FACULTY OF ENGINEERING AND BUILT ENVIRONMENT

3rd YEAR INTEGRATED PROJECT

(CHEMICAL ENGINEERING
PROGRAMME) SEMESTER II
2020/2021

TITLE: PRODUCTION OF PENICILIN G FROM PENICILIUM

CHRYSOGENUM
COURSE CODES: KKKR3853, KKKR3733, KKKR3723, KKKR3513

LECTURERS: COURSE LECTURERS:

KKKR3583: PROF. MADYA TS. EN. NOORHISHAM BIN TAN KOFLI
TS. DR. AHMAD RAZI OTHMAN
KKKR3733: PROF. MADYA IR. DR. HASSIMI BIN ABU HASAN
PROF. MADYA TS. EN. NOORHISHAM BIN TAN KOFLI
DR. DARMAN NORDIN
DR. MUHAMMAD ZULHAZIMAN MAT SALLEH
KKKR3723: PROF. MADYA IR. DR. SHUHAIDA HARUN
IR. DR. TEOW YEIT HAAN
KKKR3713: PROF. MADYA DR. NORLIZA ABD. RAHMAN
DR. JARINAH MOHD ALI
GROUP: KK10

SUBMISSION 24TH JUNE 2021

DATE:
MEMBERS: ENGKU AHMAD FARIZ BIN ENGKU AHMAD
KHAIRUL ANUAR (A168003)
AHMAD TAUFEAQ ISMADI BIN AHMAD ZAKKI (A169159)
AMIR FITRI BIN MUHAMMAD SHAFIQ POH (A168739)
MUHAMMAD ASYRAF SHAFIE (A167520)
 ii

DECLARATION

We hereby declare that the works in this Integrated Project are out own work except
for quotation and summaries which have been fully acknowledged.

24TH JUNE 2021

ENGKU FARIZ
ENGKU AHMAD FARIZ BIN ENGKU AHMAD KHAIRUL ANUAR

(A168003)

TAUFEQ ISMADI
AHMAD TAUFEQ ISMADI BIN AHMAD ZAKKI

(A169159)

AMIR FITRI
AMIR FITRI BIN MUHAMMAD SHAFIQ POH

(A168739)

ASYRAF SHAFIE
MUHAMMAD ASYRAF BIN SHAFIE

(A167520)
 iii

ACKNOWLEDGEMENT

All the group members from KK10 would like to explicit out thankfulness and
gratefulness because this integrated project was able to be carried out successfully. In
performing this integrated project, all the group members had to take the help and
guideline of some respected persons, who deserve the greatest gratitude. The
completion of this integrated project gives all the group members much pleasure. All
the group members of KK10 would like to show our gratitude to all the lectures, Prof.
Madya En. Noorhisham Tan KOFLI, Dr. Ahmad Razi Othman and prof. Dr. Mohd
Sahaid hj. Kalil (KKKR3853), Prof. Madya Dr. Norliza Binti Abd Rahman and Dr.
Jarinah Mohd Ali (KKKR3733), IR. Dr. Shuhaida Harun and Dr. Nur Tantiyani Ali
Othman (KKKR3723) and Prof. Madya Ir. Dr. Hassimi Bin Abu Hasan and Dr.
Muhammad Zulhaziman Bin Mat Salleh (KKKR3513) from Universiti Kebangsaan
Malaysia for giving our group a good guideline for this project throughout numerous
consultations. All the group members would also like to expand our deepest gratitude
to all those who have directly and indirectly guided us in writing this assignment. In
addition, a thank you to about all the seniors who offered help and advice all this time,
and not to forget the authors, technical papers and articles from internet as it helps us
to get many information and reference, without all these helpful sources, it might be
difficult to complete of this project. All the group members also thank the authority of
the National University of Malaysia for giving the best facilities and comfortable
environment to complete this project. Many people, especially classmates and team
members itself, have made valuable comment suggestions on this project which gave
an inspiration to improve this project. All the group members from KK10 would like
to thank all the people for their help directly and indirectly to complete this integrated
project.
 iv

EXECUTIVE SUMMARY

Group KK10 were assigned to do the integrated project on the production of Penicilin
G from Penicillium chrysogenum. This is an alternative way of producing isoprene
using biotechnology instead of using petroleum. This integrated project including
introduction of Penicilin G, economic analysis, environmental issues, production
process, mass and energy balance, Biochemistry and Biomolecular Engineering,
Separation Process II, Utility and Pressure Vessel Design, and Dynamic and Process
Control. The main purpose for this integrated project is to design a suitable production
plant that can produce Isoprene. Group members from KK10 started working on this
integrated project since the 14th of February 2021. All the members had come to a
final decision to select Penicillium chrysogenum as the microbial host. After getting
all the information and though research, a plant with a plant capacity of 2,292,000
kg/year had been designed. It is said to be able to fulfil 9.9% of the global demand.
The heat released by reaction is -36,058,355kJ/batch. This show that the reaction is an
exothermic reaction. The manufacturing process consists of fermentation, solvent
extraction, stripping and condensation. The process produces Penicilin G along with
four other by-products which are wastewater, Butyl Acetate, carbon dioxide and
biomass. Mass balance from manual calculation is also compared with Superpro®
Simulation. Kremser equation used to estimate the number of stages for selected
separation part. Utility unit and pressure vessel were design and detailed drawing is
being drawn. A P&ID which shows the piping and process equipment together with
control device is used to monitor the process. All the members from KK10 hope to
gain more knowledge through this integrated project and learn through the mistakes
we made. The knowledge gained through this integrated project will surely be helpful
and useful to all of us in the future.
 v

TABLE OF CONTENT

Page

DECLARATION i

ACKNOWLEDGEMENT ii

EXECUTIVE SUMMARY iii

TABLE OF CONTENTS iv

LIST OF TABLES viii

LIST OF ILLUSTRATIONS x

CHAPTER I INTRODUCTION

1.1 Background of Penicilin G 1

1.2 Background of Penicillium Chrysogenum 2

1.3 Production Method Description 2

1.4 Chemical And Physical Properties of Product 3

1.5 Usage of Penicilin G 4

CHAPTER II PRODUCTION PROCESS

2.1 Process Block Diagram 5

2.2 Process Description 5

2.3 Process Flow Diagram 6

CHAPTER III ECONOMIC ANALYSIS

 vi

3.1 Supply and Demand of Isoprene 7

3.2 Production Capactiy 9

3.3 Future Marketing Potential 10

3.4 Companies Producing Isoprene 11

CHAPTER IV ENVIRONMENTAL ISSUES

4.1 Waste Generation 12

4.2 Waste Treatment 14

4.3 Environmental Act 13

4.4 Safety Issues and Precautions 14

vii
CHAPTER V MASS AND ENERGY BALANCE

5.1 Stoichiometry Equation 16

5.2 Overall Mass Balance 18

5.3 Mass Balance on Each Unit 22

5.4 Energy Balance 22

5.4 Manual and Superpro® calculation comparison 26

CHAPTER VI BIOCHEMISTRY AND

BIOMOLECULARENGINEERING

6.1 Main Enzmye Used in Production 27

6.2 Metabolic Pathway 28

6.3 Enhancement of Isoprene Production 29

CHAPTER VII SEPARATION PROCESS II

7.1 Introduction 31

7.2 Separation Unit And Mechanism 32

7.3 Mass Balance Of Separation Unit 33

7.4 Design Of Absorption Column 34

CHAPTER VIII UTILITY AND PRESSURE VESSEL DESIGN

8.1 Utility Design 40

8.1.1 Introduction to Utility Design 40

8.1.2 Utility Design Procedure 40

8.1.3 Design Of Condenser E-201 41

8.1.4 Design Of Shell and Tube Exchanger E-101 43

8.2 Design Of Pressure Vessel 45

8.2.1 Design Of Fermenter, F-201 45

 viii

8.2.2 Design Of Absorption Column C-101 62

8.3 Design Specification Of Pressure Vessel 45

8.3.1 Design Specification Of Fermenter, F-101 45

8.3.2 Design Specification Of Absorption Column C-101 63

8.4 Design Pressure, Minimum Wall Thickness, MAWP 43

8.4.1 Design Pressure, Minimum Wall Thickness, MAWP

ofFermenter F-101
49

8.4.2 Design Pressure, Minimum Wall Thickness, MAWP 65

Of Absorption Column C-101

8.5 Thickness Uniformity And Nominal Thickness Correction

8.5.1 Thickness Uniformity And Nominal Thickness

Correction Of Main Fermenter, R-2

8.5.2 Thickness Uniformity And Nominal Thickness

Correction Of Extraction Column E-1

8.6 Combines Loading Analysis

8.6.1 Combine Loading Analysis For Main Fermenter, R-2

8.6.2 Combine Loading Analysis For Extraction Column, E-

1

8.7 Analysis Of Elastic Stability

8.7.1 Analysis Of Elastic Stability Of Main Fermenter, R-2

8.7.2 Analysis Of Elastic Stability Of Extraction Column,

E-2

8.8 Vessel Support Analysis

8.8.1 Skirt Thickness Of Main Fermenter, R-2

8.8.2 Skirt Thickness Of Extraction Column, E-1

8.9 Base Ring And Anchor Bolt Design

 ix

8.9.1 8.7.1 Base Ring And Anchor Bolt Design Of Main

Fermenter R-2

8.9.2 8.7.1 Base Ring And Anchor Bolt Design Of Extraction

Column, E-1

8.10 Mechanical Drawing 77

CHAPTER IX DYNAMIC AND PROCESS CONTROL

9.1 Introduction 78

9.2 Seed Fermenter F-101 79

9.3 Main Fermenter F-201 82

9.4 Absorption Column C-101 86

9.5 Flash Vessel V-101 88

9.6 P&ID 90

CONCLUSION 91

REFERENCES 92
 xii

APPENDICES

Appendix A Mass Balance For Each Unit 95

Appendix B Separation Process II 100

Appendix C Pressure Vessel Design 107

Appendix D MSDS and SUPERPRO® report 128

 xiii

LIST OF TABLES

Table 1.1 Chemical and Physical Properties of Isoprene 3

Table 4.1 Waste Generated 11

Table 4.2 Waste Treatment 12

Table 4.3 Safety Precaution of Raw Material and Product 13

Table 5.1 Molecular weight of each component 15

Table 5.2.1 Concentration and Mass flowrate of Inlet and Outlet

Components of overall process
20

Table 5.2.2 Molar and Mass flowrate of inlet and outlet components of
overall process
20

Table 5.4.1 Specific heat capacity of individual elements 22

Table 5.4.1 Specific heat capacity of individual elements 22

Table 5.4.2 Specific heat capacity calculation for glucose 22

Table 5.4.3 Specific heat capacity calculation for biomass 23

Table 5.4.4 Specific heat capacity calculation for isoprene 23

Table 5.4.5 Coefficient Values for Heat Capacity of the Components 24

Table 5.4.6 Enthalpy of each component 24

Table 5.5 comparison between manual calculation and Superpro®

calculation
26

Table 7.1 Comparison of Mass Balance by Material Balance and

Supepro® Simulation
33

Table 7.2 Comparison of Mass Balance by Material Balance and

Supepro® Simulation
34
 xiv

Table 7.3 K-Values of components at distillate (stream 17) and bottom

(stream 16)
35

Table 7.4 Molar flow rate and mole fraction of components in feed stream 36

Table 8.1 Operating condition of Fermenter, F-201 46

 xv

Table No. Page

Table 8.2 Dimension of Fermenter, F-201 47

Table 8.3 Inlet component of Fermenter, F-201 47

Table 8.4 dimension of fermenter, F-201 48

Table 8.5 Design pressure, Minimum Wall thickness and MAWP vessel of
Fermenter, F-201
44

Table 8.6 tmin, MAWP part and MAWP vessel of each part of fermenter
after thickness uniformity and nominal thickness correction of
Fermenter, F-201

54

Table 8.7 Parameter used in the analysis of primary stress for fermenter F-
201
54

Table 8.8 Specification in skirt thickness for Fermenter F-201 58

Table 8.9 The resultant stresses in the skirt design of Fermenter F-201 58

Table 8.10 Base Ring and Anchor Bolt design specification for Fermenter
F-201
60

Table 8.11 Design shape of Absorption column, C-101 64

Table 8.12 Design Specification of C-101 64

Table 8.13 Operating Conditions of C-101 65

Table 8.14 MAWP vessel Calculation Summary 67

Table 8.15 Summary of Thickness Uniformity and Nominal Thickness

Correction
69

Table 8.16 Parameter in analysis of primary stress 70

Table 8.17 Specification in skirt thickness of Absorption column C-101 73

Table 8.18 The resultant stresses in the skirt design of Absorption column
C-101
74
 xvi

Table 8.19 Base Ring and Anchor Bolt design specification 77

Table 9.1 Control action taken on main fermenter 81

Table 9.2 Control action taken on heat exchanger 85

Table 9.3 Control action taken on absorption column 87

Table 9.4 Control action taken on flash vessel 90

 xvii

LIST OF
ILLUSTRATIONS

Figure No. Page

Figure 1.1 2D structure of Isoprene 1

Figure 1.2 Escherichia Coli 2

Figure 2.3 Process Block Diagram 5

Figure 3.1 Market Size of Isoprene from year 2018 to 2030 8

Figure 3.2 Supply and demand of Isoprene from year 2018 to 2030 9

Figure 5.1 Overall Process 18

Figure 5.2 Main Fermenter F-102 23

Figure 5.3 Plant drawn using Superpro® 26

Figure 6.1 Illustration of enzyme isoprene synthase (IspS) 27

Figure 6.2 MVA (exogenous mevalonate) pathway 28

Figure 6.3 MVA pathway after oprimization 29

Figure 7.1 Absorption unit C-101 32

Figure 7.2 Absorption unit C-101 33

Figure 8.1 Main Fermenter, F-201 45

Figure 8.2 Absorption unit C-101 63

Figure 8.3 Basic schematic of C-101 63

Figure 9.1 Control configuration of seed fermenter, F-101 81

Figure 9.2 Control configuration of main fermenter, F-201 85

Figure 9.3 Control configuration of Absorption Column 86

Figure 9.4 Control configuration of flash vessel 89

 xviii

CHAPTER I

INTRODUCTION

1.1 BACKGROUND OF PENICILLIUM CHRYSOGENUM

Penicillium Chrysogenum is a species of fungus and can be widely found in indoor

environment, particularly with high humidity, dampness or suffered from water
damage. In the industrial field, P. chrysogenum is used to produce penicillin which
brought by the discovery of penicillin by Sir Alexander Fleming in 1928 where
antibiotics are then derived from microorganisms. This relates to our Integrated
Project in producing the key compound in amoxicillin. A crucial characteristic of P.
chrysogenum is the fact that it only produces penicillin when it is under stress.
However, this species grows well in a wide range of conditions including indoor
environments, damp areas, in the soils and on decaying vegetation (Ward, 2018)

.Figure1.1: Penicillium Chrysogenum

 xix

1.1.1 Background of Penicillin G

Penicillin G is a class of antibiotics that comes from mold (fungi). Penicillin G,

discovered by Alexander Fleming in 1928, was first available commercially in 1942.
Penicillin G is natural penicillin which still maintains activity against other gram-
positive cocci and bacilli and gram-negative cocci. Staphylococci, initially sensitive to
penicillin G, are now considered highly resistant. Penicillin G also is an antibiotic
used to treat numerous of bacterial infections. The chemical formula for penicillin G is
C16H18N2O4S with molecular weight 334.39 g/mol. Penicillin G is available in
crystalline, procaine and benzathine forms. It is most effective against gram-positive
organisms, but it is susceptible to β-lactamase (Pediatric Dentistry Sixth Edition,
2019).

Figure 1.2 The chemical structure of Penicillin G

Source: Researchgate, 2013

 xx

Table 1.1 A Summary on Properties of all the Substances Involved

Name of Chemical Molar Density Boiling Melting Solubility in

substance Formula Mass (kg/m3) point Point water
(g/mol) (°C) (°C)
Phenylacetic C8H8O2 136.15 1080 265.5 76-77 Soluble
acid
Glucose C6H12O6 180.156 1560 146.111 146.11 Soluble
Ammonium (NH4)2SO4 132.14 1770 235-280 Soluble
Sulphate
Butyl acetate C6H12O2 116.16 882 126 -78 Soluble
Penicilin G C16H18N2O4S 334.3901 1410 55 209-212 Soluble
(Source: Pubchem Open Chemistry DataBase, 2005)

1.2 CHEMICAL AND PHYSICAL PROPERTIES

Name of Chemical Molar Mass Density Boiling Melting Solubility in

substance Formula (g/mol) (kg/m3) point Point water
(°C) (°C)

Phenylacetic C8H8O2 136.15 1080 265.5 76-77 Soluble

acid

Glucose C6H12O6 180.156 1560 146.111 146.11 Soluble

Ammonium (NH4)2SO4 132.14 1770 235-280 Soluble

Sulphate

Butyl acetate C6H12O2 116.16 882 126 -78 Soluble

Penicilin G C16H18N2O4S 334.3901 1410 55 209-212 Soluble

1.3 USAGE OF PENICILLIN G

Penicillin are a type of antibiotic produced from Penicillium fungi. An antibiotic is a

type of medicine that inhibits the growth of, or kills, bacteria. Penicillin G (also called
benzylpenicillin) was discovered by accident in 1928. Alexander Fleming, a Scottish
physician-scientist was growing a type of bacteria called Staphylococcus Aureus on
an uncovered petri dish when it became contaminated with mold spores. He observed
 xxi

that the areas of bacteria near the mold were dying. He isolated the substance from the
mold that was killing the bacteria and called it penicillin (Fookes 2019). Penicillin are
antibiotics used to treat bacterial infections that are derived from the antibiotic
penicillin (Omudhome, 2020).

Penicillin may be used to treat a wide range of infections caused by susceptible

bacteria, such as dental abscess, ear infections (e.g., otitis media), gonorrhoea,
pneumonia, respiratory tract infections, rheumatic fever, scarlet fever, skin infections,
urinary tract infections (Fookes 2019). Today, many derivatives of penicillin have
been developed that inhibit more types of bacteria than the original life-saving drug.
Penicillin itself is active against streptococci (including Streptococcus pneumoniae),
Listeria, Neisseria gonorrhoeae, Clostridium, Peptococcus, and Peptostreptococcus.
However, most staphylococci now are resistant to penicillin. Other penicillin
antibiotics are effective against H. influenzae, E. coli, pneumococci, certain strains of
staphylococci, Salmonella, Shigella, Pseudomonas aeruginosa, and many other types
of bacteria. Penicillin antibiotics are used to treat many types of infections caused by
susceptible bacteria. They are used to treat infections of the middle ear, sinuses,
stomach and intestines, bladder, and kidney (Omudhome, 2020).
 xxii

CHAPTER II

PROCESS DESCRIPTION

2.1 PROCESS DESCRIPTION

2.1.1 PROCESS DESCRIPTION

The Penicillin G production bioprocess is carried out with strains of Penicillium

chrysogenum where it involves upper stream and downstream process. The upper
stream process mainly consists of fermentation process while the downstream process
involves recovery and purification of the penicillin G (da Cruz et al., 1997).

2.1.2 Preparing inoculum in seed fermenter, R-1

The production of penicillin G carried out with strains of Penicillium chrysogenum

which is a strictly aerobic fungus (Cruz et al. 1997). Inoculum, is prepared by growing
the fungus in an optimum condition in which the temperature is set to be 25˚C and the
medium use is glucose and ammonium sulphate (Chun C.D et al. 2010). Since the
Penicillium chrysogenum is strictly aerobic fungus therefore oxygen is needed and
being supplied into the seed fermenter for the preparation of inoculum.

2.1.3 Production of penicillin G in batch fermenter, R-2

The main fermentor, R-2 are operate in batch operation where the temperature, pH
strictly control. The temperature and pH are control and maintain at 25℃ and 6.5
respectively (Gordon et al., n.d.). Besides, the regulation of dissolve oxygen are vital
in this fermentation as the microbes are highly aerobic fungus. The concentration of
dissolved oxygen will set to be close to saturation. However with an increasing of
 xxiii

biomass with respect to time, the fermentation broth will become more viscous and
the dissolve oxygen level will decrease rapidly. Therefore the batch fermentation will
operates with air flow rate of 12.11 kg/h with a total feed of 848 kg of air and the
dissolve oxygen value are maintain in excess of 85% throughout the course of the
fermentation by increasing agitator speed in a range of 500 to 700 rpm (Goudar &
Strevett, n.d.).

Carbon source are essential for the microbial growth, where it play a
big roles in biosynthesis and energy generation with carbohydrates being the
usual carbon source for microbial fermentation processes (Ward 1991; Stanbury et al.
1995). The Penicillium chrysogenum will be feed with glucose as it source of carbon.

Besides carbon source, nitrogen source also play a vital role in the growth
were as it represents an important nutritional factor in for Penicillium chrysogenum
due to its function in protein synthesis and sugar transport. For the production of
penicillin G, ammonium sulphate will be used as the nitrogen source (Lorenz et al.,
2007).

Finally, the fermentation process will be fed with a small amount of phenyl
acetic acid for Penicillium chrysogenum to utilize it as a side chain precursor in
penicillin G biosynthesis (Hillenga et al., 1995).

The bioreaction take place in the main fermenter, R-2:

C6H12O6 + 1.792O2 + 0.716(NH4)2SO4 + 0.899C8H8O2 → 0.485C16H18N2O4S

+ 2.89CH1.64O0.52N0.16S0.08 + 5.725H2O + 2.539CO2

C6H12O6 = Glucose; O2 = Oxygen; (NH4)2SO4 = Ammonium sulphate; C8H8O2

= Phenyl acetic acid; C16H18N2O4S = Penicillin G; CH1.64O0.52N0.16S0.08 = Biomass;
H2O = Water; CO2 = Carbon dioxide
 xxiv

The yield of pencillin G is 90% (Meštrović,T n.d.) and the yield of biomass
will be 40% (Anon n.d.) with a specific growth rate of 0.02 h-1 (Pirt & Righelato,
1967) when the optimum condition are achieve.Filtration in centrifugal stack disc, C-1

The downstream process will begin with filtration in centrifugal stack disc, C-1. The
fermenter output will enter the centrifugal stack disc to filter out the solid biomass.
The denser solid biomass are subjected to the centrifugal forces and will move
outwards towards the rotating bowl wall while the less dense fluids will move towards
the center and thus separating the biomass from the liquid broth. The liquid broth will
enter the extraction column, E-1 and further purified.

2.1.4 Extraction of Penicilin G in Extraction Column, E-1

The downstream process is continued in extraction column, E-1. Before entering the
extraction column, E-1 the liquid broth from the C-1 is first cooled down to 4°C in
order to stabilize the penicillin G (Aliwarga et al., 2019) and will enter pump, P-1 as
to recover the pressure drop due to centrifugal separation process in C-1. The
extraction processes. The extraction will operate in a batch process of liquid-liquid
extraction method. The pH value of the broth is control in a range of 2 to 5 as the
operation much favorable in acidic environment (Aliwarga et al., 2019). The solvent
use to dissolve the penicillin G is n-butyl acetate the yield for the extraction is 95.51%
with partition coefficient on penicillin 21.47 (Aliwarga et al., 2019). The penicillin-
rich solution with butyl acetate will be found on the upper product and recovery of the
penicillin G are continue in regeneration column, E-2.
 xxv

2.1.5 Recovery of Penicilin G in regeneration column, E-2

The upper product E-1 that consist of penicillin-rich solution with butyl acetate will
enter E-2 and it being introduce with water and phosphate buffer that will bring the
solution into an alkaline condition of pH 7 where the penicillin stability is high. This
condition are vital in recovery the penicillin as it will separate the penicillin from the
butyl acetate. This extraction method is known as solvent transfer method
(Antibiotics-Exhibition 2016 Penicillin, n.d.). The penicillin G will enter the watery
phase and will be found in the bottom product of E-2. The addition of water will
usually take one fifth of the volume of the original broth and the addition of phoshpate
buffer will usually take one-tenth of the volume of the broth (Gordon et al., n.d.).

2.1.6 Converting Penicilin G into Penicilin salt in CSTR reactor, R-3

The bottom product of extraction column, E-2 in stream will enter the continuous
stirrer tank reactor (CSTR), R-3 and will react with sodium bicarbonate to form
penicillin salt. The chemical reaction are given:

C16H18N2O4S + NaHCO₃ → C16H17N2NaO4S + CO2 + H2O

Where,

C16H18N2O4S = Penicillin; NaHCO₃ = Sodium bicarbonate; C16H17N2NaO4S =

Penicillin salt; CO2 = Carbon dioxide; H2O = Water.

This reaction are critical in order to allow the penicillin to be stored in a stable
powder form at a room temperature.
 xxvi

2.1.7 Extracting Penicilin salt in centrifugal basket, C-2

The product from the R-3 will enter the centrifugal basket, C-2. This allow the
penicillin salt to be extracted from the liquid material product of R-3. C-2 are being
employ because the high concentration of the solid content (penicillin salt) of the
suspension is higher than the liquid. The penicillin salt will accumulate and compress
on the C-2 wall as effect of the centrifugal force. The applied centrifugal force created
by the angular velocity of a rotating basket are larger than the capillary force, liquid in
the capillary tubes are spontaneously removed from the filter cake (Solid-Liquid
Separation, n.d.). The purification is continue as the cake containing the penicillin salt
enter the freeze drying crystallizer, F-1.

2.1.8 Drying Penicilin salt in freeze drying crystallizer, F-1

The down process is continue in the freeze drying crystallizer, F-1. Before the cake
containing penicillin salt enter F-1, the pressure and temperature of the cake are first
must to be reduce to create an environment with the optimal conditions for the
sublimation phase, to turn a frozen material immediately into gas. The temperature are
reduce slowly to about -50°C as it will contribute in to a bigger of the size crystals that
being produce and the larger the amount of remaining Penicillin sodium salt (Lorenz
et al., 2007). The freeze drying method are being employ as it the only method to dry
the penicillin without the loss of activity and will enable to keeping them chemically
stable without the need of refrigeration (E. P. Abraham et al., 1941).

The process will be a two steps process which are primary-drying and
secondary-drying. The primary drying will be conduct in a very low pressure to the
range of milibars. In this primary drying phase, about 95% of the frozen water
molecules in the material are sublimated (Lorenz et al., 2007).
 xxvii

The secondary drying aims to remove unfrozen water molecules. The temperature is set to be higher than the primary drying and the
pressure is set to be lower than the primary drying in a range of microbars. This condition are important to break the bond between the water
molecules and the frozen material and thus vaporize the unfrozen water from the solid materials. At the end of the operation, the final residual
water content in the product is extremely low, around 1% to 4%. (Lorenz et al., 2007).

The final product is stabilize powder of penicillin salt with estimate purity of 98

2.2 PROCESS FLOW DIAGRAM

xxviii
 xxix

CHAPTER III

ECONOMIC ANALYSIS

3.1 SUPPLY AND DEMAND OF PENICILLIN G PRODUCTION

Penicillin G is currently produced by only four companies in the world. It is because

the business that makes penicillin G makes a small profit. The business that made it
only managed to generate around 20% of what they were capable of. This is due to the
fact that the cost of the medicine that is being sold is a low-cost option. In 2017, there
was a penicillin G shortage in Portugal. When the former European supplier changed,
Portugal had to ask China to supply penicillin G due to a shortage. They need the
demand from China because Laboratories, a Portuguese pharmaceutical company,
claims that it lacks the complete collection of documents required by European
regulators.

Because if a product is delayed or fails, it will affect the entire planet, a small
number of manufacturers will jeopardize global dependence. Due to a lack of
availability, sick people will have to resort to less cost-effective and more expensive
medicine. A three-year shortage of benzathine penicillin hit Brazil in the midst of a
Cupid's disease epidemic, a disease linked to serious malformation in infants. The
antibiotic is the only medication that can destroy the Cupid's disease bacterium in the
craniate.

Maria Luiza Bezerra Menezes as Medical organizer of hospital Cisam in

metropolis, explains that the ministry of health worked very hard to provide the
substitute medication when shortage of Penicillin G occurs. Substitute medicines may
also be costlier. Whereas penicillin g prices between $2.30 to $ 3.20 (R$7 to R$10) in
Brazil, a dose of the antibiotic Mefoxin prices double (Guimaraes 2017).
 xxx

Demand of antibiotic increased by 65% between 2000 and 2015, from 21.1 to

44.8 million kg and in 2022 the demand is expected to reach about 58.2 billion
kg. While the demand of amoxicillin antibiotic which is one of the most common
antibiotics consumed is 39% of total kg which is 23.572 million kg in 2015 and in
2022 the demand is expected to reach about 48.56 million kg. The CAGR is estimated
in 4.8% annually (Eili Y. Klein et al. 2018).

While the production of penicillin G from China to export to the world is about

13.6 million kg in 2015 and in 2017 is about 18.7 million kg and the
production rate is about 14.5% annually. The production of penicillin G is expected to
reach about 25.64 million kg (Anon 2019). The figure 2.1 below show the graph of
global supply and demand for Penicillin G.

Figure 2.1: Global supply and demand for Penicillin G

 xxxi

3.2 PRICE AND FUTURE MARKETING POTENTIAL OF PENICILLIN G

3.2.1 Market Size

The global antibiotics market size was valued at USD 40.7 billion in 2020 and is
expected to expand at a compound annual growth rate (CAGR) of 4.5% from 2021 to
2028. High usage of antibiotics and inappropriate prescription behaviour of antibiotic
drugs worldwide are the major factors anticipated to drive the market. Moreover,
rising awareness among patients and healthcare professionals and increasing
involvement of regulatory bodies in the R&D activities of new therapies to treat
infectious diseases are expected to propel the market growth over the forecast period.
The demand for antibiotics is significantly increasing owing to the increasing
incidence of infectious diseases. The high prevalence of infectious diseases, such as
lower respiratory infections, pneumonia, malaria, and tuberculosis, is also fuelling the
market growth (Anon 2019). The figure 2.2 show U.S. antibiotics market size from
2017 – 2028.

Figure 2.2: U.S. antibiotics market size from 2017 – 2028

 xxxii

3.2.2 Penicillin Market Share

Four companies manufacture the active ingredient for penicillin - a drug that changed
modern medicine 76 years ago and because the medicine deals little profit, those
companies keep production levels low. As penicillin has been used to treat diseases, such
as syphilis and rheumatic heart disease, which extremely affect poorer countries, the
extent of the demand for the drug also is not always clearly apprehended. Over the past
decade, a few companies have left the market looking for more profitable products. Three
of the four companies that still produce the active pharmaceutical ingredient for
benzathine penicillin G are located in China.

These are North China Pharmaceutical Group Semisyntech Co. Ltd, CSPC
Pharmaceuticals Group Ltd. and Jiangxi Dongfeng Pharmaceutical Co. The fourth
company is Austria-based Sandoz GmbH. These companies manufacture only 20 percent
of what they could because benzathine penicillin G is off patent, offers little profit and
because demand data is extremely restricted according to the WHO. The medicine's cheap
selling expense also makes manufacturers reluctant to enter the market. Even India, a
leading pharmaceutical producer, outsources almost all production of penicillin G to
China, according to the Indian Drug Manufacturers' Association (IDMA) (Guimaraes
2017).
 xxxiii

3.3 LIST OF COMPANIES PRODUCING PENICILLIN G

There are only four companies that still produce active compound for Penicillin G. Three
out of four company located at China and other one at Austria. Demand for this
compound extremely limited, thus those companies just produce low levels which only 20
percent what they could. Furthermore, this medicine is selling at the lower price which
makes manufactures disinclined to enter the market. Based on table 2.1 shows the list of
companies that produce Penicillin G in the world.

Table 2.1 List of company produce Penicillin G in the

world
Name of company Location Capacity (tonnes per year)

North China Pharmaceutical Group Shanghai, China 7000

Shijiazhuang,
CSPC Pharmaceutical Group China 6800

Jiangxi Dongfeng Pharmaceutical Hong Kong, China 600

Sandoz GmbH Austria 600

(Source : Guimaraes,K 2017)

 xxxiv

3.4 PLANT CAPACITY

Penicillin G has strong demand in global market, and it is expected to increase

in the next few years. The production rate of penicillin G in 2025 is estimated
as below:

Demand- Supply = 48,560,000 kg/year – 25,640,000kg/year

= 22,920,000 kg/year

Therefore, the production of penicillin G is lower than the demand in the world as
the value calculating in above is positive value and means that the demand is more
than the supply of the penicillin G.

Assume the production of penicillin G temporarily stop in 80 days in the whole year.

Production batch a year = 285 days ÷ 3 day/batch

= 95 batch

Plant capacity = 0.099 x 22,920,000 kg/year

= 2,292,000 kg/year

= 2,292,000 kg / 95 batch

= 24000 kg/batch

Therefore, our plant capacity is approximately want to produce 2,292,000

kg/year which fulfill 9.9% from the market shortage and therefore it
approximately about 24000kg per batch of production.
 xxxv

CHAPTER IV

ENVIRONMENTAL ISSUES

4.1 INTRODUCTION TO SAFETY AND ENVIRONMENTAL ISSUES

Safety issues are issues that give out short term issues whereas environmental issues give
long term effects which tend to be harmful. In general, safety issues refer to workplace
safety, health and welfare of the employees. Safety rules and regulations are practiced as
safety includes employee awareness related to knowledge of basic safety, hazards along
with its risks and implementation of hazard preventions.Workplace safety is important to
avoid accidents, minor and major losses (Tutorialspoint, 2014). On the other hand,
environmental issues arise whenever there is a change in the quality or quantity of any
environmental factor which directly or indirectly affects the health and well-being of the
population. Environmental problems can be studied by simply looking for adverse effects
to detect trends for further investigations or by studying the cause and effect relationships
which are crucial for better prediction and proper management (Scope, 2014).

4.2 SAFETY ISSUES OF THE RAW MATERIALS AND PRODUCT IN PENICILLIN

G PRODUCTION

The classification for every chemicals and materials are different, such as the properties
and the information on potential hazards.The properties and ways of handling and storing
raw materials and product of penicillin G production are shown below.

Table 5.1
Components
materials
Penicillium chrysogenum
(Source: Enzymology
Reasearch Center)
Ammonium Sulphate
(Source: LabChem 2013)
Properties and handling method of each component
Properties
▪ Causes skin, eye and
respiratory irritation.
▪ May cause allergy or
asthma symptoms or
breathing difficulties if
inhaled.
When heated to
decomposition, material
may emit toxic fumes
▪ Harmful if swallowed.
Slightly harmful by
inhalation.
▪ Causes skin and eye
irritation.
▪ Non-flammable,
including in contact
with water.
▪ Harmful to aquatic life.
▪ May form combustible
dust concentrations in
air during processing.
xxxvi
Ways to handle
▪ Keep away from sources
of ignition, spark and
open flames.
▪ Avoid breathing dust
or spray mist. Avoid
formation of dust and
aerosols.
▪ Ensure good ventilation
of the room when
handling this product.
▪ Store container in a
dry, cool place.
▪ Product is stable for 2
years (24 months) if
stored at or below 10°C
in sealed poly bags in
boxes or drums away
from sunlight and high
humidity.
▪ Provide good ventilation
in process area to prevent
formation of vapor.
▪ Do not eat, drink or
smoke when handling this
product.
▪ Avoid all unnecessary
exposure by wearing
gloves and safety
glasses.
▪ Wash contaminated
clothing before
reuse.
▪ Store at room temperature.
▪ Keep in the original
container, a well
ventilated place away
from incompatible
materials.
▪ Keep container
closed when not in
use.
 xxxvii

Phenylacetic Acid ▪ It is stable ▪ Use chemical

(Source: Cayman material under resistance safety
Chemical) normal condition. goggles.
▪ Classified under

hazardous substance in
terms of toxicity and
corrosion.
▪ May cause to damaging
fertility, unborn child
and also cause to
serious eye and
respiratory irritation.
N-Butylacetate ▪ n-Butyl acetate is a
colorless liquid.
▪ It has a sweet, fruity
odor and tastes a little
like banana. It is very
soluble in water.
▪ Direct exposure to the
eyes resulted in redness
and pain.
▪ Skin irritation has been
reported with repeated
skin exposure.
Glucose ▪ Relatively non-toxic
▪ Prolonged exposure to
nuisance dust could result
in temporary, reversible
respiratory irritation.
▪ Prolonged contact may
cause skin sensitization.
▪ Stable under normal
temperature and
pressure.
xxxviii
▪ Always wear personal
protective clothing and
equipment when
handling the substance.
▪ Always use only under a
chemical fume hood and
avoid raising and
breathing dust, and
provide adequate
ventilation.
▪ Avoid prolonged
or repeated
exposure.
▪ Keep in a dry, cool and
well-ventilated place.
Keep container tightly
closed.
▪ Provide good ventilation
in process area to prevent
formation of vapor.
▪ Do not eat, drink or
smoke when handling this
product.
▪ Avoid all unnecessary
exposure by wearing
gloves and safety
glasses.
▪ Wash contaminated
clothing before
reuse.
▪ Store at room temperature.
▪ Keep in the original
container, a well
ventilated place away
from incompatible
materials.
▪ Keep container
closed when not in
use.
▪ Safety glass and
gloves should be
worn to avoid
unnecessary contact.
▪ Wear paper breathing
mask and protective
eyewear when dumping
or mixing.
▪ If swallowed in large
amounts and the person
is conscious,
immediately give large
 xxxix

amounts water.
▪ Store in cool, dry
and ventilated
storage area.
▪ Avoid severe
temperature changes
which cause the
material to “set up”.

Penicillin G ▪ Classified as harmful ▪ Keep away from strong

(Source: Pfizer) substance. oxidizers
▪ Unreactive and ▪ Use safety glasses
stable under or goggles.
normal condition. ▪ Always use gloves
▪ It may cause allergic under gauntlet type
symptoms or Nitriles rubber gloves
breathing difficulties and wear protective
if inhaled. clothing when
▪ Fine particles such as working with large
dust and mists quantities.
potentially cause ▪ Always wear an
explosions. appropriate respirator
with a protection factor
sufficient if airborne
exposures are within or
exceed the Occupational
Exposure Band (OEB)
range.
▪ Keep container
tightly close
 xl

4.3 ENVIRONMENTAL ISSUE

4.3.1 Waste Generation

Table 5.2 Waste generation in production of Penicillin G

Type of Waste Stream no. Component Quantity (kg/h)

Solid 18 Biomass
Waste water 11, 16 Liquid waste

Gas 19 CO2

4.3.2 Waste Treatment

First and foremost, waste treatment for carbon dioxide gas is not significant because it is
produced at small amount due to being the waste product of respiratory of Penicillium
chrysogenum. Hence, it is released into the atmosphere due to no significant harm.

Aerobic wastewater treatment is usually known as a secondary treatment plant. Aerobic

treatment system is widely used in the area of wastewater removal. The aerobic treatment uses
oxygen to break down the effluent and remove the different pollutants such as phosphorus and
nitrogen. In this process, oxygen is required to form air. This air is forced via blower or
compressor to mix with the wastewater. It converts the sludge into new biomass. It makes the
wastewater to meet the environmental requirements and clean the water to discharge safely.

Aerobic wastewater treatment is a stand-alone system and by removing the BOD and TSS
from the industrial water it makes the wastewater useful. This treatment can be used specifically
to remove nitrogen as well as phosphorus. This system plays a vital role to clean the wastewater
and maintain the eco-system (SAMCO Technologies 2019).

Use Baking soda (Sodium Bicarbonate) to fully kill the fungi and then use composting
method which is a biological process where organic portion allowed to decompose under
carefully controlled conditions. Then dispose it at sanitary landfill.
 xli

4.3.3 Regulation

It is necessary for all industrial sectors in Malaysia to abide the national law on environmental
acts and regulations. The laws and regulations below have been introduced and are strictly
enforced by the ministry of environment such as Environmental Quality Act (1974) and
Industrial Effluent Regulation (2009).
 xlii

CHAPTER V

MASS BALANCE

5.1 STOICHOIMETRY EQUATION

Stoichiometric equations for biological processes are often complicated and are highly
specific. Hence, the stoichiometric coefficients of each species must be determined
specifically using the methods as shown. Phenylacetic acid is involved in the equation
as structural formula of penicillin G contains a benzene ring and phenylacetic acid
also contains a benzene ring so it plays an important role inside the stochiometric
equation.

C6H12O6 + 1.792O2 + 0.716(NH4)2SO4 + 0.899C8H8O2 → 0.485C16H18N2O4S +

2.89CH1.64O0.52N0.16S0.08 + 5.725H2O + 2.539CO2

Components Molecular formula Molecular weight

(g/mol)
Glucose C6H12O6 180
Nitrogen gas N2 28
Oxygen O2 32
Ammonium sulphate (NH4)2SO4 132
Phenylacetic acid C8H8O2 130
Penicillin G C16H18N2O4S 422
Biomass CH1.64O0.52N0.16S0.08 404
Water H2 O 98
Carbon dioxide CO2 84
The calculation are shown in Appendix A.
 xliii

5.2 MASS BALANCE

Mass balance is used to compare the value of input and output of the system based on
the Law of conservation of mass. It is stated that the mass of a closed system will
remain constant, as the processes occur in the system.

Based on our studies, the plant capacity for our plant covers 10% from the
market shortage which 2,292,000 kg per year. Since the complete fermentation
process is about 3 day per batch and total of 285 working day throughout the year,
therefore the plant is design to produce 24800kg per batch.

List of parameter:

G = Glucose; N = Nitrogen gas; O = Oxygen; A = Ammonium sulphate; H =

Phenylacetic acid; P = Penicillin G; X = Biomass; W = Water; C = Carbon dioxide;

5.2.1 Main Fermenter (R-2)

Figure 4.1 shows the main fermenter, the place where the growth of Penicillium
chrysogenum fungi from the seed fermenter will be continued. Glucose and
ammonium sulphate are supplied into the fermenter as the carbon and nitrogen source.
Large air supplies are required during the main fermentation. The temperature is
carefully controlled for about 25-30 ℃ and pH frequently adjusted 6.5. The
fermentation occurs in batch process for
about 70 hours.
 xliv

Figure 5.1: Main Fermenter, R-2

Stoichiometry equation:

C6H12O6 + 1.792O2 + 0.716(NH4)2SO4 + 0.899C8H8O2 → 0.485C16H18N2O4S +

2.89CH1.64O0.52N0.16S0.08 + 5.725H2O + 2.539CO2

General mass balance formula :

𝐴𝑐𝑐𝑢𝑚𝑢𝑙𝑎𝑡𝑖𝑜𝑛 𝐹𝑙𝑜𝑤 𝑖𝑛 𝑡ℎ𝑟𝑜𝑢𝑔ℎ 𝐹𝑙𝑜𝑤 𝑜𝑢𝑡 𝑡ℎ𝑟𝑜𝑢𝑔ℎ

[ ]=[ ]−[ ]+
𝑤𝑖𝑡ℎ𝑖𝑛 𝑠𝑦𝑠𝑡𝑒𝑚 𝑠𝑦𝑠𝑡𝑒𝑚 𝑏𝑜𝑢𝑛𝑑𝑎𝑟𝑖𝑒𝑠 𝑠𝑦𝑠𝑡𝑒𝑚 𝑏𝑜𝑢𝑛𝑑𝑎𝑟𝑖𝑒𝑠
𝐺𝑒𝑛𝑒𝑟𝑎𝑡𝑖𝑜𝑛 𝑐𝑜𝑛𝑠𝑢𝑚𝑝𝑡𝑖𝑜𝑛
[ ]−[ ]
𝑤𝑖𝑡ℎ𝑖𝑛 𝑡𝑖𝑚𝑒 𝑤𝑖𝑡ℎ𝑖𝑛 𝑡𝑖𝑚𝑒

As the system is in batch process,

𝐴𝑐𝑐𝑢𝑚𝑢𝑙𝑎𝑡𝑖𝑜𝑛 𝐺𝑒𝑛𝑒𝑟𝑎𝑡𝑖𝑜𝑛 𝑐𝑜𝑛𝑠𝑢𝑚𝑝𝑡𝑖𝑜𝑛

[ ]=[ ]−[ ]
𝑤𝑖𝑡ℎ𝑖𝑛 𝑠𝑦𝑠𝑡𝑒𝑚 𝑤𝑖𝑡ℎ𝑖𝑛 𝑡𝑖𝑚𝑒 𝑤𝑖𝑡ℎ𝑖𝑛 𝑡𝑖𝑚𝑒

𝑑𝐶
𝑉 = ±𝑟𝑉
𝑑𝑡

Material balance for cell, X

𝑑𝐶𝑋
= +𝑟𝑋
𝑑𝑡

𝑑𝐶𝑋
= 𝑟𝑋 = 𝜇𝐶𝑋
𝑑𝑡

1
𝑑𝐶 = 𝜇𝐶𝑋 𝑑𝑡
𝐶𝑋 𝑋
 xlv

Integrate both side;

ln (𝐶𝑋 /𝐶𝑋𝑜 ) = 𝜇𝑡

𝐶𝑋 = 𝐶𝑋𝑜 𝑒𝜇𝑡

Material balance for Penicillin G, 𝐶𝑃

𝑑𝐶𝑃
= +𝑟𝑃
𝑑𝑡

𝑟𝑃 = 𝑞𝑃 𝑋

𝐶𝑃
𝑋= + 𝑋𝑜
𝑌𝑃/𝑋

𝑑𝐶𝑃 𝐶𝑃
= 𝑞𝑃 ( + 𝑋𝑜 )
𝑑𝑡 𝑌𝑃
𝑋

Using first order seperable ODE and then integrate both side will give;

𝑒 𝜇𝑡
𝐶𝑃 = − 𝑌𝑃/𝑋 𝑋𝑜
𝜇

Using the same method for Carbon dioxide and water respectively;

𝑒 𝜇𝑡
𝐶𝐶 = − 𝑌𝐶/𝑋 𝑋𝑜
𝜇

𝑒 𝜇𝑡
𝐶𝑊 = − 𝑌𝑊/𝑋 𝑋𝑜
𝜇
 xlvi

Material balance for Glucose, 𝐶𝐺

𝑑𝐶𝐺
= −𝑟𝑃
𝑑𝑡

−𝑟𝑃 = −𝑞𝐺 𝑋

𝐶𝐺0− 𝐶𝐺
𝑋= + 𝑋𝑜
𝑌𝐺/𝑋

𝑑𝐶𝐺 𝐶𝐺0− 𝐶𝐺
= 𝑞𝐺 ( + 𝑋𝑜 )
𝑑𝑡 𝑌𝐺
𝑋

Using first order seperable ODE and then integrate both side will give;

𝑒 𝜇𝑡
𝐶𝐺 = 𝐶𝐺𝑜 + 𝑌𝐺/𝑋 𝑋𝑜 −
𝜇

Using the same method for Oxygen, Ammonium sulphate and Phenylacetic acid;

𝑒 𝜇𝑡
𝐶𝑂 = 𝐶𝑂𝑜 + 𝑌𝑂 𝑋𝑜 −
𝑋 𝜇

𝑒 𝜇𝑡
𝐶𝐴 = 𝐶𝐴𝑜 + 𝑌𝐴/𝑋 𝑋𝑜 −
𝜇

𝑒 𝜇𝑡
𝐶𝐻 = 𝐶𝐻𝑜 + 𝑌𝐻/𝑋 𝑋𝑜 −
𝜇
 xlvii

The fermentor volume, V is 200m3 and the fermentation time is 70 hours by applying
the mathematical model we got the value of each compenent;

Initial Final Intial Final

Component (kg/m3) (kg/m3) (kg) (kg)
Glucose 140 25 28000 5000
Oxygen 178 3 35600 600
Nitrogen 670 670 134000 134000
Ammonium Sulphate 166 10 33200 2000
Phenylacetic acid 160 17 32000 3400
Penicillin G 0 124 0 24800
Biomass 37 150 7400 30000
Water 0 153 0 30600
Carbon dioxide 0 148 0 29600
Total 1351 1300 270200 260000

Table 5.2 Mass of each component in main fermenter

The percentage error value of the calculation is about 4% which consider to be

negligible.

Stream Stream Stream Stream Stream Stream Stream

Component 5 6 7 8 11 12 13

Glucose 0 0 28000 0 0 0 5000

Oxygen 0 0 0 0 35600 600 0
Nitrogen 0 0 0 0 134000 134000 0
Ammonium
Sulphate 0 33200 0 0 0 0 2000
Phenylacetic acid 32000 0 0 0 0 0 3400
Penicillin G 0 0 0 0 0 0 24800
Biomass 0 0 0 7400 0 0 30000
Water 0 0 0 0 0 0 30600
Carbon dioxide 0 0 0 0 0 29600 0

Table 5.3 Mass of component in main fermenter respective to stream line

 xlviii

5.2.2 Centrifugal disc (C-1)

Filtration will be carried out as the bioseparation is required to remove the biomass from
the culture. In this case, centrifugal disc, F-1 will be introduce to remove the solid
biomass from the liquid component which contains the penicillin in the stream 13. The
supernatant in stream 14 will then be transferred further in the downstream process to
continue with the extraction. The solid biomass in stream 27 will be treated and used as
feterlizer for argriculture purpose.

Figure 5.2 Centrifugal Disc

The calculation are shown in Appendix A

Stream Stream Stream

Component 13 14 27
Glucose 5000 5000 0
Oxygen 0 0 0
Nitrogen 0 0 0
Ammonium Sulphate 2000 2000 0
Phenylacetic acid 3400 3400 0
Penicillin G 24800 24800 0
Biomass 30000 0 30000
Water 30600 30600 0
Carbon dioxide 0 0 0

Table 5.3 Mass of component in Centrifugal disc, C-1

 xlix

5.2.3 Extraxtion Column (E-1)

The downstream process of penicillin G production is continue with dissolving the

penicillin from the stream 16 with n-butyl acetate in the extraction column, E-1. The n-
butyl acetate will dissolve the penicillin that present in the filtrate. This extraction process
is a batch process of liquid-liquid extraction. The yield for the extraction is 95.51%,
(Aliwarga et al., 2019). Most of the penicillin-rich solution with butyl acetate will be
found in the upper product, stream 17 while the others in the watery layer in the bottom of
extraction column in stream 30 .

Figure 5.3 Extraction Column, E-1

List of parameters

G = Glucose; A = Ammonium sulphate; H = Phenylacetic acid; P = Penicillin G; W =

Water; C = Carbon dioxide; BA = n-butyl acetate;

𝑉𝑎𝑞 − 𝑞𝑃 𝑉𝑎𝑞
𝑉𝐵𝐴 =
𝑞𝑃 𝑘𝐷

Where,
VBA = Volume of n-butyl acetate kD = Partion Coefficient

Vaq = Volume of aqueous solution

qP = Fraction of Penicillin remain in aqueous solution

Mass balance for extraction column, E-1.

 l

𝑚
Volume,, 𝑉𝑇 = ∑ 𝜌

𝑡𝑜𝑡𝑎𝑙 𝑚𝑎𝑠𝑠 𝑡𝑜𝑡𝑎𝑙 𝑚𝑎𝑠𝑠 (𝑑𝑒𝑛𝑠𝑖𝑡𝑦 𝑜𝑓 𝑠𝑜𝑙𝑣𝑒𝑛𝑡)

Density, 𝜌𝑇 = =
𝑡𝑜𝑡𝑎𝑙 𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 𝑚𝑎𝑠𝑠 𝑜𝑓 𝑠𝑜𝑙𝑣𝑒𝑛𝑡

From the calculation in Appendix A, the amount needed for extracting the penicillin G
with 95.51% efficiency is 55311 kg of n-butyl acetate.

Stream Stream Stream Stream

Component 4 16 17 30
Glucose 0 5000 0 5000
Ammonium Sulphate 0 2000 0 2000
Phenylacetic acid 0 3400 0 3400
Penicillin G 0 24800 23686 1114
Water 0 30600 0 30600
n-butyl acetate 55311 0 55311 0

Table 5.4 Mass of component in extraction column, E-1

5.2.4 Regeneration column (E-2)

he recovery of penicillin G is continue in the regeneration column, E-2. Additional of

water and phosphate buffer will bring the penicillin-rich solution with butyl acetate
from the stream 17 into an alkaline condition, pH 7 where the penicillin stabilitiy is
high (Antibiotics-Exhibition 2016 Penicillin, n.d.). This condition will bring the
penicilling G into the bottom product of E-2 in stream 18 . This extraction method is
know as solvent transfer method (Antibiotics-Exhibition 2016 Penicillin, n.d.) The
addition of water will usually take one fifth of the volume of the original broth and the
addition of phoshpate buffer will usually take one-tenth of the volume of the broth
(Gordon et al., n.d.).
 li

Figure 5.4 Regeneration Column, E-2

List of parameter:

P = Penicillin; W = Water ; BA = n-butyl acetate ; PB = Phosphate buffer;

Mass balance on Regeneration Column:

Mass in = Mass out

Volume of penicillin-rich solution with butyl acetate in stream 18

𝑉𝑇 = 𝑉𝐵𝐴 + 𝑉𝑃

𝑉𝑇 = 𝑚𝐵𝐴 /𝜌𝐵𝐴 + 𝑚𝑃 /𝜌𝑃

From the calculation in Appendix A the mass of water and phosphate buffer needed is
1723kg and 8597kg respectively.
 lii

Stream Stream Stream Stream Stream

Component 2 3 17 18 28
Penicillin G 0 0 23686 23686 0
n-butyl acetate 0 0 55311 0 55311
Water 1723 0 0 1723 0
Phosphate buffer 0 8597 0 8597 0

Table 5.5 Mass of component in extraction column, E-2

5.2.5 Continuous Stir Tank Reactor (R-3)

The bottom product of extraction column, E-2 in stream 19 will enter the CSTR, R-3
and will react with the sodium bicarbonate to form penicillin salt which allow the
penicillin to be stored in a stable powder form at a room temperature.

Figure 5.5 CSTR, R-3

The chemical reaction are given:

C16H18N2O4S + NaHCO₃ → C16H17N2NaO4S + CO2 + H2O

P SB PS C W
 liii

List of parameter:

P = Penicillin; SB = Sodium bicarbonate PS = Penicillin salt; C = Carbon dioxide;

H = Water

Mass balance in mixer:

Assume the Penicillin is completely reacted with Sodium bicarbonate.

𝑚𝑖 (𝑘𝑔)
Number of mole of species i (kmol), 𝑛𝑖 = 𝑘𝑔
𝑀𝑊𝑖 ( )
𝑘𝑚𝑜𝑙

Number of mole of penicillin, nP = 23686/334 = 70.92 kmol

From stoichiometry ratio, 1 mole of Penicillin will react completely with 1 mole of
Sodium bicarbonate and will produce 1 mole of Penicillin will produce 1 mole of
Penicillin salt, Carbon dioxide and water. Therefore, the mole needed for sodium
bicarbonate to react completely with penicillin is 70.92 kmol and the number of mole
of Penicillin salt, Carbon dioxide and water produce is 70.92 kmol.

Mass of species i (kg), 𝑚𝑖 = 𝑛𝑖 (𝑀𝑊𝑖 )

nSB = 70.92(84) = 5957kg of Sodium bicarbonate

nPS = 70.92(356.4) = 25276kg of Penicillin salt

nC = 70.92(44) = 3120kg of Carbon dioxide

nW = 70.92(18) = 1276kg of Water

 liv

Stream Stream Stream Stream

Component 1 19 20 29
Penicillin G 0 23686 0 0
Water 0 1723 2999 0
Phosphate buffer 0 8597 8597 0
Sodium bicarbonate 5957 0 0 0
Penicillin salt 0 0 25276 0
Carbon dioxide 0 0 0 3120

Table 5.5 Mass of each component in CSTR, R-3

5.2.6 Centrifugal basket (C-2)

The product from the CSTR in stream 20 will enter the centrifugal basket, C-2. This
allow the penicillin salt to be extracted. The penicillin salt will accumulate and
compress on the C-2 wall as effect of the centrifugal force. The applied centrifugal
force will remove liquid spontaneously from the filter cake (Solid-Liquid Separation,
n.d.). The efficiency of centrifugal disc is 95% (Abbas et al,. 2016). The solid will be
scrap and enter the stream 21 while the liquid waste will enter the stream 26.

Figure 5.6 Centrifugal basket, E-2

Mass balance in centrifugal basket, C-2

𝑚𝑎𝑠𝑠 𝑜𝑢𝑡 𝑚
Efficiency, η = = 𝑚𝑖
𝑀𝑎𝑠𝑠 𝑖𝑛 𝑖𝑜
 lv

η = 0.95 (Abbas et al,. 2016).

The calculation are shown in the appendix

Stream 20 Stream 21 Stream 26

Component (kg) (kg) (kg)

Water 2999 150 2849

Phosphate buffer 8597 430 8167
Penicillin salt 25276 24013 1263

Table 5.5 Mass of each component in centrifugal basket, C-2

5.2.7 Freeze drying crystallizer (F-1)

The product cake from the centrifugal basket, C-2 in stream 21 will enter the freeze
drying crystallizer, F-1. F-1 will remove excess moisture content in the product cake
by turning the moisture into frozen state and turning it into gas under sublimation
phase in a very low pressure. The waste gas will exit F-1 as the upper product in
stream 24 while the solid material in bottom product in stream 25. This process will
yield final residual water content in the product in a very low range of 0.1 to 0.4%
(Lorenz et al., 2007). Therefore, the purity of the product are estimate to be 99.8%
pure.

Figure 5.7 Freeze drying crystallizer, F-1

 lvi

Stream 23 Stream 24 Stream 25

Component (kg) (kg) (kg)

Water 150 0 50

Phosphate buffer 430 430 0

Penicillin salt 24013 0 24013

The calculation are shown in the appendix

Table 5.6 Mass of each component in centrifugal basket, C-2

5.2.8 MANUAL AND SUPERPRO® CALCULATION COMPARISON

The results between the calculation made by manual calculation and the calculation
using Superpro® is being compared and shown in table 4.7. Output of Superpro®
calculation will be shown in Appendix D.

Figure 4.8 Fermenter drawn using Superpro®

Table 4.7: Comparison between manual calculation and Superpro® calculation

Overall process Manual calculation Superpro® Percentage of Error

calculation
Mass flowrate in 270200 270200 0%

Mass flowrate out 260000 260000 0%

 lvii

The value of the calculation made using Superpro® with manual calculation is
different due to the significant number being used during calculation. Another reason
can be the amount of oxygen used is different. In manual calculation, the amount of
oxygen is fixed while in Superpro®, the amount of oxygen cannot be fixe and it
changes according to the needs of the fermentation process. The error can be reduced
by adjusting the concentration of oxygen in manual calculation to higher
concentration.

5.3 ENERGY BALANCE

Energy balance is the relationship between energy in and energy out. The
First Law of Thermodynamics states that energy can neither be destroyed
nor created but can be converted from one from to another. Energy balance
is applied in order to calculate the energy that is required in the system
and the energy released or absorbed during the process. If heat is being
released to the surrounding, hence it is exothermic reaction and if heat is
being absorbed during the reaction, then it is an endothermic reaction. The
heat capacity for each of the components that are involved in the reaction
are shown in table 4.10 below.

Table 4.10 Heat Capacity for each Component

Component Cp = A + BT + CT2 + DT3 + ET4

A B C D

Oxygen 29.1 0.01158 -6.076 x 10-6 1.311 x 10-8

Nitrogen 31.15 -1.357 x 10-2 2.680 x 10-5 -1.168 x 10-8

Ammonium Sulphate 27.31 0.02383 1.707 x 10-5 -1.185 x 10-8

Water 32.24 0.01924 1.055 x 10-5 -3.596 x 10-9

Carbon Dioxide 19.80 0.07344 -5.602 x 10-5 1.7115 x 10-8

 lviii

Source: Appendix A (Thermal-Hydraulic Analysis of Nuclear Reactors)

Since Cp values of glucose, biomass, phenylacetic acid and penicillin g is

specific, it values of Cp is determined by determining Cp values of each
element. Table 4.11 shows the heat capacity for each element.
 lix

Table 4.11 Heat Capacity for each Element

Element Cp (kJ/kmol.K)

C 7.5

H 9.6

O 17.0

N 26.0

S 26.0

Source: Felder, 2005

Glucose, C6H12O6

Element Cp (kJ/kmol.K)

C 6 x 7.5 = 45.0

H 12 x 9.6 = 115.2

O 6 x 17.0 = 102.0

Total 262.2
 lx

Phenylacetic acid, C8H8O2

Element Cp (kJ/kmol.K)

C 8 x 7.5 = 60.0

H 8 x 9.6 = 76.8

O 2 x 17.0 = 34.0

Total 170.8

Penicillin G

Element Cp (kJ/kmol.K)

C 16 x 7.5 = 120

H 18 x 9.6 = 172.8

O 4 x 17.0 = 68.0

N 2 x 26.0 = 52.0

S 1 x 26.0 = 26.0

Total 438.8

Biomass

Element Cp (kJ/kmol.K)

C 1 x 7.5 = 7.5

H 1.64 x 9.6 = 15.744

O 0.52 x 17.0 = 8.84

N 0.16 x 26.0 = 4.16

S 0.08 x 26.0 = 2.08

Total 38.324
 lxi

5.3.1 Batch Fermenter (R-2)

N19E=0.3406kmol/h

N19J=2.5317kmol/h
N19D=8.1078kmol/h
N2=0.8322kmol/h
N3=0.6632kmol/h
N4=0.926kmol/h
N8=7.5238kmol/h
N11F=0.4491kmol/h
N11G=2.9437kmol/h
N11H=5.5651kmol/h

Figure 4.11 Fermenter R-2

Assumptions:

1. The process is in steady state and mass flow into or out of the
fermenter iscontinuous.
2. The fermenter’s initial temperature is at 298.15 K and the final
temperatureafter the fermentation is 299.15 K.
 lxii

Glucose (Temperature is from T1 = 298.15K to T2 = 299.15K)

299.15

^G =
∆H ∫ 262.2 dT
298.15

= 262.2 kJ/kmol

Oxygen (Temperature is from T1 = 298.15K to T2 = 299.15K)

299.15

^O = ∫
∆H 29.1 + 0.01158 − 6.076 × 10−62 + 1.311 × 10−83 dT
298.15

= 32.3657 kJ/kmol

Nitrogen (Temperature is from T1 = 298.15K to T2 = 299.15K)

299.15

^N = ∫
∆H 31.15 − 1.357 × 10−2 + 2.680 × 10−52 − 1.168 × 10−83
298.15
dT

= 29.1765 kJ/kmol

Phenylacetic acid (Temperature is from T1 = 298.15K to T2 = 299.15K)

299.15

^H
∆H =∫ 170.8 dT
298.15

= 170.8 kJ/kmol
 lxiii

Ammonium Sulphate (Temperature is from T1 = 298.15K to T2 = 299.15K)

299.15

^A = ∫
∆H 27.31 + 0.02383 + 1.707 × 10−52 − 1.185 × 10−83 dT
298.15

= 35.6337 kJ/kmol

Biomass (Temperature is from T1 = 298.15K to T2 = 299.15K)

299.15

^x
∆H ∫
= 298.15 38.324 dT

= 38.324 kJ/kmol

Carbon Dioxide (Temperature is from T1 = 298.15K to T2 = 299.15K)

299.15

^C = ∫
∆H 19.80 + 0.07344 − 5.602 × 10−52 + 1.7115 × 10−83 dT
298.15

= 37.1922 kJ/kmol

Water (Temperature is from T1 = 298.15K to T2 = 299.15K)

299.15

^W = ∫
∆H 32.24 + 0.01924 + 1.055 × 10−52 − 3.596 × 10−93 dT
298.15

= 38.8312 kJ/kmol

Penicillin G (Temperature is from T1 = 298.15K to T2 = 299.15K)

 lxiv

299.15

^P
∆H ∫
= 298.15 438.8 dT

= 438.8 kJ/kmol

Hin ^ G(nGo) + ∆H
= ∆H ^ O(nOo) + ∆H
^ N(nNo) + ∆H
^ A(nAo) + ∆H
^ H(nHo) +
^ X(nXo)
∆H

= (262.2)( 155.42) + (32.3657)( 1112.5) + (29.1765)( 4785.71) +

(35.6337)( 251.25) + 170.8(235.0349) + 38.324(276.5321)

= 276,082.98 kJ/batch

Hout = ∆H^ G(nG) + ∆H^ O(nO) + ∆H^ N(nN) + ∆H

^ A(nA) + ∆H
^ H(nH) + ∆H
^ X(nX)
^ P(nP) + ∆H
+∆H ^ W(nW) + ∆H^ C(nC)

= (262.2)(27.75) + (32.3657)( 18.75) + (29.1765)( 4785.714) +

(35.6337)( 15.14) + 170.8(24.97) + (438.8)(74.16) +
(38.324)(11121.07) + (37.1922)( 672.73) + (38.8312)(1700)

= 702,096.305 kJ/batch

Q ^ out – H
=H ^ in + r1ΔHrxn

= 702,096.305 - 276,082.98 + (47.85)( -762,473.3196)

 lxv

= - 36,058,335 kJ/batch

Since the heat is negative, the production of Penicillin G in the

reactor is an exothermic reaction where heat is being released to the
surrounding.
 lxvi

CHAPTER VI

BIOCHEMISTRY AND BIOMOLECULAR ENGINEERING

6.1 MAIN ENZYME USED IN PRODUCTION

Penicillins are a group of antibiotics originally obtained from Penicillium moulds,

most penicillins in clinical use are chemically synthesised from naturally-produced
penicillins. Penicillins were among the first medications to be effective against many
bacterial infections caused by staphylococci and streptococci. One of the techniques
used to produce penicilins is by multiplying the enzyme, the enzyme used in the
production of penicilin is the enzyme Penicilin Acylase.

Penicillin acylase enzymes (EC 3.5.1.11) are of considerable industrial importance,

primarily in the production of 6-aminopenicillanic acid (6-APA) as an intermediate in
the manufacture of semi-synthetic penicillins. Industrially, the penicillin G acylase
(PGA) from Escherichia coli is the enzyme of choice, whether recombinant or native.
Although the optimum temperature for the hydrolysis of penicillin G is 50°C, the
enzyme loses stability above 30°C and must be used in immobilized form. Other
described PGAs with higher stability are those from Alcaligenes faecalis (AfaePGA,
t1/2 of 15 min at 55°C), Bacillus badius (t1/2 of 20 min at 55°C) and Achromobacter
xylosoxidans (t1/2 of 55 min at 55°C). PA-G gene encodes a precursor polypeptide
which consists of 4 structural elements: a signal peptide, α- and β-subunits, and an
inter-subunit spacer.
 lxvii

The mature PA-G molecule is a heterodimer with a molecular weight of 86 kDa. It

consists of two subunits, α- and β-, with molecular masses of 23 and 63 kDa,
respectively. In addition, the molecule contains a bound Ca 2+ ion, which,

6.2 METABOLIC PATHWAY

Figure 6.1: . Penicilin Acylase Metabolic

These metabolism process have been critically important in penicillin acylase use, as
they are implicated in the potentially penicillin allergy. The four-membered β-lactam
ring is present in all penicillins, and was identified in several other classes of
antibiotics since the discovery of penicillin, including cephalosporins, monobactams,
and carbapenems. The β-lactam function is the key to the lethality of these antibiotics.
Despite the efficacy of penicillin G and β-lactam antibiotics, there have been concerns
over their stability. The four-membered β-lactam function exists under a tremendous
amount of ring strain, which is the fundamental cause of its molecular instability and
susceptibility to undergo ring opening. Importantly, this lack of stability contributes
greatly to β-lactam’s reactivity and therefore, promotes antibiotic activity.

The addition of the thiazolidine ring may be essential for functionality of the
β-lactam ring because the fusion of these two features causes torsional rotation of the
molecule, resulting in its non-planar structure, and contributes to large bond angles
within the β-lactam ring. Thus, the already strained β-lactam ring is even more prone
 lxviii

to cleavage when bound to thiazolidine. The β-lactam ring is highly susceptible to

hydrolytic opening under a wide range of conditions including auto-catalysis under
neutral pH, and in the presence of acids, bases, heat, UV light, and enzymatic activity.
Due to stability issues in aqueous media, penicillin G should be kept in buffered
solutions such as phosphate, acetate, or citrate, of which citrate buffer at pH 7 is the
most effective at preventing degradation. Penicillin G is most stable in a temperature
range between 0°C to 52°C, above which it will rapidly degrade.

6.3 EXPRESSION OF PENICILIN ACYLASE ENZYME

Production and maturation for Penicillin acylase (PAC) by generated antibodies

against the α- and β-subunits of the PAC codified in the Thermus thermophilus
(Tth) HB27 genome, separately produced in E. coli cells. he apparent electrophoretic
mobilities of these protein subunits are 22 and 60 kDa for the α- and β-subunits,
respectively.

Figure 2. Presence of PAC protein in Tth cells.

In order to confirm the correct maturation of Tth pro-PAC in E. coli, the

recombinant protein was purified by IMAC chromatography (Figure 3). SDS-PAGE
of the purified protein revealed two subunits of Tth PAC with relative molecular
masses (Mr) of 26.4 ± 1.0 kDa (α-subunit) and 54.3 ± 1.0 kDa (β-subunit). MALDI-
TOF and LC-MS/MS analyses were performed on each subunit. As expected, the N-
terminal residue of the β-subunit corresponded to the catalytic Ser256. The C-terminus
 lxix

of the β-subunit was evident from the stop codon, whereas, the C-terminus of the α-
subunit depends on the length of the linker peptide cleaved from the pro-Tth PAC.

Figure 3. Tth PAC purification.

However, During cell lysis these proteases are normally activated, being
probably responsible for the reduced size of the α-subunit observed in Tth cells.
Since Tth PAC has a catalytic serine it is not possible to use serine-protease inhibitors
during our protein purification protocols. Tth proteases, as most proteases from
extremophilic bacteria, are serine proteases that are stable at high temperatures even in
the presence of high concentrations of detergents and denaturing agents. Thus, the
actual molecular weight of the α-subunit cannot be deduced from the α-PAC
processed in E. coli nor from the one matured in Tth cells.
 lxx

CHAPTER VII

SEPARATION PROCESS II

7.1 INTRODUCTION

The centrifuge remains one of the most widely used unit operations for obtaining
efficient product harvesting in the production of Penicillin G from penicillium
chrysogenum. From our process flow diagram, the fermenter output will enter the
centrifugal stack disc to filter out the solid biomass. Centrifugal forces cause the
denser solid biomass to flow outwards towards the rotating bowl wall, while the less
dense fluids move towards the centre, separating the biomass from the liquid broth.

7.2 DESIGN OF CENTRIFUGAL STACK DISC

Centrifugal stack disc is able to apply a force from 4000 to 14000 times gravitational
force, thus reducing separation time. By using this centrifuge, the separation process
can be improved by distributing the flow to a number of parallel narrow channels
between conical dishes (Amaro et al., 2017).

This centrifuge's design is based on the specifications and performance

parameters outlined in Table 7.1.
 lxxi

Table 7.1: Specification and performance characteristics of centrifugal stack disc bowl
Bowl D, (mm) Speed (rpm) Maximum centrifugal Throughput/Liquid(L/min)
force x gravity
254 10000 10 000 40-150
406 6250 6250 100-570
686 4200 4200 150-1500
762 3300 3300 150-1500

Source: Bioseparations Science and Engineering, 2015

Table 7.2: Overall design parameter of tubular bowl centrifuge

Parameters Value

Volumetric flowrate, Q 29.213m3/min

Length of centrifuge, L 1m

Speed of the centrifuge, ω 6250 rev/min

Radius of outlet opening of the bowl, r 1 0.205m

Radius of bowl, r0 0.203m

7.3 MASS BALANCE AT CENTRIFUGAL STACK DISC

The mass for all components in the centrifugal stack disc is calculated as follows:

Table 7.3: Mass component of Centrifugal stack disc

 lxxii

Stream Stream Stream

Component 13 14 27
Glucose 5000 5000 0
Oxygen 0 0 0
Nitrogen 0 0 0
Ammonium Sulphate 2000 2000 0
Phenylacetic acid 3400 3400 0
Penicillin G 24800 24800 0
Biomass 30000 0 30000
Water 30600 30600 0
Carbon dioxide 0 0 0
 lxxiii

CHAPTER VIII

UTILITY AND PRESSURE VESSEL DESIGN

8.1 INTRODUCTION

The utility design and the mechanical design of the pressure vessel are two significant
components. The first portion of the presentation will focus on the design of utility
systems used in chemical and biochemical engineering, such as coolers. The second
section focuses on the basic mechanical design of pressure vessels for industrial
operations and utility units. Its purpose is to explain the mechanical design and
construction of pressure vessels in accordance with the ASME (American Society of
Mechanical Engineers) Code.

8.2 UTILITY DESIGN

In utility design, there are one unit operation that are highlighted:

a) Cooler, H-2

8.2.1 DESIGN OF COOLER (H-1)

The primary function of the cooler is to cool the mixture produced by the main
fermenter, R-2 before it enters the storage tank. Cooling water serves as both the
mixture and the cold fluid in this cooler. Because the coolant is corrosive, the coolant
is introduced through the tube, and the hot fluid combination is introduced into the
shell to save money on insulation. Below is the figure 8.2 shows the design of cooler,
H-2.
 lxxiv

Figure 8.1 Cooler, H-2

Table 8.1 Pressure drops and coefficients

Type of Heat Tube-side Shell-side Overall Tube-side Shell-side

Utilities Load, Q coefficient, hi coefficient, hs coefficient, Pressure Pressure
(kW) (W/m2.℃) (W/m2.℃) Uo drop, ∆Pt drop, ∆Ps
(W/m2.℃) (N/m2) (N/m2)
Cooler 48.244 10551.33 4725.63 909.09 2.233 x 104 8.651 x 103
*Refer to Appendix B for detailed calculation of each utility.

8.3 PRESSURE VESSEL

A pressure vessel is a closed container designed to hold gasses or liquids at a

substantially different pressure from the ambient. There are two main procedures for
the design of the vessels according to the ASME Pressure Vessel Design Code,
namely external and internal procedures.

The mechanical designs of two pressure vessels are shown. The designs are
subject respectively to internal and external procedures (Coulson & Richardson's,
2005).

8.3.1 VESSEL DESIGN UNDER INTERNAL PRESSURE

The main fermenter, R-2 are operating in batch operation where the temperature, pH
strictly control. The temperature and pH are control and maintain at 25℃ and 5 - 7
respectively (Gordon et al., n.d.). Besides, the regulation of dissolve oxygen is vital in
this fermentation as the microbes are highly aerobic fungus. The concentration of
dissolved oxygen will set to be close to saturation. However, with an increasing of
biomass with respect to time, the fermentation broth will become more viscous, and
the dissolve oxygen level will decrease rapidly. Therefore, the batch fermentation will
operate with air flow rate of 12.11 kg/h with a total feed of 848 kg of air and the
dissolve oxygen value are maintain more than 85% throughout the course of the
fermentation by increasing agitator speed in a range of 500 to 700 rpm (Goudar &
Strevett, n.d.).
 lxxv

Hence, stainless steel grade 316 which contain 18% chromium,10% nickel and
2.5% molybdenum are used for the fermenter in the production of Penicilin G. It is the
most commonly used type of fermenter for industrial operation. The molybdenum
content increase corrosion resistance and increase strength at high temperature. It is a
more susceptible to weld decay and they are particularly effective in acidic
environments which normally occurred during the accumulation of biowaste. Stainless
steel has high wear resistance, smooth surface, therefore it is ease for cleaning process
to avoid any adhere and growth of germs hence it is selected as the material to be used
for main fermenter (R K Sinnott 2007).

8.3.2 DESIGN SPECIFICATION OF FERMENTER, R-2

The schematic diagram of fermenter is illustrated as shown in figure 8.2 and the
design specification of fermenter is shown in table 8.2.

The fermenter is designed based on the height-to-diameter ratio of 2:1 because

the mixing in cell culture is gentle. A vertical vessel with 2:1 ellipsoidal head as the
top head and bottom. The material used is stainless steel grade 316 and the corrosion
allowance is set to be 4mm because the condition of fermentation will turn acidic after
fermentation process took place and waste start to accumulate and the cleaning
process require high temperature.

Table 8.2 Operating condition of Fermenter, R-2

Condition Operating selection and condition

Operating pressure, Po 2 bar
Temperature 25℃
 lxxvi

Material selection Stainless steel grade 316

Corrosion allowance, CA 4 mm
Joint efficiency, E 0.85
H:D 2:1

Table 8.3 Dimension of Fermenter, R-2

Part Type
Top head Ellipsoidal 2:1
Shell vessel Cylindrical
Bottom head Ellipsoidal 2:1

Table 8.4 Inlet component of Fermenter, R-2

Components Mass flowrate (kg) Density (kg/m3)

Glucose 28000 1560
Oxygen 35600 1429
Nitrogen 134000 1160
Ammonium Sulphate 33200 1770
Phenylacetic Acid 32000 1080
Biomass 7400 1410
Total 270200 8409
Source: Mass Balance

Table 8.5 Summary of Vessel Specification Parameter

Specification Value (ft) Value (inch) Value (mm)

Vessel height, h 33.34 400 10160
Vessel inside Diameter, Di 16.67 200 5080
Cylindrical Shell Height, hc 25 300 7620
Ellipsoidal top head height, he 4.17 50.04 1271
Ellipsoidal bottom head height, he 4.17 50.04 1271
*Refer Appendix B (2.0) for detailed calculation.

Ellipsoidal head is used on both of the top and bottom of the reactor because it is easy
to handle and maintain.
 lxxvii

8.3.3 DESIGN PRESSURE

Since the operating pressure of the reactor (2 bar) is greater than atmospheric pressure
(Po > Patm), the reactor designed under internal pressure.

Table 8.6 Design Pressure of Each Part

Type of part Operating Pressure, PD Hydrostatic Pressure, PJ

(psi) (psi)
Ellipsoidal 2:1 top head 30.82 33.90
Ellipsoidal 2:1 bottom head 43.45 47.80
Cylindrical shell 41.64 45.80
*Refer to Appendix B (2.1) for design pressure calculations.

8.3.4 MINIMUM WALL THICKNESS

The wall thickness formulas for each part of the column are from ASME Code UG-32
part (D) and UG-27 part (c) (AN INTERNATIONAL CODE ASME BOILER &
PRESSURE VESSEL CODE, 2010). *Refer to Appendix B (2.2) for minimum wall
thickness calculations.

8.3.5 MAXIMUM ALLOWABLE WORKING PRESSURE, MAWPvessel

The maximum permissible working pressure for any type of pressure vessel, MAWP,
is calculated by the measurement of all components under internal pressure. The
formula of internal design pressure for each part of the column can be found in ASME
Code UG-32 part (D) and UG-27 part (c) (AN INTERNATIONAL CODE ASME
BOILER & PRESSURE VESSEL CODE, 2010). The MAWPvessel is calculated.
*Refer to Appendix B (2.3) MAWPvessel for calculations.

Table 8.7 Value of MAWPvessel

Part of the column MAWPpart (psi) Static head, PH (psi) MAWPvessel (psi)
Ellipsoidal Top Head 30.764 1.81 28.954
Ellipsoidal Bottom Head 43.509 14.44 29.069
Circumferential 41.589 12.63 28.959
Longitudinal 41.500 12.63 28.870
 lxxviii

8.3.6 THICKNESS UNIFORMITY AND NOMINAL THICKNESS CORRECTION

OF FERMENTER. R-2

From the values of tmin calculated previously in Appendix B, a thickness of 0.245 inch
was chosen to be the uniform wall thickness for the whole vessel. Among a thickness
of 0.405, 0.54, 0.675 and 0.84 inch (Continental chemical USA 2020), a nominal
thickness of 0.405 inch was selected as it is closest to the uniform thickness.

Nominal thickness (inch) 0.405 0.54 0.675 0.84

Nominal thickness (mm) 10.287 13.716 17.145 21.336

By using the nominal thickness of 0.405 inch, the new minimum thickness can be
calculated by using equation:

tmin, new = tnominal – CA

= 0.405 – 0.1575

= 0.248 inch

8.3.7 COMBINED LOADING ANALYSIS

Table 8.9 Analysis of Primary Stress for Fermenter, R-2

Primary Stress Value (N/mm2)

Longitudinal Stress 40.317
Circumferential Stress 80.635
Direct Stress 0.00125
Bending Stress 0
*Refer to Appendix B (2.4) for detailed calculation on primary stress.
 lxxix

8.3.8 ANALYSIS OF ELASTIC STABILITY

From the analysis of the combined loading, the material chosen met both the
maximum stress intensity and the elastic stability requirements:

(Δ𝜎)𝑚𝑎𝑥 ≤ 𝑆 𝑎𝑛𝑑 (Δ𝜎)𝑐𝑜𝑚𝑝𝑟𝑒𝑠𝑠𝑖𝑣𝑒 ≤ 𝜎𝑐

Therefore, the design is safe.

*Refer to Appendix B (2.5) for detailed calculation on Analysis of Elastic Stability.

8.3.9 VESSEL SUPPORT ANALYSIS

Conical cylindrical skirt is chosen as the vessel support for fermenter R-2. Cylindrical
skirt support is suitable for tall vessels rather than other support. The skirt support is
weld at the base of the shell. Material used to fabricate support must be high in
strength, which is stainless steel (SA-240) with nominal composition 18Cr-2Mo. As
the material fulfil minimum content to avoid corrosion, which is 12% chromium, and
contain molybdenum that can improve corrosion resistance. *Refer Appendix B (2.6)
for detailed calculation of the skirt thickness.

8.3.9 BASE RING AND ANCHOR BOLT DESIGN

Table 8.10 Base Ring and Anchor Bolt Design specification for Fermenter, R-2

Parameter Value
Bolt spacing, mm 628.218
Bending moment at the base (Ms), N mm 66064
Fb, N m-1 4059.10
Lb, mm 1.16
Base ring thickness (tb), mm 1.93
*Refer Appendix B (2.7) for detailed calculation.
 lxxx

8.3.10 DESIGN OF EXTRACTION COLUMN, (E-1)

Extraction is a process in which one or more components are separated selectively

from a liquid or solid mixture. The extraction will operate in a batch process of liquid-
liquid extraction method. The feed contains n-butyl acetate, glucose, ammonium
sulphate, phenylacetic acid, penicillin G and water. The purpose of using extraction
column is to dissolve the penicillin G with n-butyl acetate.

4 17

16

30

Figure 8.3: Extraction column, E-1

8.3.11 DESIGN SPECIFICATION OF EXTRACTION COLUMN, E-1

Tables 8.11 and 8.12 summarise the vessel specifications and table 8.13 is for the
operating conditions. The top of the extraction column has a 2:1 ellipsoidal head and
the bottom has a hemispherical head, while the shell is cylindrical. It is assumed that
the cylindrical shell is a thin shell. *Refer appendix (2.8) for calculation height of each
part

Table 8.11 Design shape of Extraction column, E-1

Part Shape

Top Ellipsoidal

Shell Cylindrical

Bottom Hemispherical
 lxxxi

Table 8.12 Design specification of E-1

Design specification Information

Operation condition, kPa and °C Pressure =170kpa, T=150°C

Construction Material Carbon steel (SA-285Gr. C)

Type of used tray Sieve tray

Column Height, m 16.46

Column Diameter, m 10.61

Column Radius, m 5.31

Top head height, m 2.65

Shell height, m 8.5

Bottom head height, m 5.31

Table 8.13 Operating conditions of E-1

Operating Condition Values Remark

Operating pressure, Po 170kPa Ensures optimal condition

Operating pressure safety 10% -

factor

Operating temperature, To 150°C Ensures optimal condition

Material Carbon steel(SA-285Gr.C) Low cost and mild operating

Join Efficiency, E 0.85 Spot radiography is chosen as

inspection to reduce cost

Max allowable stress,S 15700 Value of S for Type 316SS in

compliance to ASME Code

Overcome any damage to

Corrosion Allowance, CA 4mm equipment due to high rate

of corrosion
 lxxxii

8.3.12 DESIGN PRESSURE, MINIMUM WALL THICKNESS, MAWP OF

EXTRACTION COLUMN, E-1

Table 8.14 is the summarizes of the calculated values of design pressure, PD minimum
wall thickness, tmin and the maximum allowable working pressure of the vessel.*Refer
appendix (2.9) for calculation

Table 8.14 MAWPvessel Calculation summary

Part Shape Design tmin MAWPpart Pressure MAWPvessel

Pressure, (inch) (including CA at top head (psi)

PD of 4mm),(psi) (psi)

Top 2:1 Ellipsoidal 28.43 0.4450 28.43

Head

Shell Cylindrical 61.40 0.9645 61.40

(Circumferential) 28.43 28.43

Cylindrical 61.40 0.9601 122.80

(Longitudinal)

Bottom Hemispherical 32.20 0.2523 32.20

8.3.13 THICKNESS UNIFORMITY AND NOMINAL THICKNESS

CORRECTION OF EXTRACTION COLUMN, E-1

From the values of tmin calculated at 8.3.12, a thickness 0.9645in was chosen as the
uniform wall thickness. A nominal thickness compared to the uniformed thickness. A
table of nominal thicknesses of Carbon Steel (SA-285 Gr. C) is attached in the
appendix. The new MAWPvessel calculation were summarized in Table 8.15.
Table 8.15 Summary of thickness uniformity and nominal thickness correction
 lxxxiii

Part Shape t min,new MAWPpart Pressure at top head MAWP vessel

(psi) (psi) (psi)

Top 2:1 Ellipsoidal 0.8268 52.80 52.65 52.65

Shell Cylindrical 52.65

(Circumferential)

Cylindrical 105.72

(Longitudinal)

Bottom Hemispherical 105.47

Therefore, the MAWPvessel of E-1 is at 52.65 psi with minimum thickness of 0.8268in
based on internel design. Refer appendix for further calculation. *Refer appendix
(2.10) for calculation of thickness uniformity and nominal thickness correction.

8.3.14 COMBINE LOADING ANALYSIS FOR EXTRACTION COLUMN, E-1

Table 8.16 show the parameter use in analysis of primary stress.

Table 8.16 Parameter in analysis of primary stress

Parameter Value

Longitudinal stress, (σL) 45.47N/mm2

Circumferential stress, (σH) 90.90N/mm2

Direct stress, (σW) 0.0021N/mm2

Bending stress, (σb) 1.98N/mm2

The design operating condition is safe since (Δσ)max ≤ Design stress, s which is
45.47N/mm2. *Refer appendix (2.11) for the calculation of the parameter.

8.3.15 ANALYSIS OF ELASTIC STABILITY OF EXTRACTION COLUMN

The vessel may fail due to elastic instability (buckling) if the resultant axial stress, σz
 lxxxiv

from combined loads is compressive. For steels at E=200000 N/mm2 and with a safety
factor of 12 the buckling equation is as follow, where σc is critical buckling stress.

𝑡 21
𝜎𝑐 = 2𝑥104 (𝐷 ) = 2𝑥104 (10631.1) = 39.51𝑁𝑚𝑚−2
𝑜

σcompressive = σb + σw = 0.0021 + 1.98 = 1.9821 N/mm2

Thus,

σcompressive ≤ σc

The compressive stresses, σc is bellow critical buckling stress, therefore the design is
safe.

8.3.16 SKIRT THICKNESS OF EXTRACTION COLUMN, E-1

The skirt thickness must be sufficient enough to withstand the dead weight load and
bending moment imposed on it by the vessel. Conical skirt is suitable for high vessel
with subject to wind loading (Sinnot R. K., 2005). The specification of skirt thickness
f
or extraction column is shown in table 8.17 and the resultant stress in skirt design in
table 8.18.

• Assumption:

• Straight cylindrical skirt (carbon steel)

• Maximum allowable stress for skirt material at Tamb =20°C, fs =

135Nmm-2

• Weld joint factor, J = 0.85

 lxxxv

• Base angle, Ɵs = 90°

• Young’s modulus, E = 200000

Table 8.17 Specification in skirt thickness of Extraction Column, E-1

Variables Value

Height vessel between tangent line, mm 16460

Diameter of the vessel, mm 10610.1

Skirt support height, mm 3
Design pressure, psi 52.65
Skirt thickness, mm 21

Table 8.18 The resultant stresses in the skirt design of Extraction Column, E-1

Parameter Value

Bending stress in the skirt, σbs, N/mm2 0.83

Dead weight stress in the skirt, σws, N/mm2 0.04

Maximum σs (tensile), N/mm2 0.79

σs (Compressive), N/mm2 0.87

fsJsinƟs 114.75

0.125E (ts/Ds)sinƟs 58.91

Thickness skirt, mm 12

σs (tensile) = 0.79N/mm2 < fsJsinƟs = 114.75N/mm2

σs (Compressive) = 0.87N/mm2 < 0.125E (ts/Ds)sinƟs = 58.91N/mm2

Therefore, the thickness ts = 21mm was in acceptable range and with addition of
corrosion allowance, 4mm. The final thickness of skirt is 12mm. *Refer appendix
(2.12)
for the stress in the skirt calculation.
 lxxxvi

8.3.17 BASE RING AND ANCHOR BOLT DESIGN OF EXTRACTION COLUMN,

E-1

Table 8.19 show the base ring and anchor bolt design specification for extraction
column. *Refer appendix (2.13) for base ring and anchor bolt design calculation.

• Assumption:

• Db = 3.2m

• Circumference of bolt circle =3200πmm

• fb = 125Nmm-2

• Number of bolts is 16

• Bolt spacing > 600mm

• Table 8.19 Base ring and anchor bolt design specification

• Parameter Value

Bolt spacing, mm 628.218

Bending moment at the base, Ms, Nmm 173396.62

Fb, Nm-1 2966.06

Lb, mm 0.85

Base ring thickness, tb , mm 1.57

 CROSS-SECTIONAL VIEW OF FERMENTER TOP VIEW OF CROSS-SECTION VIEW OF
FERMENTER FERMENTER

inlet stream 5
outlet stream 12
inlet stream 6

inlet stream 7
1271 mm
4 mm
10 mm 10 mm
inlet stream 8 10 mm CROSS SECTIONAL AREA CROSS-SECTIONAL AREA
OF MANHOLE
OF WELDED NECK FLANGE FACULTY OF ENGINEERING AND BUILT
ENVIRONMENT
DEPARTMENT OF CHEMICAL ENGINEERING AND
PROCESS
5000 mm
7620 mm DRAWN BY : KK10
5100 mm SUBMISSION DATE : 24TH JUNE 2021
ANCHOR BOLT CHAIR WELD FLANGE
DRAWING UNIT : FERMENTER, R-2
A E
GROUP MEMBERS :-
C
B H
1. ENGKU AHMAD FARIZ BIN ENGKU AHMAD KHAIRUL
ANUAR (A168003)
A 2. TAUFEQ ISMADI BIN AHMAD ZAKI (A169159)
B
3. AMIR FITRI BIN MUHAMMAD SHAFIQ POH (A168739)
4. MUHAMMAD ASYRAF BIN SHAFIE (A167520)
5080 mm 5. FARREL MUHAMMAD AKBAR (A184705)
1271 mm
GROUP KK10
PRODUCTION OF PENICILLIN G BY PENICILLIUM
CHRYSOGENUM

DESIGN SPECIFICATION
inlet stream 11 outlet stream 13 CLASS 150 WELD NECK FLANGE
Pressure, P : 2 bar
DIMENSION (MM)
NO PIPE SIZE A B C D E F G
Temperature, T : 25°C
5 40 125 16 65 48 60 40 73 Material : Stainless Steel Grade 316
7 40 125 16 65 48 60 40 73 Thickness, t : 10 mm
8 40 125 16 65 48 60 40 73 Corrosion Allowance, CA : 4 mm
9 40 125 16 65 48 60 40 73 Height of Vessel : 10160 mm
10 40 125 16 65 48 60 40 73 Inside Diameter of Vessel: 5080 mm
11 40 125 16 65 48 60 40 73 Head : Ellipsoidal
13 40 125 16 65 48 60 40 73 Shell : Cylindrical
15 40 125 16 65 48 60 40 73

unit measurement in mm
DRAWING NOT TO SCALE
 2650 mm

Inlet 4 Outlet 17
2 mm

Top View E-1

2 mm
Cross section E-1
2 mm UKM (2).jpg

FACULTY OF ENGINEERING AND BUILT

Cross sectional area manhole Weld neck flange ENVIRONMENT
DEPARTMENT OF CHEMICAL ENGINEERING AND
PROCESS
4 mm
8500 mm

DRAWN BY : KK10
SUBMISSION DATE : 24TH JUNE 2021
400mm

d
500 mm DRAWING UNIT : EXTRACTION, E-1
Double plate with gusset e

25mm
GROUP MEMBERS :-
f

1. ENGKU AHMAD FARIZ BIN ENGKU AHMAD KHAIRUL

ANUAR (A168003)
g

60mm
2. TAUFEQ ISMADI BIN AHMAD ZAKI (A169159)
3. AMIR FITRI BIN MUHAMMAD SHAFIQ POH (A168739)
c
4. MUHAMMAD ASYRAF BIN SHAFIE (A167520)
130mm
a
5. FARREL MUHAMMAD AKBAR (A184705)
25mm

GROUP KK10
Anchor bolt chair design
a

PRODUCTION OF PENICILLIN G BY PENICILLIUM

531mm
CHRYSOGENUM

DESIGN SPECIFICATION
b

Inlet 16 Outlet 18
HEAD : ELLIPSOIDAL
SHELL : CYLINDRICAL
5310 mm

Material : CARBON STEEL (SA-285 GR. C)

Thickness, t : 2 mm
Corrosion Allowance, CA : 4 mm
Height of Vessel : 1646 mm
Inside Diameter of Vessel: 5310 mm
SKIRT : CONICAL SKIRT
FLANGE : WELL NECK FLANGES

unit measurement in mm
DRAWING NOT TO SCALE
 CHAPTER IX

DYNAMIC AND PROCESS CONTROL

9.1 INTRODUCTION

Process is defined as a series of actions that you take in order to achieve a result, while
dynamic means having a lot of ideas and enthusiasm (Cambridge Dictionary, 2021).
Process dynamics refers to the study of transient behavior of the process while process
control refers to the use of process dynamics to alleviate the effect of undesirable
process behavior. The main objective of this chapter is to design a process control
system as to maintain the process in the system at the desired operating condition and
to select a unit operation. There are few benefits of process control system such as
control instrumentation ensures consistency, reduces labor costs, process control
instrumentation improves quality, opportunity for additional business and many more.
But, the most important things are process control system can provide safety
operation, satisfy environmental constraints and product quality requirement. In order
to achieve the goals, a control configuration must first be specified and follow by
designing a process control system which included the four main components. The
four main components for a process control system are controller, final controller
element, process and measuring device. The process control design has been applied
to the seed and main fermenter (fed batch bioreactor), absorption column and flash
vessel (Dale E. Seborg et al. 2003). PID controller is chosen and used to control all
units stated above because PID controller gives more advantage compare to P and PI
controller such as low overshoot, less oscillatory, no offset and many more. The main
objective for the choosing of PID controller is it is necessary to keep the output from
a process as close to the target or setpoint output as possible.

9.2 SEED FERMENTER R-1

The dynamic mathematical modelling for the seed fermenter will be derived based on
the following assumption:

1. The fed-batch fermenter is perfectly mixed.

2. The volume of fed batch fermenter and the liquid density are constant.

3. Heat effect is negligible and therefore the fermenter can be assumed to

operate isothermally.

4. The feed stream is sterile and thus contains no cells.

5. The cells are growing exponentially.

6. The rate of growth of cell, rg is given by

𝑟𝑔= 𝜇𝑋 (1)
𝑆
𝜇 = 𝜇 𝑚𝑎𝑥 𝐾𝑠 +𝑆
(2)

7. Rate of product formation per unit volume, 𝑟𝑃 = 𝑌𝑃/𝑋 𝑟𝑔

The unsteady component balance for cell, substrate and product are shown below.

Cells:
𝑑(𝑋𝑉)
= 𝑉𝑟𝑔
𝑑𝑡
𝑑𝑋
𝑉 = 𝑉𝑟𝑔
𝑑𝑡
𝑑𝑋
= 𝑟𝑔 = μX
𝑑𝑡
𝑑𝑋 𝑆
= 𝜇𝑚𝑎𝑥 𝑋 (4)
𝑑𝑡 𝐾𝑠 + 𝑆

Substrate:
𝑑(𝑆𝑉) 1
= 𝐹𝑆𝑓 − 𝑉𝑟
𝑑𝑡 𝑌𝑋/𝑆 𝑔
𝑑𝑆 1
𝑉 = 𝐹𝑆𝑓 − 𝑉𝑟
𝑑𝑡 𝑌𝑋/𝑆 𝑔
𝑑𝑆 𝐹 1
= 𝑆𝑓 − 𝑟
𝑑𝑡 𝑉 𝑌𝑋 𝑔
𝑆
𝑑𝑆 𝐹 1 𝑆
= 𝑆𝑓 − 𝜇𝑚𝑎𝑥 𝑋 ( 5)
𝑑𝑡 𝑉 𝑌𝑋 𝐾𝑠 + 𝑆
𝑆

Product:
𝑑(𝑃𝑉)
= 𝑉𝑟𝑝
𝑑𝑡
𝑑𝑃
𝑉 = 𝑉𝑟𝑝
𝑑𝑡
𝑑𝑃
= 𝑟𝑝
𝑑𝑡
𝑑𝑃
= 𝑌𝑃/𝑋 𝑟𝑔
𝑑𝑡
𝑑𝑃 𝑆
= 𝑌𝑃/𝑋 𝜇𝑚𝑎𝑥 𝑋 (6)
𝑑𝑡 𝐾𝑠 + 𝑆

Where,
rg = cell growth rate g/dm3 s
X = concentration of cell, g/dm3 µ = specific growth rate, s-1
µmax = maximum specific growth reaction rate, s-1
Ks = Monod constant, g/dm3
S = substrate concentration, g/dm3
V = volume of fermenter, dm3
F = mass feed flow, dm3/s
Sf = glucose concentration, g/dm3
rp = rate of production formation, g/dm3 s
P = product concentration, g/dm3

Degree of freedom analysis:

5 parameters: 𝑉, 𝜇𝑚𝑎𝑥 , 𝐾𝑆 , 𝑌𝑃/𝑋 , 𝑌𝑋/𝑆

5 variables (NV = 5): S, X, P, F, Sf
3 equations (NE = 3): Equations (4), (5), and (6)

Thus, degree of freedom, NF = NV – NE = 5 – 3 = 2.

3 outputs : S, X, P

2 inputs : F, Sf

From 2 inputs,
1 disturbance: mass feed flow, F
1 manipulated variable: medium feed rate, Sf.

Figure 9.1 show the P&ID diagram of seed fermenter, R-1:

Table 9.1 Control action taken on seed fermenter R-1

Type of Objective Control Manipulated Control Action Type of

controller variables variables valve

pH To control the pH of seed Inlet The transmitter will Globe valve

controller pH of seed fermenter, flowrate of measure the pH and
fermenter, R-1. The set nutrient send signal to the - control the
R-1 point pH is medium controller. If the pH is flowrate of
pH 6~6.5. higher than the liquid more
setpoint, pH 6.5 then precisely
the valve opening is
decreased to allow
less mediumflow into
the fermenter. If the
pH is below the
setpoint, the valve
opening is increased
 to allow more
nutrient medium into
the fermenter to allow
the temperature to
settle to its desired set
point.

Flowrate To control the Flowrate of Amount of The transmitter will Globe valve
controller flowrate of nutrient nutrient measure the flowrate
nutrient medium into medium used and send signal to the
medium into the seed for controller. If the - control the
the seed fermenter, R- cultivation of flowrate is higher flowrate of
fermenter, 1. The set Penicillium than the setpoint, then liquid more
point is to chrysogenum the valve opening is precisely
R-1
produce in seed decreased to allow
37kg/m3 fermenter. less nutrient medium
concentration flow into seed
of microbe. fermenter. If the
flowrate is below the
setpoint, the valve
opening is increased
to allow more
medium flow into the
seed fermenter to
settle to its desired set
point.

Temperature
Temperat To control the Inlet flowrate The transmitter will Globe valve
of seed
ure temperature of cooling measure the
fermenter, R-
controller of fermenter, water temperature and send
1. The set
R-1. signal to the
point
controller. If the
temperature is - control the
temperature is higher
25oC. flowrate of
than the setpoint,
25oC then the valve liquid more
opening is increased precisely
to allow more cooling
water to reduce the
temperature. If the
temperature is below
the setpoint, the valve
opening is decreased
to reduce the amount
of cooling water into
the fermenter to allow
the temperature to
settle to its desired set
point.
9.3 MAIN FERMENTER R-2

The dynamic mathematical modelling for the seed fermenter will be derived based on
the following assumption:

1. The fed-batch fermenter is perfectly mixed.

2. The volume of fed batch fermenter and the liquid density are constant.
3. Heat effect is negligible and therefore the fermenter can be assumed to operate
isothermally.
4. The feed stream is sterile and thus contains no cells.
5. The cells are growing exponentially.
6. The rate of growth of cell, rg is given by

𝑟𝑔= 𝜇𝑋 (1)
𝑆
𝜇 = 𝜇 𝑚𝑎𝑥 (2)
𝐾𝑠 +𝑆

7. Rate of product formation per unit volume, 𝑟𝑃 = 𝑌𝑃/𝑋 𝑟𝑔

The unsteady component balance for cell, substrate and product are shown below.

Cells:
𝑑(𝑋𝑉)
= 𝑉𝑟𝑔
𝑑𝑡
𝑑𝑋
𝑉 = 𝑉𝑟𝑔
𝑑𝑡
𝑑𝑋
= 𝑟𝑔 = 𝜇𝑋
𝑑𝑡
𝑑𝑋 𝑆
= 𝜇𝑚𝑎𝑥 𝐾 +𝑆 𝑋 (4)
𝑑𝑡 𝑠

Substrate:
𝑑(𝑆𝑉) 1
= 𝐹𝑆𝑓 − 𝑉𝑟
𝑑𝑡 𝑌𝑋 𝑔
𝑆
 𝑑𝑆 1
𝑉 = 𝐹𝑆𝑓 − 𝑉𝑟
𝑑𝑡 𝑌𝑋 𝑔
𝑆

𝑑𝑆 𝐹 1
= 𝑆𝑓 − 𝑟
𝑑𝑡 𝑉 𝑌𝑋 𝑔
𝑆

𝑑𝑆 𝐹 1 𝑆
= 𝑆𝑓 − 𝜇𝑚𝑎𝑥 𝑋 (5)
𝑑𝑡 𝑉 𝑌𝑋 𝐾𝑠 + 𝑆
𝑆

Product:
𝑑(𝑃𝑉)
= 𝑉𝑟𝑃
𝑑𝑡
𝑑𝑃
𝑉 = 𝑉𝑟𝑃
𝑑𝑡
𝑑𝑃
= 𝑟𝑃
𝑑𝑡
𝑑𝑃
= 𝑌𝑃 𝑟𝑔
𝑑𝑡 𝑋

𝑑𝑃 𝑆
= 𝑌𝑃 𝜇𝑚𝑎𝑥 𝑋 (6)
𝑑𝑡 𝑋 𝐾𝑠 + 𝑆

Where,
rg = cell growth rate g/dm3 s
X = concentration of cell, g/dm3 µ = specific growth rate, s-1
µmax = maximum specific growth reaction rate, s-1
Ks = Monod constant, g/dm3
S = substrate concentration, g/dm3
V = volume of fermenter, dm3
F = mass feed flow, dm3/s
Sf = glucose concentration, g/dm3
rp = rate of production formation, g/dm3 s
P = product concentration, g/dm3
Degree of freedom analysis:

5 parameters: 𝑉, 𝜇𝑚𝑎𝑥, 𝐾𝑆, 𝑌𝑃/𝑋, 𝑌𝑋/𝑆

5 variables (NV = 5): S, X, P, F, Sf
3 equations (NE = 3): Equations (4), (5), and (6)

Thus, degree of freedom, NF = NV – NE = 5 – 3 = 2.

3 outputs: S, X, P
2 inputs: F, Sf

From 2 inputs,
1 disturbance: mass feed flow, F
1 manipulated variable: medium feed rate, Sf.

Figure 9.2 show the P&ID diagram of main fermenter, R-2:

Ammonium Sulphate

Air
 Table 9.2 Control action taken on main fermenter

Type of Objective Control Manipulat Control Action Type of valve

controller variables ed
variables
pH To control the pH of seed Inlet The transmitter will Globe valve
controller pH of seed fermenter, flowrate of measure the pH and
fermenter, R-1. The set Ammoniu send signal to the - control the
R-1 point pH is m controller. If the pH is flowrate of
pH 6~6.5. Sulphate higher than the liquid more
setpoint, pH 6.5 then precisely
the valve opening is
decreased to allow
less ammonium
sulohate flow into the
fermenter. If the pH is
below the setpoint,
the valve
opening is increased
to allow more
ammonium into the
fermenter to allow the
temperature to settle
to its desired set
point.

Flowrate To control the Flowrate of Amount of The transmitter will Globe valve
controller flowrate of air air into the air used measure the flowrate
into the main main for and send signal to the
fermenter, fermenter, fermentati controller. If the - control the
R-2 R-2. The set on of flowrate is higher flowrate of
point is to Penicilliu than the setpoint, then liquid more
supply air m the valve opening is precisely
flow rate of chrysogen decreased to allow
12.11 kg/h. um in main less nutrient medium
fermenter. flow into seed
fermenter. If the
flowrate is below the
setpoint, the valve
opening is increased
to allow more
medium flow into the
main fermenter to
settle to its desired set
point.
 Temperat To control the Temperature Inlet The transmitter will Globe valve
ure temperature of main flowrate of measure the
controller of main fermenter, cooling temperature and send
fermenter, R- R-1. The set water signal to the - control the
2. point controller. If the flowrate of
temperature temperature is higher liquid more
is 25oC. than the setpoint, precisely
25oC then the valve
opening is increased
to allow more cooling
water to reduce the
temperature. If the
temperature is below
the setpoint, the valve
opening is decreased
to reduce the amount
of cooling water into
the fermenter to allow
the temperature to
settle to its desired set
point.

9.4 EXTRACTION COLUMN E-1

The dynamic mathematical modelling for the extraction will be derived based on the
following assumption:

1. The density of fluid, ρ and the specific capacity of fluid, Cp remain constant
in the system.
2. Heat effect is small so can be neglected.

Rate of change of mass = Total mass into - Total mass

out
Accumulated in control volume control volume control
volume
𝑑(𝜌𝑣)
= ∑𝜌𝐹𝑖𝑛 − ∑𝜌𝐹𝑜𝑢𝑡
𝑑𝑡
𝑑ℎ
𝐴 [ 𝑑𝑡 ] = 𝜌4 𝐹4 + 𝜌16 𝐹16 − 𝜌17 𝐹17 (1)

Energy Balance:
Rate of change of = Energy into - Energy out + Energy
energy accumulated control control associated with
in control volume volume volume reaction
 𝑑𝑇
𝑉𝜌𝐶𝑣 [ ] = ∑𝐹𝑖 𝜌𝐶𝑝 𝑇𝑖 − ∑𝐹0 𝜌𝐶𝑝 𝑇 − 𝑈𝐴[𝑇 − 𝑇𝑐 ] + 𝑄
𝑑𝑡
𝑑𝑇
𝑉𝜌𝐶𝑣 [ ] = ∑𝐹4 𝜌𝐶𝑝 𝑇4 + ∑𝐹16 𝜌𝐶𝑝 𝑇16 − ∑𝐹17 𝜌𝐶𝑝 𝑇17 (2)
𝑑𝑡

Degree of freedom analysis:

3 parameters: 𝜌, V, Cp
6 variables (Nv =6): F4, F16, F17, T4, T16, T17
2 equations (NE=2): [Equation (1) and (2)]

Degree of freedom, NF = Nv - NE = 6 – 2 =4
2 specified variables:
F4, T4
Now, degree of freedom = 2
Number of process control required: 2

Figure 9.3 shows the P&ID diagram of extraction column E-1:

Table 9.3 Control action taken on extraction column E-1

Type of Objective Control Manipulated Control Action Type of

controller variables variables valve
 Level To control level Level of fluid Flow rate of The transmitter will Globe valve
controller of fluid in in extraction liquid out of measure the level and - control the
extraction column, E-1. extraction send signal to the flowrate of
column, E-1. The set point column, controller. If the level liquid more
level of fluid stream 17. is higher than the precisely
is set as 70% setpoint, 10m then the
of the valve opening is
extraction increased to allow
column. more liquid flow out
from the extraction
column to reduce the
level of fluid. If the
level is below the
setpoint, the valve
opening is decreased
to reduce the amount
of fluid flow out from
the extraction column
to allow the level to
set point
settle to its desired set
pH To control the The pH Flowrate of The transmitter will
controller pH of the broth value of the the broth measure the pH and
from stream 16 broth is into the send signal to the
flow into the control in a extraction controller. If the pH is
extraction range of 2 to column higher than the
5 as the setpoint, pH 5 then the
operation valve opening is
much decreased to allow
favorable in less broth flow into
acidic the extraction column.
environment If the pH is below the
(Aliwarga et setpoint, the valve
al., 2019) opening is increased
to allow more broth
flow into the
extraction column to
allow the temperature
to settle to its desired
set point.

9.5 REGENARATION COLUMN E-2

The dynamic mathematical modelling for the regeneration will be derived based on
the following assumption:

1. The density of fluid, ρ and the specific capacity of fluid, Cp remain constant
in the system.
 2. Heat effect is small so can be neglected.

Rate of change of mass = Total mass into - Total mass

out
Accumulated in control volume control volume control
volume
𝑑(𝜌𝑣)
= ∑𝜌𝐹𝑖𝑛 − ∑𝜌𝐹𝑜𝑢𝑡
𝑑𝑡
𝑑ℎ
𝐴 [ 𝑑𝑡 ] = 𝜌2 𝐹2 + 𝜌3 𝐹3 + 𝜌17 𝐹17 − 𝜌18 𝐹18 − 𝜌28 𝐹28
(1)

Energy Balance:
Rate of change of = Energy into - Energy out + Energy
energy accumulated control control associated with
in control volume volume volume reaction

𝑑𝑇
𝑉𝜌𝐶𝑣 [ ] = ∑𝐹𝑖 𝜌𝐶𝑝 𝑇𝑖 − ∑𝐹0 𝜌𝐶𝑝 𝑇 − 𝑈𝐴[𝑇 − 𝑇𝑐 ] + 𝑄
𝑑𝑡
𝑑𝑇
𝑉𝜌𝐶𝑣 [ ] = ∑𝐹2 𝜌𝐶2 𝑇2 + ∑𝐹3 𝜌𝐶3 𝑇3 + ∑𝐹17 𝜌𝐶𝑝 𝑇17 − ∑𝐹18 𝜌𝐶𝑝 𝑇18
𝑑𝑡
− ∑𝐹28 𝜌𝐶𝑝 𝑇28 (2)

Degree of freedom analysis:

3 parameters: 𝜌, V, Cp
10 variables (Nv =10): F2, F3, F17, F18, F28, T2, T3, T17, T18, T28
2 equations (NE=2): [Equation (1) and (2)]

Degree of freedom, NF = Nv - NE = 10– 2 = 8

6 specified variables:
F2, F3, F17, T2, T3, T17,
Now, degree of freedom = 2
Number of process control required: 2
 3

2
28

17

18

Table 9.1 Control action taken on regeneration column E-2

Type of Objective Control Manipulated Control Action Type of

controller variables variables valve
Level To control level Level of fluid Flow rate of The transmitter will Globe valve
controller of fluid in in liquid out of measure the level and - control the
regeneration regeneration extraction send signal to the flowrate of
column, E-2. column, E-2. column, controller. If the level liquid more
The set point stream 17. is higher than the precisely
level of fluid setpoint, 10m then the
is set as 70% valve opening is
of the increased to allow
extraction more liquid flow out
column. from the regeneration
column to reduce the
level of fluid. If the
level is below the
setpoint, the valve
opening is decreased
to reduce the amount
of fluid flow out from
the regenaration
column to allow the
level to settle to its
desired set poin
pH To control the The pH value Flowrate of The transmitter will
controll pH of the of the broth is the broth measure the pH and
er broth from control in a into the send signal to the
stream 16 range of the regeneration controller. If the pH
flow into the solution into column is higher than the
regeneration an alkaline setpoint, pH 7 then
column, E-2 condition of the valve opening is
pH 7 where decreased to allow
the penicillin less broth flow into
stability is the regeneration
high. column. If the pH is
below the setpoint,
the valve
opening is increased
to allow more broth
flow into the
regenaration column
to allow the
temperature to settle
to its desired set
point.
 R-1 R-2 B-1 A-1 C-1 H-1 P-1 E-1 E-2 H-2 R-3 C-2 T-1 H-3 F-1
Seed fermenter Main fermenter Blower Air filter Centrifugal disc Cooler Pump Extraction column Regeneration column Cooler CSTR Centrifugal basket Turbine Cooler Freeze dryer crystallizer

1
FAKULTI KEJURUTERAAN DAN
ALAM BINA
Sodium bicarbonate 2 JABATAN KEJURUTERAAN KIMIA
DAN PROSES

Phosphate buffer 3 28
TITLE: PFD OF PRODUCTION OF
4 PENICILLIN G
Water 17
Butyl acetate

Butyl acetate 5 LT
16

LC pHI
NAME OF LECTURERS:-
Phenylacetic acid 6
pHIC pHIC E-2
• PM. IR. DR. SHUHAIDA BINTI
E-1 HARUN
12 pHI 29
Ammonium sulphate 7 • PM. IR. DR. HASSIMI BIN ABU
HASAN
Exhaust gas •
LC LT
Glucose 8 PM. DR. NOORHISHAM BIN TAN
pHI KOFLI
pHIC
19 • PM. DR. NORLIZA BINTI ABD
Medium R-1
RAHMAN
R-3
•
FC FT TT pHI
DR. MUHAMMAD ZULHAZIMAN
TC pHIC 18 BIN MAT SALLEH
TT
Cooling
water in 20 21
TC

Cooling water out

Cooling H-2
water in
25
24
Cooling R-2 C-2
water out 22
GROUP MEMBERS:-
23
10 11
9
T-1
Wet air • MUHAMMAD ASYRAF BIN
13 14 15 SHAFIE (A1697520)
Air B-1 A-1 H-3 • ENGKU AHMAD FARIZ BIN
FC FT
ENGKU AHMAD KHAIRUL
F-1 25
C-1 H-1 P-1 ANUAR (A168003)
• AMIR FITRI BIN MUHAMMAD
Penicillin G salt SHAFIQ POH (A168739)
26
• AHMAD TAUFEQ ISMADI BIN
AHMAD ZAKKI (A169159)
• FARREL MUHAMMAD AKBAR
27 Liquid waste
(A184705)

Biomass INPUT

OUTPUT

STREAM NUMBER
 CONCLUSION

Amoxicillin is the one of antibiotic that widely used to treat infection of bacteria like
tonsillitis and stomach ulcers affected by Helicobacter pylori infection. The chemical
formula and molecular weight for this antibiotic is C16H19N3O5S and 419.45g/mol.

This antibiotic can be produced by using Penicillin G which originated from the
fungi that called as Penicillium chrysogenum. This producer cell is chosen because of the
high fermentation efficiency is high than other fungi cell. It also has the high antibiotic
activity, and this fungus is easy to deal.

Global demand for this antibiotic is increase by 65% between 2000 to 2015 and
expected to reach about 48,560,000kg/year in 2025. For the proposal production of year is
expected to produce 2,292,000 kg/year which fulfill 9.9% from the market shortage.

Penicillium chrysogenum is chosen as producer cell because the fermentation

efficiency is the highest compared to Penicillium nalgiovense and Penicillium
griseofulvum. Other than that, the optimal the optimal temperature and pressure for P.
chrysogenum growth are 25℃ and 6.5 respectively (Gordon et al., n.d.) which are easily to
control and achieve.

The waste production in this plant is carbon dioxide (CO2), oxygen (O2), wastewater
and biomass. All this waste product must be treated before release to environment. This
waste must be handling and treat according to suitable treatment because it maybe contains
some harmful substances and can give effect to environment and human.

For the separation part, the chosen unit to be determine the process design is
centrifugal stack disc. The calculation for flowrate Q in tubular bowl centrifuge was
calculated which is 29.213m3/min. The sedimentation velocity under gravity, Vg is 1.99x10-
7
m/min. The angle chosen typically between 40° to 55° and the number of discs used is
between 50 to 150 disc. Radius of bowl, ro is estimated to be 0.203m and length of
centrifuge is 1m. The speed of centrifuge, ω is set at 6250rpm for safety purpose. By using
all these parameters, we can get the factor of centrifuge, ∑ which is 1.468x108m2 and the
radius of outlet opening of the bowl, r1 0.205m is obtained.

For the dynamic part, the conclusion is that the mass balance and energy balance
calculation is fundamental in any unit process because there are a lot of things need to be
considered in certain process in industry. For our project, after the calculation of the degree
of freedoms, we knew that some of our unit operators need to be inserted with controllers
for them to operation efficiently. For example, the level controller and pH controller need
to be inserted into our extraction column, E-1 and pH controller, temperature controller and
flowrate controller need to be inserted into both our seed and main fermenter, R-1, and R-
2.

For the utility and pressure vessel design part, the utility that has been chosen is a
heat exchanger, H-2. Based on the calculation using the Kern’s method to find the thermal
analysis, the head load, Q is 48.244 kW. The tube-side coefficient, hi is 10551.33 W/m2.℃
and for the shell-side coefficient, hs is 4725.63 W/m2.℃. The value that we have gotten
from tube-side coefficient and shell-side coefficient, we can calculate the overall
coefficient, Uo which we get 909.09 W/m2.℃. For the tube-side pressure drop, ∆Pt and
shell-side pressure drop, ∆Ps, the value that have been calculated are 2.233 x 104 N/m2 and
8.651 x 103 N/m2 respectively. Next, we have two pressure vessel that have been chosen
which are fermenter, R-2 and extraction column, E-1. The fermenter, R-2 is calculated
under internal pressure procedure while for extraction column, E-1 is calculated under
external pressure procedure. After all the calculation that had been done on both the
pressure vessels, mechanical drawings have then been designed on the AutoCAD software
to show roughly of the schematic diagram of the pressure vessel that will be used in our
production.
 REFERENCES

Aliwarga, L., Susanto, H., Reynard, R., & Victoria, A. V. (2019). UNIFAC Model for
Liquid-Liquid Phase Equilibrium of Penicillin G and 6-APA System. Jurnal Kimia
Valensi, 5(2), 185–193. https://doi.org/10.15408/jkv.v5i2.9869

Amaro, H. M., Sousa-Pinto, I., F.X., M. & Caterina, G. A. (n.d.). Disc Stack Centrifuge - an
overview | ScienceDirect Topics. Microalgal fatty acids - From harvesting until
extraction. https://www.sciencedirect.com/topics/engineering/disc-stack-centrifuge [24
June 2021].

Angelo, K. 2020. Disinfectants Can Kill the Coronavirus, but Can Also Harm Health | UMass
Lowell. UMass Lowell. https://www.uml.edu/news/stories/2020/disinfectingsafely.aspx
[21 January 2021].

ASTM A285 Grade C Carbon Steel Pressure Vessel and Boiler Steel Plate Stockists in
Mumbai India. (n.d.). https://www.csecplates.com/astm-a285-grade-c-plate-stockists-
suppliers.html [24 June 2021].

Bauweleers HMK, G. D. (2014). Genes useful for the industrial production of citric acid.

Canzani, D. and Aldeek, F. (2017) ‘Penicillin G ’ s function , metabolites , allergy , and

resistance .’, 1(1).
Ciriminna, R., Meneguzzo, F., Delisi, R. & Pagliaro, M. 2017, March 8. Citric acid:
Emerging applications of key biotechnology industrial product. Chemistry Central
Journal. BioMed Central Ltd. doi:10.1186/s13065-017-0251-y

Cole, M., Kenig, M. D. and Hewitt, V. A. (1973) ‘Metabolism of penicillins to penicilloic

acids and 6-aminopenicillanic acid in man and its significance in assessing penicillin
absorption.’, Antimicrobial agents and chemotherapy, 3(4), pp. 463–468. doi:
10.1128/AAC.3.4.463.

Da Cruz, A. J. G., Hokka, C. O., & Giordano, R. C. (1997). A SIMULATION OF THE

PENICILLIN G PRODUCTION BIOPROCESS APPLYING NEURAL
NETWORKS. Brazilian Journal of Chemical Engineering, 14(4).
https://doi.org/10.1590/S0104-66321997000400004
Gordon, J. J., Grenfell, E., Knowles, E., Legge, B. J., Mcallister, R. C. A., & White, T. (n.d.).
Method& of Penicillin Production in Submerged Culture on a Pilot-Plant Scale.

Goudar, C. T., & Strevett, K. A. (n.d.). Biochemical Eiigiieefhg hmnl. Estimating growth
kinetics of Penicillium chrysogenum by nonlinear regression.

Grimwood, G. C. & Ainsworth, A. 2015. Batch centrifuge basket design. International Sugar
Journal.

Hillenga, D. J., Versantvoort, H. J. M., van der Molen, S., Driessen, A. J. M., & Konings, W.
N. (1995). Penicillium chrysogenum Takes up the Penicillin G Precursor Phenylacetic
Acid by Passive Diffusion. In APPLIED AND ENVIRONMENTAL MICROBIOLOGY
(Vol. 61, Issue 7).

Laurence Brundrett. 2012. External Pressure – Pressure Vessel Engineering. Pressure Vessel
Engineering. https://www.pveng.com/home/asme-code-design/external-pressure-
methods/ [24 June 2021].

Lorenz, Th., Diekmann, J., Frueh, K., Hiddessen, R., Moeller, J., Niehoff, J., & Schügerl, K.
(2007). Online measurement and control of penicillin V production in a tower
loop reactor. Journal of Chemical Technology & Biotechnology, 38(1).
https://doi.org/10.1002/jctb.280380105

Ortho-Nitrophenyl-β-galactoside - an overview | ScienceDirect Topics. (n.d.).

https://www.sciencedirect.com/topics/computer-science/fermentation-broth [24 June
2021].

Papagianni. (2007). Advances in citric acid fermentation by Aspergillus niger: biochemical

aspects, membrane transport and modeling. Biotechnol Adv. 25, 244–263.

Penicillium Species - Doctor Fungus. 2017. https://drfungus.org/knowledge-base/penicillium-

species/ [24 June 2021].

Pirt, S. J., & Righelato, R. C. (1967). Effect of Growth Rate on the Synthesis of Penicillin by
Penicillium chrysogenum in Batch and Chemostat Cultures. Applied Microbiology,
15(6), 1284–1290. https://doi.org/10.1128/aem.15.6.1284-1290.1967

Pradhan, D., Biswasroy, P., Kumar Naik, P., Ghosh, G. & Rath, G. 2020, July 1. A Review of
Current Interventions for COVID-19 Prevention. Archives of Medical Research.
 Elsevier Inc. doi:10.1016/j.arcmed.2020.04.020

Report, G. V. R. (n.d.). Antibiotics Market Size, Growth & Trends Report, 2021-2028.
https://www.grandviewresearch.com/industry-analysis/antibiotic-market [23 April
2021].

Richardson, A. & Walker, J. 2018. Continuous Solids Discharging centrifugation to Solve

Clarifying High-Cell-Density Mammalian Cell Cultures. BioProcess International.
https://bioprocessintl.com/downstream-processing/separation-purification/continuous-
solids-discharging-centrifugation-a-solution-to-the-challenges-of-clarifying-high-cell-
density-mammalian-cell-cultures/ [24 June 2021].

Srirangan, K. et al. (2013) ‘Biotechnological advances on Penicillin G acylase:

Pharmaceutical implications, unique expression mechanism and production
strategies’, Biotechnology Advances. Elsevier Inc., 31(8), pp. 1319–1332. doi:
10.1016/j.biotechadv.2013.05.006.

Tarleton, S. & Wakeman, R. (n.d.). 6.3: Design Equations for Centrifugal Filter Cycles |
Engineering360. https://www.globalspec.com/reference/26324/203279/6-3-design-
equations-for-centrifugal-filter-cycles [24 June 2021].

Tishkov, V. et al. (2010) ‘Protein Engineering of Penicillin Acylase’,

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3347563/#:~:text=Penicillin%20acy
lase%20(PA%2C%20EC%203.5,called%20N%2Dterminal%20nucleophilic%20hyd
rolases.

Virden, R. (1990) ‘Structure, processing and catalytic action of penicillin acylase’,

Biotechnology and Genetic Engineering Reviews, 8(1), pp. 189–218. doi:
10.1080/02648725.1990.10647869.

Walle, G. Van De. 2019. What Is Citric Acid, and Is It Bad for You? Healthline. Retrieved
from https://www.healthline.com/nutrition/citric-acid

Why is the world suffering from a penicillin shortage? | Health News | Al Jazeera. (n.d.).
https://www.aljazeera.com/features/2017/5/21/why-is-the-world-suffering-from-a-
penicillin-shortage [23 April 2021].
 APPENDIX A

1.1 STOICHIOMETRY EQUATION

Yield of penicillin=0.90 (Meštrović,T n.d.)

α4(334)/180 = 0.90

α4 = 0.485

Yield of biomass = 0.43(Anon n.d.)

α5(26.76)/180 = 0.43

α5 = 2.890

Yield of oxygen = 1.35(Shuler 2002)

α5(26.76)/ α1(32) = 1.35

α1=1.792

Element balance for each component

C: -6 -8α3+16α4 + α5 + α7 =0

H: -12-8α2 -8α3 + 1.64α5 + 2α6 +18 α4 =0

N: -2α2 + 2α4 +0.16α5 =0

S: -α2 + α4 +0.08α5 =0

O: -6-2α1 -4α2 - 2α3 + 4α4 +0.52α5 + α6 +2 α7 =0

By using Gauss elimination method, the stoichiometric equation is solve.

C6H12O6 + 1.792O2 + 0.716(NH4)2SO4 + 0.899C8H8O2 → 0.485C16H18N2O4S +

2.89CH1.64O0.52N0.16S0.08 + 5.725H2O + 2.539CO2
1.2 MASS BALANCE

1.2.1 Main fermenter

List of parameter

G = Glucose; N = Nitrogen gas; O = Oxygen; A = Ammonium sulphate; H = Phenylacetic

acid; P = Penicillin G; X = Biomass; W = Water; C = Carbon dioxide;

From the experimental data of Penicillium chrysogenum.

𝜇 = 0.02 ℎ−1 ( S.J. Pirt, 1967 )

Theoritical value from the stoichiometry.

YG/X = 2.370576 qG = 0.04741152

YO/X = 0.754542 qO = 0.01509084
YA/X = 1.244924 qA = 0.02489848
YH/X = 1.610545 qH = 0.0322109
YP/X = 2.134045 qP = 0.0426809
YW/X = 1.35595 qW = 0.02711899
YC/X = 1.469978 qC = 0.02939957

Mass of Glucose, G

𝑒 𝜇𝑡
𝐶𝐺 = 𝐶𝐺𝑜 + 𝑌𝐺/𝑋 𝑋𝑜 −
𝜇

𝑒 0.02(70)
𝐶𝐺 = 140 + 2.370576(37) −
0.02

kg
𝐶𝐺 = 25
m3

𝑚𝐺 = 5000kg 𝑚𝐺𝑜 = 28000kg

Mass of oxygen, O

𝑒 𝜇𝑡
𝐶𝑂 = 𝐶𝑂𝑜 + 𝑌𝑂/𝑋 𝑋𝑜 − 𝜇

𝑒 0.02(70)
𝐶𝑂 = 178 + 0.75454(37) −
0.02

kg
𝐶𝑂 = 3
m3

𝑚𝑂 = 600kg 𝑚𝑂𝑜 = 35600kg

Mass of amminium sulphate, A

𝑒 𝜇𝑡
𝐶𝐴 = 𝐶𝐴𝑜 + 𝑌𝐴/𝑋 𝑋𝑜 −
𝜇

𝑒 0.02(70)
𝐶𝐴 = 166 + 1.24492(37) −
0.02

kg
𝐶𝐴 = 10
m3

𝑚𝐴 = 2000kg 𝑚𝐴𝑜 = 33200kg

Mass of phenyl acetic acid, H

𝑒 𝜇𝑡
𝐶𝐻 = 𝐶𝐻𝑜 + 𝑌𝐻/𝑋 𝑋𝑜 −
𝜇

𝑒 0.02(70)
𝐶𝐻 = 160 + 1.61(37) −
0.02

kg
𝐶𝐻 = 17
m3

𝑚𝐻 = 3400kg 𝑚𝐻𝑜 = 32000kg

Mass of biomass, X

𝐶𝑋 = 𝐶𝑋𝑜 𝑒 𝜇𝑡

𝐶𝑋 = 37𝑒 0.02(70)

𝐶𝑋 = 150kg

Mass of penicillin G, P

𝑒 𝜇𝑡
𝐶𝑃 = − 𝑌𝑃/𝑋 𝑋𝑜
𝜇

𝑒 0.02(70)
𝐶𝑃 = − (2.134)(37)
0.02

kg
𝐶𝑃 = 124
m3

𝑚𝑃 = 24800 kg

Mass of Carbon dioxide, C

𝑒 𝜇𝑡
𝐶𝐶 = − 𝑌𝐶/𝑋 𝑋𝑜
𝜇

𝑒 0.02(70)
𝐶𝐶 = − (1.46998)(37)
0.02

kg
𝐶𝐶 = 148
m3

𝑚𝐶 = 29600 kg

Mass of water, W

𝑒 𝜇𝑡
𝐶𝑊 = − 𝑌𝑊/𝑋 𝑋𝑜
𝜇
 𝑒 0.02(70)
𝐶𝑊 = − (1.35595)(37)
0.02

kg
𝐶𝑊 = 153
m3

𝑚𝑊 = 30600 kg

1.2.2 Extraction column (E-1)

List of parameter

G = Glucose; A = Ammonium sulphate; H = Phenylacetic acid; P = Penicillin G; W =

Water; C = Carbon dioxide; BA = n-butyl acetate;

𝑉𝑎𝑞 − 𝑞𝑃 𝑉𝑎𝑞
𝑉𝐵𝐴 =
𝑞𝑃 𝑘𝐷

Where,
VBA = Volume of n-butyl acetate kD = Partion Coefficient

Vaq = Volume of aqueous solution

qP = Fraction of Penicillin remain in aqueous solution

Mass balance for extraction column, E-1.

𝑚
Volume, 𝑉𝑇 = ∑ 𝜌

𝜌𝐺 = 1560 𝑘𝑔/𝑚3 𝜌𝐴 = 1130 𝑘𝑔/𝑚3 𝜌𝐻 = 1080 𝑘𝑔/𝑚3

𝜌𝑃 = 1000 𝑘𝑔/𝑚3 𝜌𝑊 = 997 𝑘𝑔/𝑚3

 5000 2000 3400 24800 30600
𝑉𝑇 = + + + + = 63.61𝑚3
1560 1130 1080 1000 997

The mass of organic solvent needed to dissolve the Penicillin G is;

𝑉𝑇 − 𝑞𝑃 𝑉𝑇
𝑚𝐵𝐴 = 𝑉𝐵𝐴 𝜌𝐵𝐴 = 𝜌𝐵𝐴
𝑞𝑃 𝑘𝐷

kD = 21.47 (Aliwarga et al., 2019)

63.31 − 0.0449(63.61)
𝑚𝐵𝐴 = (882) = 55311𝑘𝑔
0.0449(21.47)

1.2.3 Regeneration column (E-2)

List of parameter:

P = Penicillin; W = Water ; BA = n-butyl acetate ; PB = Phosphate buffer;

Mass balance on Regeneration Column:

Mass in = Mass out

Volume of penicillin-rich solution with butyl acetate in stream

𝑉𝑇 = 𝑉𝐵𝐴 + 𝑉𝑃

𝑉𝑇 = 𝑚𝐵𝐴 /𝜌𝐵𝐴 + 𝑚𝑃 /𝜌𝑃

Where,

𝜌𝑃 = 1000𝑘𝑔/𝑚3 𝜌𝑃𝐵 = 995𝑘𝑔/𝑚3

𝜌𝐵𝐴 = 882𝑘𝑔/𝑚3 𝜌𝑤 = 997𝑘𝑔/𝑚3

 55311 23686
𝑉𝑇 = + = 86.40𝑚3
882 1000

𝑉𝑃𝐵 = 0.1𝑉𝑇

𝑉𝑃𝐵 = 0.1(86.40) = 8.64𝑚3

𝑚𝑃𝐵 = 𝑉𝑃𝐵 𝜌𝑃𝐵

𝑚𝑃𝐵 = (8.64)(995) = 8596.8𝑘𝑔

𝑉𝑤 = 0.2𝑉𝑇

𝑉𝑤 = 0.2(86.40) = 1.728𝑚3

𝑚𝑊 = 𝑉𝑊 𝜌𝑊

𝑚𝑃𝐵 = (1.728)(997) = 1722.816𝑘𝑔

1.2.4 Centrifugal basket (C-2)

Mass of water in stream 26

𝑚𝑊26 = η𝑚𝑊26

𝑚𝑊26 = 0.95(2999)

𝑚𝑊26 = 2849 kg

Mass of phosphate buffer in stream 26

𝑚𝑃𝐵26 = η𝑚𝑃𝐵26

𝑚𝑃𝐵26 = 0.95(8597)
𝑚𝑃𝐵26 = 8167 kg

Mass of penicillin salt in stream 26

𝑚𝑃𝑆26 = η𝑚𝑃𝑆26

𝑚𝑃𝑆26 = 0.05(25276)

𝑚𝑃𝑆26 = 1263 kg

By applying the law of conservation of mass the output mass in stream 21 are calculated.

Mass of water in stream 21

𝑚𝑊21 = 𝑚𝑊𝑜 − 𝑚𝑊21

𝑚𝑊21 = 2999 − 2849

𝑚𝑊21 = 150 kg

Mass of phosphate buffer in stream 21

𝑚𝑃𝐵21 = 𝑚𝑃𝐵𝑜 − 𝑚𝑃𝐵21

𝑚𝑃𝐵21 = 8597 − 8167

𝑚𝑃𝐵21 = 430 kg

Mass of penicillin salt in stream 26

𝑚𝑃𝑆21 = 𝑚𝑃𝑆𝑜 − 𝑚𝑃𝑆21

𝑚𝑃𝑆21 = 25276 − 1263

𝑚𝑃𝑆21 = 24013 kg
 APPENDIX B

SEPARATION PROCESS II

Table 1: Mass balance in centrifugal stack disc

Stream Stream Stream

Component 13 14 27
Glucose 5000 5000 0
Oxygen 0 0 0
Nitrogen 0 0 0
Ammonium Sulphate 2000 2000 0
Phenylacetic acid 3400 3400 0
Penicillin G 24800 24800 0
Biomass 30000 0 30000
Water 30600 30600 0
Carbon dioxide 0 0 0

The fermenter out, glucose, ammonium sulphate, phenylacetic acid, penicillin

G, biomass and water will enter the centrifugal stack disc to filter out the solid
biomass.

Mass flowrate of biomass = 73.0325kg/h

Convert the mass flowrate to volumetric flowrate by use the biomass density
(ρ=150kg/m3)

The calculation for flowrate, Q in tubular bowl centrifuge is given as below:

𝑄 = 𝑉𝑔 ∑

Where ∑ = factor for the centrifuge

Vg = sedimentation velocity under gravity

Volumetric flowrate of biomass, Q = 29.213m3/min

The equation for settling velocity under gravity is;

2𝛼2 (𝜌𝑠 −𝜌0 )𝑔

Vg= 9µ

Where α = radius of cell

ρs = density of cell

ρ0 = density of broth

g = gravitational acceleration

µ = viscosity of the broth

Size of penicillium chrysogenum, α = 5µm (Barreiro. C, 2019)

Density of cell, ρs = 37kg/m3

Density of broth, ρ0 = 1032.02kg/m3 (Devrim. B, 2017)

Viscosity of broth, µ = 0.25 Pa.s (Goudar. CT, 1999)

Sedimentation velocity under gravity, Vg

2𝛼2 (𝜌𝑠 −𝜌0 )𝑔

Vg = 9µ

2(5𝑥10−6 )2 (37 − 1032.02)(9.81)

=
9(0.25)

=1.99x10-7m/min
The factor of centrifuge;

2πn(𝑟0 2 − 𝑟1 2 )𝜔2
∑= cotƟ
3g

Where n = number of disc stack

r0 = radius of bowl

r1 = radius outlet opening of the bowl

ω = speed of the centrifuge

29.213𝑚3 /𝑚𝑖𝑛
∑ = 1.99𝑥10−7 𝑚/𝑚𝑖𝑛

= 1.468x108m2

Factor of centrifuge, ∑

The angle chosen typically between 40° to 55° and the number of disc used is between
50 to 150. (Perry, 1973)

The speed is set as 6250rpm for safety purpose.

2π(100)x(0.2032 − 𝑟1 2 )𝑥 (6250𝑟𝑒𝑣/ min 𝑥2𝜋𝑟𝑎𝑑/𝑟𝑒𝑣)2

∑= cot (44)
3𝑥9.81

Based on Table 7.1 above, since the volumetric flowrate of biomass is 29.213m3/min,
estimated bowl diameter, r0 is 0.203m and length of centrifuge, L is 1m, speed of
centrifuge, ω is set as 6250rpm for safety purpose. We can calculate the radius of outlet
opening of the bowl, r1 using the formula of factor of centrifuge, ∑. Therefore, we get
r1 is 0.205m
 APPENDIX C

UTILITY AND PRESSURE VESSEL DESIGN CALCULATION

1. Heat Exchanger, H-2

Figure 8.2 Heat Exchanger, H-2

Mainstream contains water and penicillin which in shell part whereas steam is also used
in tube part of the heat exchanger. Tm is the mean temperature for in and out both shell
and tube. Table below shows physical properties of component that come in and out of
the heat exchanger.

Table 8.2 Temperature for in and out cooler (H-2)

Temperature (℃) Shell (Mixture) Temperature (℃) Tube (Steam)
t1 4 T1 68
t2 28 T2 48
Mean Temperature, Tm 16 Mean Temperature, Tm 58

Table 8.3 Properties for stream in and out cooler (H-2)

Physical Properties Water (Steam)

Specific Heat Capacity, Cp (kJ/kg.℃) 4.2
Viscosity, µ (kg/m.s) 4.81 x 10-4
Density, ρ (kg/m3) 984.2
Thermal Conductivity, k (W/m.℃) 0.649

For thermal design calculation, Kern’s method is being used.

Mass flow rate of water, mw = 1723 kg/hr

1723
Heat Load, Q = 3600 (4.2)(28 − 4) = 48.244 𝑘𝑊

48.244
Flow of hot water = 4.2(20−3) = 0.676 𝑘𝑔/𝑠

Log mean difference temperature:

 (𝑇1 − 𝑡2 ) − (𝑇2 − 𝑡1 ) (68 − 28) − (48 − 4)
∆𝑇𝑙𝑚 = = = 41.97℃
(𝑇 − 𝑡2 ) (68 − 28)
𝑙𝑛 1 𝑙𝑛
(𝑇2 − 𝑡1 ) (48 − 4)

(𝑇1 − 𝑇2 ) (68 − 48)

𝑅= = = 0.83
(𝑡2 − 𝑡1 ) (28 − 4)

(𝑡2 − 𝑡1 ) (28 − 4)
𝑆= = = 0.38
(𝑇1 − 𝑡1 ) (68 − 4)

From formula (12.8) of Chemical Engineering Design by Gavin Towler & Ray Sinnot,

Ft = 0.95

∆𝑇𝑚 = 𝐹𝑡 ∆𝑇𝑙𝑚 = (0.95)(41.97) = 39.87℃

U is assumed to be 800 W/m2.℃

𝑄 48244
Provisional Area, A = 𝑈∆𝑇 = (800)(39.87) = 1.513 𝑚2
𝑙𝑚

Table 8.4 Dimension of tubes

Tubes
Outer Diameter 20 mm
Inner Diameter 16 mm
Length 4.83 m

Area of one tube, A = πDoL = π(20 x 10-3)(4.83) = 0.303 m2

𝑃𝑟𝑜𝑣𝑖𝑠𝑖𝑜𝑛𝑎𝑙 𝐴𝑟𝑒𝑎, 𝑚2 1.513

Number of tubes, Nt = 𝑆𝑢𝑟𝑓𝑎𝑐𝑒 𝐴𝑟𝑒𝑎 𝑜𝑓 𝑂𝑛𝑒 𝑇𝑢𝑏𝑒, 𝑚2 = 0.303 = 5 𝑡𝑢𝑏𝑒𝑠

Use triangular pitch, Pt = 1.25do = 1.25(20) = 25 mm

1
𝑁𝑡 1 5 2.207
Tube bundle diameter, Db = 𝑑𝑜 (𝐾 )𝑛1 = (20) (0.249) = 77.86 𝑚𝑚
1

Use a split-ring floating head type:

Diameter clearance = 55 mm

Shell diameter, Ds = 77.86 + 55 = 132.86 mm

Tube-side Coefficient

68+48
Mean Temperature, Tm = = 58℃
2

𝐷𝑖2 162
Cross sectional area = 𝜋 =𝜋 = 201.1 𝑚𝑚
4 4

Tubes per pass = 5/2 = 2.5

Tube side flowrate = 2.5(201.1 x 10-6) = 5.03 x 10-4 m2

𝑤𝑎𝑡𝑒𝑟 𝑓𝑙𝑜𝑤 0.676

Mass velocity = = = 1343.94 𝑘𝑔/𝑠𝑚2
𝑓𝑙𝑜𝑤𝑟𝑎𝑡𝑒 5.03 ×10−4

𝑚𝑎𝑠𝑠 𝑣𝑒𝑙𝑜𝑐𝑖𝑡𝑦 1343.94

Linear velocity, Ut = 𝑤𝑎𝑡𝑒𝑟 𝑑𝑒𝑛𝑠𝑖𝑡𝑦 = = 1.366 𝑚/𝑠
984.2

𝐶𝑝 µ (4.2×103 )(4.81×10−4 )
Prandlt number, Pr = = = 3.11
𝑘𝑓 0.649

𝜌𝑢𝑡 𝑑𝑖 (984.2)(1.366)(16×10−3 )
Reynolds number, Re = = = 4.4721 × 104
µ (4.81×10−4 )

µ
By neglecting µ and from graph jh = 4.0 x 10-3
𝑤

𝑘𝑓 0.33 µ
Tube-side heat transfer coefficient, hi = 𝑗 𝑅𝑒𝑃𝑟 (µ )
𝑑𝑖 ℎ 𝑤

0.649
= (16×10−3 )(4.0x10-3)(4.4721x104)(3.11)0.33

= 10551.33 W/m2.℃

Shell-side Coefficient

By taking 0.2 baffle spacing shell diameter for higher heat transfer coefficient,

Baffle spacing, Ib = 0.2Ds = 0.2(132.86) = 26.57 mm

Tube pitch, Pt = 1.25do = 1.25(20) = 25mm

(𝑃𝑡−𝑑𝑜 )𝐷𝑠 𝑙𝑏 (25−20)(132.86)(26.57×10−6 )

Cross-flow area, As = = = 7.06 × 10−4 𝑚2
𝑃𝑡 25

𝑊𝑠 1723 1
Shell-side mass velocity, Gs = = 3600 (7.06×10−4) = 677.92 𝑘𝑔/𝑠𝑚2
𝐴𝑠
 𝐺𝑠 677.92
Shell-side linear velocity, Us = = 1272.9 = 0.53 𝑚/𝑠
𝑝

1.10 1.10
Shell equivalent diameter, de = (Pt2 – 0.917do2) = (252 – 0.917(202)) = 14.2 m
𝑑𝑜 20

4+28
Mean shell-side water temperature, Tavg = = 16℃
2

𝜌𝑈𝑠 𝑑𝑒 (984.2)(0.53)(14.2×10−3 )
Reynolds Number, Re = µ
= 4.81×10−4
= 15399

𝐶𝑝 µ (4.2×103 )(4.81×10−4 )
Prandlt Number, Pr = 𝑘𝑓
= 0.649
= 3.11

1
ℎ𝑠 𝑑𝑒 µ
Nusselt Number, Nu = 𝑘𝑓
= 𝑗ℎ 𝑅𝑒𝑃𝑟 3 (µ )0.14
𝑤

µ
By neglecting µ and taking baffle cut as 25%, from graph jh = 4.6 x 10-3
𝑤

1
(0.649)(4.6×10−3 )(15399)(3.11)3
hs = = 4725.63
(14.2×10−3 )

Mean temperature difference across all resistance = 58 – 16 = 42℃

𝑈 800
Mean temperature difference across water = ℎ (∆𝑇) = 4725.63 (42) = 7.11℃
𝑜

Mean tube wall temperature = 58 – 7.11 = 50.89℃

Overall Coefficient

Assumption:

1) Thermal Conductivity of cupro-nickel alloys, Kw = 50 W/m2.℃

2) Take fouling factors as 0.0003 for river water
20
1 1 1 (20×10−3 )ln ( ) 20 1 20 1
16
= 4725.63 + 3000 + + 16 (3000) + 16 (10551.33) = 0.0011
𝑈 2×50

U = 909.09 W/m2.℃

Pressure Drop

Tube-side
From graph, jh = 4.0 x 10-3 by taking Re = 44721 and neglect the viscosity term.

𝐿 µ 𝜌𝑈𝑡 2
∆Pt = Np[8𝑗ℎ (𝑑𝑖 ) (µ )−𝑚 + 2.5] ( )
𝑤 2

4.83×103 984.2(1.3662 )
= 2 [8(4.0 × 10−3 ) ( ) + 2.5] ( )
16 2

= 2.233 x 104 N/m2

Shell-side

From graph, jh = 4.6 x 10-3 by taking Re = 15399

𝐷𝑠 𝐿 𝜌𝑈𝑠 2
∆Ps = 8𝑗ℎ (𝑑𝑒) (𝑙𝑏) ( )
2

132.86 4.83×103 984.2(0.532 )

= 8 (4.6 × 10−3 [( )( )( )])
14.2 26.57 2

= 8.651 x 103 N/m2

 PRESSURE VESSEL DESIGN (INTERNAL PRESSURE)

2.0 CALCULATION ON FERMENTER, R-2

ρ inlet = (28000/270200)(1560) + (35600/270200)(1429) + (134000/270200)(1160) +

(33200/270200)(1770) + (32000/270200)(1080) + (7400/270200)(1410)

= 1309.22 kg/m3

Volume required of fermenter for a fermentation of 70 hours is calculated using

equation, V= m/ρ

V = 270200/1309.22 = 206.38 m3

There is one main fermenter used in the production,

Diameter of fermenter calculated using this formula:

𝜋
V = 4 𝐷𝑖2 ℎ , 2Di = h

𝜋
206 = 𝐷𝑖2 (2𝐷𝑖 )
4

Di = 5.08 m = 16.67 ft

h = 2Di = 2(5.08) = 10.16 m = 33.34 ft

For ellipsoidal head and bottom, the ratio of the ellipsoidal head is 2:1,

he = Di/4 = 16.67/4 = 4.17 ft

For cylindrical shell height,

hc = h – 2(he) = 33.34 – 2(4.17) = 25 ft

2.1 DESIGN PRESSURE

Since the operating pressure of the reactor (2 bar) is greater than atmospheric pressure
(Po > Patm), the reactor designed under internal pressure. Firstly, the design pressure is
determined for each section of the vessel, using equation:
 𝑃𝐷 = 𝑃𝑜 + 0.433(ℎ)

where, h = height of each part

Po = operating pressure (gage value)

Hence,

1) PD, top = 29.01 + 0.433(4.17) = 30.82 psi

2) PD, bottom = 29.01 + 0.433(25 + 4.17 + 4.17) = 43.45 psi
3) PD, shell = 29.01 + 0.433(25 + 4.17) = 41.64 psi

A safety factor about 10% is added to design pressure for safety purpose. So, the new
design pressure is PJ = 1.10PD and is tabulated in the Table 8.6.

Hydrostatic pressure,

PH = 0.433(h)

PH, top = 0.433(4.17) = 1.81 psi

PH, bottom = 0.433(25 + 4.17 +4.17) = 14.44 psi

PH, shell = 0.433(25 + 4.17) = 12.63 psi

2.2 MINIMUM WALL THICKNESS

The wall thickness formulas for each part of the column are from ASME Code UG-32
part (D) and UG-27 part (c).

Ellipsoidal Top Head,

𝑃𝐷 (30.82)(200)
𝑡= = = 0.181 𝑖𝑛𝑐ℎ = 4.597 𝑚𝑚
2𝑆𝐸 − 0.2𝑃 2(20000)(0.85) − 0.2(30.82)

Ellipsoidal Bottom Head,

𝑃𝐷 (43.45)(200)
𝑡= = = 0.256 𝑖𝑛𝑐ℎ = 6.502 𝑚𝑚
2𝑆𝐸 − 0.2𝑃 2(20000)(0.85) − 0.2(43.45)

Cylindrical Shell,

i. Circumferential stress
 𝑃𝑅 (41.64)(100)
𝑡= = = 0.245 𝑖𝑛𝑐ℎ = 6.233 𝑚𝑚
𝑆𝐸 − 0.6𝑃 (20000)(0.85) − 0.6(41.64)

ii. Longitudinal stress

𝑃𝑅 (41.64)(100)
𝑡= = = 0.122 𝑖𝑛𝑐ℎ = 3.099 𝑚𝑚
2𝑆𝐸 + 0.4𝑃 2(20000)(0.85) + 0.4(41.64)

Considering the corrosion allowance of 4 mm, thus the new thickness for each part of
the vessel:

Ellipsoidal Top Head,

tnew = 4 mm + 4.597 mm = 8.597 mm

Ellipsoidal Bottom Head,

tnew = 4 mm + 6.502 mm = 10.502 mm

Cylindrical Shell,

i. Circumferential stress

tnew = 4 mm + 6.233 mm = 10.233 mm

ii. Longitudinal stress

tnew = 4 mm + 3.099 mm = 7.099 mm

2.3 MAXIMUM ALLOWABLE WORKING PRESSURE

In any form of pressure vessel, the maximum permissible working pressure, MAWP is
calculated by measuring each component under internal pressure. The internal design
pressure formula for each part of the column can be found in Part UG-32 of the ASME
Code (D) and part UG-27 (c).

Ellipsoidal Top Head,

2𝑆𝐸𝑡 2(20000)(0.85)(0.181)
Internal pressure, 𝑃 = 𝐷+0.2𝑡 = 200+0.2(0.181)
= 30.764 𝑝𝑠𝑖

Static head, PH = 1.81 psi

MAWPvessel = internal – static head = 28.954 psi

Ellipsoidal Bottom Head,

2𝑆𝐸𝑡 2(20000)(0.85)(0.256)
Internal pressure, 𝑃 = 𝐷+0.2𝑡 = = 43.509 𝑝𝑠𝑖
200+0.2(0.256)

Static head, PH = 14.44 psi

MAWPvessel = internal – static head = 29.069 psi

Cylindrical Shell,

i. Circumferential stress

𝑆𝐸𝑡 (20000)(0.85)(0.245)
Internal pressure, 𝑃 = 𝑅+0.6𝑡 = = 41.589 𝑝𝑠𝑖
100+0.6(0.245)

Static head, PH = 12.63 psi

MAWPvessel = internal – static head = 28.959 psi

ii. Longitudinal stress

2𝑆𝐸𝑡 2(20000)(0.85)(0.122)
Internal pressure, 𝑃 = = = 41.500 𝑝𝑠𝑖
𝑅−0.4𝑡 100−0.4(0.122)

Static head, PH = 12.63 psi

MAWPvessel = internal – static head = 28.870 psi

The internal pressure and MAWPvessel value are obtained, and the result is displayed
in the table 8.7

2.4 COMBINED LOADING ANALYSIS

In addition to pressures, the pressure vessels are always exposed to other loads and must
be designed to withstand the maximum load combination without failure.

Primary Stress

Po = 29.01 psi = 0.2 N/mm2

Di = 200 inch = 5080 mm

tnew, nominal = 6.30 mm

Calculation of Longitudinal Stresses (𝜎𝐿) and Circumferential Stresses (𝜎𝐻)

𝑃𝐷𝑖 (0.2)(5080)
𝜎𝐿 = = = 40.317 𝑁 𝑚𝑚−2
4𝑡 4(6.30)

𝑃𝐷𝑖 (0.2)(5080)
𝜎𝐻 = = = 80.635 𝑁 𝑚𝑚−2
2𝑡 2(6.30)

Calculation of Direct Stresses (𝜎W)

Direct stress (𝜎W) can be calculated as follows,

𝑊
𝜎W = 𝜋(𝐷𝑖+𝑡)𝑡 , W = total weight of the shell

Direct stress of this due to the weight of vessel, its content and attachment. Assumption
for factor 𝐶𝑣 = 1.15 for fermenter.

Hv = 10.16 m

t = 0.0063 m

Dm = Di + t = 5.08 + 0.0063 = 5.0863 m

W = 240𝐶𝑣𝐷𝑚(𝐻𝑣 + 0.8𝐷𝑚)t

= 240(1.15)(5.0863)(10.16 + 0.8 x 5.0863)(0.0063)

= 125.84 N

(125.84)
𝜎W = 𝜋(5080+6.3)(6.3) = 0.00125 𝑁 𝑚𝑚−2

Calculation of Bending Stress, (𝜎b)

Bending stress is neglected because earthquake in Malaysia does not occur. The vessel
also does not have any additional equipment or attachment on it. Therefore, the bending
stress can be neglected.

Calculation of Torsional Shear Stress, τ

Torsional shear stress, 𝜏 resulting from torque caused by load offset from the vessel
axis. These loads will normally be small and can be neglected in preliminary vessel
design
Therefore, 𝜏 = 0

Analysis of Principle Stress

1
σ1 = [(𝜎ℎ + 𝜎𝑧 + √(𝜎ℎ −𝜎𝑧 ) + 4𝜋 2 )] = 𝜎ℎ
2

1
σ2 = [(𝜎ℎ + 𝜎𝑧 − √(𝜎ℎ −𝜎𝑧 ) + 4𝜋 2 )] = 𝜎𝑧
2

σ3 = 0 (𝑁𝑒𝑔𝑙𝑖𝑔𝑖𝑏𝑙𝑒 𝑓𝑜𝑟 𝑡ℎ𝑖𝑛 𝑣𝑒𝑠𝑠𝑒𝑙)

With bending stress and torsional shear stress = 0

Total longitudinal stresses for upwind and downwind can be calculated as follow,

Upwind, 𝜎𝑧 = 𝜎𝐿 + 𝜎𝑊 + 𝜎𝑏 = 35.780 + 0.00125 + 0 = 35.781 𝑁 𝑚𝑚−2

Downwind, 𝜎𝑧 = 𝜎𝐿 + 𝜎𝑊 + 𝜎𝑏 = 35.780 + 0.00125 − 0 = 35.781 𝑁 𝑚𝑚−2

(Δ𝜎)𝑚𝑎𝑥 = 𝜎𝐻 − 𝜎𝑧,𝑢𝑝𝑤𝑖𝑛𝑑 = 80.635 − 35.781 = 44.854 𝑁 𝑚𝑚−2

(Δ𝜎)𝑚𝑎𝑥 = 𝜎𝐻 − 𝜎𝑧,𝑑𝑜𝑤𝑛𝑤𝑖𝑛𝑑 = 80.635 − 35.781 = 44.854 𝑁 𝑚𝑚−2

Maximum allowable stress intensity (Δ𝜎)𝑚𝑎𝑥 is at 44.854 N/mm2

From Chemical Engineering Design, Sinnott, R. & Towler, G. (2009), maximum

allowable stress for stainless steel grade 316 is 137.89515 N/mm2.

Therefore, (Δ𝜎)𝑚𝑎𝑥 < S

The thickness of the vessel's wall is sufficient to ensure that the maximum stress
intensity at any point does not exceed the design stress for the construction material.
Hence, the design operating condition is safe.

2.5 ANALYSIS OF ELASTIC STABILITY

Under conditions where the resultant axial stress, 𝜎𝑧 due to the combined loading is
compressive, the vessel may fail by elastic instability (buckling). For steels at T = 25℃,
the Young’s Modulus, E=200,000 N/mm2 and the poisson’s ratio is 0.3, the buckling
equation is as follow, where 𝜎𝑐 is critical buckling stress.
 𝑡 6.30
𝜎𝑐 = 0.6𝐸 ( ) = 0.6(20000) ( ) = 14.88 𝑁 𝑚𝑚−2
𝐷𝑖 5080

𝜎𝑐𝑜𝑚𝑝𝑟𝑒𝑠𝑠𝑖𝑣𝑒 = 𝜎𝑏 + 𝜎𝑊 = 0 + 0.00125 = 0.00125 𝑁 𝑚𝑚−2

Hence,

(Δ𝜎)𝑐𝑜𝑚𝑝𝑟𝑒𝑠𝑠𝑖𝑣𝑒 ≤ 𝜎𝑐

The compressive stresses, 𝜎𝑐 is below critical buckling stress, hence the design is safe.

2.6 SKIRT THICKNESS

Assume the skirt diameter (Ds) is equal to the inner wall diameter (Di) and the skirt
thickness is equal to the nominal thickness (ts = tnominal = 0.405 in =10 mm).

4(3.36×108 )
Bending stress in the skirt, 𝜎𝑏𝑠 = 𝜋(5080.0126+10)(10)(5080.0126) = 1.6545 𝑁 𝑚𝑚−2

Dead weight of vessel:

Cv = 1.5

Mineral wool density = 130 𝑘𝑔/𝑚3

𝑊𝑣 = 𝐶𝑣𝜋𝜌𝑚𝐷𝑚𝑔(𝐻𝑣 + 0.8𝐷𝑚)𝑡 × 10−3

= 1.5π(130)(5.0863)(9.81)(10.16 + 0.8 x 5.0863)(6.30) x 10-3

= 2740.13 N

2740.13
Dead weight stress in the skirt, 𝜎𝑤𝑠 = = 0.017 𝑁 𝑚𝑚−2
𝜋(5080+10)(10)

Therefore, resultant stresses

Maximum 𝜎𝑠 (tensile) = 𝜎𝑏𝑠 − 𝜎𝑤𝑠 = 1.65 − 0.017 = 1.6375 𝑁/𝑚𝑚2

𝜎𝑠 (compressive) = 𝜎𝑏𝑠 + 𝜎𝑤𝑠 = 1.65 + 0.017 = 1.6715 𝑁/𝑚𝑚2

The skirt thickness under worst condition of wind and dead weight loading should not
exceed the following design criteria

𝜎𝑠 (tensile) < 𝑓𝑠𝐽𝑠𝑖𝑛𝜃𝑠

𝜎𝑠 (compressive) < 0.125E (𝑡𝑠/𝐷𝑠)𝑠𝑖𝑛𝜃s
Assumptions:

• Straight cylindrical skirt (carbon steel)

• Maximum allowable stress for skirt material at Tamb = 20℃, 𝑓𝑠 = 135 𝑁/𝑚𝑚2
• Weld joint factor, J = 0.85
• Base angle, 𝜃𝑠 = 90°
• Young’s modulus, E = 200000

Thus,

𝑓𝑠𝐽𝑠𝑖𝑛𝜃𝑠 = 135(0.85) sin(90°) = 114.75 𝑁/𝑚𝑚2

10
0.125E(𝑡𝑠/𝐷𝑠)𝑠𝑖𝑛𝜃𝑠 = 0.125(200000) ( ) sin(90°) = 49.213 𝑁/𝑚𝑚2
5080

2.7 BASE RING AND ANCHOR BOLT DESIGN

The anchor bolt is assumed to share the overturning load equally and the bolt area
required:

1 4𝑀𝑠
𝐴𝑏 = ( − 𝑊)
𝑁𝑏 𝑓𝑏 𝐷𝑏

With, Ab = area of one bolt at the root of the thread, 𝑚𝑚2

Nb = number of bolts

fb = maximum allowable bolt stress, N mm-2

Ms = bending moment at the base, Nm

W = weight of the vessel, N

Db = bolt circle diameter, m

Assumption:

• Bolt circle diameter, Db = 3.2 m = 3200 mm

• Circumference of bolt circle = 3200 𝜋 𝑚𝑚 = 10053.09 mm
• Maximum allowable bolt stress, 𝑓𝑏 = 125 𝑁/𝑚𝑚2
 • Number of bolts as 16
• Bolt spacing > 600 mm

𝐶𝑖𝑟𝑐𝑢𝑚𝑓𝑒𝑟𝑒𝑛𝑐𝑒 𝑜𝑓 𝑏𝑜𝑙𝑡 𝑐𝑖𝑟𝑐𝑙𝑒 10053.09

Bolt spacing = = = 628.218 𝑚𝑚
𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑏𝑜𝑙𝑡𝑠 16

Bolt spacing > 600 mm, therefore the bolt spacing is satisfactory.

Thickness skirt = 25 mm

Wind loading = 0.00128 N 𝑚𝑚−2 (1280 N/𝑚2)

The bending moment at the skirt base,

𝑤𝑥 2
𝑀𝑠 =
2

Where, w = wind loading, N mm-2

x = height of vessel, mm

(0.00128)(10160)2
Ms = = 66064 𝑁 𝑚𝑚
2

The weight of the vessel, W = 12761.46 N

1 4𝑀𝑠
𝐴𝑏 = ( − 𝑊)
𝑁𝑏 𝑓𝑏 𝐷𝑏

1 4(66064)
𝐴𝑏 = ( − 12761.46) = 34.909 𝑚𝑚2
(16)(125) 3.2

Use M24 bolts (BS44190:1967) root area = 353 𝑚𝑚2 (Towler & Sinnott 2013)

Total compressive load on the base ring per unit length, F𝑏

4𝑀 𝑊
Fb = [𝜋𝐷 𝑠2 + 𝜋𝐷 ]
𝑠 𝑠

4(66064) 12761.46
Fb = [𝜋(5.080)2 + 𝜋(5.080) ]

Fb = 4059.10 N m-1

Taking the bearing pressure as 3.5 N/𝑚𝑚2, the minimum width of the base rings, 𝐿b

𝑓𝑏 1
𝐿𝑏 = × 3
𝑓𝑐 10
 4059.10 1
𝐿𝑏 = × 3 = 1.16 𝑚𝑚
3.5 10

The required thickness for thickness base ring found by treating the rings as cantilever
beam:

3𝑓𝑐′
𝑡𝑏 = 𝐿𝑟 √
𝑓𝑟

Where, tb = base ring thickness, mm

Lr = The distance from the edge of the skirt to the outer edge of the ring, mm

fc’ = Actual bearing pressure, N/mm2

fr = Allowable design stress in the ring of material, fr = 140 N/mm2

Take Lr = 76 mm,

Actual width required = Lr + ts + 50 mm

= 76 + 10+ 50 mm

= 136 mm

𝑓𝑏 4059.10
𝑓𝑐 ′ = 3
= = 0.03 𝑁/𝑚𝑚2
136 × 10 136 × 103

3𝑓 ′ 3(0.03)
Base ring thickness, 𝑡𝑏 = 𝐿𝑟 √ 𝑓 𝑐 = 76√ = 1.93 𝑚𝑚
𝑟 140
 PRESSURE VESSEL DESIGN (EXTERNAL PRESSURE)

2.8 CALCULATION ON EXTRACTION COLUMN, E-1

Calculation height of each part:

Top head (Ellipsoidal 2:1)

Htop = D/4

Htop = 34.81/4 = 8.7ft

Bottom head (Hemispherical)

Hbtm = L(radius of Column)

Hbtm = 17.41ft

Shell (Cylindrical)

Hs = 54 (8.7+17.41)

Hs = 27.89ft

2.9 CALCULATION OF DESIGN PRESSURE, PD, MINIMUM WALL

THICKNESS, tmin AND MAWPvessel

Hydrostatic pressure

Ph = 0.433h

Phtop = 0.433(8.7) = 3.77psi

Phbtm = 0.433(17.41) = 7.54psi

Phs = 0.433(27.89) = 12.08psi

Design pressure, PD:

PD = Po + Ph

PDtop = 24.66 + 3.77 = 28.43psi

PDbtm = 24.66 + 7.54 = 32.20psi

PDs = 24.66 + 36.74 = 61.40psi

Minimum wall thickness, tmin:

Ellipsoidal head

𝑃𝐷
𝑡=
2𝑆𝐸 − 0.2𝑃

(28.43)(417.72)
𝑡=
2(15700)(0.85) − 0.2(28.43)

𝑡 = 0.4450𝑖𝑛

Hemispherical head

𝑃𝐿
𝑡=
2𝑆𝐸 − 0.2𝑃

(32.20)(209.06)
𝑡=
2(15700)(0.85) − 0.2(32.20)

𝑡 = 0.2523𝑖𝑛
Cylindrical shell

Circumference stress

Condition: t<R/2

𝑃𝑅
𝑡 = 𝑆𝐸−0.6𝑃

(61.40)(209.06)
𝑡 = (15700)(0.85)−0.6(61.40)

𝑡 = 0.9645𝑖𝑛

Longitudinal stress

𝑃𝑅
𝑡 = 𝑆𝐸+0.4𝑃

(61.40)(209.06)
𝑡 = (15700)(0.85)+0.4(61.40)

𝑡 = 0.9601𝑖𝑛

MAWPEllipsoidal

2𝑆𝐸𝑡
𝑃 = 𝐷+0.2𝑡

2(15700)(0.85)(0.4450)
𝑃= 417.72+0.2(0.4450)

𝑃 = 28.43𝑝𝑠𝑖

MAWPHemispherical

2𝑆𝐸𝑡
𝑃 = 𝐿+0.2𝑡
 2(15700)(0.85)(0.2523)
𝑃= 209.06+0.2(0.2523)

𝑃 = 32.20𝑝𝑠𝑖

MAWPShell

Circumference stress

𝑆𝐸𝑡
𝑃 = 𝑅+0.6𝑡

(15700)(0.85)(0.9645)
𝑃= 209.06+0.6(0.9645)

𝑃 = 61.40𝑝𝑠𝑖

Longitudinal stress

2𝑆𝐸𝑡
𝑃 = 𝑅−0.4𝑡

2(15700)(0.85)(0.9601)
𝑃= 209.06−0.4(0.9601)

𝑃 = 122.80𝑝𝑠𝑖

2.10 THICKNESS UNIFORMITY AND NOMINAL THICKNESS

CORRECTION
 (Source: Champak Steel & Engg.co, 2017)

tmin,new = tnominal -CA

=0.9843-0.15748

=0.8268in

New MAWP calculation

MAWPEllipsoidal

2𝑆𝐸𝑡
𝑃 = 𝐷+0.2𝑡
 2(15700)(0.85)(0.8268)
𝑃= 417.72+0.2(0.8268)

𝑃 = 52.80𝑝𝑠𝑖

MAWPHemispherical

2𝑆𝐸𝑡
𝑃 = 𝐿+0.2𝑡

2(15700)(0.85)(0.8268)
𝑃= 209.06+0.2(0.8268)

𝑃 = 105.47𝑝𝑠𝑖

MAWPShell

Circumference stress

𝑆𝐸𝑡
𝑃 = 𝑅+0.6𝑡

(15700)(0.85)(0.8268)
𝑃= 209.06+0.6(0.8268)

𝑃 = 52.65𝑝𝑠𝑖

Longitudinal stress

2𝑆𝐸𝑡
𝑃 = 𝑅−0.4𝑡

2(15700)0(0.85)(0.8268)
𝑃= 209.06−0.4(0.8268)

𝑃 = 105.72𝑝𝑠𝑖

2.11 COMBINE LOADING ANALYSIS

Calculation of Circumferential Stresses, (σH)

𝑃𝐷𝑖
σ𝐻 = 2𝑡

0.36(10610.1)
= 2(21)

= 90.90𝑁𝑚𝑚−2

Calculation of Longitudinal Stresses, (σL)

𝑃𝐷𝑖
σ𝐿 = 4𝑡

0.36(10610.1)
= 4(21)

= 45.47𝑁𝑚𝑚−2

Calculation of Direct Stresses, (σW)

This is subjected to direct stress as a result of the vessel's weight, contents, and
attachments. Assumption for factor Cv = 1.15 for extraction column.

Hv=16.46m

t =0.021m

Dm =Di + t

=10.61 + 0.021

= 10.631m

W = 240CvDm(Hv + 0.8Dm)t
 = 240(1.15)(10.631)[16.46 + 0.8(10.631)](0.021)

= 1499.20N

𝑊
σ𝑊 = 𝜋(𝐷 +𝑡)𝑡
𝑖

1499.20
σ𝑊 = 𝜋(10610.1+21)(21)

σ𝑊 = 0.0021𝑁/𝑚𝑚−2

Calculation of bending stress, (σb)

𝐷𝑒𝑓𝑓 = 𝐷𝑖 + 2𝑡

𝐷𝑒𝑓𝑓 = 10610.1 + 2(0.021)

𝐷𝑒𝑓𝑓 = 10610.14𝑚𝑚

W = PwDeff

Pw = 0.00128N/mm-2

W = 0.00128(10610.14)

W = 13.58

𝑊𝐻𝑣 2
𝑀𝑥 = 2

13.58(16460)2
𝑀𝑥 = 2

𝑀𝑥 = 1.84 𝑥 109 𝑁𝑚𝑚

 𝜋
𝑙𝑣 = 64 (𝐷𝑜 4 − 𝐷𝑖 4 )

𝜋
𝑙𝑣 = (10631.14 − 10610.14 )
64

𝑙𝑣 = 4.94𝑥1012 𝑛𝑚4

𝑀 𝐷𝑖
σb = ( 2 + 1)
𝑙𝑣

1.84𝑥109 10610.1
σb = ( + 1)
4.94𝑥1012 2

σb = 1.98𝑁𝑚𝑚−2

Upwinnd: σz = σL + σW + σb = 47.45Nmm-2

Downwind: σz = σL + σW – σb = 45.47Nmm-2

(Δσ)max = σH - σzUpwind = 90.94 – 47.45 = 43.49Nmm-2

(Δσ)max = σH - σzDownwind = 90.94 – 45.47 = 45.47Nmm-2

2.12 STRESS IN THE SKIRT CALCULATION

Assume the skirt diameter (Ds) is the same as the inner wall diameter (Di) and the skirt
thickness is equal to the nominal thickness (tnominal = 0.9843in)

4(1.84𝑥109 )
Bending stress in the skirt, 𝜎𝑏𝑠 = 𝜋(10610.1+25)(25)(10610.1)

𝜎𝑏𝑠 = 0.83 𝑁𝑚𝑚−2

ρm = 130kg/m3

𝑊𝑣 = 𝐶𝑣 𝜋𝜌𝑚 𝐷𝑚 𝑔(𝐻𝑣 + 0.8𝐷𝑚 )𝑡 𝑥 10−3

 𝑊𝑣 = 1.5𝜋(130)(10.631)(9.81)[16.46 + 0.8(10.631)](21)𝑥10−3

𝑊𝑣 = 33494.62𝑁

33494.62
Dead weight stress in the skirt, 𝜎𝑤𝑠 = 𝜋(10610.1+25)(25)

𝜎𝑤𝑠 = 0.04 𝑁𝑚𝑚−2

Therefore, resultant stresses:

Maximum σs (tensile) = σbs – σws = 0.83 – 0.04 = 0.79Nmm-2

σs (compressive) = σbs + σws = 0.83 + 0.04 = 0.87Nmm-2

σs (tensile) < fsJsinƟs

σs (compressive) < 0.125E (ts/Ds)sinƟs

Assumption:

• Straight cylindrical skirt (carbon steel)

• Maximum allowable stress for skirt material at Tamb =20°C, fs = 135Nmm-2
• Weld joint factor, J = 0.85
• Base angle, Ɵs = 90°
• Young’s modulus, E = 200000

fsJsinƟs = 135(0.85)(sin90°)

= 114.75Nmm-2

25
0.125E (ts/Ds)sinƟs = 0.125(200000) (10610.1) sin (90°)
 = 58.91Nmm-2

2.13 BASE RING AND ANCHOR BOLT DESIGN CALCULATION

The anchor bolt is assumed to share the overturning load equally and the bolt area
required:

1 4𝑀𝑠
𝐴𝑏 = ( − 𝑊)
𝑁𝑏 𝑓𝑏 𝐷𝑏

Where,

Ab = area of one bolt at the root of the thread, mm2

Fb = maximum allowable bolt stress, Nmm-2

Ms = bending moment at the base, Nm

W = weight moment at the base, Nm

Db = bolt circle diameter, m

Assumption:

• Db = 3.2m

• Circumference of bolt circle =3200πmm

• fb = 125Nmm-2

• Number of bolts is 16

• Bolt spacing > 600mm

 𝐶𝑖𝑟𝑐𝑢𝑚𝑓𝑒𝑟𝑒𝑛𝑐𝑒 𝑜𝑓 𝑏𝑜𝑙𝑡 𝑐𝑖𝑟𝑐𝑙𝑒
𝐵𝑜𝑙𝑡 𝑠𝑝𝑎𝑐𝑖𝑛𝑔 =
𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑏𝑜𝑙𝑡𝑠

3200𝜋
𝐵𝑜𝑙𝑡 𝑠𝑝𝑎𝑐𝑖𝑛𝑔 =
16

𝐵𝑜𝑙𝑡 𝑠𝑝𝑎𝑐𝑖𝑛𝑔 = 628.218𝑚𝑚

Bolt spacing, 628.218mm>600mm, thus the bolt spacing is acceptable.

The bending moment at the skirt base, (Ms)

𝑤𝑥 2
𝑀𝑠 = 2

Where,

w = wind loading, Nmm-1

x = height of vessel, mm

Wind loading, w = 0.00128Nmm-2

0(.001280)(16460)2
𝑀𝑠 = 2

𝑀𝑠 = 173396.22𝑁𝑚𝑚

The weight of the vessel, W = 33494.62N

Therefore,

1 4𝑀
𝐴𝑏 = ( 𝐷 𝑠 − 𝑊)
𝑁𝑏 𝑓𝑏 𝑏
 1 4(173396.22)
𝐴𝑏 = (16𝑥125)
[ − 33494.62]
3.2

𝐴𝑏 = 91.63𝑚𝑚2

Use M24 bolts (BS44190:1967) root area = 353mm2 (Towler & Sinnott, 2013)

Total compressive load on the base ring per unit length, Fb

4𝑀 𝑊
𝐹𝑏 = [𝜋𝐷 22 + ]
𝑠 𝜋𝐷𝑠

4(173396.22) 33494.62
𝐹𝑏 = [ + ]
𝜋(10.61)2 𝜋(10.61)

𝐹𝑏 = 2966.06𝑁𝑚−1

Taking the bearing pressure,fc as 3.5Nmm-2, the minimum width of the base rings, Lb

𝑓 1
𝐿𝑏 = [ 𝑓𝑏 𝑥 103 ]
𝑐

2966.06
𝐿𝑏 = 3.5𝑥103

𝐿𝑏 = 0.85𝑚𝑚

The required thickness for thickness base ring found by treating the rings as cantilever
beam:

3𝑓 ′ 𝑐
𝑡𝑏 = 𝐿𝑟 √
𝑓𝑟

Where,

tb = base ring thickness, mm

 Lr = the distance from the edge of the skirt to the outher edge of the ring, mm

f’c = actual bearing pressure, Nmm-2

fr = allowable design stress in the ring of the material = 140Nmm-2

Lr = 76mm

Actual width required = Lr + ts + 50mm

= 76 + 10 +50

= 136mm

𝑓𝑏
𝑓𝑐 = 136𝑥103

2966.06
𝑓𝑐 = 136𝑥103

𝑓𝑐 = 0.02𝑁𝑚𝑚−2

3𝑓 ′ 𝑐
Base ring thickness, 𝑡𝑏 = 𝐿𝑟 √ 𝑓𝑟

3(0.02)
𝑡𝑏 = 76√ 140

𝑡𝑏 = 1.57𝑚𝑚
 SAFETY DATA SHEET Page: 1 of 5

Penicillin G (potassium salt) Revision: 10/16/2017

according to Regulation (EC) No. 1907/2006 as amended by (EC) No. 1272/2008

Section 1. Identification of the Substance/Mixture and of the Company/Undertaking

1.1 Product Code: 21615
Product Name: Penicillin G (potassium salt)
Synonyms: (2S,5R,6R)-3,3-dimethyl-7-oxo-6-[(2-phenylacetyl)amino]-4-thia-1-azabicyclo[3.2.0]heptane-2-ca
rboxylic acid, monopotassium salt; Benzylpenicillin; NSC 131815; PEN-G;

1.2 Relevant identified uses of the substance or mixture and uses advised against:
Relevant identified uses: For research use only, not for human or veterinary use.

1.3 Details of the Supplier of the Safety Data Sheet:

Company Name: Cayman Chemical Company
1180 E. Ellsworth Rd.
Ann Arbor, MI 48108
Web site address: www.caymanchem.com
Information: Cayman Chemical Company +1 (734)971-3335
1.4 Emergency telephone number:
Emergency Contact: CHEMTREC Within USA and Canada: +1 (800)424-9300
CHEMTREC Outside USA and Canada: +1 (703)527-3887

Section 2. Hazards Identification

2.1 Classification of the Substance or Mixture:
Skin Sensitization, Category 1
2.2 Label Elements:

GHS Signal Word: Warning

GHS Hazard Phrases:
H317: May cause an allergic skin reaction.
GHS Precaution Phrases:
P261: Avoid breathing {dust/fume/gas/mist/vapors/spray}.
P272: Contaminated work clothing should not be allowed out of the workplace.
P280: Wear {protective gloves/protective clothing/eye protection/face protection}.
GHS Response Phrases:
P302+352: IF ON SKIN: Wash with plenty of soap and water.
P333+313: If skin irritation or rash occurs, seek medical advice/attention.
P362+364: Take off contaminated clothing and wash it before reuse.
GHS Storage and Disposal Phrases:
Please refer to Section 7 for Storage and Section 13 for Disposal information.
2.3 Adverse Human Health Material may be irritating to the mucous membranes and upper respiratory tract.
Effects and Symptoms: May be harmful by inhalation, ingestion, or skin absorption.
May cause an allergic skin reaction.
May cause eye, skin, or respiratory system irritation.
To the best of our knowledge, the toxicological properties have not been thoroughly investigated.

Multi-region format
 SAFETY DATA SHEET Page: 2 of 5

Penicillin G (potassium salt) Revision: 10/16/2017

Section 3. Composition/Information on Ingredients

CAS # / Hazardous Components (Chemical Name)/ Concentration EC No./ GHS Classification
RTECS # REACH Registration No. EC Index No.
113-98-4 4-Thia-1-azabicyclo3.2.0heptane-2-carboxylicacid, 100.0 % 204-038-0 Skin Sens. 1: H317
XH9700000 3,3-dimethyl-7-oxo-6-(phenylacetyl)amino-2S-(2.al NA

Section 4. First Aid Measures

4.1 Description of First Aid
Measures:
In Case of Inhalation: Remove to fresh air. If not breathing, give artificial respiration or give oxygen by trained personnel.
Get immediate medical attention.
In Case of Skin Contact: Immediately wash skin with soap and plenty of water for at least 15 minutes. Remove contaminated
clothing. Get medical attention if symptoms occur. Wash clothing before reuse.
In Case of Eye Contact: Hold eyelids apart and flush eyes with plenty of water for at least 15 minutes. Have eyes examined
and tested by medical personnel.
In Case of Ingestion: Wash out mouth with water provided person is conscious. Never give anything by mouth to an
unconscious person. Get medical attention. Do NOT induce vomiting unless directed to do so by
medical personnel.

Section 5. Fire Fighting Measures

5.1 Suitable Extinguishing Use alcohol-resistant foam, carbon dioxide, water, or dry chemical spray.
Media: Use water spray to cool fire-exposed containers.
Unsuitable Extinguishing A solid water stream may be inefficient.
Media:
5.2 Flammable Properties andNo data available.
Hazards:
No data available.
Flash Pt: No data.
Explosive Limits: LEL: No data. UEL: No data.
Autoignition Pt: No data.
5.3 Fire Fighting Instructions: As in any fire, wear self-contained breathing apparatus pressure-demand (NIOSH approved or
equivalent), and full protective gear to prevent contact with skin and eyes.

Section 6. Accidental Release Measures

6.1 Protective Precautions, Avoid raising and breathing dust, and provide adequate ventilation.
Protective Equipment and As conditions warrant, wear a NIOSH approved self-contained breathing apparatus, or respirator,
Emergency Procedures: and appropriate personal protection (rubber boots, safety goggles, and heavy rubber gloves).
6.2 Environmental Take steps to avoid release into the environment, if safe to do so.
Precautions:
6.3 Methods and Material For Contain spill and collect, as appropriate.
Containment and Cleaning Transfer to a chemical waste container for disposal in accordance with local regulations.
Up:

Section 7. Handling and Storage

7.1 Precautions To Be Taken Avoid breathing dust/fume/gas/mist/vapours/spray.
in Handling: Avoid prolonged or repeated exposure.
7.2 Precautions To Be Taken Keep container tightly closed.
in Storing: Store in accordance with information listed on the product insert.

Multi-region format
 SAFETY DATA SHEET Page: 3 of 5

Penicillin G (potassium salt) Revision: 10/16/2017

Section 8. Exposure Controls/Personal Protection

8.1 Exposure Parameters:
8.2 Exposure Controls:
8.2.1 Engineering Controls Use process enclosures, local exhaust ventilation, or other engineering controls to control airborne
(Ventilation etc.): levels below recommended exposure limits.
8.2.2 Personal protection equipment:
Eye Protection: Safety glasses
Protective Gloves: Compatible chemical-resistant gloves
Other Protective Clothing: Lab coat
Respiratory Equipment NIOSH approved respirator, as conditions warrant.
(Specify Type):
Work/Hygienic/Maintenan Do not take internally.
ce Practices: Facilities storing or utilizing this material should be equipped with an eyewash and a safety shower.
Wash thoroughly after handling.
No data available.

Section 9. Physical and Chemical Properties

9.1 Information on Basic Physical and Chemical Properties
Physical States: [ ] Gas [ ] Liquid [ X ] Solid
Appearance and Odor: A crystalline solid
pH: No data.
Melting Point: No data.
Boiling Point: No data.
Flash Pt: No data.
Evaporation Rate: No data.
Flammability (solid, gas): No data available.
Explosive Limits: LEL: No data. UEL: No data.
Vapor Pressure (vs. Air or mm No data.
Hg):
Vapor Density (vs. Air = 1): No data.
Specific Gravity (Water = 1): No data.
Solubility in Water: No data.
Solubility Notes: ~10 mg/ml in PBS (pH 7.2);
Octanol/Water Partition No data.
Coefficient:
Autoignition Pt: No data.
Decomposition Temperature: No data.
Viscosity: No data.
9.2 Other Information
Percent Volatile: No data.
Molecular Formula & Weight: C16H17N2O4S • K 372.5

Multi-region format
 SAFETY DATA SHEET Page: 4 of 5

Penicillin G (potassium salt) Revision: 10/16/2017

Section 10. Stability and Reactivity

10.1 Reactivity: No data available.
10.2 Stability: Unstable [ ] Stable [ X ]
10.3 Stability Note(s): Stable if stored in accordance with information listed on the product insert.
Polymerization: Will occur [ ] Will not occur [ X ]
10.4 Conditions To Avoid: No data available.
10.5 Incompatibility - Materials acids
To Avoid: heavy metal salts
oxidizing agents
10.6 Hazardous carbon dioxide
Decomposition or carbon monoxide
Byproducts: potassium oxides
nitrogen oxides
sulfur oxides

Section 11. Toxicological Information

11.1 Information on The toxicological effects of this product have not been thoroughly studied.
Toxicological Effects: Penicillin G (potassium salt) - Toxicity Data: Oral LD50 (rat): 8900 mg/kg; Intraperitoneal LD50
(rat): >2 gm/kg; Subcutaneous LD50 (rat): 11250 mg/kg; Oral LD50 (mouse): 6257 mg/kg;
Intraperitoneal LD50 (mouse): >2 gm/kg;
Chronic Toxicological Penicillin G (potassium salt) - Investigated as a drug and mutagen.
Effects: Only select Registry of Toxic Effects of Chemical Substances (RTECS) data is presented here.
See actual entry in RTECS for complete information.
Penicillin G (potassium salt) RTECS Number: XH9700000

CAS # Hazardous Components (Chemical Name) NTP IARC ACGIH OSHA

113-98-4 4-Thia-1-azabicyclo3.2.0heptane-2-carboxylicacid, n.a. n.a. n.a. n.a.
3,3-dimethyl-7-oxo-6-(phenylacetyl)amino-2S-(2.al

Section 12. Ecological Information

12.1 Toxicity: Avoid release into the environment.
Runoff from fire control or dilution water may cause pollution.
12.2 Persistence and No data available.
Degradability:
12.3 Bioaccumulative No data available.
Potential:
12.4 Mobility in Soil: No data available.
12.5 Results of PBT and vPvB No data available.
assessment:
12.6 Other adverse effects: No data available.

Multi-region format
 SAFETY DATA SHEET Page: 5 of 5

Penicillin G (potassium salt) Revision: 10/16/2017

Section 13. Disposal Considerations

13.1 Waste Disposal Method: Dispose in accordance with local, state, and federal regulations.

Section 14. Transport Information

14.1 LAND TRANSPORT (US DOT):
DOT Proper Shipping Name: Not dangerous goods.
DOT Hazard Class:
UN/NA Number:
14.1 LAND TRANSPORT (European ADR/RID):
ADR/RID Shipping Name: Not dangerous goods.
UN Number:
Hazard Class:
14.3 AIR TRANSPORT (ICAO/IATA):
ICAO/IATA Shipping Name: Not dangerous goods.
Additional Transport Transport in accordance with local, state, and federal regulations.
Information:

Section 15. Regulatory Information

EPA SARA (Superfund Amendments and Reauthorization Act of 1986) Lists
CAS # Hazardous Components (Chemical Name) S. 302 (EHS) S. 304 RQ S. 313 (TRI)
113-98-4 4-Thia-1-azabicyclo3.2.0heptane-2-carboxylicacid No No No
,
3,3-dimethyl-7-oxo-6-(phenylacetyl)amino-2S-(2.a
l

CAS # Hazardous Components (Chemical Name) Other US EPA or State Lists

113-98-4 4-Thia-1-azabicyclo3.2.0heptane-2-carboxylicacid CAA HAP,ODC: No; CWA NPDES: No; TSCA: Yes -
, Inventory; CA PROP.65: No
3,3-dimethyl-7-oxo-6-(phenylacetyl)amino-2S-(2.a
l

Regulatory Information This SDS was prepared in accordance with 29 CFR 1910.1200 and Regulation (EC)
Statement: No.1272/2008.

Section 16. Other Information

Revision Date:
Additional Information About
This Product:
Company Policy or Disclaimer:
10/16/2017
No data available.
DISCLAIMER: This information is believed to be accurate and represents the best information
currently available to us. However, we make no warranty of merchantability or any other warranty,
express or implied, with respect to such information, and we assume no liability resulting from its
use. Users should make their own investigations to determine the suitability of the information for
their particular purposes.

Multi-region format
 SODIUM BICARBONATE
www.natrium.com Safety Data Sheet Page 1 of 3
_________________________________________________________________________________________
1. IDENTIFICATION
Product name: Sodium bicarbonate
Synonyms: Sodium hydrogen carbonate; Baking soda; Bicarbonate of soda; Sodium acid carbonate; Carbonic
acid, monosodium salt.
Manufacturer: Telephone numbers:
Natrium Products, Inc. General inquiries: (607) 753-9829
58 Pendleton Street Emergencies (US and Canada ):
Cortland, NY 13045 CHEMTREC (Customer Number 724993)
USA (800) 424-9300 or 703-527-3887 (collect)
Recommended uses:
Food additive; pharmaceutical ingredient; water treatment; raw material for paper and chemical
manufacturing; animal feed additive; pH control.
2. HAZARD IDENTIFICATION
There are no appreciable health or environmental effects associated with this material.
Hazard classification: Not classified Label elements: No applicable labeling
Other potential health effects:
Eyes: Direct contact may cause irritation due to abrasion.
Skin: Not a skin irritant.
Inhalation: No known effects.
3. COMPOSITION/INFORMATION ON INGREDIENTS

Chemical name: Sodium hydrogen carbonate Chemical formula: NaHCO3

Synonyms: Sodium bicarbonate; Baking soda; Bicarbonate of soda; Sodium acid carbonate; Carbonic acid,
monosodium salt.
CAS Number: 144-55-8
Concentration (% by Weight): 100%
4. FIRST AID MEASURES
Eye contact: Irrigate with flowing water immediately and continuously for 15 minutes. Consult a physician if
necessary.
Skin contact: Wash off in flowing water or shower. If necessary, consult physician.
Ingestion: Do not induce vomiting. Seek medical attention immediately if overdose is taken.
Note to physician: Large doses, particularly in patients with renal insufficiency, have produced systemic
alkalosis and/or expansion in the extra-cellular fluid volume with edema.
Inhalation: Remove to fresh air. Seek medical attention if discomfort persists.
5. FIRE-FIGHTING MEASURES
Product is non-combustible. Thermal decomposition products are carbon dioxide and sodium carbonate (soda
ash). Carbon dioxide is an asphyxiant, and soda ash is an irritant.
Protective equipment: Self- contained breathing apparatus is necessary if large quantities are involved.
Extinguishing media: Use extinguishing material that is appropriate for fire in the surrounding area.
 SODIUM BICARBONATE
www.natrium.com Safety Data Sheet Page 2 of 3
_________________________________________________________________________________________
6. ACCIDENTAL RELEASE MEASURES
Sweep up into clean, dry containers for salvage or disposal. Wash away uncontaminated residue with water.
7. HANDLING AND STORAGE
Avoid contact with eyes and skin. Keep separated from acids. Store in a cool, dry place.
8. EXPOSURE CONTROLS/PERSONAL PROTECTION
Exposure limits: Not established.
Engineering controls: Provide general and/or local exhaust ventilation to control airborne dust.
Personal Protection:
Eyes & Face: Safety glasses for dusty conditions.
Respiratory: NIOSH approved dust mask.
Miscellaneous: Full cover clothing, general purpose gloves.
9. PHYSICAL AND CHEMICAL PROPERTIES
Appearance: White crystalline powder or granules.
Flammability: None.
Upper/lower flammability/explosive limits: Not applicable.
Odor: None.
Odor threshold: Not applicable.
Vapor pressure: Not applicable.
Vapor density: Not applicable.

pH of 0.1 M solution (0.84% w/v): 8.3 @ 25°C

3
Density: 2.2 g/cm .
Melting point: Not applicable (thermal decomposition occurs on heating).
Solubility in water: 86 g/L @ 20°C.
Boiling point: Not applicable.
Flash point: Not applicable.
Evaporation rate: Not applicable.
Partition coefficient, n-octanol/water: No data available.
Auto-ignition temperature: Not applicable.

Decomposition temperature: Starts to decompose when heated above 50°C (122°F).

Viscosity: Not applicable.
10. STABILITY AND REACTIVITY
Reactivity: Hazardous reactions or polymerization will not occur under normal conditions.
Chemical stability: Stable under recommended handling and storage conditions. (See Section 7.)
Conditions to avoid: Temperatures above 50°C (122°F).
Incompatible materials: Reacts with acids, releasing carbon dioxide.
Hazardous decomposition products: Carbon dioxide and sodium carbonate (soda ash).
 SODIUM BICARBONATE
www.natrium.com Safety Data Sheet Page 3 of 3
_________________________________________________________________________________________
11. TOXICOLOGICAL INFORMATION

Acute Oral: LD50 (rat) > 4000 mg/kg. Acute Inhalation: LC50 (rat) > 4.74 mg/L.
Eyes: Minimally irritating (rabbit, EPA TSCA 40 CFR 798.4500); Irritating (rabbit, Draize test, dose of 220 mg).
Skin: Slightly irritating (rabbit).
Carcinogenicity: Not listed as a carcinogen or potential carcinogen by the National Toxicology Program (NTP),
the International Agency for Research on Cancer (IARC), or the U.S. Occupational Safety and Health
Administration (OSHA).
12. ECOLOGICAL INFORMATION
Aquatic toxicity:
Fish: LC50 = 7700 mg/L (Rainbow trout, 96-hr. exposure).
Fish: LC50 = 7100 mg/L (Bluegill sunfish, 96-hr. exposure).
Invertebrates: EC50 > 1000 mg/L (Daphnia magna, 48-hr. exposure).
Persistence/Bioaccumulation potential: Not expected to persist or bioaccumulate in the environment.
Biodegradation: Not applicable.
Mobility: High potential for movement from soil to groundwater is expected based on aqueous solubility.
13. DISPOSAL CONSIDERATIONS
Not a hazardous material. Dispose in a landfill in accordance with pertinent federal, state and local regulations.
Empty containers may be incinerated or discarded as ordinary waste.
14. TRANSPORT INFORMATION
Not regulated by the U.S. Department of Transportation.
15. REGULATORY INFORMATION
CERCLA (40 CFR 302.4): Not a hazardous substance.
RCRA (40 CFR 261): Not a hazardous waste.
TSCA (40 CFR 710): Listed.
OSHA (29 CFR 1910.1200): Not hazardous.
SARA. Title III Sections 302 (40 CFR 355), 313 (40 CFR 372): Not a hazardous or toxic chemical.
European Inventory (EINECS): 205-633-8.
Japanese Inventory (MITI): 1-164.
U.S. Food and Drug Administration: Generally recognized as safe (GRAS) direct food additive
(21 CFR 184.1736).
16. OTHER INFORMATION
Maximum use level for drinking water corrosion and scale control: 100mg/L per NSF/ANSI 60 – 2014a.

Issue Date: 2/14/2017 Supersedes: 5/1/2015

This Safety Data Sheet is offered solely for your information, consideration, and investigation. Natrium Products, Inc. provides no
warranties, either expressed or implied, and assumes no responsibility for the accuracy or the completeness of the data contained herein.
 SAFETY DATA SHEET Page: 1 of 5

Phosphate Buffer Revision: 11/14/2018

Supersedes Revision: 06/26/2013

according to Regulation (EC) No. 1907/2006 as amended by (EC) No. 2015/830 and US OSHA HCS 2015

Section 1. Identification of the Substance/Mixture and of the Company/Undertaking

1.1 Product Code: 400032
Product Name: Phosphate Buffer

1.2 Relevant identified uses of the substance or mixture and uses advised against:
Relevant identified uses: For research use only, not for human or veterinary use.

1.3 Details of the Supplier of the Safety Data Sheet:

Company Name: Cayman Chemical Company
1180 E. Ellsworth Rd.
Ann Arbor, MI 48108
Web site address: www.caymanchem.com
Information: Cayman Chemical Company +1 (734)971-3335
1.4 Emergency telephone number:
Emergency Contact: CHEMTREC Within USA and Canada: +1 (800)424-9300
CHEMTREC Outside USA and Canada: +1 (703)527-3887

Section 2. Hazards Identification

2.1 Classification of the Substance or Mixture:
2.2 Label Elements:

GHS Signal Word: None

GHS Hazard Phrases:
Based on evaluation of currently available data this substance or mixture is not classifiable according to GHS.
GHS Precaution Phrases:
No phrases apply.
GHS Response Phrases:
No phrases apply.
GHS Storage and Disposal Phrases:
Please refer to Section 7 for Storage and Section 13 for Disposal information.
2.3 Adverse Human Health Material may be irritating to the mucous membranes and upper respiratory tract.
Effects and Symptoms: May be harmful by inhalation, ingestion, or skin absorption.
May cause eye, skin, or respiratory system irritation.
To the best of our knowledge, the toxicological properties have not been thoroughly investigated.

Section 3. Composition/Information on Ingredients

CAS # / Hazardous Components (Chemical Name)/ Concentration EC No./ GHS Classification
RTECS # REACH Registration No. EC Index No.
7778-77-0 Potassium phosphate, Monobasic 100.0 % 231-913-4 No GHS classifications apply.
TC6615500 01-2119490224-41 NA

Multi-region format
 SAFETY DATA SHEET Page: 2 of 5

Phosphate Buffer Revision: 11/14/2018

Supersedes Revision: 06/26/2013

Section 4. First Aid Measures

4.1 Description of First Aid
Measures:
In Case of Inhalation: Remove to fresh air. If not breathing, give artificial respiration or give oxygen by trained
personnel. Get immediate medical attention.
In Case of Skin Contact: Immediately wash skin with soap and plenty of water for at least 20 minutes. Remove
contaminated clothing. Get medical attention if symptoms occur. Wash clothing before reuse.
In Case of Eye Contact: Hold eyelids apart and flush eyes with plenty of water for at least 20 minutes. Have eyes
examined and tested by medical personnel.
In Case of Ingestion: Wash out mouth with water provided person is conscious. Never give anything by mouth to an
unconscious person. Get medical attention. Do NOT induce vomiting unless directed to do so by
medical personnel.

Section 5. Fire Fighting Measures

5.1 Suitable Extinguishing Use alcohol-resistant foam, carbon dioxide, water, or dry chemical spray.
Media: Use water spray to cool fire-exposed containers.
Unsuitable Extinguishing A solid water stream may be inefficient.
Media:
5.2 Flammable Properties andNo data available.
Hazards:
No data available.
Flash Pt: No data.
Explosive Limits: LEL: No data. UEL: No data.
Autoignition Pt: No data.
5.3 Fire Fighting Instructions: As in any fire, wear self-contained breathing apparatus pressure-demand (NIOSH approved or
equivalent), and full protective gear to prevent contact with skin and eyes.

Section 6. Accidental Release Measures

6.1 Protective Precautions, Avoid raising and breathing dust, and provide adequate ventilation.
Protective Equipment and As conditions warrant, wear a NIOSH approved self-contained breathing apparatus, or respirator,
Emergency Procedures: and appropriate personal protection (rubber boots, safety goggles, and heavy rubber gloves).
6.2 Environmental Take steps to avoid release into the environment, if safe to do so.
Precautions:
6.3 Methods and Material For Contain spill and collect, as appropriate.
Containment and Cleaning Transfer to a chemical waste container for disposal in accordance with local regulations.
Up:

Section 7. Handling and Storage

7.1 Precautions To Be Taken Avoid breathing dust/fume/gas/mist/vapours/spray.
in Handling: Avoid prolonged or repeated exposure.
7.2 Precautions To Be Taken Keep container tightly closed.
in Storing: Store in accordance with information listed on the product insert.
Other Precautions: Hygroscopic.

Multi-region format
 SAFETY DATA SHEET Page: 3 of 5

Phosphate Buffer Revision: 11/14/2018

Supersedes Revision: 06/26/2013

Section 8. Exposure Controls/Personal Protection

8.1 Exposure Parameters:
8.2 Exposure Controls:
8.2.1 Engineering Controls Use process enclosures, local exhaust ventilation, or other engineering controls to control airborne
(Ventilation etc.): levels below recommended exposure limits.
8.2.2 Personal protection equipment:
Eye Protection: Safety glasses
Protective Gloves: Compatible chemical-resistant gloves
Other Protective Clothing: Lab coat
Respiratory Equipment NIOSH approved respirator, as conditions warrant.
(Specify Type):
Work/Hygienic/Maintenan Do not take internally.
ce Practices: Facilities storing or utilizing this material should be equipped with an eyewash facility and a safety
shower.
Wash thoroughly after handling.
No data available.

Section 9. Physical and Chemical Properties

9.1 Information on Basic Physical and Chemical Properties
Physical States: [ ] Gas [ ] Liquid [ X ] Solid
Appearance and Odor: A crystalline solid
pH: No data.
Melting Point: No data.
Boiling Point: No data.
Flash Pt: No data.
Evaporation Rate: No data.
Flammability (solid, gas): No data available.
Explosive Limits: LEL: No data. UEL: No data.
Vapor Pressure (vs. Air or mm No data.
Hg):
Vapor Density (vs. Air = 1): No data.
Specific Gravity (Water = 1): No data.
Solubility in Water: No data.
Solubility Notes: Soluble in: water;
Octanol/Water Partition No data.
Coefficient:
Autoignition Pt: No data.
Decomposition Temperature: No data.
Viscosity: No data.
9.2 Other Information
Percent Volatile: No data.
Molecular Formula & Weight: KH2PO4 136.1

Multi-region format
 SAFETY DATA SHEET Page: 4 of 5

Phosphate Buffer Revision: 11/14/2018

Supersedes Revision: 06/26/2013

Section 10. Stability and Reactivity

10.1 Reactivity: No data available.
10.2 Stability: Unstable [ ] Stable [ X ]
10.3 Stability Note(s): No data available.
Polymerization: Will occur [ ] Will not occur [ X ]
10.4 Conditions To Avoid: No data available.
10.5 Incompatibility - Materials strong oxidizing agents
To Avoid:
10.6 Hazardous phosphorus oxides
Decomposition or potassium oxides
Byproducts:

Section 11. Toxicological Information

11.1 Information on The toxicological effects of this product have not been thoroughly studied.
Toxicological Effects: Potassium phosphate, Monobasic - Toxicity Data: Oral LDLo (rat): 4,640 mg/kg;
Chronic Toxicological Only select Registry of Toxic Effects of Chemical Substances (RTECS) data is presented here.
Effects: See actual entry in RTECS for complete information.
Potassium phosphate, Monobasic RTECS Number: TC6615500

CAS # Hazardous Components (Chemical Name) NTP IARC ACGIH OSHA

7778-77-0 Potassium phosphate, Monobasic n.a. n.a. n.a. n.a.

Section 12. Ecological Information

12.1 Toxicity: Avoid release into the environment.
Runoff from fire control or dilution water may cause pollution.
12.2 Persistence and No data available.
Degradability:
12.3 Bioaccumulative No data available.
Potential:
12.4 Mobility in Soil: No data available.
12.5 Results of PBT and vPvB No data available.
assessment:
12.6 Other adverse effects: No data available.

Section 13. Disposal Considerations

13.1 Waste Disposal Method: Dispose in accordance with local, state, and federal regulations.

Section 14. Transport Information

14.1 LAND TRANSPORT (US DOT):
DOT Proper Shipping Name: Not dangerous goods.
DOT Hazard Class:
UN/NA Number:
14.1 LAND TRANSPORT (European ADR/RID):
ADR/RID Shipping Name: Not dangerous goods.
UN Number:
Hazard Class:

Multi-region format
 SAFETY DATA SHEET Page: 5 of 5

Phosphate Buffer Revision: 11/14/2018

Supersedes Revision: 06/26/2013
14.3 AIR TRANSPORT (ICAO/IATA):
ICAO/IATA Shipping Name: Not dangerous goods.
Additional Transport Transport in accordance with local, state, and federal regulations.
Information:

Section 15. Regulatory Information

EPA SARA (Superfund Amendments and Reauthorization Act of 1986) Lists
CAS # Hazardous Components (Chemical Name) S. 302 (EHS) S. 304 RQ S. 313 (TRI)
7778-77-0 Potassium phosphate, Monobasic No No No

CAS # Hazardous Components (Chemical Name) Other US EPA or State Lists

7778-77-0 Potassium phosphate, Monobasic CAA HAP,ODC: No; CWA NPDES: No; TSCA: Yes -
Inventory; CA PROP.65: No

Regulatory Information This SDS was prepared in accordance with 29 CFR 1910.1200 and Regulation (EC)
Statement: No.1272/2008.

Section 16. Other Information

Revision Date:
Additional Information About
This Product:
Company Policy or Disclaimer:
11/14/2018
No data available.
DISCLAIMER: This information is believed to be accurate and represents the best information
currently available to us. However, we make no warranty of merchantability or any other warranty,
express or implied, with respect to such information, and we assume no liability resulting from its
use. Users should make their own investigations to determine the suitability of the information for
their particular purposes.

Multi-region format
 Page 1/8
Safety Data Sheet
acc. to OSHA HCS
Printing date 11/09/2020 Revision date 11/09/2020

1 Identification
· Product identifier
· Trade name: Phenylacetic Acid
· Synonym
Benzeneacetic acid
NSC 125718
Phenylethanoic acid
α-Toluic acid
· Article number: 18709
· CAS Number:
103-82-2
· EC number:
203-148-6
· Application of the substance / the mixture For research use only, not for human or veterinary use.
· Details of the supplier of the safety data sheet
· Manufacturer/Supplier:
Cayman Chemical Co.
1180 E. Ellsworth Rd.
Ann Arbor, MI 48108
USA
· Information department: Product safety department
· Emergency telephone number:
During normal opening times: +1 (734) 971-3335
US/CANADA: 800-424-9300
Outside US/CANADA: 703-741-5970

2 Hazard(s) identification
· Classification of the substance or mixture

~
d GHS07

Eye Irrit. 2A H319 Causes serious eye irritation.

· Label elements
· GHS label elements
The substance is classified and labeled according to the Globally Harmonized System (GHS).
· Hazard pictograms

~
d
GHS07

· Signal word Warning

· Hazard statements
Causes serious eye irritation.
· Precautionary statements
Wash thoroughly after handling.
(Contd. on page 2)
US

51.0.15
 Page 2/8
Safety Data Sheet
acc. to OSHA HCS
Printing date 11/09/2020 Revision date 11/09/2020

Trade name: Phenylacetic Acid

(Contd. from page 1)

Wear eye protection / face protection.
If in eyes: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy
to do. Continue rinsing.
If eye irritation persists: Get medical advice/attention.
· Classification system:
· NFPA ratings (scale 0 - 4)

Health = 2
1 Fire = 1
2 0 Reactivity = 0
· HMIS-ratings (scale 0 - 4)
HEALTH 2 Health = 2
FIRE 1 Fire = 1
REACTIVITY 0 Reactivity = 0

· Other hazards
· Results of PBT and vPvB assessment
· PBT: Not applicable.
· vPvB: Not applicable.

3 Composition/information on ingredients
· Chemical characterization: Substances
· CAS No. Description
103-82-2 Phenylacetic Acid
· Identification number(s)
· EC number: 203-148-6

4 First-aid measures
· Description of first aid measures
· After inhalation: Supply fresh air; consult doctor in case of complaints.
· After skin contact: Generally the product does not irritate the skin.
· After eye contact:
Rinse opened eye for several minutes under running water. If symptoms persist, consult a doctor.
· After swallowing: If symptoms persist consult doctor.
· Information for doctor:
· Most important symptoms and effects, both acute and delayed
May cause anemia, cough, CNS depression, drowsiness, headache, heart damage, lassitude
(weakness, exhaustion), liver damage, narcosis, reproductive effects, teratogenic effects.
No further relevant information available.
· Indication of any immediate medical attention and special treatment needed
No further relevant information available.

5 Fire-fighting measures
· Extinguishing media
· Suitable extinguishing agents:
CO2, extinguishing powder or water spray. Fight larger fires with water spray or alcohol resistant foam.
· Special hazards arising from the substance or mixture No further relevant information available.
(Contd. on page 3)
US

51.0.15
 Page 3/8
Safety Data Sheet
acc. to OSHA HCS
Printing date 11/09/2020 Revision date 11/09/2020

Trade name: Phenylacetic Acid

(Contd. from page 2)

· Advice for firefighters
· Protective equipment: No special measures required.

6 Accidental release measures

· Personal precautions, protective equipment and emergency procedures Not required.
· Environmental precautions: Do not allow to enter sewers/ surface or ground water.
· Methods and material for containment and cleaning up: Pick up mechanically.
· Reference to other sections
See Section 7 for information on safe handling.
See Section 8 for information on personal protection equipment.
See Section 13 for disposal information.
· Protective Action Criteria for Chemicals
· PAC-1: Substance is not listed.
· PAC-2: Substance is not listed.
· PAC-3: Substance is not listed.

7 Handling and storage

· Handling:
· Precautions for safe handling
No special precautions are necessary if used correctly.
Avoid breathing dust/fume/gas/mist/vapours/spray.
Avoid prolonged or repeated exposure.
Keep away from sources of ignition.
Take precautionary measures against static discharge.re.
· Information about protection against explosions and fires: No special measures required.
· Conditions for safe storage, including any incompatibilities
Keep container tightly closed.
Store in accordance with information listed on the product insert.
· Storage:
· Requirements to be met by storerooms and receptacles: No special requirements.
· Information about storage in one common storage facility: Not required.
· Further information about storage conditions: Keep receptacle tightly sealed.
· Specific end use(s) No further relevant information available.

8 Exposure controls/personal protection

· Additional information about design of technical systems: No further data; see item 7.
· Control parameters
· Components with limit values that require monitoring at the workplace: Not required.
· Additional information: The lists that were valid during the creation were used as basis.
· Exposure controls
· Personal protective equipment:
· General protective and hygienic measures:
Keep away from foodstuffs, beverages and feed.
Immediately remove all soiled and contaminated clothing.
Wash hands before breaks and at the end of work.
Avoid contact with the eyes.
Avoid contact with the eyes and skin.
(Contd. on page 4)
US

51.0.15
 Page 4/8
Safety Data Sheet
acc. to OSHA HCS
Printing date 11/09/2020 Revision date 11/09/2020

Trade name: Phenylacetic Acid

(Contd. from page 3)

· Breathing equipment: Not required.
· Protection of hands:
The glove material has to be impermeable and resistant to the product/ the substance/ the preparation.
Due to missing tests no recommendation to the glove material can be given for the product/ the
preparation/ the chemical mixture.
Selection of the glove material on consideration of the penetration times, rates of diffusion and the
degradation
· Material of gloves
The selection of the suitable gloves does not only depend on the material, but also on further marks of
quality and varies from manufacturer to manufacturer.
· Penetration time of glove material
The exact break through time has to be found out by the manufacturer of the protective gloves and has
to be observed.
· Eye protection:

R
_ Tightly sealed goggles

9 Physical and chemical properties

· Information on basic physical and chemical properties
· General Information
· Appearance:
Form: Crystalline
Color: Not determined.
· Odor: Characteristic
· Structural Formula C8H8O2
· Molecular Weight 136.2 g/mol
· Odor threshold: Not determined.
· pH-value: Not applicable.
· Change in condition
Melting point/Melting range: 76–77 °C (168.8–170.6 °F)
Boiling point/Boiling range: Undetermined.
· Flash point: >93 °C (>199.4 °F)
· Flammability (solid, gaseous): Product is not flammable.
· Decomposition temperature: Not determined.
· Auto igniting: Not determined.
· Danger of explosion: Product does not present an explosion hazard.
· Explosion limits:
Lower: Not determined.
Upper: Not determined.
· Vapor pressure: Not applicable.
· Density at 20 °C (68 °F): 0.54 g/cm³ (4.5063 lbs/gal)
· Relative density Not determined.
· Vapor density Not applicable.
(Contd. on page 5)
US

51.0.15
 Page 5/8
Safety Data Sheet
acc. to OSHA HCS
Printing date 11/09/2020 Revision date 11/09/2020

Trade name: Phenylacetic Acid

(Contd. from page 4)

· Evaporation rate Not applicable.

· Solubility in / Miscibility with
Water: Not determined.
· Partition coefficient (n-octanol/water): Not determined.
· Viscosity:
Dynamic: Not applicable.
Kinematic: Not applicable.
SOLUBILITY ~10 mg/ml in PBS (pH 7.2); ~1 mg/ml in DMF
· Other information No further relevant information available.

10 Stability and reactivity

· Reactivity No further relevant information available.
· Chemical stability
· Thermal decomposition / conditions to be avoided:
No decomposition if used according to specifications.
· Possibility of hazardous reactions No dangerous reactions known.
· Conditions to avoid No further relevant information available.
· Incompatible materials: strong bases, strong oxidizing agents, strong reducing agents
· Hazardous decomposition products: carbon oxides

11 Toxicological information
· RTECS Number AJ2430000
· Information on toxicological effects
· Acute toxicity:
· LD/LC50 values that are relevant for classification:
Oral LD50 2,250 mg/kg (mouse)
2,250 mg/kg (rat)
Intraperitoneal LD50 2,270 mg/kg (mouse)
1,600 mg/kg (rat)
Subcutaneous LD50 1,500 mg/kg (mouse)
· Primary irritant effect:
· on the skin: No irritant effect.
· on the eye: Irritating effect.
· Sensitization: No sensitizing effects known.
· Additional toxicological information:
· Carcinogenic categories
· IARC (International Agency for Research on Cancer) Substance is not listed.
· NTP (National Toxicology Program) Substance is not listed.
· OSHA-Ca (Occupational Safety & Health Administration) Substance is not listed.

12 Ecological information
· Toxicity
· Aquatic toxicity: No further relevant information available.
(Contd. on page 6)
US

51.0.15
 Page 6/8
Safety Data Sheet
acc. to OSHA HCS
Printing date 11/09/2020 Revision date 11/09/2020

Trade name: Phenylacetic Acid

(Contd. from page 5)

· Persistence and degradability No further relevant information available.
· Behavior in environmental systems:
· Bioaccumulative potential No further relevant information available.
· Mobility in soil No further relevant information available.
· Additional ecological information:
· General notes:
Water hazard class 1 (Assessment by list): slightly hazardous for water
Do not allow undiluted product or large quantities of it to reach ground water, water course or sewage
system.
· Results of PBT and vPvB assessment
· PBT: Not applicable.
· vPvB: Not applicable.
· Other adverse effects No further relevant information available.

13 Disposal considerations
· Waste treatment methods
· Recommendation:
Must not be disposed of together with household garbage. Do not allow product to reach sewage
system.
· Uncleaned packagings:
· Recommendation: Disposal must be made according to official regulations.

* 14 Transport information
· UN-Number
· DOT, IMDG, IATA UN3335
· UN proper shipping name
· DOT, IATA Aviation regulated solid, n.o.s. (Phenylacetic Acid)
· IMDG Aviation regulated solid, n.o.s.
· Transport hazard class(es)
· DOT, IMDG, IATA

ó
p̀
c
d
· Class 9 Miscellaneous dangerous substances and articles
· Label 9
· Packing group
· DOT, IMDG, IATA III
· Environmental hazards: Not applicable.
· Special precautions for user Warning: Miscellaneous dangerous substances and
articles
· Hazard identification number (Kemler code): 90
· Transport in bulk according to Annex II of
MARPOL73/78 and the IBC Code Not applicable.
(Contd. on page 7)
US

51.0.15
 Page 7/8
Safety Data Sheet
acc. to OSHA HCS
Printing date 11/09/2020 Revision date 11/09/2020

Trade name: Phenylacetic Acid

(Contd. from page 6)

· Transport/Additional information:
· DOT
· Quantity limitations On passenger aircraft/rail: 400 kg
On cargo aircraft only: 400 kg
· IMDG
· Limited quantities (LQ) -
· Excepted quantities (EQ) Code: -
· IATA
· Remarks: When sold in quantities of less than or equal to 1 mL,
or 1 g, with an Excepted Quantity Code of
E1, E2, E4, or E5, this item meets the De Minimis
Quantities exemption, per IATA 2.6.10.
Therefore packaging does not have to be labeled as
Dangerous Goods/Excepted Quantity.
· UN "Model Regulation": UN 3335 AVIATION REGULATED SOLID, N.O.S., 9,
III

15 Regulatory information
· Safety, health and environmental regulations/legislation specific for the substance or mixture
· Sara
· Section 355 (extremely hazardous substances): Substance is not listed.
· Section 313 (Specific toxic chemical listings): Substance is not listed.
· TSCA (Toxic Substances Control Act): ACTIVE
· Hazardous Air Pollutants Substance is not listed.
· Proposition 65
· Chemicals known to cause cancer: Substance is not listed.
· Chemicals known to cause reproductive toxicity for females: Substance is not listed.
· Chemicals known to cause reproductive toxicity for males: Substance is not listed.
· Chemicals known to cause developmental toxicity: Substance is not listed.
· Carcinogenic categories
· EPA (Environmental Protection Agency) Substance is not listed.
· TLV (Threshold Limit Value established by ACGIH) Substance is not listed.
· NIOSH-Ca (National Institute for Occupational Safety and Health) Substance is not listed.
· Chemical safety assessment: A Chemical Safety Assessment has not been carried out.

16 Other information
This information is based on our present knowledge. However, this shall not constitute a guarantee for
any specific product features and shall not establish a legally valid contractual relationship.
· Department issuing SDS: Environment protection department.
· Contact: -
· Date of preparation / last revision 11/09/2020 / -
· Abbreviations and acronyms:
IMDG: International Maritime Code for Dangerous Goods
DOT: US Department of Transportation
IATA: International Air Transport Association
ACGIH: American Conference of Governmental Industrial Hygienists
EINECS: European Inventory of Existing Commercial Chemical Substances
(Contd. on page 8)
US

51.0.15
 Page 8/8
Safety Data Sheet
acc. to OSHA HCS
Printing date 11/09/2020 Revision date 11/09/2020

Trade name: Phenylacetic Acid

(Contd. from page 7)

CAS: Chemical Abstracts Service (division of the American Chemical Society)
NFPA: National Fire Protection Association (USA)
HMIS: Hazardous Materials Identification System (USA)
LC50: Lethal concentration, 50 percent
LD50: Lethal dose, 50 percent
PBT: Persistent, Bioaccumulative and Toxic
vPvB: very Persistent and very Bioaccumulative
NIOSH: National Institute for Occupational Safety
OSHA: Occupational Safety & Health
TLV: Threshold Limit Value
PEL: Permissible Exposure Limit
REL: Recommended Exposure Limit
Eye Irrit. 2A: Serious eye damage/eye irritation – Category 2A
· * Data compared to the previous version altered.
US

51.0.15
 Safety Data Sheet
according to 29CFR1910/1200 and GHS Rev. 3
Effective date : 12.28.2014 Page 1 of 7
Ammonium Sulfate,

SECTION 1 : Identification of the substance/mixture and of the supplier

Product name : Ammonium Sulfate,
Manufacturer/Supplier Trade name:
Manufacturer/Supplier Article number: S25176A
Recommended uses of the product and uses restrictions on use:
Manufacturer Details:
AquaPhoenix Scientific
9 Barnhart Drive, Hanover, PA 17331

Supplier Details:
Fisher Science Education
15 Jet View Drive, Rochester, NY 14624

Emergency telephone number:

Fisher Science Education Emergency Telephone No.: 800-535-5053

SECTION 2 : Hazards identification

Classification of the substance or mixture:

Irritant
Skin irritation, category 2
Eye irritation, category 2A
Acute toxicity (oral, dermal, inhalation), category 3

Eye irrit. cat 2

Skin Sens, cat 2
STOT SE 3
AcTox Oral 4
Hazards Not Otherwise Classified - Combustible Dust

Signal word :Warning

Hazard statements:
Harmful if swallowed
Causes skin irritation
Causes serious eye irritation
May cause respiratory irritation
Precautionary statements:
Wash … thoroughly after handling
Do not eat, drink or smoke when using this product
Avoid breathing dust/fume/gas/mist/vapours/spray
Use only outdoors or in a well-ventilated area
Wear protective gloves/protective clothing/eye protection/face protection
Specific treatment (see supplemental first aid instructions on this label)
Rinse mouth
Take off contaminated clothing and wash before reuse
IF SWALLOWED: Call a POISON CENTER or doctor/physician if you feel unwell
IF ON SKIN: Wash with soap and water
IF INHALED: Remove victim to fresh air and keep at rest in a position comfortable for breathing

Created by Global Safety Management, Inc. -Tel: 1-813-435-5161 - www.gsmsds.com

 Safety Data Sheet
according to 29CFR1910/1200 and GHS Rev. 3
Effective date : 12.28.2014 Page 2 of 7
Ammonium Sulfate,

If skin irritation occurs: Get medical advice/attention

If eye irritation persists get medical advice/attention
IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses if present and easy to do.
Continue rinsing
Store locked up
Store in a well ventilated place. Keep container tightly closed
Dispose of contents/container to …

Combustible Dust Hazard: :

May form combustible dust concentrations in air (during processing).

Other Non-GHS Classification:

WHMIS
NFPA/HMIS

NFPA SCALE (0-4) HMIS RATINGS (0-4)

SECTION 3 : Composition/information on ingredients

Ingredients:

CAS 7783-20-2 Ammonium Sulfate,ACS >95 %

Percentages are by weight

SECTION 4 : First aid measures

Description of first aid measures
After inhalation: Move exposed individual to fresh air. Loosen clothing as necessary and position individual in
a comfortable position.Seek medical advice if discomfort or irritation persists.If breathing difficult, give oxygen.
After skin contact: Wash affected area with soap and water. Rinse/flush exposed skin gently using water for
15-20 minutes. Seek medical advice if discomfort or irritation persists.
After eye contact: Protect unexposed eye. Rinse/flush exposed eye(s) gently using water for 15-20 minutes.
Remove contact lens(es) if able to do so during rinsing. Seek medical attention if irritation persists or if
concerned.
After swallowing: Rinse mouth thoroughly. Do not induce vomiting. Have exposed individual drink sips of
water. Seek medical attention if irritation, discomfort or vomiting persists.

Most important symptoms and effects, both acute and delayed:

Irritation, Nausea,Headache, Shortness of breath.;
Indication of any immediate medical attention and special treatment needed:
If seeking medical attention, provide SDS document to physician.

Created by Global Safety Management, Inc. -Tel: 1-813-435-5161 - www.gsmsds.com

 Safety Data Sheet
according to 29CFR1910/1200 and GHS Rev. 3
Effective date : 12.28.2014 Page 3 of 7
Ammonium Sulfate,

SECTION 5 : Firefighting measures

Extinguishing media
Suitable extinguishing agents: If in laboratory setting, follow laboratory fire suppression procedures. Use
appropriate fire suppression agents for adjacent combustible materials or sources of ignition
For safety reasons unsuitable extinguishing agents:
Special hazards arising from the substance or mixture:
Combustion products may include carbon oxides or other toxic vapors.Thermal decomposition can lead to
release of irritating gases and vapors.Avoid generating dust; fine dust dispersed in air in sufficient
concentrations, and in the presence of an ignition source is a potential dust explosion hazard.
Advice for firefighters:
Protective equipment: Use NIOSH-approved respiratory protection/breathing apparatus.
Additional information (precautions): Move product containers away from fire or keep cool with water
spray as a protective measure, where feasible.Use spark-proof tools and explosion-proof equipment.

SECTION 6 : Accidental release measures

Personal precautions, protective equipment and emergency procedures:
Wear protective equipment. Transfer to a disposal or recovery container.Use spark-proof tools and explosion-
proof equipment.Use respiratory protective device against the effects of fumes/dust/aerosol. Keep unprotected
persons away. Ensure adequate ventilation.Keep away from ignition sources. Protect from heat.Stop the spill, if
possible. Contain spilled material by diking or using inert absorbent.
Environmental precautions:
Prevent from reaching drains, sewer or waterway. Collect contaminated soil for characterization per Section 13
Methods and material for containment and cleaning up:
If in a laboratory setting, follow Chemical Hygiene Plan procedures.Place into properly labeled containers for
recovery or disposal. If necessary, use trained response staff/contractor.Dust deposits should not be allowed to
accumulate on surfaces, as these may form an explosive mixture if they are released into the atmosphere in
sufficient concentration. Avoid dispersal of dust in the air (i.e., clearing dust surfaces with compressed air).
Collect solids in powder form using vacuum with (HEPA filter)
Reference to other sections:

SECTION 7 : Handling and storage

Precautions for safe handling:
Minimize dust generation and accumulation. Wash hands after handling. Avoid dispersal of dust in the air (i.e.,
clearing dust surfaces with compressed air). Routine housekeeping should be instituted to ensure that dusts do
not accumulate on surfaces. Dry powders can build static electricity charges when subjected to the friction of
transfer and mixing operations. Follow good hygiene procedures when handling chemical materials. Do not eat,
drink, smoke, or use personal products when handling chemical substances. If in a laboratory setting, follow
Chemical Hygiene Plan.Use only in well ventilated areas.Avoid generation of dust or fine particulate.Avoid
contact with eyes, skin, and clothing.
Conditions for safe storage, including any incompatibilities:
Store in a cool location. Provide ventilation for containers. Avoid storage near extreme heat, ignition sources or
open flame. Store away from foodstuffs. Store away from oxidizing agents.Store in cool, dry conditions in well
sealed containers. Keep container tightly sealed.Store with like hazards

SECTION 8 : Exposure controls/personal protection

Created by Global Safety Management, Inc. -Tel: 1-813-435-5161 - www.gsmsds.com

 Safety Data Sheet
according to 29CFR1910/1200 and GHS Rev. 3
Effective date : 12.28.2014 Page 4 of 7
Ammonium Sulfate,

Control Parameters: , , OSHA PEL TWA (Total Dust) 15 mg/m3 (50 mppcf*)
, , ACGIH TLV TWA (inhalable particles) 10 mg/m3
Appropriate Engineering controls: Emergency eye wash fountains and safety showers should be available in
the immediate vicinity of use/handling.Provide exhaust ventilation or
other engineering controls to keep the airborne concentrations of vapor
or dusts (total/respirable) below the applicable workplace exposure limits
(Occupational Exposure Limits-OELs) indicated above.Use under a fume
hood. It is recommended that all dust control equipment such as local
exhaust ventilation and material transport systems involved in handling
of this product contain explosion relief vents or an explosion suppression
system or an oxygen deficient environment.Ensure that dust-handling
systems (such as exhaust ducts, dust collectors, vessels, and processing
equipment) are designed in a manner to prevent the escape of dust into
the work area (i.e., there is no leakage from the equipment).
Respiratory protection: Not required under normal conditions of use. Use suitable respiratory
protective device when high concentrations are present. Use suitable
respiratory protective device when aerosol or mist is formed. For spills,
respiratory protection may be advisable.
Protection of skin: The glove material has to be impermeable and resistant to the product/
the substance/ the preparation being used/handled.Selection of the glove
material on consideration of the penetration times, rates of diffusion and
the degradation.
Eye protection: Safety glasses with side shields or goggles.
General hygienic measures: The usual precautionary measures are to be adhered to when handling
chemicals. Keep away from food, beverages and feed sources.
Immediately remove all soiled and contaminated clothing. Wash hands
before breaks and at the end of work. Do not inhale
gases/fumes/dust/mist/vapor/aerosols. Avoid contact with the eyes and
skin.

SECTION 9 : Physical and chemical properties

Appearance (physical Explosion limit lower: Not Determined

Colorless Solid
state,color): Explosion limit upper: Not Determined

Odor: Odorless Vapor pressure: Not Determined

Odor threshold: Not Determined Vapor density: Not Determined

pH-value: 5-6 (5% aq. sol.) Relative density: 1.8

Melting/Freezing point: 280 C Solubilities: Material is water soluble.

Boiling point/Boiling Partition coefficient (n-

Not Determined n-octanol/water: log Pow: -5.1
range: octanol/water):

Flash point (closed Auto/Self-ignition

Not Determined Not Determined
cup): temperature:

Created by Global Safety Management, Inc. -Tel: 1-813-435-5161 - www.gsmsds.com

 Safety Data Sheet
according to 29CFR1910/1200 and GHS Rev. 3
Effective date : 12.28.2014 Page 5 of 7
Ammonium Sulfate,

Decomposition
Evaporation rate: Insignificant 350 C
temperature:

Flammability a. Kinematic:Not Determined

Not Determined Viscosity:
(solid,gaseous): b. Dynamic: Not Determined

Density: Not Determined

SECTION 10 : Stability and reactivity

Reactivity:Nonreactive under normal conditions.

Chemical stability:No decomposition if used and stored according to specifications.
Possible hazardous reactions:None under normal processing
Conditions to avoid:Store away from oxidizing agents, strong acids or bases.Incompatible Materials.excess
heat.Dust generation.
Incompatible materials:Strong acids.Strong bases.Strong oxidizing agents.
Hazardous decomposition products:sulfur dioxide.nitrogen.Ammonia.ammonium bisulfate.

SECTION 11 : Toxicological information

Acute Toxicity:

Oral: 2840mg/kg APS: LD50 orl-rat

Chronic Toxicity: No additional information.

Corrosion Irritation: No additional information.

Sensitization: No additional information.

Single Target Organ (STOT): No additional information.

Numerical Measures: No additional information.

Carcinogenicity: No additional information.

Mutagenicity: No additional information.

Reproductive Toxicity: No additional information.

SECTION 12 : Ecological information

Ecotoxicity Persistence and degradability: Readily degradable in the environment.

Bioaccumulative potential:
Mobility in soil:
Other adverse effects:

SECTION 13 : Disposal considerations

Waste disposal recommendations:

Product/containers must not be disposed together with household garbage. Do not allow product to reach
sewage system or open water.It is the responsibility of the waste generator to properly characterize all waste
materials according to applicable regulatory entities (US 40CFR262.11). Consult federal state/ provincial and
local regulations regarding the proper disposal of waste material that may incorporate some amount of this
product.

Created by Global Safety Management, Inc. -Tel: 1-813-435-5161 - www.gsmsds.com

 Safety Data Sheet
according to 29CFR1910/1200 and GHS Rev. 3
Effective date : 12.28.2014 Page 6 of 7
Ammonium Sulfate,

SECTION 14 : Transport information

UN-Number
Not Dangerous Goods
UN proper shipping name
Not Dangerous Goods
Transport hazard class(es)
Packing group:Not Dangerous Goods
Environmental hazard:
Transport in bulk:
Special precautions for user:

SECTION 15 : Regulatory information

United States (USA)

SARA Section 311/312 (Specific toxic chemical listings):
None of the ingredients is listed
SARA Section 313 (Specific toxic chemical listings):
7783-20-2 Ammonium Sulfate
RCRA (hazardous waste code):
None of the ingredients is listed
TSCA (Toxic Substances Control Act):
All ingredients are listed.
CERCLA (Comprehensive Environmental Response, Compensation, and Liability Act):
None of the ingredients is listed

Proposition 65 (California):

Chemicals known to cause cancer:

None of the ingredients is listed
Chemicals known to cause reproductive toxicity for females:
None of the ingredients is listed
Chemicals known to cause reproductive toxicity for males:
None of the ingredients is listed
Chemicals known to cause developmental toxicity:
None of the ingredients is listed

Canada

Canadian Domestic Substances List (DSL):

All ingredients are listed.
Canadian NPRI Ingredient Disclosure list (limit 0.1%):
None of the ingredients is listed
Canadian NPRI Ingredient Disclosure list (limit 1%):
None of the ingredients is listed

SECTION 16 : Other information

Created by Global Safety Management, Inc. -Tel: 1-813-435-5161 - www.gsmsds.com

 Safety Data Sheet
according to 29CFR1910/1200 and GHS Rev. 3
Effective date : 12.28.2014 Page 7 of 7
Ammonium Sulfate,

This product has been classified in accordance with hazard criteria of the Controlled Products Regulations and the
SDS contains all the information required by the Controlled Products Regulations.Note:. The responsibility to
provide a safe workplace remains with the user.The user should consider the health hazards and safety information
contained herein as a guide and should take those precautions required in an individual operation to instruct
employees and develop work practice procedures for a safe work environment.The information contained herein is,
to the best of our knowledge and belief, accurate.However, since the conditions of handling and use are beyond
our control, we make no guarantee of results, and assume no liability for damages incurred by the use of this
material.It is the responsibility of the user to comply with all applicable laws and regulations applicable to this
material.
GHS Full Text Phrases:

Abbreviations and acronyms:

IMDG: International Maritime Code for Dangerous Goods
PNEC: Predicted No-Effect Concentration (REACH)
CFR: Code of Federal Regulations (USA)
SARA: Superfund Amendments and Reauthorization Act (USA)
RCRA: Resource Conservation and Recovery Act (USA)
TSCA: Toxic Substances Control Act (USA)
NPRI: National Pollutant Release Inventory (Canada)
DOT: US Department of Transportation
IATA: International Air Transport Association
GHS: Globally Harmonized System of Classification and Labelling of Chemicals
ACGIH: American Conference of Governmental Industrial Hygienists
CAS: Chemical Abstracts Service (division of the American Chemical Society)
NFPA: National Fire Protection Association (USA)
HMIS: Hazardous Materials Identification System (USA)
WHMIS: Workplace Hazardous Materials Information System (Canada)
DNEL: Derived No-Effect Level (REACH)

Effective date : 12.28.2014

Last updated : 03.19.2015

Created by Global Safety Management, Inc. -Tel: 1-813-435-5161 - www.gsmsds.com