Pharmaceutical Chemistry Journal, Vol. 53, No. 8, November, 2019 (Russian Original Vol. 53, No.

8, September, 2019)

DESIGN OF EXPERIMENTS IN PHARMACEUTICAL

DEVELOPMENT

Abhishek S. Dhoot,1 Gasper J. Fernandes,1 Anup Naha,1

Mahalaxmi Rathnanand,1 and Lalit Kumar1,*

Original article submitted July 18, 2019.

In order to develop high-quality pharmaceutical products, a traditional approach based the univariate or trial
and error method was used in the past that led to several problems like non-reproducible, high-cost, and time
consuming methods. To overcome these drawbacks, a new concept of the Design of Experiment (DoE) was
introduced. DoE is a statistical element of the Quality by Design (QbD) approach introduced by British statis-
tician Sir Ronald Fisher in 1925. The basic objectives of DoE are screening, optimization, and robustness. It
involves the execution of experimental design on the basis of suitable variables along with statistical evalua-
tion of obtained responses and exploration of the design space using mathematical or graphical approach. The
statistical evaluation empowers to build up the quality of finished products and helps to meet the increasing
demands for product of superior quality and standards. This article mainly focuses on the applications of DoE
in pharmaceutical product development along with its objectives, design, and selection criteria.
Keywords: Design of Experiment (DoE); Quality by Design (QbD); design; pharmaceutical product develop-
ment.

1. INTRODUCTION
The concept of experimental design was introduced in
the early 20th century by famous statistician Sir Ronald
Fisher [1]. Books written by Sir Fisher such as “Statistical
Methods for Research Workers” [2], “Arrangements of field
experiments” [3], and “The design of experiments” [4] serve
as the ground work in the sphere of modern statistics. He in-
troduced the concept of applying statistical analysis at early
stages of a research work rather than at the final stages [5].
Various pharmaceutical products like tablets, capsules, emul-
sions, suspensions, gels, patches and creams are formulated
for effective delivery of drugs and ensuring safety and effi-
cacy.
Various technologies are involved in the formulation of a
pharmaceutical dosage form. Formulation scientists encoun-
ter several complex technical challenges during formulation
development and thus Design of Experiments (DoE) and re-
lated statistical analysis have been applied widely in process
optimization and validation to obtain a high-quality product
[6]. A good formulation should be easily manufacturable,
1 tested in products but it should be built in products by design
Department of Pharmaceutics, Manipal College of Pharmaceutical Sci-
ences, Manipal Academy of Higher Education, Madhav Nagar 576104,
Manipal, Udupi, Karnataka, India.
*
e-mail: lk.kundlas@gmail.com
chemically stable, and have good shelf life, high safety. and
good bioavailablity. In addition, many quality and special re-
quirements should be met as both patient safety and product
efficacy are related to the quality of pharmaceutical products
[6, 7].
In pharmaceutical industry, there is continuous demand
for the development of new products and processes, and the
improvement of existing products and processes to meet the
increasing requirements to quality and standards. Using
DoE, one can study the effect of each formulation variable
on its response and also the interaction between factors so as
to determine the critical variables. Further, the formulation
can be optimized using DoE by selecting the levels of critical
variables. By using this approach, manufacturing process can
also be optimized hence it contributes to a successful and ef-
ficient scale-up and process validation and results in high
product quality [6]. The “Quality by Design” (QbD) is the
approach which helps in achieving the product of required
standards and quality and is accepted by various regulatory
bodies [7 – 10]. The term “Quality by Design” was coined by
Joseph M. Juran [8] according to which the quality cannot be
and pre-planning, which was further also recognised by the
regulatory agencies [9, 10]. DoE is a tool of QbD which
plays a prime role to achieve Total Quality Product Profile

730
0091-150X/19/5308-0730 © 2019 Springer Science+Business Media, LLC
Design of Experiments in Pharmaceutical Development 731

(QTPP). It helps to analyze the obtained responses and to
achieve the goal of study in an acceptable form [8, 11]. It can
be ascertained that DoE proved to be the most effective and
efficient tool throughout many stages of formulation process
by help in making intelligent decisions and obtaining a prod-
uct of superior design, performance and stability [6].
Thus, DoE is a crucial tool to deploy QbD in pharmaceu-
tical product development [12]. As per USFDA, DoE is a
structured, organized and pre-planned method for determin-
ing the association between factors (inputs) influencing a
process and the output of the process also known as “formal
experimental design.” The classical method used in DoE is
One Factor at a Time (OFAT) accepted as a univariate ap-
proach, in which one factor is accessed at a time keeping
other factors constant, thus making it very time consuming.
This method has several disadvantages and is incapable of
studying the interaction of factors and simultaneous study of
all the factors, non-reproducible, time consuming, expensive,
and covering only a small fraction of the total possible space
of factors [5, 7, 13]. Hence, when dealing with multiple fac-
tors and their potential interactions, DoE is effective and effi-
cient approach to understand the formulation system and to
optimize the formulation [6].
2. ADVANTAGES OF DESIGN OF EXPERIMENT
DoE has a number o advantages over OFAT, in particular
[5, 6, 13]:
– Provides more accurate and precise information
– Multiple potential factors can be studied simulta-
neously, systematically and quickly
– Factors interaction can be studied
– Strong relationship of critical processing parameters
(CPPs) and/or critical material attributes (CMAs) with criti-
cal quality attributes (CQA) can be established
– DoE can be helpful for screening of variables. It means
out of various factors, significant factors can be determined
– Variables with least and/or highly significant effect of
responses can be determined with the help of statistical anal-
ysis
– DoE makes the process and product more robust
– Effective design helps to build up the quality in the
pharmaceutical dosage forms
– High research efficiency, enhanced product quality and
safety
– Quantifies the relationship between factors (input) and
responses (output)
– Can helps to reduce the production cost
– DoE helps to achieve the target or objectives in short
duration
3. OBJECTIVES OF DESIGN OF EXPERIMENT
DoE has three main objectives [7]:
(a) Screening: identification of critical factors and their
levels.
(b) Optimization: identification of optimum input vari-
ables (factors) with their levels to achieve the optimum re-
sponse.

TABLE 1. Various Types of Design for Screening and Optimization

Experimental
No. of Factors Levels Model Purpose Applications/ Advantages
design type
Plackett-Burman De- 7 to 32 2 Linear and interac- Screening Used when large number of factors are
sign (PBD) tion model to be studied and Fractional Factorial
Design (FFD) is not applicable.
Fractional Factorial 3 to 6 2 or 3 Linear and interac- Screening and/or optimi- Used when large number of factors are
Design (FFD) tion model zation to be studied.
Full Factorial Design £6 2 or 3 Linear and interac- Screening and/or optimi- To examine main and interactive ef-
(FD) tion model zation fects.
Central Composite 2 to 6 3 or 5 Quadratic and inter- Optimization To examine main and interactive ef-
Design (CCD) action model fects. Lesser experimental runs than
FFD/FD.
Box-Behnken Design 3 to 5 3 to 5 Quadratic and inter- Optimization Non linear effects can be studied.
(BBD) action model
Mixture Design – 2 to 4 3 Mixture model Optimization Study of multiple components or factors
Simplex Lattice or material attributes.
Mixture Design – 3 to 5 3 Mixture model Optimization Study of multiple components or factors
Simplex Centorid or material attributes.
Mixture Design – 2 to 6 3 Mixture model Optimization Study of multiple components or factors
Constrained Mixture or material attributes.
D-optimal Design ³3 ³3 Custom made model Large level product or To minimize the variance of parameters.
process optimization
732 Abhishek S. Dhoot et al.

TABLE 2. Some Recent Pharmaceutical Applications of Design of Experiments

No. Object of study Drug name Formulation Design name Application Reference
1 Oral Drug Delivery Baclofen Oral bioadhesive Central Compos- To identify critical factors, their interactions [18]
bilayer tablet of ite Design and ideal process conditions that accomplishes
narrow absorp- the targeted responses.
tion window
drug
2 Oral Drug Delivery Diclofenac Dispersible tab- Factorial Design Implementation of QbD approach in formula- [19]
lets tion development.
3 Transdermal Drug Lornoxicam Transdermal Box-Behnken Optimization of micro-emulsion formulation. [20]
Delivery patch Design
4 Transdermal Drug Tacrine hydro- Patch develop- Central Compos- Study the impact of formulation and electronic [21]
Delivery chloride ment by ite Design variables on the Tacrine permeation.
iontophoretic
drug delivery
5 Oral Drug Delivery Doxycycline hy- Emulsion Factorial Design Optimize the emulsion for electro spinning. [22]
drochloride and D-optimal
Design
6 Oral Drug Delivery Ibuprofen and Immediate re- Fractional Facto- Examining the relative impact of active phar- [23]
Theophyline lease tablet rial Statistical maceutical ingredient (API) properties, pro-
Design cessing methods, and excipients variability on
drug product quality attributes.
7 Topical Drug Deliv- Silver Drug loaded Full Factorial To optimize the formulation. [24]
ery sulfadiazine microsponge in- Design
corporated in gel
base
8 Ocular Drug Deliv- Dexibuprofen PEGylated Central compos- To optimize the formulation and analyse the [25]
ery PLGA ite factorial de- effect of factors on the responses.
nanospheres sign
9 Analytical Method Valsartan Nanoparticles Full Factorial Optimization, development and validation of [26]
Development Design HPLC method for estimation of valsartan in
nanoparticles.
10 Oral Drug Delivery Gefitinib Polymeric nano Full Factorial Application of QbD approach to study the ef- [27]
suspension Design fect of CMAs and CPPs on critical quality at-
tributes (CQAs) and to improve the quality
and safety of formulation.
11 Oral Drug Delivery Bromopride Extended release Full Factorial Preparation and scale-up of extended release [28]
tablets Design low dose tablets of bromopride.
12 Oral Drug Delivery Piroxicam Co-crystals Full Factorial Utilising the design of experiment approach to [29]
oro-dispersible Design formulate, evaluate and co-crystal of
tablets piroxicam.
13 Nanopharmaceutics Rifampicin Solid Lipid Full Factorial Utilising Design of Experiments approach to [30]
Nanoparticles Design investigate the influence of pre-freezing con-
ditions on the powder respirability.
14 Pharmaceutical Pro- Lactose Excipients Full Factorial Effect of grinding pressure, injector pressure [31]
cesses monohydrate Micronization Design and feed rate on the particulate attributes of
micronized powders procured from the differ-
ent size grades.
15 Oral Gastroretentive Clarithromycin Bi-dependent Box-Behnken Optimization of formulation. [32]
Drug Delivery Gastroretentive Design
Tablets
16 Bioanalytical Valsartan Oral Drug Sus- Full Factorial Statistical optimization of extraction process [33]
Method Develop- pension Design for quantification of valsartan in rabbit
ment plasma.
17 Oral drug delivery API Tablet Mixture Design Optimization of Formulation. [34]
18 Oral drug delivery Brivanib Film coated tab- Full Factorial To assess formulation ruggedness and opti- [35]
Alaninate lets Design mize composition of excipients.
Design of Experiments in Pharmaceutical Development 733

No. Object of study Drug name Formulation Design name Application Reference
19 Analytical method Abacavir, – Response surface Optimization, sensitivity analysis and robust- [36]
development Lamivudine and methodology ness study of analytical method
Dolutegravir
20 Drug release study Dexamethasone PLGA Central compos- To optimize drug loading, burst release and [37]
microspheres ite design lag phase
21 Biodegradation Amoxicillin – Central compos- Optimization of parameters for biodegradation [38]
study and ite design of APIs to achieve high degree biodegradation
Cephalexin efficiency
22 Analytical study Isoprenaline – Fractional Facto- Screening and optimization of factors [39]
rial design and
Box-Behnken
design
23 Pharmaceutical pro- Risperidone – Full Factorial Optimization of instrumental parameters for [40]
cesses Design spray drying of risperidone nanosuspension

(c) Robustness: identification of sensitivity of response
to small changes in the factors.
DoE can be used for number of purpose like comparison
between number of materials and suppliers, variables/factors
screening, system optimization, robustness and relation be-
tween input variables/factors and output responses [14].
4. TYPES OF DESIGN
DoE designs are broadly classified into two categories
[7], screening and optimal designs. The selection of these de-
signs is based on number of factors or variables and their
number of levels. The selection criteria of these factors or
variables are briefly explained in Table 1.
5. METHODOLOGY OF CONDUCTING DESIGN OF
EXPERIMENTS
Procedure of conducting DoE can be summarised in fol-
lowing steps [14]:
1. Determination of objective: There should be clear ob-
jective of the experiment for appropriate selection of experi-
mental design, factors or variables, levels and responses.
2. Selection of critical quality attributes (CQAs): CQAs
are the outcomes or responses of performed experiments,
which may vary based on the objectives. CQAs help to
achieve the targets of study.
3. Selection of factors and their levels: Factors are stud-
ied to determine their effect and intensity of the effect on the
responses. Factors can be of two types such as material at-
tributes and/or processing parameters. Level is the range
which has to be selected for a specific factor. Usually two or
more values within the range can be selected.
4. Selection of experimental design: Based on the objec-
tives, factors and their levels; and suitable experimental de-
sign can be selected. Various experimental designs and their
purpose and advantages are shown in Table 1.
5. Conduct of experiments: After selection of appropri-
ate design, experimental plan is obtained, based on that the
experiments are performed and responses are obtained. Here
the responses are collected with the help of CQAs.
6. Analysis of data or obtained responses: The obtained
responses are statistically analyzed using statistical methods
such as regression analysis, Student’s t-test and analysis of
variance (ANOVA).
7. Conclusion: Once the data is analyzed, suitable con-
clusion is made based on which recommendations are taken.
8. Design space: On the basis of criteria and desirability
values, software suggested compositions are preformed prac-
tically. Further confirmatory tests are performed to validate
the conclusion and recommendations.
6. PHARMACEUTICAL APPLICATIONS OF DESIGN
OF EXPERIMENTS
Figure 1 represents various areas of DoE in pharmacy.
The first DoE application in design of pharmaceutical dosage
form was considered by Marlow and Shangraw in 1967 [15],
after which there was a steady increasing trend of DoE im-
plementation of pharmaceutical research [5]. Some recent
examples of various types of designs applied in development
of pharmaceutical products and processes are given in Ta-
ble 2.
7. SOFTWARES AVAILABLE FOR DESIGN
OF EXPERIMENTS
There are number of software available used for imple-
mentation of DoE. Use of software for DoE makes the task
solution easier, faster and economical. DoE helps one to en-
hance the accuracy, avoid wastage of chemicals, and also
helps one to minimize the time spent for developing new
products and methods. Some commonly used software pack-
ages are Design Expert, Minitab, Statistica, JMP, OPTIMA,
CAMO, R-Tutorial, SigmaXL, Statgraphics Centurion, and
ReliaSoft. Some of them are freely available and some are
paid [7, 16, 17].
734
Fig. 1. Pharmaceutical applications of design of experiments.
8. INDUSTRIAL APPLICATIONS OF DESIGN OF
EXPERIMENTS
There are several applications of DoE in industry. The
main applications are listed below:
(1) DoE can help in reucing the production and/or
method development time.
(2) DoE can help in increasing the quality of products.
(3) DoE can help in improving the stability and efficacy
of newly developed pharmaceutical formuations.
(4) DoE can help in meeting the regulatory requiremets.
(5) DoE helps in controlling the cost of production
and/or method development.
(6) DoE can help in saving the energy by reducing the
use of insturements and/or power for new formulation devel-
opment.
(7) DoE can help in reducing the wastage of water.
Thus, DoE is becoming an important tool for the phar-
maceutical industry in developing roubst, efficient, sensitive
and simple analytical and/or bioanalytical methods at mini-
mum expenses and time. DoE not only helps in the produc-
Abhishek S. Dhoot et al.
tion and/or analytical method development, but it can also
help meeting the regulatory requirements with QbD ap-
proach.
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