Spectrophotometer PDF - Medical Lab Tech Lec. 9

Med. Lab. Tech. Dep. Medical Instruments and equipments Lec.9/ 1st year...

Med. Lab. Tech. Dep. Medical Instruments and equipments Lec.9/ 1st year Lect Assistance Muhja Sattar Spectrophotometer: Introduction Spectrophotometry is a method to measure how much a chemical substance absorbs light (at one or more wavelengths) by measuring the intensity of light as the beam of light passes through a sample solution. This depends on the concentration of that chemical substance. The basic principle is that each compound absorbs light over a certain range of wavelength. ‫تعريف‬ Spectrophotometer is an instrument that measures the intensity of light after it passes through a sample solution. ‫جهاز يقيس كمية الضوء الذي يخترق العينه‬ ‫مبدأ العمل‬ ‫يقيس الفوتونات‬. Principle of Working: A spectrophotometer consists of two words as ‘spectrometer’ for producing light of selected wavelength and ‘photometer’ for measuring the intensity of light. Various spectra are used in spectrophotometers such as ultraviolet and visible spectra. A clear specimen will yield 100% transmittance, while a turbid sample will deflect a considerable portion of the light rays and will have a lower percent transmittance. When light is transmitted through a ‫ يتكون مقياس الطيف الضوئي من كلمت= باسم‬:‫مبدأ العمل‬ solution, some of it may be absorbed. ‫نتاج‬B‫"مطياف" ل‬ If the absorption occurs (in‫ضوء‬ the‫ ال‬ultraviolet ‫ء لقياس شدة‬or ‫ضو‬infrared ‫مقياس ال‬regions )‫حدد و‬H‫ا‬of‫ي‬the ‫وج‬H‫ا‬electromagnetic ‫ضوء الطول‬ spectrum, the solution will appear colorless. But if the absorption occurs in the visible region of the spectrum, the solution will appear colored. The transmission of a sample proportional to thickness and concentration decreases, exponentially. Absorbance of a sample is proportional to thickness of the sample and ‫حفيد الش?ل‬ concentration increases, linearly.  Med. Lab. Tech. Dep. Medical Instruments and equipments Lec.9/ 1st year Lect Assistance Muhja Sattar ‫طلوبه تذكرها ويا الوحدة‬C‫ ا‬uv ‫ شكد ال‬-١ Depending on the range of wave length of the light source, spectrophotometers can be classified into two different types: ‫ الرقم والوحدة‬1-UV range spectrophotometer: Uses light over the ultraviolet range, and wave length ranges between /‫ مطلوب‬185 - 400 nm. ‫ستخدامات مالتهن مهمة‬H‫ا‬ 2-Visible range spectrophotometer: Uses a tungsten light range and wave length ranges between 400 - 700 nm. Kj In an experiment, three different solutions are prepared in three tubes and are referred to as ‘Blank’, ‘Test’, and ‘Standard’ and marked by the first capital letter of each word as B, T, and S, respectively. Sample ( test ): is made from the biological fluid being analyzed and subjected to all steps of the analysis to determine the concentration of an analytic in it. Standard solution (ST): are defined as the solutions that contain a known accurate amount (i.e. concentration) of a substance or element. ‫انواعه ممكن تجي تعداد‬ ‫او شرح كل واحد بيهن‬  Med. Lab. Tech. Dep. Medical Instruments and equipments Lec.9/ 1st year Khattab Lect Assistance Muhja Sattar These solutions are used to compare a known concentration of a substance with the unknown concentration of the same substance in the sample. Blank solution (B): Is a solution that contains all of the reagents needed in analysis of a substance except the substance under test or the standard. Applications of Spectrophotometer: ‫ثال‬-‫مطلوب ا‬ 1-For estimation of biochemical samples, like plasma, serum, urine. ‫استخدامات‬ ‫كلهن مهمات‬ 2-Determination of the concentration of a compound or particles in solution such as: blood glucose, blood urea, serum creatinine, serum proteins, serum cholesterol. 3-Used/ by the food industry. 4- Spectrophotometry is one of the most useful methods of quantitative analysis in various fields such as chemistry, physics, biochemistry, forensic science, agriculture, material and chemical engineering and clinical applications..‫صل والبول‬9‫زما وا‬6‫ لتقدير العينات البيوكيميائية مثل الب‬-1 ،‫ جلوكوز الدم‬:‫حلول مثل‬9‫تحديد تركيز مركب أو جزيئات في ا‬-2.‫ الكولسترول في الدم‬،‫صل‬9‫ بروتينات ا‬،‫ في الدم‬J‫ الكرياتين‬،‫يوريا الدم‬. ‫ يستخدم في الصناعات الغذائية‬-3 ‫ت مختلفة مثل‬Y‫ يعد القياس الطيفي من أكثر طرق التحليل الكمي فائدة في مجا‬-4 ‫واد‬9‫مثل الكيمياء والفيزياء والكيمياء الحيوية وعلوم الطب الشرعي والزراعة وهندسة ا‬.‫والكيمياء والتطبيقات السريرية‬ The translation was done by khattab alshallal