UNIVERSITI TEKNOLOGI MALAYSIA Dote : as lob l>olY
Contents
1. 20 30 3l CGM LOC SPE PDMS PBS 3D GOD MPC Ag AgCl Pt MEMS WE CE RE DC USB FYPI FYP2 mg dl Xl LIST OF ABBREVIATIONS Continuous Glucose Monitoring Lab On Chip Screen Printed Electrode Polydimethyl Siloxane Phosphate Buffer Solution 3 Dimensional Glucose Oxidase Methacryloyloxyethyl Phosphorylcholine Silver Silver Chloride Platinum Micromechanical System Working Electrode Counter Electrode Reference Electrode Direct Current Universal Serial Bus Final Year Project 1 Final Year Project 2 Miligram Deci liter XII LIST OFSYMBOLS micro xiii LIST OF APPENDICES APPENDIX TITLE PAGE A Coding 37 B Data Sheet 40 CHAPTER 1 INTRODUCTION This chapter gives the explanation about the project background problem statement objectives as well as project scope 1 1 Introduction Glucose is an important aspect in human body where its level in the blood is controlled by the secretion of insulin in the negative feedback system For a diabetic patient the blood glucose concentrations usually fall outside the normal range of 70 180mg dl This happened because their body does not secrete insulin or maybe there is derangement in both insulin secretion and action 1 Usually in surgical procedures before a patient undergo any surgery glucose measurement must be taken several times per day in the perioperative environment 2 The measurement must be taken a week before the operat2. Counter clectrode Reaction chamber Figure 2 1 Screen Printed Electrode 12 2 2 2 Glucose Sensor Based on Functional Polymers H Kudo T Sawada E Kazawa et al 10 provide a glucose sensor using functional polymer such as polydimethylsiloxane PDMS and methacryloyloxyethyl phosphorylcholine MPC as its material with Pt and Ag AgCl as its electrodes through a process called micromechanical system MEMS The sensor showed a better flexibility such as shown in Figure 2 2 since it was fabricated using functional polymer It also showed a good sensitivity towards different glucose concentration However the performance decreases after rinsing the electrode with potassium ferricyanade as electron mediator The decrease is due to the loss of electrons during rinsing Ag AgC electrode P nt Pt electrode Figure 2 2 Glucose sensor based on functional polymer 10 23 Lab on A Chip Lab on chip is a microfluidic device that uses to integrate miniaturized laboratory function such as separation and analysis of mixture components on a single microprocessor chip using extremely small volume of fluids 13 2 3 1 Methods of Moving Fluids inside Microfluidic Devices The fluidic transport system is an important element that needs to be considered in microfluidic devices B Weigl R Bardell C Cabrera 14 have proposed several techniques for fluidic transport such as pressure driven flow electroosmotic flow electroki
3. EN ENL MEME Created by Cytron Technologies Sdn Bhd All Rights Reserved 7 ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield Reset button User can press this button to restart Arduino program Motor connectors Connect DC motor here External power PWRIN connector Connect external power source here PWR LED Motor power Indicator Power source selection User can choose power source either from external PWRIN or internal VIN External power PWRIN reverse polarity protection circuit In case user wrongly connect the external power source polarity this circuit will protect the shield from broken L298P driver motor IC SHIELD 2AMOTOR uses L298P driver motor IC Motor control mode selection User can choose motor control mode either Signed Magnitude or Locked Anti Phase Test switch When button A is pressed current flows from output A to B and motor will turn CW or CCW depending on the connection When button B is pressed current flows from output B to A and motor will turn CCW or CW depending on the connection LSS05 connector LSS05 can connect directly to this shield and can be interfaced with Arduino Arduino pinout Other Arduino shield can be stacked on top of this shield Created by Cytron Technologies Sdn Bhd All Rights Reserved 8 ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield 6 HARDWARE This section shows th
4. Arduino Uno microcontroller a potentiostat and SPE as a detector for glucose In order to control the speed of peristaltic pump Arduino Uno microcontroller as well as the motor driver has been used The Arduino Uno microcontroller board has been program using Arduino software The program are then transferred into the board using USB cable Additional 9V battery power supply has been used so that the Arduino Uno can standalone without the need to be connected to any machine Figure 3 3 shows the hardware implementation to run the peristaltic pump Motor Driver Program the Ard mii gps TER pumps as well as V w battery via wires to the Arduino software D channel of motor driver Arduino Uno board with motor driver Ardumo Uno microcontroller board Figure 3 3 Hardware implementation to run the peristaltic Pumps 13 3 3 1 Peristaltic Pump This project uses 2 peristaltic pumps such as in Figure 3 4 in order to pump in sample volume of glucose oxidase and glucose solution into inlet 1 and inlet 2 of lab on a chip Figure 3 4 Peristaltic pump Table 3 1 also shows the specification of the peristaltic pump The peristaltic pumps have silicon tubing that can move the fluid from one place to another place safely Furthermore it also operate at motor voltage of 12V DC with 300 mA motor current which is quite small compared to other DC motor The maximum flow rate that can be handle by this peristaltic pump is up to 100mL mi
5. Peutjapun AO peA4es84 ff pex4eu suornonaisur 40 Saunyes Aue jo SJNSTJSJIPJPUD 40 e2uesqe sy uo f e4 JOU YSNW 49Uuojsnj ul e2nou ijnouirm Yewn Aue je suondria sep ionpoad pue suoneoriji2eds o sabueyd eeu Mew OUTNPay ISOdand UYTINIILUYd Y 404 SS3NIIJ HO ALITISUINUHOH3H 40 S3ILNURHHUM GSITdWI ANY OL G3LIWIT LON LNG SNIGQONI fS129n 08Hd 9NIQHUS3H QSIIdWI HO SS38dX3 S3ILNUHHUM YSHLO T1U SWIBTISIG ournpag SLINGS 1Y HLIM ONY SI SU Q3 InOdd Ang subrseg e2ue248j8at N NO LO CO N 3 C2 G 8H JTXQT ONS no AND AT INA Z non r 9 10S81SZTTTdON TN GNI AT Nd d8ZEVv9AWLY 0dd axuy LGd GX1 ZGd 01NI dd 1LNI vad 0L sad 9ad 0NIv Gd LNIV 0Od 05Gv AND LOd LOGV DON L9 Zod zoav og od coqv ANDV net vod roav OOV SOd sSOqv TH CY Lc ZHNSL BU EGNOWOTAILSO u3 100g gsn 08d d1 LEd LOO ZHad SS TIVLX dd ISOW vad OSIN G8d MOS 902709 GZGT S l _ l ONS i fe Y OO G 8H 41 8 ONS OO C za QS Og Os ONS ONS OF ungage 986Zzd1 SE 4 Of nT YAIMOd NG OND OGH4 0LNI 8000 avd F bad LNI 0NIV ZGd ZLNI LNIV L OXY anon Cd LNI LX L d vAd ENIV SLNI a Sad zLLNIOd ENIV MOX DDAN 9Gd 9 LNI GNIV S Li dvon ZGA LLNI OL 9NIV SMH S LI o G ZOd LLLNIOd ZNIV AND N EN vOd 0LLNIOd 99 NO GOd GLOO ELNIOd 9Od 81NI
6. from gold material while the reference electrode were made up from the silver material This type of material were chosen since it provide higher stability towards the glucose reaction compare to other material 29 Figure 4 2 shows the hardware implementation of CGM system Figure 4 2 Hardware implementation of CGM system 43 Data Analysis Four different concentration samples of glucose solution were used to react with the glucose oxidase solution The different concentration were prepared according to the standard glucose level in the human body Normally the glucose level in human body falls in the range of 70 180mg dl which is approximately 4mmol L to 8mmol L Therefore glucose concentration that is 4mmol L 5mmol L 6mmol L and 8mmol L were prepared as samples The measurement of each of the concentration were recorded and analyzed The graph that has been produced in the PS Trace program were exported and combined in Microsoft Excel 4 3 1 Cyclic Voltametry CV Technique Figure 4 3 shows the cyclic voltametry graph of different concentration of glucose solution Cyclic voltametry is an electrochemical technique that measure 30 the current value that has been develop in electrochemical cell Besides that it is also being used to study about the mechanism and rates of oxidation and reduction process Using cyclic voltamery technique the time dependent potential signal were supplied to the working electrode by Emstat
7. im E k 1 EN1 Sf Ss JS DIR1 A HII e Wi m iL Si TET Magnitude F k c Le 8 5 4 8 dl k e a m v v p T np Je Ai AZ AI Ad AS D3 G1 07 07 04 D3 C ari LIGA lA 1 Ix SHIELD 2AMOTOR 2 5xmini jumper Created by Cytron Technologies Sdn Bhd All Rights Reserved 4 ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield 3 PRODUCT SPECIFICATION AND LIMITATIONS Absolute Maximum Rating Parameter Power Input Voltage Motor supply voltage Imax Maximum Continuous Motor Current Irak Peak Motor Current Vion Logic Input High Level Vio Logic Input Low Level Maximum PWM Frequency Must not exceed 10 seconds Created by Cytron Technologies Sdn Bhd All Rights Reserved 5 ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield 4 DIMENSION d EL TT 7 Ig DIR2 Fi a EN in 4 ENa SE E Ht JS DIR1 Ta TITT TT S 1 E Locked Anti Ph ase 1 k Signed Hagnitude VA T MET Ll W be es bs T e 55mm Ou G og 1020704 08 C Taka AA b ETT EEE 1 i me i Created by Cytron Technologies Sdn Bhd All Rights Reserved 6 ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield 5 BOARD LAYOUT EXITIUM i engel fr RY HJEP Su si i Jalotas 17 BETALERE PA AZ A3 A4 AS OZ LOL 02050405 C H ak c
8. system specification 3 1 Project Work Flow This project is divided into three main stages which are designed lab on a chip that is suitable to be used in a CGM system using Solidworks 2013 fabrication process of lab on a chip using PDMS material and lastly assembly of the hardware together and analyze the data using PSTrace 10 3 2 Block Diagram Figure 3 1 shows the block diagram for CGM system A sample of glucose oxidase and glucose will be pumped into the inlet 1 and inlet 2 of lab on chip using peristaltic pump and motor driver that has been controlled by the Arduino Uno microcontroller Three electrodes of potentiostat that is connected to the PSTrace programmed inside the computer will be connected to the three electrodes of SPE which is working electrode WE counter electrode CE and a reference electrode RE that has been integrated with the lab on a chip The PSTrace program will analyze the output measured from the potentiostat Figure 3 1 Block diagram of CGM process 11 3 2 1 Process Flow Chart Figure 3 2 shows the process flow chart for the CGM system Start pin sam ples mto lab on Chip Detect the reaction of sam ple by using integrated SPE Measure the glucose level using potentiostat Analyse the output data using PSTrace program Figure 3 2 Process flow chart for the CGM System 12 3 3 Hardware Implementation This project requires two peristaltic pumps one motor driver
9. Od VI2O E 49d 09x19 LdOl E LND OON a Ol pt I Odd OLNIOd SS LIVLX T z LAd LLNIOd MTOS TONY i zad zLNiod ISOW IQd 00d Z1VLX dena AZZ dd ELNIOd OSIW Odd QugagcG Z 898 JNSW AN CX vad vINIOd LL GSd SLNIOd 9gd 91NIOd 48d 2120 v090 41NIOd MQ LOd LASSY d reNd A UNS no AGT BIN ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield Cytron Technologie SHIELD 2AMOTOR Cytron 2A Motor Driver Shield User s Manual V1 1 Feb 2015 Information contained in this publication regarding device applications and the like is intended through suggestion only and may be superseded by updates It is your responsibility to ensure that your application meets with your specifications No representation or warranty is given and no liability is assumed by Cytron Technologies Incorporated with respect to the accuracy or use of such information or infringement of patents or other intellectual property rights arising from such use or otherwise Use of Cytron Technologies s products as critical components in life support systems is nol authorized except with express written approval by Cytron Technologies No licenses are conveyed implicitly ar otherwise under any intellectual property rights Created by Cytron Technologies Sdn Bhd All Rights Reserved l ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield INDEX l Introduction 3 2 Packing List 4 3 Pr
10. PSZ 19 16 Pind 1 13 UNIVERSITI TEKNOLOGI MALAYSIA d DECLARATION OF THESIS UNDERGRADUATE PROJECT PAPER Author s full name NURUL ATIRAH BINTI MUSTAFFA Date of birth 09 FEBRUARY 1992 Title CONTINUOUS GLUCOSE MONITORING CGM USING LAB ON CHIP LOC Academic Session 2014 2015 declare that this thesis is classified as CONFIDENTIAL Containing confidential information under the Official Secret Act 1972 RESTRICTED Containing restricted information as specified by the organisation where the research was done OPEN ACCESS agree that my thesis be published and accessed online full text at I acknowledge that Universiti Teknologi Malaysia reserves the right as follows 1 The thesis is the property of Universiti Teknologi Malaysia 2 The Library of Universiti Teknologi Malaysia has the right to make copies for academic purposes Certified by SIGNATURE SIGNATURE OF SUPERVISOR 920209 11 5530 DR SUHANA BINTI MOHAMED SULTAN NEW IC NO PASSPORT NO NAME OF SUPERVISOR Date 24 fob 2016 Date 24 o 20 5 NOTES If the thesis is CONFIDENTAL or RESTRICTED please attach the letter from the organisation concerned stating the reason s and duration for the confidentiality or restriction I hereby declare that I have read this final year project report and in my opinion this final year project report is sufficient In terms of scope and quality for the purpose of awarding the degree of Bache
11. RENCES 35 Appendices A B 37 56 1X LIST OF TABLES TABLE NO TITLE PAGE 3 1 Specification of peristaltic pump 13 3 2 Specification and limitation of motor driver 14 313 Gantt chart FYPI 23 3 4 Gantt chart FY P2 24 55 Cost estimation for lab on chip fabrication 25 3 6 Cost estimation for solution preparation 25 3 7 Cost estimation for hardware implementation 26 3 8 Cost estimation for CGM system 26 4 1 Result of glucose concentration samples and current reading 32 FIGURE NO 2 1 2 2 2 3 2 4 Jal 0 2 3 3 3 4 3 5 3 6 3 7 3 8 3 9 3 10 3 11 3 12 3 15 4 1 4 2 4 3 4 4 LIST OF FIGURES TITLE Screen Printed Electrode Glucose sensor based on functional polymer Model based design of microfluidic biochip PDMS casting Block diagram of CGM process Process flow chart for the CGM System Hardware implementation to run the peristaltic Pumps Peristaltic pump Motor driver Screen printed electrode Potentiostat Solidworks design of lab on chip LOC Fabrication process of LOC Flowchart of programming in Arduino software Implementation of Emstat and PSTrace program PSTrace program using cyclic voltametry measurement technique PSTrace program using amperometric detection measurement technique Lab on chip Hardware implementation of CGM system Cyclic voltametry graph of various concentration of glucose solution Amperometric detection of various glucose concentration graph 21 22 28
12. T ABSTRACT ABSTRAK TABLE OF CONTENTS LIST OF TABLES LIST OF FIGURES LIST OF ABBREVIATIONS LIST OF SYMBOLS INTRODUCTION 1 1 Introduction 1 2 Problem Statement 1 3 Objectives 1 4 Research Scope 1 5 Thesis Outline LITERATURE REVIEW 2 1 Continuous Glucose Monitoring 2 2 Glucose Detection 2 24 2 22 Screen Printed Electrode Glucose Sensor Based on Functional Polymers 2 3 Lab on A Chip 234 Methods of Moving Fluids inside Mi crofluidic Devices 2 5 2 Pattern of Microfluidic Devices vii PAGE ii iii IV V1 Vil Xl xli DN N N N E n A A N Vill 23 3 Fabrication Process of Microfluidic De vice 2 4 Conclusion 8 3 RESEARCH METHODOLOGY ox Project Work Flow 2 2 Block Diagram 10 202 4 Process Flow Chart 11 3 3 Hardware Implementation 12 3 3 1 Peristaltic Pump 13 3312 Motor Driver 14 3 3 3 Screen Printed Electrode SPE 15 3 3 4 Potentiostat 15 3 4 Experiment Flow 16 3 4 1 Fabrication of Lab on Chip LOC 16 3 4 2 Preparation of Solution 18 3 4 3 Arduino Programming 19 3 4 4 Emstat And PSTrace Program 20 3 5 Project Management 22 2495 4 Gantt Chart 23 552 Cost Estimation 25 3 6 Conclusion 26 4 RESULTS AND DISCUSSION 27 4 1 Lab on Chip LOC 27 4 2 CGM System 28 4 3 Data Analysis 29 4 3 1 Cyclic Voltametry CV Technique 29 4 3 2 Amperometric Detection Technique 31 4 4 Conclusion 32 5 CONCLUSION AND RECOMMENDATION 33 5 1 Conclusion 33 5 2 Limitation 34 513 Recommendation 34 REFE
13. a clear understanding about type of glucose sensors that is suitable to be used in CGM system methods of moving fluids inside microfluidic devices and pattern of microfluidic device that usually use in designing lab on a chip 2 1 Continuous Glucose Monitoring Continuous glucose monitoring CGM is a way to measure the glucose level throughout a long period of time CGM provides all information about the magnitude duration direction and causes of fluctuation in the glucose level 7 2 2 Glucose Detection The glucose detection is based on an amperometric approach of enzymatic oxidation between glucose and glucose oxidase GOD such as shown in Equation 2 1 10 Glucose O gt GOD Gluconolactane H O5 H O Os 2Ht i2e 2 1 The enzyme was immobilized onto the surface of working electrode which react with glucose to produce electric current 11 2 2 1 Screen Printed Electrode A research by H Xu G Li J Wu et al 12 provide a screen printed electrode as glucose detector such in Figure 2 1 The sensor use carbon and Ag AgCl ink to form a working electrode WE counter electrode CE and reference electrode RE GOD was immobilized onto the carbon electrode of WE and CE While silver were printed onto the three electrodes as a conductive lead The sensor provides better response current which gives linear relationship with the increment of glucose concentration Silver lcad Working electrode F
14. agnitude Arduino Pin Arduino Pin Created by Cytron Technologies Sdn Bhd All Rights Reserved 12 ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield 6 2 LSS05 PIN LSS05 can connect directly to this shield and can be interfaced with Arduino Below is a table showing the pins connection between LSS05 and Arduino on 2A motor driver shield T EN NN Created by Cytron Technologies Sdn Bhd All Rights Reserved 13 ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield 7 SOFTWARE Arduino library and example code can be downloaded from the SHIELD 2AMOTOR product page at Cytron s website Created by Cytron Technologies Sdn Bhd All Rights Reserved 14 ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield 8 WARRANTY Product warranty 1s valid for 12 months Warranty only applies to manufacturing defect Damaged caused by misuse is not covered under warranty Warranty does not cover freight cost for both ways Prepared by Cytron Technologies Sdn Bhd No 16 Jalan Industri Ringan Permatang Tinggi 2 Kawasan Industri Ringan Permatang Tinggi 14100 Simpang Ampat Penang Malaysia Tel 604 504 1878 Fax 604 504 0138 URL www cytron com my Email support cytron com my sales cytron com my Created by Cytron Technologies Sdn Bhd All Rights Reserved 15
15. ation Basically FYP1 involves more on understanding the topic by doing research and literature review based on the journals It also includes the design thinking of the whole CGM system and overall project flow While FYP2 involve more on fabrication and hardware implementation in order to get the output data to be analyzed by the PSTrace program 25 3 5 2 Cost Estimation Table 3 5 shows the cost estimation for lab on chip fabrication Table 3 5 Cost estimation for lab on chip fabrication Table 3 6 shows the cost estimation to prepare the glucose and glucose oxidase solution Table 3 6 Cost estimation for solution preparation LIST QUANTITY UNIT PRICE RM AMOUNT RM D Glucose Anhyrdous 93 00 93 00 Glucose Oxidase From Aspergillus Niger 362 00 362 00 TOTAL 455 00 26 Table 3 7 shows the cost estimation for hardware implementation to run the peristaltic pump and to detect the glucose level Table 3 7 Cost estimation for hardware implementation LIST QUANTITY UNIT PRICE AMOUNT RM Peristaltic Pump 43 00 86 00 TOTAL 335 50 Table 3 8 shows the cost estimation for the whole CGM system where the total cost for the CGM system is RM1190 15 Table 3 8 Cost estimation for CGM system PART SUBTOTAL RM Lab on chip 399 65 Solution preparation 455 00 Hardware Implementation 335 50 TOTAL 1190 15 3 6 Conclusion In short the continuous glucose monitoring CGM system focus on t
16. e example of using SHIELD 2AMOTOR with Arduino Uno as the main controller to control dual brush DC motor However other Arduino main board controller can be used Leonardo Mega Due Created by Cytron Technologies Sdn Bhd All Rights Reserved 9 ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield Figure above shows that SHIELD 2AMOTOR is stacked on the Arduino UNO Move 4 mini jumpers to Signed Magnitude or Locked Anti Phase The default setting is Signed Magnitude Make sure correct power source is selected VIN or PWRIN VIN will connect to the Arduino power source while PWRIN needs external power source connected to the PWRIN terminal s Ea HE Da p zd Created by Cytron Technologies Sdn Bhd All Rights Reserved 10 ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield Connect motor to the MOTORI and MOTOR2 terminal block Upload SHIELD 2AMOTOR example code to Arduino Or you can test directly by using test switch Created by Cytron Technologies Sdn Bhd All Rights Reserved 11 ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield SHIELD 2AMOTOR also Include with LSS05 connector Means you can plug in LSS05 directly to Arduino 6 1 MOTOR CONTROL PIN Choose either Locked Anti Phase or Signed Magnitude mode by moving the 4 mini jumpers on shield Below is a table to summarize pin used for motor control Locked Anti Phase Signed M
17. er electrode CE and reference electrode RE It is used as a detector to detect the presence of glucose through the reaction of glucose and glucose oxidase inside Lab on a chip In this project screen printed electrode that are made up of gold material for its working and counter electrode as well as silver material for its reference electrode were used This type of material were selected since its has more stability towards glucose reaction 4 SI SK P ta lt SN Figure 3 6 Screen printed electrode 3 3 4 Potentiostat This project uses an Emstat potentiostat as in Figure 3 7 to measure the glucose level It is connected to the working electrode WE counter electrode CE and reference electrode RE of SPE The glucose level is a measure of the voltage difference between WE and RE as well as current flow between WE and CE The Emstat is powered via USB and controlled by the PSTrace program that has been installed inside the computer Emstat has eight current ranges that is from InA to 100mA with minimun resolution of IpA 16 Figure 3 7 Potentiostat 3 4 Experiment Flow In this proJect the experimental flow Include the fabrication process of lab on chip using the PDMS material preparation of solution for both glucose and glucose oxidase as well as programming using Arduino Uno software to control the the speed of peristaltic pumps in order to pump in the solution samples into the LOC 3 4 1 Fab
18. gh outlet of lab on chip 34 5 2 Limitation The whole CGM system and process can be applied during the surgery However in this experiment the real blood is not being tested Instead the glucose and glucose oxidase has been used as samples to replace the blood The result may differ and more accurate if using the real blood sample 5 3 Recommendation The CGM system is able to measure the glucose level continuously but in invasive way which need to damage the blood vessel Therefore for future recommendation instead of measuring the glucose level invasively make a smaller lab on chip that can measure the glucose level non invasively and can be used not only during the surgery but also in the normal condition 10 11 12 13 REFERENCES G Sparacino F Zanderigo S Corozza et al Glucose Concentration Can Be Predicted Ahead In Time From Continuous Glucose Monitoring Sensor Time Series 2007 54 5 931937 M Rice A Pitkin D Coursin Glucose Measurement In The Operating Room More Complicated Than It Seems Anesthesia and Analgesia 2010 110 4 1056 65 A Trial Annals of Internal Medicine Article Intensive Intraoperative Insulin Therapy versus Conventional Glucose Management during Cardiac Surgery 2007 J Marks Perioperative Management of Diabetes 2003 4 6 A Facchinetti S Favero G Sparacino et al Modeling The Glucose Sensor Error 2014 61 3 620 629 M Chu K Mitsubaya
19. he fabrication of lab on chip Arduino programming and glucose level measurement techniques by using Emstat and PSTrace program that has been implemented in the specific time schedule CHAPTER 4 RESULTS AND DISCUSSION This chapter will discuss about the design and functionality of the project Therefore it will give more explanation about the applications of the CGM system and the suitability of the lab on chip design with the CGM system This chapter also include the result obtained from the reactions of different concentration of glucose solution with the glucose oxidase solution in terms of cyclic voltametry and amperometric detection graph 4 1 Lab on Chip LOC Using Solidworks 2013 a mould of LOC has been designed The designed LOC needs to have dimension of 50mm x 20mm with an inlet diameter of 2mm to pump in sample volume of glucose and glucose oxidase The fluid flow is in spiral shape with Imm diameter An outlet with diameter 2mm also included in order to pump out the waste The channel of LOC is designed in Y shape inlets that are made to pump in two different samples and spiral shape channel to give more time for the different samples to mix completely in order to get maximum output current Although it might will be time consuming but in this project time is not being a concern since the glucose level needs to be measured continuously through a long period The end product of LOC is shown such as in Figure 4 1 T
20. he LOC has been successfully fabricated that allows the two different samples of fluids to flow and mix inside the channel without leakage 28 Figure 4 1 Lab on chip 4 2 CGM System Basically this CGM system aims to be used during the surgery to monitor the glucose level of a patient continuously Therefore the microfluidic device which 1s lab on chip has been introduced in order to allow the blood of patient to flow through the channel of lab on chip The lab on chip also has been used as a medium for glucose detection to measure the concentration of glucose by using integrated screen printed electrode For this project the real blood sample is not being used since the focus is more on the design and fabrication of lab on chip Therefore the glucose and glucose oxidase has been used as samples to replace the blood As in Equation 2 1 the glucose level is measured amperometrically by the reaction of glucose and its enzyme which 1s glucose oxidase in the present of oxygen The screen printed electrode has been used to detect the glucose level 1n this project Basically the screen printed electrode has been widely used in the intermittent measurement of glucose such as finger prick Therefore by interfacing the screen printed electrode with the lab on chip the glucose level can be measured continuously through a period of time For this project the working electrode and counter electrode of screen printed electrode were made up
21. iew will take place that give an overview about previous project that are related with the CGM system or lab on chip fabrication This chapter help to give a clear picture about the working principles of CGM system types of sensor that are usually used in CGM system methods of moving fluids inside microfluidic devices and pattern of microfluidic device that usually use in designing lab on chip In third chapter a brief explanation about project work flow block diagram software implementation In designing mold of lab on chip steps fabrication of lab on chip using PDMS materials and hardware implementation for CGM system The fourth chapter deals with the functionality of the system that analyze the result obtained based on the hardware and software implementation as well as the reaction from different concentration of glucose and glucose oxidase inside the channel of lab on chip that detected by SPE The result based on cyclic voltametry and amperometric detection graph also included in this chapter The fifth chapter gives a brief explanation about the project management Where in this chapter project schedule and project costing has been included Lastly chapter six gives conclusion about the result obtained and give a brief explanation about the limitation and recommendation for future works CHAPTER 2 LITERATURE REVIEW This chapter gives a brief review about the previous or similar projects of CGM system It provides
22. ing agent is cast on top of the silicon layer After cross linking process the elastomer is peeled off from the substrate Then access is created frequently with a part holder After that the part is transferred into another piece of PDMS This technique gives property that makes the PDMS easy to close because it places the open structure on a flat substrate and easy to be stripped as PDMS is an elastomer Furthermore this technique is preferred to be done in a laboratory that did not have advanced technology tools as it uses material such as PDMS ar Kd Figure 2 4 PDMS casting 16 2 4 Conclusion Based on the research that has been made for this project the CGM system in this project will use screen printed electrode SPE as a detector because of its good sensitivity towards different glucose level Furthermore since this project only focus to detect and measure the glucose level therefore the pattern of lab on a chip will be designed by having two inlets for injection of glucose and GOD enzyme together with one outlet to pump out the waste CHAPTER 3 RESEARCH METHODOLOGY This chapter gives a brief explanation about project work flow block diagram experimental flow that involves in fabrication process of lab on chip using PDMS materials preparation of glucose and glucose oxidase solution as well as software and hardware implementation in designing CGM system Besides that this chapter also explain about the
23. ion so that the insulin regimen can be adjusted according to the condition of the patient 3 This is an important aspect that has to be considered especially in diabetic patient because they face more challenges during surgery compare to the non diabetic patients Many of the challenges occur due to inabilities to maintain a balance between insulin and its counter regulatory hormones 4 Nowadays several sensors have been developed in order to measure the glucose concentration continuously in the human body With the continuous glucose monitoring CGM the glucose concentration in the human body can be measured continuously for a long period of time 5 Compare to the intermittent measurement such as finger prick 6 CGM provides more information about the glucose level such as frequency magnitude direction duration and cause of fluctuation in glucose level 7 This information is useful if applied during the surgery because it can continuously measure the glucose level of patients and give a sign if there are any problems that happened during the surgery 1 2 Problem Statement Based on the application in the operation room the intermittent measurement that has been applied to measure the glucose level is very inconvenient 8 because the measurement must be taken several times in order to get the pattern of the glucose level in the patient s blood 4 The measurement is taken only before the surgery and it is not being m
24. is chapter give conclusion about the result and the whole CGM process Recommendation for the future work also included in this chapter 5 1 Conclusion A CGM system has been designed so that 1t can measure the glucose level continuously during the surgery The design include the whole CGM system as well as the fabrication of Lab on Chip The Lab on Chip was designed with two inlets and one outlet that has Y shape and spiral channel in Solidworks program Then the Lab on Chip was fabricated using PDMS material that consist of silicon elastomer from base and curing agent in ratio of 10 1 The end product of lab on chip that has been integrated with screen printed electrode shows that the samples were able to flow and mix through the channel without leakage In addition the screen printed electrode is able to detect the amperometric reaction between both samples Based on the result the current depicts an increment with the increasing concentration of glucose which is as expected since high concentration of glucose produce more hydogen peroxide and more electron as stated in Equation 2 1 The whole CGM process also can be applied in the surgery The peristaltic pump can pump out blood from the blood vessel into the lab on chip which then the integrated screen printed electrode inside the lab on chip will detect the glucose concentration of blood amperometrically The blood will then flow back into the blood vessel of the patient body throu
25. lor of Electrical Engineering Electric Electronic Signature l Name DR SUHANA BINTI MOHAMED SULTAN Date June 24 2015 CONTINUOUS GLUCOSE MONITORING CGM USING LAB ON CHIP LOC NURUL ATIRAH BINTIMUSTAFFA A final year project report submitted in fulfilment of the requirements for the award of the degree of Bachelor of Electrical Engineering Electric Electronic Faculty of Electrical Engineering Universiti Teknologi Malaysia JUNE 2015 ll I declare that this final year project report entitled CONTINUOUS GLUCOSE MONITORING CGM USING LAB ON CHIP LOC is the result of my own research except as cited in the references The final year project report has not been accepted for any degree and is not concurrently submitted in candidature of any other degree Signature Name NURUL ATIRAH BINTI MUSTAFFA Date June 24 2015 Special thanks to My beloved parents and family that always keep their prayers for me My supervisor that guide me through hardship My friends who always assists and give me moral support And also to Everyone that help me through this research iii IV ACKNOWLEDGEMENT Greatest thanks and grateful to Allah s w t with all His majesty that give me an opportunities to complete this thesis and gift me a good health that makes me able to think and solve problem Alhamdulillah A special thanks to my family especially my parents Mustaffa bin Hamzah and Fauziah binti Wahid that a
26. lways give motivation and keep their prayers to me Without their motivation and prayers it would be hard for me to complete this project I would also express my deepest gratitude towards my supervisor Dr Suhana binti Mohamed Sultan for her care and supervision that provide me guidance throughout the entire research process She also always check my mistake and help me to improve myself for the future challenge In addition I want to express my sincere appreciation to Dr Fauzan Khairi that help me to generate ideas and solve the problem regarding this project Besides that I want to express my thankfulness to Sherrin and Aldi Michi for their kindness that help me during this project Last but not least I deliver my sincere gratitude to my friends that had give me their support either physically or mentally and also to everyone that involved in this project ABSTRACT Glucose is an Important parameter that has to be considered before undergoing any surgery Variation of glucose level in body will affect the decisions during surgery Utilizing lab on chip LOC glucose measurement can be taken continuously during operation time in order to give any sign if a problem occurs LOC with channel of two inlets and one outlet can be implemented using polydimethyl siloxane PDMS that is designed using Solidworks program A screen printed electrode SPE is placed in the middle of the channel inside the LOC to measure the amperometric reactio
27. n Table 3 1 Specification of peristaltic pump Specifications 14 3 3 20 Motor Driver This project requires one motor driver in order to control speed of the pump to pump in the sample volume into lab on chip The motor driver used 1s the shield type that allows the motor driver to be stacked on top of Arduino Uno as in Figure 3 5 Furthermore the motor driver used are compatible with arduino Uno and can be used to control two DC motor up to 2A for each channel Since the peristaltic pump operate at 300mA therefore by using 2A motor driver it can be used to drive the peristaltic pumps I am s 1 L fig ii I ESI CE NEN SURT DR L EN1 E Easo Pea uiii nm as erret teris fete gc Figure 3 5 Motor driver Table 3 2shows the specification and limitation of the motor driver The motor driver can operate at maximum 2A current value and can be supplied with minimum SV to 26V input power Table 3 2 Specification and limitation of motor driver Power Input Voltage Motor supply volage 5 26 v Tax Maximum Continuous Motor Corren 2 A dgeukPekMeoeCumm 2 fa VionGogicInput HighLeve 33 SS V x Vis eehwlovlee 0 o os V Maximum PWM Fregueney 10 RH 15 33 3 Screen Printed Electrode SPE Screen printed electrode SPE such as in Figure 3 6 contain three electrodes such as working electrode WE count
28. n using Emstat between glucose and glucose oxidase that is pumped in through both inlets Glucose buffer solution and glucose oxidase is prepared using combination of phosphate buffer saline PBS and deionized water The system performance is measured using PS Trace program The result shows that the current detected by the SPE is proportional to the concentration of glucose V1 ABSTRAK Glukosa adalah parameter penting yang perlu dipertimbangkan sebelum menjalani sebarang pembedahan Perubahan paras glukosa dalam badan akan memberi kesan kepada keputusan semasa pembedahan Menggunakan makmal pada cip LOC pengukuran glukosa boleh diambil secara berterusan sepanjang waktu pembedahan untuk memberi sebarang tanda Jika terdapat masalah LOC dengan dua saluran masuk dan satu salur keluar boleh dilaksanakan menggunakan polydimethyl siloksan PDMS yang direka menggunakan program Solidworks Elektrod bercetak skrin SPE diletakkan di tengah tengah saluran dalam LOC untuk mengukur reaksi amperometric menggunakan Emstat antara glukosa dan glukosa oksidase yang disuntik melalui kedua dua salur masuk Cairan penampan glukosa dan glukosa oksidase disediakan menggunakan kombinasi garam fosfat penampan PBS dan air ternyahion Prestasi sistem diukur menggunakan program PSTrace Hasil menunjukkan arus yang dikesan oleh SPE adalah berkadar terus dengan kepekatan glukosa CHAPTER TABLE OF CONTENTS TITLE DECLARATION DEDICATION ACKNOWLEDGEMEN
29. netic flow control and electrokinetic mixing Thus it concludes that electroosmotic flow provides very sensitive to the variations in both liquid decomposition and channel wall coating It requires limited use of generic pumping solution while pressure driven flow significantly less sensitive towards viscosity of fluids 2 3 2 Pattern of Microfluidic Devices Y Atalay P Verboven S Vermeir et al in 15 provide model based design for microfluidic biochip that uses to detect multiple analytes The design such in Figure 2 3 consists of five inlets and one outlet Where in this case inlet 1 is used to inject a buffer solution inlet 2 and 3 for sample volume while inlet 4 and 5 used to inject enzyme Therefore it stated that the design was used for fast quantification of multiple components in the multiplexed microchannel reactor Furthermore since this model is designed to detect multiple analytes thus it 1s quite complex and not necessarily considered in detecting one sample of analytes such as glucose EJ 4 E a E E z J z E i Cia te De d WA Figure 2 3 Model based design of microfluidic biochip 15 2 3 3 Fabrication Process of Microfluidic Device P Abgrall A Gue 16 show the fabrication of microfluidic device based on the PDMS casting process As shown in Figure 2 4 the process begins by manufacturing the mould by bulk machining silicon frequently Then a mixture of silicon rubber and cross link
30. nsor P Sample This method is used to check the noise level Pp t Current range 1 1 10 100 1 10 100 1 10 0 57 n n n pA yA uA m mA Pretreatment settings a 5 1 E condition 12 V lt t condition 0 8 E deposition 02 V 1 07 t deposition 0 3 x O Amperometric Detection settings tequilibration 0 8 aL Ed c 42 V vL m t interval 05 s x trun 60 0 s L X CJ Measure OCP Post measurement T r 4 IP Cell on after measurement 20 le 0 10 20 30 40 50 Expected duration 00 01 00s x3 EA Timels 121 datapoints is Figure 3 13 PSTrace program using amperometric detection measurement technique By using amperometric detection technique the voltage bias is set to 0 2V with 60 second running time 3 5 Project Management Project management is a project development that help to achieve the of the objectives of the projects Project management include the gantt chart for the project planning and cost estimation for the whole system 23 3 5 1 Gantt Chart The Gantt chart in Table 3 3 and Table 3 4 shows the overview of the project implementation during Final Year Project 1 FYP1 and Final Year Project 2 FYP2 Table 3 3 Gantt chart FYP1 pe jer pw pu e we pr px pes pe EE ee 24 TET 3 4 Gantt chart FYP2 s 6 v s 9 e far Tu us ua 5 e Em TT TITT Integrate SPE with Lab on Chip Glucose and Enzyme Solution Assemble Hardware Result Exhibition Prepar
31. oduct Specification And Limitations 5 4 Dimension 6 5 Board Layout 7 6 Hardware 9 7 Software 14 8 Warranty 15 Created by Cytron Technologies Sdn Bhd All Rights Reserved 2 ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield 1 INTRODUCTION SHIELD 2AMOTOR is an Arduino shield for controlling dual DC motor up to 2A for each channel It is compatible with Arduino Uno Arduino Leonardo Arduino Mega Arduino Due and possibly other pin compatible main boards SHIELD 2AMOTOR uses L298P SMD IC and support for both signed magnitude and locked anti phase SHIELD 2AMOTOR shield has stackable side headers which allows for more Arduino shields to be stacked on top of it SHIELD 2AMOTOR come with this features e Bi directional control for 2 brushed DC motor support motor voltage ranges from 5V to 26V Maximum current up to 2A continuous per channel 3 3V and 5V logic level input Stackable I O header pin Selectable pins for Signed Magnitude and Locked Anti Phase Test switch for both channel External voltage polarity protector Created by Cytron Technologies Sdn Bhd All Rights Reserved 3 ROBOT HEAD to TOE Product User s Manual 2A Motor Driver Shield 2 PACKING LIST Please check the parts and components according to the packing list If there are any parts missing please contact us at sales cytron com my immediately L i p a E Cae dl j6 oIR2 76 k Z EN2
32. onitored during the operation 9 This may provide risk to the patient as there is no indication in fluctuation of glucose level during the operation 1 3 Objectives This project aims to i Design a system that can measure the glucose level continuously ii To fabricate lab on chip using PDMS materials iii Measure the system performance by using PSTrace program 1 4 Research Scope In order to achieve the objectives stated in 1 3 several scopes have been outlined The scope includes Solidworks 2013 as a platform to design 3D image that will be used to make mold of lab on a chip While Polydimethylsiloxane PDMS will be used as material to fabricate the lab on a chip Moreover the screen printed electrode SPE will be used as a detector and the glucose level will be measured using the Emstat Lastly the PSTrace program will be used to analyze the performance of the CGM system 1 5 Thesis Outline This thesis consists of six chapters that include the introduction literature review research methodology result and discussion project management conclusion as well as future works The first chapter gives the brief explanation about the project background as well as the problem statement that are related to the project Based on the problem statement several objectives have been introduced that aim to solve the problem issues with the guideline from the research scope In second chapter discussion about literature rev
33. onnect 4 pin on SHIELD 2 AMOTOR to Signed Magnitude mode Shield2 AMotor motor SIGNED MAGNITUDE signed int leftSpeed rightSpeed int 1 void setup void loop Rotate 360 degree CW ford 0 1 lt 50 i motor control 100 0 rotate motor delay 10 motor control 100 0 rotate motor1 motor control O 100 stop motor delay 10 motor control O 100 stop motor delay 10 motor control O 100 stop motor motor control 100 0 rotate motor 1 delay 10 motor control 100 0 rotate motor 1 delay 10 motor control 100 0 rotate motor1 motor control O 100 stop motor delay 10 motor control O 100 stop motor delay 10 motor control O 100 stop motor motor control 100 0 rotate motor delay 10 delay 1000 Rotate 360 degree CCW ford 0 i lt 50 i APPENDIX B DATA SHEET 1 Schematic diagram of Arduino Uno 2 2A Motor Driver Shield hor og ure j ue 22 outnpa4e nnn 7 diiu UlIM iuer duo2 eq isnu eueu ONINGYY sui j0 esf JJEWSPEI pea4eistbaea4 e ST QNINGYY UOTIEWAOJUT STU YIM UDISSp e ezi eut YOU OG e 2nou 1NOYIIM ebueu 0 i2efqns ST s ertasaie 4O STS GSM eui uo uoneua4ojur ionpoad ul UuSuy O sebueu 4N N uo DUTSTJE senijigiyedwoDbut 40 Si2I jUOD 40j JeSASOSJEUM fur tq rsuodsa4 ou a8Aeu eus pue UOTIIUTJSP e4ninj 40 SSeU dcc 9ENH co UZZ GENU 7 S A4 S 4 OUINPAY
34. phate buffer solution that 1s need to be added in order to get the desired dilute molarity of glucose concentration Another sample which is glucose oxidase solution were prepared by mixing Smg of the enzyme powder with 200uL 0 1M phosphate buffer solution Both of the samples were kept in temperature below 10 C to ensure the stability of the solution 3 4 3 Arduino Programming In this project two peristaltic pumps were used in order to pump in two different samples inside lab on chip Therefore to control the speed of the peristaltic pump Arduino Uno microcontroller as well as motor driver were used The motor driver that been used is the shield type that can be placed on top of the Arduino Uno The Arduino software were used to program the Arduino Uno microcontroller thus to drive the motor driver to control the speed of the peristaltic pumps Figure 3 10 shows the process flowchart of programming in arduino software 20 Define motori Rotate motorl 360 cw Stop motor Rotate motor Figure 3 10 Flowchart of programming in Arduino software In the Arduino software programming two motors that is motor and motor2 were defined When the switch is turned on motorl will start to rotate 360 clockwise After motor complete a 360 rotation it will stop and motor2 will take place and rotate 360 clockwise When motor2 complete a 360 rotation it will stop and the process will be repeated un
35. r to make the second and third layer that Is use to make interface with SPE Figure 3 9 shows fabrication steps of LOC 18 design the mould paste the sticker on cut the mould the petri dish using solidworks design using cutter pour PDMS onto petri FM cut the PDMS in Fuse dish and let it dry 50mmx20mm Figure 3 9 Fabrication process of LOC 3 42 Preparation of Solution Different concentration of glucose solution were prepared using the phosphate buffer solution PBS and anhydrous glucose powder The available high concentration of 1 0M of phosphate buffer solution were diluted using deionized water to 0 1M using ratio 1 10 Then the glucose stock solution were prepared by mixing 700mg of anhydrous glucose powder into 1L of 0 1M phosphate buffer solution which then produce 1 0M glucose stock solution Since in this project the standard glucose level in human body are being referred therefore samples of 4mmol 5mmol 6mmol and Smmol glucose concentration were prepared To prepare the more diluted glucose concentration from 1 0M glucose stock solution the 0 1M of phosphate buffer solution were added into 1 0M glucose stock solution according to the Equation 3 1 below 19 MiM MV 3 1 Basically M refers to molarity of glucose stock solution Vi refers to the volume of glucose stock concentration while M refers to the desired dilute glucose concentration and V5 refers to the volume of 0 1M phos
36. rication of Lab on Chip LOC In this process before fabricating LOC using PDMS material the Solidworks program were used to design the mould of LOC The mold is designed in 50mm x 20mm dimension with Imm diameter of Y and spiral shape channel At the end of the spiral shape channel 6mm circular diameter is designed that use to place the integrated SPE Figure 3 8 shows the mold design of LOC using Solidworks Figure 3 8 Solidworks design of lab on chip LOC 17 After Solidworks design the mould has been cut using electronic cutter on a sticker paper Then the sticker paper that has been cut was placed onto a petri dish to be used as mould In order to develop channel After that the PDMS material that consist of Silicon elastomer base and curing agent has been mixed in ratio of 10 1 Basically to fabricate a layer of PDMS that has 1 5mm thickness only 10g of Silicon elastomer base and 1g Silicon elastomer curing agent were needed The mixture were then stirred and poured onto the petri dish The petri dish was then placed inside the vacuum chamber for 30 minutes In order to suck out trapped air bubbles After 30 minutes the petri dish is let to be dry In room temperature for one day Then the dry PDMS was repealed from the petri dish using Isopropanol solution and cut into dimension of 50mmx20mm using cutter After that inlets and outlet of the mould were punched using puncher Then the process was repeated two times in orde
37. s negative value of current since the amperometric detection measure the reduction reaction of glucose and glucose oxidase Amperometric Detection of Various Concentration of Glucose ED 70 I 10 zu wi 2 F Ammol pA S5mmol p amp mmol pA Bmmol p current uA Time 5 Figure 4 4 Amperometric detection of various glucose concentration graph Table 4 1 shows the result of current value for four different concentration of glucose solution Based on the reading 8mmol L produce higher current value which is 0 0014 while the lowest concentration which is 4mmol L produce lowest current value which is 0 0007 uA 32 Table 4 1 Result of glucose concentration samples and current reading Glucose Concentration mmol L Current LA 4 4 Conclusion Based on the cyclic voltametry and amperometric detection techniques it shows that the current values increases as the concentration of glucose increases Therefore in normal condition of glucose level the current value do not fall outside of range 0 0007A to 0 0014j A This current measurement condition can be applied during surgery Therefore when the current value falls outside this range the doctor can make preparation to adjust the insulin regimen of the patient blood Furthermore the whole CGM system was successfully implemented which has the advantage of smaller size and mobile CHAPTER 5 CONCLUSION AND RECOMMENDATION Th
38. shi Soft Contact lens Sensor for Monitoring Tear Sugar as Novel Wearable Device of Body Sensor Network M Minimed R City Continuous Glucose Monitoring 2005 28 5 N Sharma S Singh Designing a Non Invasive Blood Glucose Measurement sensor 2012 12 14 R Plodkowski S Edelman Pre Surgical Evaluation of Diabetic Patients 2001 19 2 92 95 H Kudo T Sawada E Kazawa et al A flexible and Wearable Glucose Sensor Based on Functional Polymers With Soft MEMS Techniques Biosensors amp Bioelectronics 2006 22 4 558 62 K Louchis S Driscoll Fundamental Sensing Limit of Electrochemical Glucose Sensors 2011 1 7670 7673 H Xu G Li J Wu et al A Glucose Oxidase Sensor Based On Screen printed Carbon Electrodes Modified By Polypyrrole 2005 1917 1920 Manz et al Miniaturized Total Chemical Analysis Systems 2007 99 108 14 15 16 36 B Weigl R Bardell C Cabrera Lab on a chip For Drug Development Advanced Drug Delivery Reviews 2003 55 3 349 377 Y Atalay P Verboven S Vermeir et al Model based Design and Optimization of a Multiplexed Microfluidic Biochip for Multi analyte Detection 2008 482 485 P Abgrall A Gue Lab on chip Technologies Making a Microfluidic Network and Coupling It Into a Complete Microsystem 15 APPENDIX A ARDUINO UNO SOURCE CODE CODE FLOW TO CONTROL THE SPEED OF PERISTALTIC PUMPS include Shield2 AMotor h C
39. tammetry d i Scan 5 Gidi Boo d P 1 Scan 1 Sensor Testsensor o8 Scan 2 Sample This method is used to check the nose level p eB ram Sf Scan 4 Current range e scan5 1 0 KE 0 10 1 10 n n n yA pA pA m mA Pretreatment settings E condition 12 V m t condition 0 s E E deposition 40 V t deposition 0 8 x O Cyclic Voltammetry settings t equilibration 0 s 4 L E start 00 V Y E vertex1 45 V E vertex2 05 V L X E step 0 005 V Scan rate 01 Ws id Number of scans 5 l 94 92 00 02 04 Expected duration 00 01 40s aya x WA Potential V 401 datapoints PSSI un Connected Underload E 0 028 V Results for Scan 5 47 Underloads 0 Overloads Figure 3 12 PSTrace program using cyclic voltametry measurement technique 22 By using cyclic voltametry techniques the voltage range has been set from 0 5V until 0 5V with 100mV s scan rate which give complete cycle graph Figure 3 13 shows amperometric detection graph based on one concentration of glucose solution in PSTrace program ar a O ag x Mode f3 Method Curve Measure Plot Peaks Script Tools Help PalmSens 9 e F ahhh E Autosave settings D New Connected EmStat2 mana Pot Da Menuai Con 6mmol psmethod AA CN hy Measurement info Peaks Levels vy Available curves E x 2 Technigue Amperometric Detection v M Curve ghe Z Show al e Cure Sensor Testse
40. that change its potential in accordance to the fixed potential of reference electrode While the voltage potential 1s being supplied the current flow between the working and counter electrode are being measured which then translated into current against voltage graph For glucose and glucose oxidase reaction to get the complete cycle graph the voltage is supplied in the range of 0 5V to 0 5V This range gives complete graph cycle with oxidation and reduction peak If smaller range are used it may give distortion to the graph As illustrated 1n the graph the oxidation and reduction peak of the glucose concentration increases with increasing concentration value Cyclic Voltametry of Various Concentration of Glucose e ga 4mmol pA C 1 Smmol yA 106585 0 6 6mmol pA o amp mm ol yA voltage V Reduction Peak Figure 4 3 Cyclic voltametry graph of various concentration of glucose solution 31 4 3 2 Amperometric Detection Technique Amperometry is the changing in electric current due to the detection of ion present in the solution Therefore Amperometric detection technique is used to measure the current value based on the detection of ion that present in the glucose with the enzyme Figure 4 4 shows the amperometric detection graph based on various concentration of glucose level The glucose level also shows an increament in the current value with the increasing concentration The graph show
41. til the switch 1s turned off 3 44 Emstat And PSTrace Program Emstat comes together with the PSTrace program that need to be installed through its driver The Emstat USB cable that has three crocodile clips were then connected to the three electrode of screen printed electrode SPE that has been integrated with the lab on chip Figure 3 11 shows the implementation of Emstat and PSTrace program 21 connect to the integrated SPE E REPE via USB gt PS Trace program cable Figure 3 11 Implementation of Emstat and PSTrace program When the samples of glucose oxidase and glucose solution were pumped into lab on chip the screen printed electrode will detect the rate of reaction between the samples Meanwhile Emstat will measure the reaction by cyclic voltametry and amperometric detection techniques The different measurement technique can be set in the PSTrace program The PSTrace program will produce cyclic voltametry and amperometric graph based on the reaction between the samples Figure 3 12 shows the cyclic voltametry graph of one sample of glucose concentration in PS Trace program Mode 9 Method Curve Measure Plot Peaks Script Tools Help 05560 9 amp F ahh E Autosave settings D New Connected EmStat2 Plot Data Manual Cont ni 6mmol psmethod Manual peak v Je R Hn Ej m m I R AA Available curves Measurement Info Peaks 1 P Technique Cyclic Vot
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