با سرعت جریان 00/1 میلی لیتر در دقیقه است. طول موج 238 نانومتر انتخاب شد. روش به طور کامل اعتبارسنجی شد و پارامترهای اعتبارسنجی عبارت بودند از: محدوده خطی 00/25-15/0میلی گرم بر لیتر و ضریب همبستگی 999/0 برای همه نمونه ها. مقادیر حد تشخیص داروی بنیدیپین هیدروکلرید به ترتیب 46/0، 20/1 و 30/8 میکرو گرم بر لیتر در استونیتریل، سرم و اورین انسان یافت شد. مقادیر حد کمی سازی در استونیتریل، سرم و اورین انسان به ترتیب 50/1، 80/3 و 00/27 میکرو گرم بر لیتر بود. دقت بین روز و درون روز، که با انحراف استاندارد نسبی (%RSD) نشان داده شده است، به ترتیب 13/0 و 27/0 درصد در محلول بافر/استونیتریل بدست آمد. میانگین مقادیر بازیابی نسبی بین 00/97٪ و 00/105٪ بود. بنابراین، روش پیشنهادی سریع و دقیق است و می‌تواند با موفقیت در مطالعات فارماکوکینتیک و عملکرد بالینی معمول استفاده شود
کلیدواژه‌ها
موضوعات

عنوان مقاله English

Development and Step-by-Step Optimization of the HPLC Method with Dilute-and-Shoot Sample Preparation Technique for Determination of Benidipine Hydrochloride in Pharmaceutical and Biological Samples

نویسنده English

Fariba Mollarasouli
Faculty of Basic Sciences, Department of Chemistry, Yasuj University, Yasuj, Iran
چکیده English

This research employed a method using high-performance liquid chromatography with a visible-ultraviolet detector (HPLC-UV-Vis) to measure Benidipine Hydrochloride in pharmaceutical, human urine, and serum samples. The proposed method was validated based on the guidelines of the International Conference on Harmonisation (ICH) after optimizing various chromatography conditions and other experimental parameters.
Optimal results were obtained using an ACE 5µm C18 column (150 mm × 6.4 mm; 0.5µm) at 45 °C. The mobile phase consisted of acetonitrile: buffer (40 mM ammonium acetate) in a ratio of 75:25 v/v% adjusted to a pH of 6.75 at a flow rate of 0.1 mL/min. A wavelength of 238 nm was selected. The method was fully validated, and the validation parameters included a linear range of 0.15-25.00 mg/L and a correlation coefficient of 0.999 for all samples. The detection limits of Benidipine Hydrochloride were found to be 0.46, 1.20, and 8.30 µg/L in acetonitrile, serum, and human urine, respectively. The quantification limits in acetonitrile, serum, and human urine were 1.50, 3.80, and 27.00 µg/L, respectively. The precision, between-day and within-day, represented by the relative standard deviation (%RSD), was found to be 0.13% and 0.27% respectively in the buffer/acetonitrile solution. The average relative recovery values ranged between 97.00% and 105.00%. Thus, the proposed method is rapid and precise, and it can be successfully employed in pharmacokinetic studies and routine clinical performance.

کلیدواژه‌ها English

High-performance liquid chromatography
Benidipine Hydrochloride
Blood serum sample
human urine sample
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