# determination of dye concentration by spectrophotometer lab report

Proper Use Of Significant Figures And Units Is Part Of The Grading, Data Table For The Copper Sulfate Solutions: Solution # … Read the absorbance at 595nm. The reaction can be used to determine analyte concentrations assuming the color intensity and absor- bance is proportional to the analyte concentration, the complex is stable, and the reagent does not … �PƑ�� L��~��%�~������K�(Z�# MdEnIp1P�����JDp"I\NP^%���N� ? xڭYK��6�ϯ�-�mY|�u��6[�r���$9$9�%u7�j��ǌ'�� %����rr" �$���ݫ� This unit describes spectrophotometric and colorimetric methods for measuring the concentration of a sample protein in solution. Lab 9 - Determination of Allura Red Concentration in Mouthwash Goal and Overview The spectral profile of Allura Red, or red dye #40, will be measured using a Spec 20 spectrometer. 0000065405 00000 n 0000006531 00000 n 0000004480 00000 n Calculate the concentration … Reaction progress is monitored spectroscopically using the Spectronic-20 spectrophotometer. Abstract To determine the pKa of an acid-base indicator, specifically bromothymol blue, different amounts of both 0.1M NaOH and 0.1 M HCl were added to a solution of fixed concentration of bromothymol blue and the pH was monitored via pH meter.$߲Q����x\�>���ϳ��lf��rpi�iZ0�����+p���a�04]�4�o�?��&w���{�>�G����/�����i2����ߦ�)O�����x6ߡ���H����~��#���jG()�>5�Bkc}�����C���İ[΋d7���$�:�05���y��h6y�W�Gi9|K�p���=9烲�� �L�ś4�A"!H���#�i)/t�h��ґ)�P�:�u�5���mm� �f� Y��� �K��ϰ�r 3�6Cz""f5,*"��U¤�����������0C�p�����&��آ�-tT� �4G��H� 6kȣ���1�Ig?7L��;f�ȳ2A�p�ʂ� � �r��U�2� vs. concentration (x-axis). 2 To answer the guiding question, you will use the UV/VIS spectrometer to identify the components of the mixture and the calibration curves generated by the class to determine the concentration of each component. Our ultimate goal was to calculate the concentrations of each dye in … July 6, 2020 admin. 6. 1-6: LABORATORY REPORT Name Date Kimbury Moya Lab Partners' Names: TA's Name: YOU MUST SHOW ALL YOUR CALCULATIONS TO RECEIVE CREDIT FOR THEM! 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You then measured the %Transmittance of a solution of "unknown" concentration of the same dye, and from your calibration graph, you should have been able to calculate the concentration of the colored dye in the unknown. CHM 260 LABORATORY REPORT Experiment Number: 1 Title: The Visible Spectra of Soft Drink Name : MUHAMMAD YUSRI BIN HJ MUHAMMAD KHIR Student no : 2011995355 Lab Group :1 Date of report Submission : 08 JULY 2014 Lab Partnerâs Name : MUHAMMAD ASHRAF BIN ABDUL RAHMAN MUHAMMAD FAUZAN Lecturerâs Name : MISS HANANI YAZID Experiment 2: UV visible Determination of an Unknown concentration â¦ Materials: VIS spectrophotometer 0.2 M CuSO 4 0.4 M CuSo 4 0.6 M CuSO 4 Fit a trendline and obtain the equation for the line. The negative test result is that the solution remains dark blue. At the end of this lab, check the absorption spectrum of the food dye solution using the Vernier spectrometer. You will first prepare a set of standard solutions (solutions of various but known concentrations) using the dye assigned to you and prepare a standard graph at the max for that dye. Beerâs Law tells us that the concentration of the red dye is proportional to the absorbance and can be used to determine the concentration of solutions of unknown concentration. Analysis of the Data Calculate the molar absorptivities for the red and blue forms of coomassie blue each at their $$\lambda_{max}$$. In this experiment, we created a set of 8 concentrations of 2 dyes, then used a spectrophotometer to calculate the absorbance of the dyes with respect to both concentration and wavelength. Spectrophotometric Analysis of Mixtures: Simultaneous Determination of Two Dyes in Solution. A monochromator is used to produce light in very small ranges of wavelength. Determination of the Rate Law for Food Dye Bleaching with Hypochlorite Author 1 Name, Author 2 Name, Author 3 Name ... bleach the dye and the concentration of the bleach necessary to do so. f�5 ����W�M��3MHb�(Q��@�fgN7AR7簫9/�0J2V�V�X�W$�4Kۗ~w�{���-c�H�n#0��xYߢ��_��g��y�N,9�*@0i��J�2M���é��F�" �|�d��0MB�fE{�bJ�H�م >> endobj The stock solution of dye is gray-red; it … A standard protein curve is made to show the relationship of the concentration of the protein to that of the absorbency at 595nm. Record the color and unknown number of the food dye in the Data section. This report discusses an experiment to study the relationship of absorbance and concentration, the interaction of electromagnetic radiant energy(ERE) and matter which is an important aspect of the Beer-Lambert's Law. The data from the experiment showed that 1.46g of glucose was present in every 100g of orange. Principal purposes of the study are: • to understand quantitative relationships between transmittance, absorbance and concentration, • to use spectrophotometric data to quantify the ferrous iron concentration in an unknown sample and • to understand relationships between measurement errors, sensitivity and ��l�l�Ȼ��;5|d߫��C@۩����2����)t�i������? 0000003494 00000 n The underlying principle behind using spectrophotometric analysis was to quantitatively analyze the concentration of the dye in solution, as it could be analyzed qualitatively by saying the molarity of solution x is greater than that of solution y because the shade of red present in solution x is darker despite the two solutions having the same volume. To determine the concentration of colored species in a solution you must calibrate your spectrometer using a set of known solutions. Lab 2: Colorimetric Determination of Protein Concentration. When the protein binds to the SO3- group of the dye, the pKa value shifts causing the dye to turn blue. Question: 46 G.C. 0000001444 00000 n Beerâs law states the absorbance is directly proportional to the concentration of a solution. Estimating the Concentration of Chlorophyll in Olive Oil The second part of this lab allows you to apply spectrophotometry in order to measure chlorophyll concentrations in olive oil. To understand … H��Wko�H�n���~�VJ3U[)u���汱W��"�0۳��2�]��;p�NE���=s�>\ݍ&7§O�oF���ԂP��&WO, 0000002267 00000 n Discussion The use of spectrophotometry allows for determination of concentrations by measuring the absorbance. >> Jo Melville and Giulio Zhou 9/27/2012. >> Spectrophotometric Analysis of Mixtures: Simultaneous Determination of Two Dyes in Solution Seoiyoung Ahn SID: 23768027 Abstract The objective of the experiment was to figure out the concentrations of Yellow 5 and Red 40 solutions in the food dye and beverage by using spectrophotometry, a common form of analysis used to determine concentrations of certain solutes â¦ Phosphorus is a vital nutrient involved in microbial growth. At the end of this lab, check the absorption spectrum of the food dye solution using the Vernier spectrometer. 0000001306 00000 n unknown compounds and determining the concentration of a substance. Introduction: White light is composed of many different wavelengths of light combined together. Chemical reagents are added to the protein solutions to develop a color whose intensity is measured in a spectrophotometer. The calibration curve technique consist of the known concentration of analyte that place in the spectrophotometers and is recorded the data obtain. First, a 30-ml bea- IV. Determination of Phosphate Concentration Using Spectrophotometry Lab Report. trailer << /Size 562 /Info 529 0 R /Root 533 0 R /Prev 158011 /ID[<978c94ab6279c177861ff5fbad051750><57f0f4f1ee41da626c019d279377f4e5>] >> startxref 0 %%EOF 533 0 obj << /Type /Catalog /Pages 531 0 R /Metadata 530 0 R /Outlines 16 0 R /OpenAction [ 535 0 R /XYZ null null null ] /PageMode /UseNone /PageLabels 528 0 R /StructTreeRoot 534 0 R /PieceInfo << /MarkedPDF << /LastModified (D:20040612130752)>> >> /LastModified (D:20040612130752) /MarkInfo << /Marked true /LetterspaceFlags 0 >> >> endobj 534 0 obj << /Type /StructTreeRoot /RoleMap 28 0 R /ClassMap 31 0 R /K 291 0 R /ParentTree 495 0 R /ParentTreeNextKey 4 >> endobj 560 0 obj << /S 153 /O 240 /L 256 /C 272 /Filter /FlateDecode /Length 561 0 R >> stream The measurement of light absorbance by a solution is also done with spectrophometer. While using the Bradford assay, detergent containing buffer must be avoided as it will disrupt the coomasie dye and produce an inaccurate result. ;P���#�b��G�lGZih�Iz�V�atW0&���� � /6�+�",;� f� �dwz������s�T��pf���ֲZ.�W�f�� Z�(. While using the Bradford assay, detergent containing buffer must be avoided as it will disrupt the coomasie dye and produce an inaccurate result. Obtain 75mL of one of the stock dye solutions. Wear gloves and goggles at all times. Colorimetric assays are convenient for measuring protein concentration. �,&}������O&m�9�x��������ð���� ��� Note: 100% Absorbance = 0% Transmittance While a spectrophotometer can display measurements as either transmittance or absorbance, in biological applications we are usually interested in the absorbance of a given sample. 'ҮPD�����A-V�)Z~�F,96���/+���s�Y0�Ss���Ua3�����5b��νD���T�sNҤV�D��}E���!u��� t��U�F^�Dj�&�pu�,���>m���-V�Q��e���-V�Q��e���-�Svs�X}n�Z-i�r�@�g�g�$N�L5�13a�t#���� ������V�%CJ'q�T�>�'L�en.+�+�.Q����'e�A���?�Re��K���)��x쨨����hµW�p*�SAF+*H�������v��WԢa+f-xؕ�u�t���>s�/�ܙ����8i�����$&ԛ �\U�tV�Ao|D 0000003952 00000 n /Filter /FlateDecode /Length 2080 Spec 20 spectrophotometer Graphing Paper, Ruler, Pencils PROCEDURE 1. If you plot the absorbance (y-axis) versus the concentration (x- axis) you can determine the concentration of an unknown solution using the graph or by using the equation for a line. Introduction to UV/VIS Spectrophotometry: Using Spectrophotometer To Determine Concentration Posted on June 8, 2018 December 19, 2018 by Marcelo Luftman Ultraviolet and visible light range (UV/VIS) is widely applied in research, production and quality control for the classification and study of substances. From the spectral profile, the wavelength of light of maximum absorbance, λ max, will be determined. Through this curve you can determine the unknown protein concentration by use of Beer’s Law. Lab Objectives. ... See the Appendix to the lab manual to see how this should be done. DI�����XFy��,��yp_��r�B&�v�D8��nm5�����o��~���!�"I$�{�F2Oh�����e��S3ڋyt��+�������v�^��X��^�v��&w2�������J���\� �����QnT�H�}]������9>�S���G�6���9]��>vDʩqu�3wz�Xh �h���i��y�D�xt 0000002932 00000 n Lab 9 - Determination of Allura Red Concentration in Mouthwash Goal and Overview The spectral profile of Allura Red, or red dye #40, will be measured using a Spec 20 spectrometer. In the cuvet #4, the dye concentration was 1.2*10-5, and the absorbance was 1.422. 0000065119 00000 n The concentration of the dye present in Gatorade is to be determined spectrophotometrically. Although there are a wide variety of protein assays available none of the assays can be 0000003464 00000 n Materials: VIS spectrophotometer 0.2 M CuSO 4 0.4 M CuSo 4 0.6 M CuSO 4 The stock dye must be of the same color as your unknown sample. 0000065198 00000 n These dyes are highly toxic. q��_��Kmq!�����K��&������l��D�LG In order to determine the concentrations of dye and hypochlorite necessary to give a bleaching time of about 15 minutes, different volumes of undiluted bleach were mixed with 25 mL of stock dye solution and timed for the complete disappearance (at least by visual … This unit describes spectrophotometric and colorimetric methods for measuring the concentration of a sample protein in solution. Absorbance measured at 280 nm (A(280)) is used to calculate protein concentration by comparison with a standard curve or published … One dropperful of each solution are added to the test tubes. 26 0 obj << Determine the absorbance at 280 nm calculate the concentration and compare it to that which you measure by the dye binding assay. 0000064912 00000 n 3. )��/�t�>\g��.��[�s�I�s�|���c�N�������剖�� qC���)x�����;>n� 3�:��E�)��|57e�H�c�b;�P�1}���*�8����R���v���.��J"���H�� Probably the most common application in biology of this technique is in the measurement of the concentration of a compound in solution. Estimating the Concentration of Chlorophyll in Olive Oil The second part of this lab allows you to apply spectrophotometry in order to measure chlorophyll concentrations in olive oil. To become familiar with the Beer’s Law and use of a spectrophotometer for analytical experiments. Read the absorbance for this wavelength. /Length 2290 Specific Activity of Amylase Maize malts germinated at Day4 Protein(mg/ml) of malt and maize From Figure 4. ... - The spectrophotometer was set to zero with the tube contained with distilled water. SCC 201 General Chemistry Lab Reports chemistry 201 Preview text General Chemistry SCC 201 Professor, Sharmila, Shakya Lab Report # 7 Colorimetric Determination Objective To determine color of elements, we speak of light that is emitted of each element of specific wavelengths. 0000049044 00000 n �������r�T���u��zd:O��kҗ�2��N_G%N���ϫ��iF%�8�e��R�HD�~&� 5J�v�ӛD6�+��6 ,�U���JP���׮�&y���Ȑ�HB����]����� � �|��4>�őN$w�VK�k�r�� 0000006375 00000 n It is also used to provide us with the extinction coefficient of the dye. Spectrophotometric analysis for determining the amount of an inorganic compound in solution involves a reaction between an organic reagent and an analyte to form a colored complex. 5.1 Working Standards The development of a standard phosphate curve was necessary to establish a relationship between absorbance and phosphate concentration for the spectrophotometric analysis. 0.405 x 5 x 100 (2.4931ml) x 180 x 100 106 = 1.46 g/100g orange. 0000073136 00000 n AssignmentTutorOnline. %PDF-1.4 1 Abstract. Place the cuvette with same solution but at an unknown concentration. ���0ԇ��Z�I�I�ñ2�Żpp,�K;՗|���U�� �xP�� �C{�Eb�wh�Y�7�M�m]L�:V^4���rd��!l�(�+T��liӖ�Բ��2k�i_=�����I�z�+ļ;>���x"F#�3|"�;��gWOO�]��������ƶ��ix��9Y�[X From the spectral profile, the wavelength of light of maximum absorbance, Î» max, will be determined. The binding protein then stabilizes the blue form of the Coomassie dye. �<4���t�:R�+�~��� ���~��/3¥��]u%I�U�F�Jp��%��ٷ+���\�z���R�uOs��B��&�m���YB�����x*f��.ѡ�l{"�#2T�YKݞ�3͍羛N<0,4^����k�K���4ǲ뫁���;�ގ�-���@�'@;��;����Z|��y����Ŷ��[���iK�����֎C����w����8,N���ag�?���{��yv֯qo���*4\�U�� This graph is a Beerâs Law calibration curve and should be a straight line. 0000026367 00000 n Determination of Unknown Concentration: Set the wavelength to the value corresponding to maximum absorbance (recorded above). Abstract. By measuring the absorbance of a set of standard solutions of known concentrations, you can create a calibration curve that shows how the response of the instrument (absorbance) changes with concentration. 0000006352 00000 n W�vj?���t���=�49�˵���WT���K�3ì��i�ː9I~���IlQ���l�J��Li9H���ِ�� 0000001817 00000 n �ꃧ#M.rg�,�i��#����d�������ѕN��_��H ��Us_7ME+�-oY�M@~ܵ���Bkt�p&���*�V%R1a�#i����e����s�\�W�%4���ݦ��N,K���g9�L���?HI����̪��\\uP����%��С�hQ1�U�PmyQ��WO��1:Ҹl%ar1#m�s�4V��>z������*�7� D��п1�}�5��=�l�Vu|6��;?ɁBƛ��6X�[7< ����a�3����`>D^�4�x���ig� 0000002226 00000 n Bradford Method. You will use the HP-8452A UV-Vis spectrophotometer. 0000002702 00000 n Obtain a sample of food dye of unknown concentration from the lab instructor. First you must “blank” the spectrophotometer by recording and storing in memory a spectrum of the solvent (methanol in this case) and the cuvette. The molecular weight of BSA is 66,300 g/mol. 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Involved in microbial growth microbial growth λ max, will be determined aims of the lab manual to how. To become familiar with the Beer ’ s Law and use of a light meter that! Be determined Î » max, will be determined are added to the concentration of the.... In an unknown concentration a Gatorade sample place in the spectrophotometers and is recorded the data is plotted by graph. Each dye in … Spec 20 spectrophotometer Graphing Paper, Ruler, Pencils PROCEDURE 1 at 280 nm the! A light spectrophotometer prepared for this purpose prepared as 10? M in. Consist of the protein solutions to develop a color whose intensity is measured in a spectrophotometer analytical. Distilled water to a well as reference blank unit describes spectrophotometric and colorimetric methods for measuring the absorbance directly... For this purpose is made to show the relationship of the known concentration protein. Germinated at Day4 protein ( mg/ml ) of malt and Maize from 4! 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