i

RESEARCH PROJECT

PREPARATION OF ANTIBACTERIAL BENTONITE – β LACTAM

ANTIBIOTIC COMPOSITE

Diajukan oleh :

Gisela Anke Hanelin 5203014001

Lukius Denny Wiyanto 5203014026

DEPARTMENT OF CHEMICAL ENGINEERING

FACULTY OF ENGINEERING

WIDYA MANDALA CATHOLIC UNIVERSITY

SURABAYA

2017

ii

LETTER OF APPROVAL

The research entitled :

Preparation of Antibacterial Bentonite – β Lactam Antibiotic Composite

Which was conducted and submitted by :

Nama : Gisela Anke Hanelin

NRP : 5203014001

Has been approved and accepted as one of the requirement for Bachelor of

Engineering degree in Chemical Engineering Department, Faculty of

Engineering, Widya Mandala Surabaya Catholic University, by following

supervisor/s and has been examined by the commitees on May, 23rd 2017.

Surabaya, May 31st 2017

Supervisor Co-Supervisor

Suryadi Ismadji, Ph,D Felycia Edi Soetaredjo, Ph.D.

NIK. 521. 93.0198 NIK. 521.99.0391

The Commitees

Chairman Secretary

Wenny Irawaty, Ph.D. Suryadi Ismadji, Ph,D

NIK. 521.97.0284 NIK. 521. 93.0198

Member Member

Sandy Budi Hartono, Ph.D Dra. Adriana Anteng Anggorowati, M.Si.

NIK. 521.99.0401 NIK. 521.86.0124

Authorized by

Dean of Head of Chemical

Engineering Faculty Engineering Department

Suryadi Ismadji, Ph,D Sandy Budi Hartono, Ph.D

NIK.521.93.0198 NIK. 521.99.0401

iii

LETTER OF APPROVAL

The research entitled :

Preparation of Antibacterial Bentonite – β Lactam Antibiotic Composite

Which was conducted and submitted by :

Nama : Lukius Denny Wiyanto

NRP : 5203014026

Has been approved and accepted as one of the requirement for Bachelor of

Engineering degree in Chemical Engineering Department, Faculty of

Engineering, Widya Mandala Surabaya Catholic University, by following

supervisor/s and has been examined by the commitees on May, 23rd 2017.

Surabaya, May 31st 2017

Supervisor Co-Supervisor

Suryadi Ismadji, Ph,D Felycia Edi Soetaredjo, Ph.D.

NIK. 521. 93.0198 NIK. 521.99.0391

The Commitees

Chairman Secretary

Wenny Irawaty, Ph.D. Suryadi Ismadji, Ph,D

NIK. 521.97.0284 NIK. 521. 93.0198

Member Member

Sandy Budi Hartono, Ph.D Dra. Adriana Anteng Anggorowati, M.Si.

NIK. 521.99.0401 NIK. 521.86.0124

Authorized by

Dean of Head of Chemical

Engineering Faculty Engineering Department

Suryadi Ismadji, Ph,D Sandy Budi Hartono, Ph.D

NIK.521.93.0198 NIK. 521.99.0401

iv

COPY RIGHT AGREEMENT

In order to support development of science and technology, I am as the

student of Widya Mandala Surabaya Catholic University :

Nama : Gisela Anke Hanelin

NRP : 5203014001

Agree to transfer the copyright of my thesis/paper :

Title :

PREPARATION OF ANTIBACTERIAL BENTONITE – β LACTAM

ANTIBIOTIC COMPOSITE

To be published in internet or other media (Digital Library of Widya Mandala

Surabaya Catholic University) for academic purposes according to copyright law

in Indonesia.

Surabaya, May 31st 2017

Author,

Gisela Anke Hanelin

NRP. 5203014001

v

vi

vii

LETTER OF DECLARATION

I declare that this research was my own work and does not contain any

material that belongs to the others, unless it was stated in the references.

Should it is known that this research belongs to others. I aware and accept the

consequences that this research cannot be used as a requirement to achieve a

Bachelor of Engineering degree.

Surabaya, May 31st 2017

Student,

Lukius Denny Wiyanto

NRP. 5203014026

viii

PREFACE

The authors would like to thank God for His blessing that the Research

Project entitled Preparation of Antibacterial Bentonite – β Lactam Antibiotic

Composite has been accomplished. This report is a prerequisite in achieving

Bachelor of Engineering degree in Chemical Engineering.

The authors realize that the completion of this report is achieved by the

help of many people. There for, the authors would like to thank the persons

below:

1. Suryadi Ismadji, Ph.D as Principal Supervisor and Felycia Edi

Soetaredjo, Ph.D as Co-Supervisor

2. Wenny Irawaty, Ph.D as Head of the Committees, Sandy Budi

Hartono, Ph.D and Dra. Andriana Anteng Anggorowati, M.Si as

members of committees

3. Felycia Edi Soetaredjo, Ph.D as the Head of Chemical Engineering

Process laboratory, Dra. Adriana Anteng Anggorowati, M.Si. as the

Head of Chemical Analysis Laboratory, and Ery Susiany R., ST.,

MT as the Head of Bioprocess Laboratory.

4. Mr. Novi as laborant of Chemical Engineering Process Laboratory,

Mr. Pudjo as laborant of Chemical Engineering Operation

Laboratory, and Mr. Agus as laborant of Bioprocess Laboratory.

5. Sandy Budi Hartono, Ph.D as members of committees as Head of

Chemical Engineering Department

6. Suryadi Ismadji, Ph.D as Dean of Engineering Faculty

7. Our parents and family who have given a lot of help and support,

both materially and morally

8. Our lecturers, friends and also those who are too many to be listed

by name that had contributed their kind assistance

ix

The authors realize that this report is far from perfect, therefore any

critics and comments which will better improve the research is gladly

accepted. Lastly the authors hope that the report will be useful to all readers

who need information regarding the research of the report.

Surabaya, May 31st 2017

The authors

x

ABSTRACT

Antibiotic potential for people who are injured get infection are

high. First aid to evade the infection use antibiotics. Antibiotics have ability

to inhibit or kill bacteria that cause infection. Tetracycline is antibiotic that

usually uses for first treatment because of tetracycline easy to obtain. There

are six classification of carbapenem antibiotic such as imipenem, ertapenem,

mesopenem, and doripenem. Doripenem is one of the classification of

carbapenem that can be obtained easily. Doripenem can be used for first aid

if infection is happened. Silver usually uses too for avoiding infection but it

is not easily to degrade. It is necessary to find antibacterial composites that

easily to find and biodegradable.

The purpose of this research is to study the effect of pH, kinetic, and

isotherm adsorption of doripenem into bentonite. The optimum condition of

doripenem adsorption will be determined. After knowing the optimum

condition, the antibacterial composite will be tested on nutrient agar.

This research has been done in 5 steps, purification of bentonite

before it is used for adsorption using H2O2, tested the effect of pH doripenem

on adsoption has been done and from the results show that the optimum pH

to adsorp doripenem onto bentonite is at pH 3 (26.8035 mg/g), adsorption

kinetics using bentonite as adsorbents fitted better with pseudo – first order

model (R2=0.9935) and it takes 48h until the doripenem cannot be adsorped

onto bentonite surface. This mean doripenem loading into bentonite

dominated by physiosorption, Isotherm adsorption of doripenem into

bentonite and antibacterial composite that tested in nutrient agar makes wider

area if the doripenem concentration is higher.

xi

ABSTRAK

Potensi penggunaan antibiotik untuk orang yang terkena infeksi

sangatlah tinggi. Pertolongan pertama untuk menangani infeksi adalah

menggunakan antibiotik. Tetrasiklin adalah antibiotik yang biasanya

digunakan untuk pertolongan pertama karena tetrasiklin mudah untuk

ditemukan. Ada empat penggolongan dari antibiotik carbapenem, yakni

imipenem, ertapenem, mesopenem, dan doripenem. Doripenem adalah salah

satu anggota carbapenem yang dapat ditemukan dengan cukup mudah.

Doripenem dapat digunakan untuk pertolongan pertama pada infeksi. Silver

yang biasanya ada pada antibakterial sangatlah sulit untuk terdegradasi. Maka

diperlukan antibakterial yang dapat dengan mudah terdegradasi.

Tujuan dari penelitian ini adalah untuk mempelajari efek pH,

kinetik, dan isoterm adsorpsi dari adsorpsi doripenem ke bentonite. Kondisi

optimum untuk adsorpsi doripenem juga telah ditentukam. Setelah

mengetahui kondisi optimum, antibakterial komposit ini akan diuji ke nutrien

agar.

Penelitian ini terbagi menjadi lima tahap, purifikasi bentonite

sebelum digunakan menggunakan peroksida, tes pH untuk adsorpsi

doripenem dan hasilnya adalah pada pH 3 (26.8035 mg/g), kinetika adsorpsi

menunjukkan bahwa hasil mendekati persamaan pseudo-first dengan

R2=0.9935 dan membutuhkan 48 jam agara doripenem teradsorpsi sempurna

ke bentonite. Hal ini menunjukkan bahwa adsorpsi doripenem ke bentonite

termasuk adsorpsi fisika. Hasil tes nutrien agar menunjukkan bahwa semakin

besar konsentrasi doripenem akan makin besar pula area bersih di sekitar well

nutrie agar.

xii

CONTENTS

RESEARCH PROJECT ............................................................................. i

LETTER OF APPROVAL ....................................................................... ii

COPY RIGHT AGREEMENT ............................................................... iv LETTER OF DECLARATION .............................................................. vi

PREFACE ................................................................................................ viii

ABSTRACT ............................................................................................... x

ABSTRAK ................................................................................................. xi

CONTENTS .............................................................................................. iv

LIST OF FIGURE ..................................................................................... v

LIST OF TABLE ...................................................................................... vi

CHAPTER I ................................................................................................ 1

I.1 Background.......................................................................... 1

I.2 Objective ............................................................................. 2

CHAPTER II .............................................................................................. 3

II.1 Antibiotic ............................................................................. 7

II.2 Adsorption ........................................................................... 7

II.3 Bentonite ............................................................................. 9

II.4 Point of Zero Charge ......................................................... 11

II.5 Cation Exchange Capacity…………………… 12

II.6 Boehm Titration ................................................................. 12

II.7 Doripenem ......................................................................... 12

CHAPTER III ........................................................................................... 13

III.1 Experimental Design ......................................................... 13

III.2 Materials ............................................................................ 18

III.3 Instruments ........................................................................ 19

III.4 Variables ............................................................................ 19

III.5 Research Methodology ...................................................... 20

III.6 Characteristics ................................................................... 22

CHAPTER IV .......................................................................................... 23

IV.1 pH Optimum for Adsorption ............................................ 23

IV.2 Characterization ................................................................. 23

IV.3 Drug Loading..................................................................... 27

IV.4 Kinetic Desorption ............................................................. 31

IV.5 Antibacterial Test............................................................... 33

CHAPTER V ............................................................................................ 36

xiii

REFERENCES ......................................................................................... 37

APPENDIX A .......................................................................................... 39

APPENDIX B .......................................................................................... 49

APPENDIX C .......................................................................................... 51

APPENDIX D .......................................................................................... 53

APPENDIX E ........................................................................................... 55

APPENDIX F ........................................................................................... 57

xiv

LIST OF FIGURES

Figure II.1. Structure of Montmorillonite .................................................. 11

Figure III.1. Preparation of Bentonite ......................................................... 14

Figure III.2. Effect of pH on adsorption ..................................................... 15

Figure III.3. Isotherm Adsorption ............................................................... 16

Figure III.4. Kinetics Adsorption ................................................................ 17

Figure III.5. Well-diffusion method............................................................ 18

Figure IV.1. pH optimum of Bentonite ....................................................... 23

Figure IV.2. pHpzc of Purified Bentonite .............................................. 24

Figure IV.3. pHpzc of Non-Purified Bentonite ............................................ 24

Figure IV.4. FTIR of Bentonite and Doripenem ......................................... 25

Figure IV.5. Pseudo First Order of Doripenem 200 ppm ........................... 27

Figure IV.6. Freundlich and Langmuir of Doripenem 200 ppm ................. 30

Figure IV.7. Kinetica Drug Release at pH 3 ............................................... 31

Figure IV.8. Kientica Drug Release at ....................................................... 32

Figure IV.9. Antibacterial Inhibition Area Test (2.8cm) ............................ 33

Figure IV.10. Antibacterial Inhibition Area Test (2cm) ............................. 34

Figure IV.11. Antibacterial Inhibition Area Test (2.6cm) .......................... 34

Figure IV.12. Antibacterial Inhibition Area Test (3cm) ............................. 35

Figure IV.13. Antibacterial Inhibition Area Test (3.1cm) .......................... 35

Figure B.1. FTIR Characterization of Bentonite......................................... 49

Figure B.2. FTIR Characterization of Bentonite-Doripenem ..................... 50

Figure C.1. Standard Curve of Doripenem using NaCl .............................. 51

Figure C.2. Standard Curve of Doripenem using Aquadest ........................ 52

Figure D.1. Pseudo first order for doripenem 200 ppm .............................. 54

Figure D.2. Pseudo second order for doripenem 200 ppm ......................... 54

Figure E.1. Langmuir Plot of Doripenem 200 ppm .................................... 55

Figure E.2. Freundlich Plot for Doripenem 200 ppm ................................. 56

Figure F.1. Pseudo first order at pH 3 ......................................................... 59

Figure F.2. Pseudo second order at pH 3 .................................................... 59

Figure F.3. Pseudo first order at pH 7 ......................................................... 60

Figure F.4. Pseudo second order at pH 7 .................................................... 60

xv

LIST OF TABLES

Table II.1. Comparison of Antibiotic Composite ...................................... 4

Table IV.1. Comparison of functional group on bentonite-doripenem ...... 26

Table IV.2. Boehm Result ......................................................................... 26

Table IV.3. Adsorption Kinetics Parameters ............................................. 28

Table IV.4. Adsorption Isotherm Parameters ............................................ 30

Table IV.5. Drug Release Kinetics Parameters.......................................... 32

Table A.1. Doripenem at various pH ......................................................... 47

Table C.1. Concentration vs Absorbance Doripenem Standard ................. 51

Table C.2. Concentration vs Absorbance Doripenem Standard ................. 52

Table D.1. Values of qt for Initial Conc. Doripenem 200 ppm ................ 53

Table E.2. Isotherm Adsorption of Doripenem 200 ppm .......................... 56

Table F.1. Values of qt at pH 3 ................................................................ 57

Table F.2. Values of qt at pH 7 ................................................................ 58