Project 3 Optical Method for Detection and Analysis

Project #3: Optical Method for Detection and Analysis of Biological Molecules Participants: Heather K. Cooper, Chemistry, University of Cincinnati, Senior Kyle A. Frank, Chemical Engineering, University of Cincinnati, Sophom Graduate Student Mentor: Srivasthan Ravi, Chemical Engineering, University of Cincinnati Faculty Mentor: Dr. Anastasios Angelopoulos, Chemical Engineering, University of Cincinnati Wednesday, December 15, 2021 1

Introduction Bradykinin � Hereditary Angioedema � Photo Source: http: //api. ning. com/files/l. Ts. GW 1 Tpdp. RTPV TM 1 gt. Nqdfyr. DJO 3 PY 6 rd. Un. ZEM 73 b. Zyecx n. Szuz 2 h 73 vq. VRL 90 Vfd. PJ 4 x. Mv. REjn 3 Wo mj 6 x. EGFx. Gy. Dt. Lixf 0/Myrtle. HAE 9108 comp. jpg Photo Source: http: //upload. wikimedia. org/wikipedia/commons/ 5/51/Bradykinin_updated. png 2

Photo Source: http: //www. uwgb. edu/heuerc/2 D/Color. Wheel 1. jpg Objective Use optical spectroscopy to detect and quantify Bradykinin � Understand principles of color � Learn to use UV-Visible Spectrophotometer � Concentrate on visible region to permit use of inexpensive and portable LED's as a light source � Photo Source: http: //www. dnr. sc. gov/ael/personals/pjpb/lecture/spectrum. gif 3

Approach Perflurosulfonic acid (Nafion®) membrane as catalysts � Specific reaction between analyte and reactive dye molecule within the membrane � Optical response monitored using visible light spectroscopy � Analyte Reactive dye molecule (resorcinol) UV UV Nafion membrane Product Visible Region Prior Work Fig 1. Catalytic optodes after exposure to . indicated TMA concentrations Fig 2. Catalytic optode exposed to formaldehyde 4

Approach Cont. �Proposed Sensing Reaction: Preparing the 0. 5 Resorcinol Solution Placing the Membrane in the Solution 5

Methods �Sample Preparation ◦ PSA Membranes ◦ Immobilize BK ◦ Immobilize Resorcinol �Thermo Conducting the Reaction at 90⁰C on a Hot Plate activation ◦ Reaction at 90⁰C ◦ UV-Visible Spectrometer 6

Materials & Instrumentation UV-Visible Spectrometer Resorcinol Solutions Bradykinin 10 mg Stock Solution Light Path from UV-Visible Spectrometer 7

Procedure Pretreatment of Nafion Membrane Prepare Stock Solutions Immobilize Bradykinin Immobilize Resorcinol Record Spectra Heat at 90⁰C Compare/Analyze Graphs 8

Procedure � 10 mg BK in 100 m. L Water � 4 Resorcinol Solutions: ◦ ◦ 1 g/L 0. 5 g/L 0. 3 g/L 0. 2 g/L 9

Results Pretreated (Control) Resorcinol Membranes before heating at 90 degrees Celsius 1 Absorbance 1 g/L 0. 5 g/L 0, 5 0. 3 g/L 0. 2 g/L 0 200 300 400 500 Wavelength, nm 600 700 800 10

Results Resorcinol Membranes heated at 90 degrees Celsuis Absorbance 1 0, 5 1 g/L 0. 5 g/L 0. 3 g/L 0 200 300 400 500 Wavelength, nm 600 700 800 11

Results Bradykinin 10 mg Samples Absorbance 1 Sample 1 0, 5 Sample 2 Sample 3 Sample 4 0 200 300 400 500 Wavelength, nm 600 700 800 12

Results Bradykinin 10 mg Samples with Resorcinol before heating at 90⁰ Celsius Absorbance 1 Sample 1 with 1 g/L Res 0, 5 Sample 2 with 0. 5 g/L Res Sample 3 with 0. 3 g/L Res Sample 4 with 0. 2 g/L Res 0 200 300 400 500 Wavelength, nm 600 700 800 13

Results Bradykinin 10 mg Samples with Resorcinol heated at 90 degrees Celsius Absorbance 1 • Background and sample membranes pretreated in boiling acid to ensure cleanliness • Baselines corrected to background signal at 300 nm (no reaction) • Statistics of sample variation assessed 0, 5 Sample 1 with 1 g/L Res Sample 2 with 0. 5 g/L Res Sample 3 with 0. 3 g/L Res 0 200 300 400 500 Wavelength, nm 600 700 800 14

Goals �Obtain Maximum Absorbance Point 0, 1 0, 09 0, 08 Absorbance 0, 07 0, 06 Experimental Data 0, 05 Hypothesis 0, 04 Control 0, 03 0, 02 0, 01 0 0 0, 2 0, 4 0, 6 0, 8 Resorcinol Concentration 1 1, 2 15

Tasks �Low Concentrations Preparing a Resorcinol Solution Placing a Membrane in a Resorcinol Solution 16

Timeline – Week 1 Monday Tuesday Wednesday Thursday Friday Research Training Literature Research Background Briefing Progress Meeting Workshop Enrichment Seminar Progress Report 17

Timeline – Week 2 Monday Tuesday Wednesday Thursday Friday Research Training Literature Research Background Briefing Progress Meeting Workshop Presentation Preparation Progress Report Field Trip Conducting Research Biweekly Presentation Technical Paper 18

Timeline – Week 3 Monday Tuesday Wednesday Thursday Friday Literature Research Workshop Progress Meeting Field Trip Conducting Research Progress Report Presentation Preparation Technical Paper Poster Preparation 19

Timeline – Week 4 Monday Tuesday Wednesday Thursday Friday Literature Research Progress Meeting Field Trip Conducting Research Progress Report Presentation Preparation Technical Paper Poster Preparation Enrichment Seminar Biweekly Presentation 20

Timeline – Week 5 Monday Tuesday Wednesday Thursday Friday Literature Research Progress Meeting Conducting Research Progress Report Presentation Preparation Technical Paper Poster Preparation 21

Timeline – Week 6 Monday Tuesday Wednesday Thursday Friday Literature Research Progress Meeting Conducting Research Progress Report Presentation Preparation Technical Paper Poster Preparation Field Trip Biweekly Presentation Workshop 22

Timeline – Week 7 Monday Tuesday Wednesday Thursday Friday Literature Research Progress Meeting Conducting Research Progress Report Presentation Preparation Technical Paper DRAFT Poster Preparation Enrichment Seminar 23

Timeline – Week 8 Monday Tuesday Wednesday Thursday Friday Presentation Preparation Technical Paper FINAL Poster Preparation FINAL Progress Meeting Final Presentation FINAL PRESENT 24

Questions? 25