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Concurrent process validation; A case study for Artesunate and Amodiaquine

tablets

Article · August 2020

DOI: 10.22271/tpi.2020.v9.i7a.5042

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 The Pharma Innovation Journal 2020; 9(7): 16-25

ISSN (E): 2277- 7695

ISSN (P): 2349-8242
NAAS Rating: 5.03 Concurrent process validation; A case study for
TPI 2020; 9(7): 16-25
© 2020 TPI Artesunate and Amodiaquine tablets
[Link]
Received: 10-05-2020
Accepted: 12-06-2020 Tuani YT, Amo-Koi Seth, Gordon-Jackson Andrew, Angela Asor and
Tuani YT David Mingle
Letap Pharmaceuticals Limited-
Accra, Ghana
Abstract
Process validation is a requirement of Current Good Manufacturing Practices (cGMP). The essence of
Amo-Koi Seth
United States Pharmacopeia,
process validation is to ascertain the quality of a product throughout its production life cycle. The various
Ghana steps in manufacturing processes must be validated as it is essential for quality to be built into the
manufacturing processes of drugs. In this study, we discuss the Process validation of Artesunate (Art)
Gordon-Jackson Andrew tablets and Amodiaquine (Amod) tablets. The following critical tests were performed on the blend;
Department of Science Assay, blend uniformity studies, blend characteristics and flowability properties (Bulk Density, Tap
Laboratory Technology, Accra Density, Compressibility Index, Hausner Ratio), resistance to Segregation studies, Loss on Drying and
Technical University, Accra, Blend hold-time studies. The following analytical tests were performed on the compressed tablets;
Ghana Identification, weight variation, Disintegration, Hardness, Average weight, Loss on Drying, Friability,
Assay, Dissolution, Thickness and Uniformity of Dosage Unit. The following were performed on the
Angela Asor packaged finished products; leak test, Print quality, carton seal integrity. Percentage yield was also
Department of Chemistry, determined. The following statistical tools were used in the data evaluation; F-test, Shapiro-Wilk test,
Michigan State University, East
Kurtosis, Skewness, T-test emanating from regression analysis, Tests for Normality and Tests for
Lansing, USA
Significant differences. Results indicated an acceptable level of homogeneity within a batch and a high
David Mingle
level of consistencies between batches. The manufacturing process was concluded to be capable and
Department of Pharmaceutical stable to assure quality and safe products.
Sciences, College of Pharmacy,
University of Tennessee Health Keywords: Artesunate, Amodiaquine, Statistics, Process Validation
Science Centre, Memphis, USA
1. Introduction
The US FDA in its January 2011 Guidance for Industry (Process Validation: General
Principles and Practices) defines Process Validation as the collection and evaluation of data,
from the process design stage through commercial production, which establishes scientific
evidence that a process is capable of consistently delivering quality product.
Effective process validation contributes significantly to assuring drug quality. The basic
principle of quality assurance is that a drug should be produced that is fit for its intended use.
This principle incorporates the understanding that quality, safety, and efficacy are designed or
built into the product.
The guidance describes process validation activities in three stages.
 Stage 1: Process Design: The commercial manufacturing process is defined during this
stage based on knowledge gained through development and scale-up activities.
 Stage 2: Process Qualification: During this stage, the process design is evaluated to
determine if the process is capable of reproducible commercial manufacturing.
 Stage 3: Continued Process Verification: Ongoing assurance is gained during routine
production that the process remains in a state of control [1].

Process validation is a requirement of current cGMP. The essence of process validation is to

establish through scientific/designed data collection and analysis that the defined
manufacturing process is capable of reliably and repeatedly rendering a product of the required
quality that consistently meets all quality and design specifications. cGMP has identified three
different ways of validating a process; Prospective, Concurrent and Retrospective validations.
Revalidation is also performed after significant changes in a manufacturing process.
According to cGMP, Qualification and validation should establish and provide documentary
Corresponding Author: evidence that a specific process will consistently produce a product meeting its predetermined
Tuani YT specifications and quality attributes (process validation or PV, also called performance
Letap Pharmaceuticals Limited-
Accra, Ghana
qualification or PQ) [2].
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1.1 Types of process validation of desired characteristics from a review of historical data,
1.1.1 Prospective Validation based on the precondition that composition, procedures and
This form of validation which should be performed for new or equipment remain unchanged, and that facility, experience
substantially modified manufacturing processes, is essential and the results from in-process and final control tests are
for restricting the risk of errors occurring in advance; e.g. the evaluated.
preparation of injectable products requires this form of
validation. 1.1.4 Revalidation
This is needed to ensure that changes in the process and/or in
1.1.2 Concurrent Validation the process environment, whether introduced intentionally or
This is the form of validation carried out during the normal unintentionally, do not adversely affect process characteristics
production of a product to ensure that a process produces and product quality [3].
products with desired characteristics as the manufacturing
process is implemented. 2. Materials and Methods
The following production and analytical equipment were
1.1.3 Retrospective Validation utilized during production and quality control processes;
It is the form of validation which demonstrates process Moisture Analyzer, Analytical Balance, Dissolution Tester,
consistency and involves looking back into past experiences Disintegration Tester, Weighing Balance, Vernier Caliper,
obtained during production; i.e. establishment of documented Friability Tester, Tablet Compression Machine, Mechanical
evidence that a process will continuously produce a product Mixer, Dispensing Booth, Stop Watch.

Fig 1: Schematic Diagram of Manufacturing Process

(Wet granulation)

Table 1: Product Compositions

Raw Material/Ingredient
Artesunate Tablet Amodiaquine Tablet
Artesunate BP Amodiaquine hydrochloride BP
Lactose granules Super Tab 24 AN BP Lactose BP
Sodium lauryl sulphate BP Starch BP
Magnesium stearate BP P.V.P. K30 BP
Starch BP Nipasept sodium BP
Sodium starch glycolate BP Sodium starcy glycolate BP
Magnesium stearate BP
Cab-O-sil BP
Stearicacid BP
Talc BP

2.1 Study Design were identified as T5 and those from the bottom were
The validation design was based on the study of the effect of represented by B5. Samples represented by T1, T2, T3 and T4
critical operation variables on the quality of intermediate and were sampled about 6 cm away from the walls of the blender
finished products at identified critical control points. The and equidistance to each other and the middle sampling
design considered sampling from ten points. points. Samples from the bottom region were represented as
The blender employed was U-shaped. Cluster sampling was follows; B5, B1, B2, B3 and B4.
adopted for the blend. Top samples from the middle region

Fig 2: Sampling plan for Top region Fig 3: Sampling Plan for Bottom region
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NB: The following figures; 1,2,3,4,5,6,7,8,9 and 10 were used and end of each process. The blister packaging machine was
to represent the following cluster sampling points qualified and all packing materials were pretested. 100 tablets
respectively; T1,T2,T3,T4,T5,B1,B2,B3,B4 and B5, for the were taken at initial, midway and closing stages of blister
purposes of statistical analysis. For compression and packaging operations. Complete quality control tests were
packaging, the expected operations and performance of the performed on the blistered and packed samples. Results
compression machine was verified. 100 tablets for each batch obtained were evaluated and suitable conclusions were drawn.
were sampled during compression at the beginning, middle

Table 2: Critical Control Variables

Identified Critical Control Variables
No. Critical Test
Control Point Variable Defined Value
Mixing times after
After Mixing and 3 minutes
Lubrication
lubrication;
Mixer capacity 5 L – 1200 L
Artesunate Blend Uniformity and Characteristic Study
Mixing speed Low speed
1. (Assay, Appearance, Bulk density, tap density, hausner ratio,
Mixing times after
3 minutes compressibility, index, LOD)
Lubrication
Amodiaquine
Mixer capacity 5 L – 1200 L
Mixing speed Low speed
Transfer of blend
2. - - Blend segregation study (Assay)
into holding drums
oC
After Holding Storage Temp NMT 30
Blend Hold-Time Study (Assay, Appearance, Bulk density, Tap density,
3. blend for one
Storage Hum. NMT 75 % RH hausner ratio, compressibility, index, LOD)
month
Compression speed 36 rpm
Punch size 12 mm
Tablets
Lower punch =
Compression;
‘Break line’
Artesunate Embossment
Upper punch = ‘LS
Compression Process Quality and Consistency
100’
4. (Assay, content uniformity, dissolution, Weight variation, LOD, DT,
Compression speed 36 rpm
friability, hardness, diameter, thickness)
Punch size 12 mm
Lower punch =
Amodiaquine
‘Break line’
Embossment
Upper punch = LQS
300
Output 240 blisters/minute
Sealing and Blistered product Quality
Forming/Film
5. Blister 172 oC (Leak test, Print/embossment quality, Assay, dissolution, LOD, DT,
temperature
friability)
Sealing Temperature 225 oC
Carton seal integrity (Print/embossment quality, Assay, dissolution, LOD,
6. Packaging - -
DT, friability, Average weight, Hardness, Weight variation, Identification)

2.2 Statistical Tools

Statistical tools used are shown in table using SPSS 13 [4].

Table 3: Formulas for Statistical Analysis

No. Tool Formula
1. Mean Sum of sample/number of sample

2. Standard deviation

3. T-test

F = explained variance
4. F-test
unexplained variance
5. Cpk USL – LSL/6*SD, USL: upper specification limit, LSL: lower specification limit, SD: standard deviation
6. CpU USL – µ/3*SD, USL: upper specification limit, LSL: lower specification limit, SD: standard deviation; µ: Mean
7. CpL µ – LSL/3*SD, USL: upper specification limit, LSL: lower specification limit, SD: standard deviation; µ: Mean

8. Skewness

9. ketosis
Confidence
10. Mean ± t * s / √N; s = standard deviation; N = sample size; t = Constant from t-distribution table
Interval
11. Total Yield (%) Actual Yield/Expected Yield * 100

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3. Results and Discussions

3.1 Blend Analysis
3.1.1 Blend Uniformity Studies

Table 4: Blend uniformity studies

Artesunate: 90.0 – 110.0 %; Amodiaquine: 90.0 – 110.0 %
Mixing Time = 3 minutes
No. Sampling Point Artesunate Amodiaquine
Batch 1 Batch 2 Batch 3 Batch 1 Batch 2 Batch 3
1 T1 97.91 103.92 98.49 98.47 100.65 99.73
2 T2 99.78 101.01 101.18 99.23 101.87 99.73
3 T3 102.35 101.38 96.98 98.47 97.72 99.58
4 T4 103.2 99.68 98.4 100.18 101.67 99.44
5 T5 102.02 106.23 97.85 98.93 100.23 99.33
6 B1 102.94 106.99 106.31 99.29 98.57 99.47
7 B2 100.6 97.97 97.95 98.42 98.52 99.87
8 B3 102.24 98.54 104.97 98.77 98.73 100.16
9 B4 103.99 104.5 97.35 98.48 98.74 99.82
10 B5 98.92 104.99 102.88 98.60 101.12 99.36
CI at 99 % 101.40 102.52 ± 100.24 98.88 99.78 99.65
Probability level ± 2.05 3.32 ± 3.48 ± 0.57 ± 1.54 ± 0.27

[Link] Graphical Representation of Regression Analysis for Blend Uniformity Studies

Fig 4: Regression analysis for Batch 1(Art) Fig 5: Regression analysis for Batch 1(Amod)

Regression Analysis (Cubic) and Test for Normality on each 3.0. If Kurtosis is greater than 3.0, it is heavily tailed. If
batch was performed. Null Hypothesis: The data are normally Kurtosis is less than 3.0, data set is slightly tailed.
distributed. The null hypothesis is rejected if the p-value is Skewness: If the skewness is between -0.5 and 0.5, the data
below 0.05 [5]. A histogram for each Batch was plotted. Also are fairly symmetrical. If the skewness is between -1 and –
included was Boxplot for each batch showing the Content 0.5 or between 0.5 and 1, the data are moderately skewed. If
Distribution of Artesunate and Amodiaquine. Skewness and the skewness is less than -1 or greater than 1, the data are
Kurtosis were determined. For normal distribution, Kurtosis = highly skewed [6].

[Link] Statistical Data for Blend Uniformity

Table 5: Statistical Descriptives of Test for Normality (Blend Uniformity of Artesunate)

Batch 1 Batch 2 Batch 3
Statistic Std. Error Statistic Std. Error Statistic Std. Error
Mean 1.0140E2 0.63164 1.0252E2 1.02152 1.0024E2 1.07215
Lower Bound 99.9661 1.0021E2 97.8106
95% Confidence Interval for Mean
Upper Bound 1.0282E2 1.0483E2 1.0266E2
5% Trimmed Mean 1.0144E2 1.0253E2 1.0008E2
Median 1.0213E2 1.0265E2 98.4450
Variance 3.990 10.435 11.495
Std. Deviation 1.99741 3.23032 3.39043
Minimum 97.91 97.97 96.98
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Maximum 103.99 106.99 106.31

Range 6.08 9.02 9.33
Interquartile Range 3.44 5.90 5.68
Skewness -0.583 0.687 -0.080 0.687 0.905 0.687
Kurtosis -0.837 1.334 -1.540 1.334 -0.746 1.334

Table 6: Tests of Normality

Specification: Null hypothesis is rejected if p-value is less than 0.05 for Shapiro-Wilk test. p-value is labeled as “sig” in SPSS.
Shapiro-Wilk
Artesunate Amodiaquine
Statistic df Sig. Statistic df Sig.
Batch 1 0.936 10 0.509 0.814 10 0.021
Batch 2 0.936 10 0.504 0.899 10 0.215
Batch 3 0.843 10 0.048 0.943 10 0.584

[Link].1 Histogram and Normal Distributive Curve showing the Content Distribution

Fig 6: Normal distributive curve for Batch 1 of (Art) Fig 7: Boxplot for Batch 1 of (Art)

Artesunate 0.690 respectively. The null hypothesis is accepted as all

The p-values for Batches 1, 2 and 3 were 0.509, 0.504 and batches are greater than 0.05 for the Shapiro-Wilk test. This
0.048 respectively. The null hypothesis is accepted for implies the data for all batches are not numerically significant.
Batches 1 and 2 as their p-values are greater than 0.05 for the In terms of skewness, data for Batches 1, 2 and 3 were 1.579,
Shapiro-Wilk test. For Batch 3, the p-value of 0.048 implies 0.170 and 0.606 respectively. Batch 1 is greater than 1 that,
the batch fell slightly short of significance (p>0.0167). the data is highly skewed. Batch 2 is between -0.5 and 0.5
In terms of skewness, data for batches 1, 2 and 3 were -0.583, implying that the data is moderately skewed. Batch 3 is
-0.080 and 0.905 respectively. Batch 3 is between 0.5 and 1 between 0.5 and 1 implying that, the data is moderately
implying that, the data is moderately skewed. Batch 1 is symmetrical.
between-1 and – 0.5 implying that the data is moderately For kurtosis, batches 1, 2 and 3 were 2.519, -1.703 and -0.091
skewed. Batch 2 is between -0.5 and 0.5 implying that, the respectively which are all less than 3 implying that, all the
data is fairly symmetrical. For kurtosis, batches 1, 2 and 3 sets of data had lighter tails.
were -0.837, -1.540 and -0746 respectively which are all less
than 3 implying that, all the sets of data had lighter tails. [Link] Inter-batch Analysis
Null Hypothesis: There are no significant differences between
Amodiaquine batches 1, 2 and 3 at 0.1 % significance level for Artesunate
The p-values for Batches 1, 2 and 3 were 0.457, 0.057 and and Amodiaquine.

Table 7: Results of inter-batch analysis of Artesunate

Specification: F– calculated for inter batch samples should be NMT 3.18 at 95 % confidence level; RSD %: NMT 5.00 %
Artesunate
No. Sampling Point
Batch 1 Batch 2 Batch 1 Batch 3 Batch 2 Batch 3
1 T1 97.91 103.92 97.91 98.49 103.92 98.49
2 T2 99.78 101.01 99.78 101.18 101.01 101.18
3 T3 102.35 101.38 102.35 96.98 101.38 96.98
4 T4 103.2 99.68 103.2 98.4 99.68 98.4
5 T5 102.02 106.23 102.02 97.85 106.23 97.85
6 B1 102.94 106.99 102.94 106.31 106.99 106.31
7 B2 100.6 97.97 100.6 97.95 97.97 97.95
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8 B3 102.24 98.54 102.24 104.97 98.54 104.97

9 B4 103.99 104.5 103.99 97.35 104.5 97.35
10 B5 98.92 104.99 98.92 102.88 104.99 102.88
SD 2.00 3.23 2.00 3.39 3.23 3.39
Variance 3.99 10.43 3.99 11.49 10.43 11.49
RSD % 1.97 3.15 1.97 3.38 3.15 3.38
F-test at 0.1 % significance level 2.62 2.88 1.10

For Artesunate, the F-calculated values of 2.62, 2.88 and 1.10 1, 2 and 3 were 1.176 ± 0.018, 1.182 ± 0.002 and 1.171 ±
were lower than the F-tabulated value of 10.11 at 0.1 % 0.002 respectively. This implies that the flow character for all
significance level indicating insignificant differences between the batches were good (1.12 – 1.18). Bulk and tap densities
the batches. were used to predict the flow and the compressibility
For Amodiaquine, the F-calculated values of 7.25, 4.50 and character of the powders.
8.32 were lower than the F-tabulated value of 10.11 at 0.1 %
significance level indicating insignificant differences between Amodiaquine
the batches. The compressibility indices of Batches 1, 2 and 3 were
11.969±0.078, 13.651±0.063 and 13.860±0.040 respectively.
[Link] Blend Characterization and Flowability Properties This implies that the compressibility characteristics of all the
A Bulk and tap density apparatus was used to determine the batches were good (11-15). Hausner ratio values for Batches
following parameters, bulk density, tap density, 1, 2 and 3 were 1.133±0.001, 1.161±0.008 and 1.140±0.011
compressibility index and hausner ratio [7]. respectively. This implies that the flow character for all the
batches were good (1.12 – 1.18). Bulk and tap densities were
Table 8: Scale of Flowability used to predict the flow and the compressibility character of
Compressibility index (%) Flow Character Hausner Ratio the powders.
≤ 10 Excellent 1.00 – 1.11
11 – 15 Good 1.12 – 1.18 [Link] Loss on Drying (LOD) (%)
16 – 20 Fair 1.19 – 1.25 LOD (%) for the granules was determined after 3 minutes of
21 – 25 Passable 1.26 – 1.34 mixing with sampling from all 10 sampling points. Data was
26 – 31 Poor 1.35 – 1.45 recorded and regression analysis was performed on the data
32 – 37 Very poor 1.46 – 1.59 obtained for each batch.
> 38 Very, very poor > 1.60 For Artesunate, the Loss on drying gave the following values
for Batches 1, 2 and 3 respectively; 1.41 ± 0.44 %, 1.30 ±
Artesunate 0.17 % and 0.98 ± 0.07 %. These values are less than the
The compressibility indices of Batches 1, 2 and 3 were 14.943 specified upper limit of 5.00 %. For Amodiaquine, the Loss
± 1.275, 15.454 ± 0.161 and 14.506 ± 0.122 respectively. This on drying gave the following values for Batches 1, 2 and 3
implies that the compressibility characteristics of all the respectively; 1.51 ± 0.10 %, 1.53 ± 0.11 % and 1.86 ± 0.07 %.
batches were good (11-15). Hausner ratio values for Batches These values are less than the specified upper limit of 2.00 %.

[Link].1 Graphical Representation for LOD (%)

Fig 8: Regression analysis of LOD (Art, Batch 1) Fig 9: Regression analysis of LOD (Amod, Batch 1)

[Link] Resistance to Segregation Studies at 99 % probability level = 9.925.

Statistical evaluation of Data obtained from holding drums as Null Hypothesis: There is no significant difference between
against data from Blend Uniformity studies was performed for the assay results obtained from the blend uniformity studies
statistical differences. T-test (Two tailed test); Critical value and those obtained from the holding drums.

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Table 9: Assay results of holding drums for Artesunate

Holding Drum Batch No. 1 Batch Bo. 2 Batch No. 3
Drum 1 99.51 101.55 104.52

Table 10: Mean values of sampling positions1 for Artesunate

Sampling Positions Batch No. 1 Batch No. 2 Batch No. 3
T1 97.91 103.92 98.49
B1 102.94 106.99 106.31
Mean ± SEM 100.43±2.52 105.46±1.53 102.40±3.91
T2 99.78 101.01 101.18
B2 100.6 97.97 97.95
Mean ± SEM 100.19±0.41 99.49±1.52 99.57±1.62
T3 102.35 101.38 96.98
B3 102.24 98.54 104.97
Mean ± SEM 102.30±0.05 99.96±1.42 100.98±4.00
T4 103.20 99.68 98.4
B4 103.99 104.5 97.35
Mean ± SEM 103.60±0.40 102.09±2.41 97.88±0.53
T5 102.02 106.23 97.85
B5 98.92 104.99 102.88
Mean ± SEM 100.47±1.55 105.61±0.62 100.37±2.51

Table 11: Statistical evaluation of Data obtained from holding drums as against data from Blend Uniformity (Artesunate)
Specification: T-critical value = 9.925 at 99 % probability level.
Artesunate Batch 1
Drum 1 T1 and B1 T2 and B2 T3 and B3 T4 and B4 T5 and B5
Mean Assay (%) 99.51 100.43 100.19 102.30 103.60 100.47
T-Test 0.414 0.884 4.886 2.208 0.540
Artesunate Batch 2
Mean Assay (%) 101.55 105.46 99.49 99.96 102.09 105.61
T-Test 1.095 0.799 0.727 0.325 1.757
Artesunate Batch 3
Mean Assay (%) 104.52 102.40 99.57 100.98 97.88 100.37
T-Test 0.506 1.203 0.647 2.443 0.882

T-test was carried out to prove that, there is no significant For Amodiaquine, T-test was carried out to prove that, there is
difference between the assay results obtained from the no significant difference between the assay results obtained
analysis of the blend transferred into the holding drums and from the analysis of the blend transferred into the holding
the assay results obtained at the various sampling positions drums and the assay results obtained at the various sampling
during the uniformity of blend studies. The t-values were less positions during the uniformity of blend studies. The t-values
than the t-critical (tabulated) value of 9.925 at 99 % were less than the t-critical (tabulated) value of 9.925 at 99 %
probability level. Hence, there is no significant difference probability level. Hence, there is no significant difference
between the mean assay results. The null hypothesis is between the mean assay results. The null hypothesis is
therefore accepted implying that, no segregation takes place therefore accepted implying that, no segregation takes place
when blends are transferred into holding drums. when blends are transferred into holding drums.

[Link] Blend Hold-Time studies

Table 12: Results from Blend hold-time studies for Artesunate

Specification: Must be within specifications for Appearance, Assay, LOD, Bulk density, tap density, Compressibility index and Hausner
ratio.
Study time: Initial Study time: 30th Day
Test Parameter
Batch No. 1 Batch No. 2 Batch No. 3 Batch No. 1 Batch No. 2 Batch No. 3
Appearance Granular Granular Granular Granular Granular Granular
Assay (%) 102.35 104.68 97.85 101.29 102.84 98.08
LOD (%) 1.20 1.18 0.99 1.17 1.21 1.08
Bulk Density 0.551 0.546 0.554 0.561 0.558 0.564
Tap Density 0.647 0.645 0.646 0.645 0.646 0.648
Comp. Index 14.835 15.217 14.285 14.845 15.381 14.981
Hausner Ratio 1.174 1.179 1.166 1.149 1.158 1.149

A simulated container was adopted for the blend holding time Hausner ratio and Appearance were the parameters tested at
studies. The blends were held for one month in a polyethylene initial and at the 30th day. Results show insignificant
rubber inserted in High Density Poly Ethylene (HDPE) differences between results obtained at initial and those
bottles. Assay, LOD, Bulk density, Compressibility index, obtained on the 30th day. Thus, holding the blend for a period
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of one month in HDPE drums and also at the required storage within specification of 9.96 – 10.02 for Artesunate and 12.76
condition (below 30 oC and NMT 60 % RH) will – 12.82 for Amodiaquine.
insignificantly affect the blend with respect to the tested
parameters [Link] Uniformity of Dosage Unit (Weight Variation
(WV))
[Link] Tablet Compression Process Performance and
Product Quality Studies Artesunate
Tests for Hardness, Thickness and Diameter were performed
Dose and ratio of Drug substance implies 25 mg and 25%
on the compressed tablets at beginning, middle and end of the [8]
compression process for all three batches of Artesunate and
Amodiaquine. With Artesunate have a specification of Not Table 13: Assay Results (Artesunate)
less than 2.5 for hardness, all tested tablets were within
specification and that of Amodiaquine were also within Batch Average weight (mg) of 20 Tablets Assay (%)
specification of Not less than 1.5. For Thickness test, the Batch No. 1 355.0 103.65
specification for Artesunate was 4.30 – 4.50 mm and all tested Batch No. 2 354.0 106.98
tablets were within specification. For diameter, results were Batch No. 3 352.0 103.72

Table 14: Sample Uniformity of Dosage Unit (Batch No. 1; 10 Determinations)

Specification: AV of first 10 dosage units is ≤ L1 %; L1 = 15
Batch No. 1 (Assay =103.65)
Beginning Middle End
Individual Weights Individual Estimated Individual Individual Estimated Individual Weights Individual Estimated
(mg) Content (%) Weights (mg) Content (%) (mg) Content (%)
Mean ± SEM 350.4 ± 1.25 105.55 ± 0.36 348.2 ± 1.36 101.66 ± 0.4 350.3 ± 0.63 102.28 ± 0.18
SD 3.95 1.15 4.29 1.25 2 0.58
RSD % - 1.13% - 1.23% - 0.57%
AV = IM-XI + ks; k = 2.4; SD = 0.41 ; AV = IM-XI + ks; k = 2.4; SD = 0.60; X =
AV = IM-XI + ks; k = 2.4; SD = 1.19 ; X = 105.55; T = 100
X = 104.89; T = 100 %; M(Case 1); X > 105.52; T = 100 %; M(Case 1); X > 101.5
%; M(Case 1); X > 101.5 %, AV = X – 101.5 + ks
101.5 %, AV = X – 101.5 + ks %, AV = X – 101.5 + ks
AV = 3.57 AV = 3.17 AV = 2.18

Fig 10: Regression analysis for Art Batch 1 (WV) Fig 11: Regression analysis for Amod Batch 1 (WV)

Artesunate Amodiaquine
Acceptance values for Batch 1 at the beginning, middle and Acceptance values for Batch 1 at the beginning, middle and
end of compression were 3.57, 3.17 and 2.18 respectively. end of compression were 2.52, 0.96 and 2.29 respectively.
Those of Batch 2 at the beginning, middle and end of Those of Batch 2 at the beginning, middle and end of
compression were 7.11, 6.96 and 5.70 respectively whereas compression were 2.17, 1.15 and 1.31 respectively whereas
Batch 3 at the beginning, middle and end of compression Batch 3 at the beginning, middle and end of compression
were 3.28, 4.01 and 3.10 respectively. The acceptance values were 2.00, 3.01 and 1.83 respectively. The acceptance values
are all less than L1 = 15.0 [8] which indicates consistent are all less than L1 = 15.0 which indicates consistent
uniformity in the dosage units of all the batches. uniformity in the dosage units of all the batches.

3.2 Finished Products Analysis

3.2.1 Quality Control Tests

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Table 15: Complete analysis of Compressed Tablets (Artesunate)

Result
No. Test Specification Reference
Batch 1 Batch 2 Batch 3
The retention time of Artesunate in Chromatograms United States
Identification
1 obtained from both standard and test solutions are Pharmacopeia Complies Complies Complies
By HPLC
comparable. (USP)
Not more than 2 of the individual masses deviates
from average mass by more than the percentage British
2 Weight variation Complies Complies Complies
deviation of 7.5 % and none deviates by more than Pharmacopeia (BP)
15 %.
3 Disintegration Not more than 15 minutes In-House (IH) 5 4 4
4 Hardness (Kp) Not Less than 2.5 IH 5.46 4.86 4.47
5 Average weight (mg) 332.50 – 367.50 (mg) IH 349 351 351
Loss on Drying (by
6 Not more than 5.00 % IH 1.68 1.61 1.20
moisture analyzer at 105 oC)
7 Friability Not more than 1.00 % BP 0.56 0.59 0.68
8 Assay 90.0 % - 110.0 % Ph. Int 106.59 106.76 103.11
9 Dissolution Not less than 75 % (Q) Ph. Int 86.49 90.96 95.28

Table 16: Complete analysis of Compressed Tablets (Amodiaquine)

Result
No. Test Specification Reference
Batch 1 Batch 2 Batch 3
Identification The spectra of Amodiaquine obtained from both test and IH
a) By UV standard solutions are comparable
1 Complies Complies Complies
Or The retention times of Amodiaquine in both chromatograms USP
b) By HPLC obtained from both standard and test solutions are comparable
Not more than 2 of the individual masses deviates from
2 Weight variation average mass by more than the percentage deviation of 7.5 % BP Complies Complies Complies
and none deviates by more than 15 %.
3 Disintegration Not more than 15 minutes USP 4 4 6
4 Hardness (Kp) Not Less than 1.5 IH 6.80 5.35 4.91
5 Average weight (mg) 455.70 – 474.30 (mg) IH 464 468 461
Loss on Drying (by
6 Not more than 2.00 % IH 1.63 1.56 1.75
moisture analyzer at 105 oC)
7 Friability Not more than 1.00 % BP 0.52 0.53 0.42
8 Assay 93.0 % - 107.0 % USP 98.79 98.56 99.48
9 Dissolution Not less than 75 % (Q) USP 96.94 97.41 94.47

3.2.2 Process Capability

Table 17: Process Capability Index of Batches 1, 2 and 3 of Artesunate and Amodiaquine with sampling at beginning middle and End of process
Specifications: Cpk< 1 -not capable; Cpk= 1 -marginally capable; Cpk> 1 -capable [9]
Artesunate Amodiaquine
Hardness Test Thickness Diameter Hardness Test Thickness Diameter
µ ± SEM 5.57 ± 0.09 4.41 ± 0.01 10.00 ± 0.01 6.68 ± 0.12 4.60 ± 0.01 12.01 ± 0.01
SD 0.89 0.10 0.10 1.14 0.11 0.08
LSL 2.5 3.00 8.5 2 3.50 11.00
3*sd 2.67 0.29 0.29 3.41 0.32 0.23
CpL 1.15 4.85 5.25 1.37 3.39 4.49

Table 18: Process capability index for Uniformity of Dosage Units and 3.57 respectively, which are greater than 1. This implies
(Art); 10 Determinations that, the manufacturing process is reproducible as well as
Art Amod capable of consistently delivering quality products.
SD 1.40 0.94
LSL 85 85 Amodiaquine
USL 115 115 Process capability indices for Hardness, Thickness, Diameter
6SD 8.40 5.64 and Uniformity of Dosage units tests were 1.37, 3.39, 4.49
Cp 3.57 5.32 and 5.32 respectively, which are greater than 1. This implies
that, the manufacturing process is reproducible as well as
Artesunate capable of consistently delivering quality products.
Process capability indices for Hardness, Thickness, Diameter
and Uniformity of Dosage units tests were 1.15, 4.85, 5.25 3.2.3 Yield Analysis

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Table 19: Yield Analysis of Artesunate

Granulation (95 % - 105 %) (IH) Compression (95 - 105 %) (IH)
Parameter
Batch 1 Batch 2 Batch 3 Batch 1 Batch 2 Batch 3
Artesunate
35.00 35.00 35.00 100000 100000 100000
Expected Yield
Amodiaquine
46.41 46.50 46.50 100000 100000 100000
Artesunate
34.84 35.06 35.08 96800 95485 97314
Actual yield
Amodiaquine
45.41 46.02 45.64 95763 96817 96623
Artesunate
99.54 100.17 100.23 96.80 95.49 97.31
% Yield / Reconciliation
Amodiaquine
97.85 98.97 98.15 95.73 96.82 96.62

The percentage yields obtained from the compression and to-statistical-applications-for-process-validation

granulation stages for all three batches were within
specification for both Artesunate and Amodiaquine.

4. Conclusion
Results obtained indicated that, there was an acceptable level
of homogeneity within a batch and consistency between
batches. All critical variables are therefore valid indicating
that, the manufacturing processes for Artesunate and
Amodiaquine tablets had been robustly designed enough to
meet predetermined standards and quality attributes. The
manufacturing process as a result is capable and stable to
assure quality and safe products. Quality control tests carried
out on the compressed tablets of Artesunate and Amodiaquine
were within the acceptance criteria for Identification, Weight
variation, Disintegration, Friability, Assay, Hardness,
Dissolution, Average weight and Loss on Drying.

5. References
1. USFDA, “Guidance for Industry, Process Validation:
General Principles and Practices”, 2011,1-18
[Link]/downloads/Drugs/GuidanceComplianceRe
gulatory; [Visited on 07/31/2020]
2. Annex 2, WHO good manufacturing practices for
pharmaceutical products: main principles1; WHO
Technical Report Series No. 986, 201
3. Ajibola A. Olaniyi, Principles of Drug Quality Assurance
and Pharmaceutical analysis. 2005; 116-120:447-469.
Published in 2000 AD by Mosuro Publishers; Reprinted
4. Statistical Package for the Social Sciences (SPSS 13.0);
initially developed by Norman H. Nie, Dale H. Bent, C.
Hadlai Hull
5. [Link]
significant-2/; [Visited on 07/31/2020]
6. [Link]://[Link]/doc/en/reporting-and-
dashboards/maql-analytical-query-language/maql-
expression-reference/aggregation-functions/statistical-
functions/predictive-statistical-use-cases/normality-
testing-skewness-and-kurtosis; [Visited on 07/31/2020]
7. USP 29-NF24; General chapter 1174, Powder Flow
8. BP; Appendix XII C. Consistency of Formulated
Preparations; Uniformity of Weight (Mass) (Ph. Eur.
Method 2.9.5); Uniformity of Content ([Link]. Method
2.9.6), Uniformity of Dosage Units ([Link]. Method
2.9.40), 2008.
9. Eugenie Khlenikova, McNeil Consumer Healthcare;
Introduction to Statistical Applications for Process
Validation;
[Link]
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