Content uploaded by Musa Autamashih
Author content
All content in this area was uploaded by Musa Autamashih on Dec 05, 2022
Content may be subject to copyright.
www.mnf-journal.com Page 1 Molecular Nutrition & Food Research
Received: 21/01/2021; Revised: 19/05/2021; Accepted: 19/05/2021
This article has been accepted for publication and undergone full peer review but has not been
through the copyediting, typesetting, pagination and proofreading process, which may lead to
differences between this version and the Version of Record. Please cite this article as doi:
10.1002/star.202100016.
This article is protected by copyright. All rights reserved.
4-in-1 multipurpose excipient from Musa acuminata fruit by alkaline-steeping/retrogradation (ASR)
in acetaminophen tablet formulation
Musa Autamashih1, Naushaad Ebrahim1, Samuel Egieyeh1, Marique Aucamp1, Paulo Cesar
Pires Rosa2, Madan S. Poka3, Jean Baptiste Ngilirabanga1 and Halima Samsodien1
1School of Pharmacy, The University of the Western Cape, Cape Town, South Africa.
2Faculty of Pharmaceutical Sciences, University of Campinas, Sao Paulo, Brazil.
3Department of Pharmacy, School of Health Sciences, University of Limpopo, South Africa
*Corresponding author:
Dr. Musa Autamashih, Discipline of Pharmaceutics, School of Pharmacy, University of the
Western Cape, South Africa. Tel: +16165897259 (USA), +27635743310 (SA).
Email: autamash@gmail.com, 3699619@myuwc.ac.za, musa-autamashih@outlook.com
Abbreviations
www.mnf-journal.com Page 2 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
ACM Acetaminophen
API Active pharmaceutical ingredients
ASR. Alkaline-steeping/retrogradation
ASS. Alkaline solution steeping
MAP Musa acuminata advanced starch polymer
MAS Musa acuminata starch
MASS Modified alkaline solution steeping
MMS Mass of Musa acuminata slurry
MCC Microcrystalline cellulose
MSR Mass of starch recovered
MRS Mass of the retrograded starch
MS Modified maize starch (Starch 1500)
IMS Initial mass of a starch
MST Magnesium stearate
NA Not available
PVP Polyvinylpyrrolidone
RMAS Retrograded Musa acuminata starch
RCFR:DT Resistance to crush-friability/disintegration time ratio
TC Talc
TGA Thermogravimetric Analysis
www.mnf-journal.com Page 3 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
Keywords: 4-in-1 multipurpose excipient; acetaminophen tablet formulation; alkaline-
steeping; Musa acuminata; retrogradation.
Abstract
The conventional approach in the tablet formulation of acetaminophen (ACM) suggests the
use of five or more different excipients in a wet granulation tableting process. The use of
many excipients in tablet formulation may negatively create excipient-excipient interactions,
excipient-drug interactions, exaggerated product side effects, and high drug load. Cutting-
edge technology would be the use of one excipient with a quadrupled functional purpose (4-
in-1) by direct compression tableting. In this study, a novel two-phase process called
‘alkaline-steeping/retrogradation’ (ASR) was employed to obtain the desired starch polymer
excipient of quadrupled functional purpose. In phase I, the biopolymer was extracted from
the unripe fruits of Musa acuminata by a modified alkaline solution steeping method, while in
phase II, 50% w/w of the extracted polymer was retrograded. The retrograded product was
re-mixed with the non-retrograded 50% w/w that was left from phase I. This gave a novel 50-
50% w/w blend named Musa acuminata advanced starch polymer (MAP). To authenticate
the efficiency of the ASR, the physicomechanical, analytical, and drug release properties of
different concentrations of MAP/ACM solid systems were characterized to ascertain the
www.mnf-journal.com Page 4 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
compatibility. The ASR method produced a unique semi-hygroscopic biopolymer excipient of
quadruple-function in ACM high-dose tablet formulation by direct compression.
1. Introduction
Efforts have been intensified in the exploration of starch to meet the increasing demand for
excipients in the pharmaceutical industry [1, 2]. Starch is a semi-crystalline composite
substrate made of two biopolymers, amylose and amylopectin. Amylose is an amorphous,
substantially linear glucan polymer with α-1,4 linked glucose residues, while amylopectin is a
highly branched molecule made of α-1,4 linked glucose residues as the backbone and 5% α-
1,6 limked glucose as branches [3]. Amyloses have good structural properties because they
pack closely to form strong, rigid, and insoluble material, thus providing good thickening
properties, unlike amylopectin, which is readily soluble in aqueous systems.
Alkaline steeping, water steeping, and acid steeping, along with enzymatic isolation, are the
main extraction processes for starch. The alkaline steeping-method has the advantage of
simplicity, while producing a high purity starch [4]. Retrogradation, may also be seen as a
process of isolation and modification of starch molecules by heating in the presence of water
(gelatinization) and then freezing. When the amylose and amylopectin chains are heated,
gelatinized starch realign themselves as the cooked starch is cooled [5].
www.mnf-journal.com Page 5 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
The starch content of the unripe fruit of Musa acuminata has been found to be 84% w/w of
the total mass [3]. This concentration is higher than those in Zea mays (73% w/w) [6],
Ipomoea batatas (80% w/w) [7], and Solanum tuberosum (60% w/w) [8]. Starch polymers
differ in morphology [1, 2] and swelling capacity [1, 7] depending on the species of the plant
and the physicochemical processes applied during extraction and purification [2, 9]. The
characteristic semi-hygroscopic nature of modified starch has been found to relate to good
swelling, binding, and disintegrating capacity [1]. It is, therefore, reasonable to combine the
individual evaluated characteristics of starch polymers in a tablet formulation and evaluate
their multipurpose function.
There are many categories of pharmaceutical excipients serving different individual functions
in tablet formulation. Commonly used excipient categories based on their functions include:
fillers, binders, disintegrants, lubricants, glidants, and solvents (mostly water) [8, 9]. The
lower the number and quantity of excipients used in a formulation, the less incompatibilities
and side effects, and the lower the product cost [1]. Developing a quadrupled-function (4-in-
1) excipient (serving as a filler, binder, solvent, and disintegrant) in tablet formulation, would
be a ground-breaking technology.
This study was aimed at applying the novel two-phase ASR method of extraction, and
characterization of the starch polymer of the unripe fruit of Musa acuminata to enable its use
as a 4-in-1 excipient in acetaminophen (ACM) tablet formulation by direct compression.
ACM is an over-the-counter analgesic and antipyretic medicine widely used in the
pharmaceutical market [8]. The procedure for the preparation and analysis of ACM is
outlined in the United States Pharmacopeia [11]. To the authors’ knowledge, no literature
www.mnf-journal.com Page 6 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
exists in the application of the dual-phase ASR method to obtain a starch polymer of
quadruple-functional purpose, serving as filler, binder, disintegrant, and solvent in the
formulation of tablets.
2. Materials and Methods
2.1. Materials
Musa acuminata starch (MAS) was obtained from the fresh unripe fruits of green banana
packed by ‘Freshmark Farms’ (batch ID# 6009517208965), Brackenfell, South Africa. ACM
was obtained from Zhengzhou United Asia, China, (code 3401961603), Aerosil® 200 (AR)
from Evonik GmbH, Germany (CAS_No. 122945-52-5 7631-86-9), and microcrystalline
cellulose (MCC) branded Avicel® 200 was acquired from FMC BioPolymer (CAS_No. 9004-
34-6), Belgium. Starch 1500® called modified maize starch (MS) was obtained from
Colorcon, Janssen (US Patent No 6,667,060). Magnesium stearate (MTS) was acquired
from Redbrain Chemicals, Birmingham, UK (ID# EG-MGC18H35022-1), polyvinylpyrrolidone
(PVP) from ChemCenter, USA, (CAS Number: 7758-79-4), and talc (TC) from Sigma-
Aldrich, USA, (CAS Number: 14807-96-6).
2.2. Methods
2.2.1. The two-phase Alkaline-steeping/retrogradation extraction - ASR
www.mnf-journal.com Page 7 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
The dual-phase extraction method called ‘alkaline-steeping/retrogradation’ (ASR) was
employed in the extraction processes of MAS to obtain a unique Musa acuminata advanced
starch polymer (MAP). This consisted of two phases of starch extraction namely, the
modified alkaline solution steeping (MASS) method, and the retrogradation (RTG) method.
2.2.1.1. Modified alkaline solution steeping extraction (MASS) method (Phase I
extraction)
The alkaline solution steeping method employed to remove unwanted materials such as
lipids and proteins, was adapted from Sun et al [12] with modifications as follows: Freshly
peeled unripe green banana fruits weighing about 3 kg were cut into small pieces and milled
into a slurry. The slurry was steeped in 2000 mL 0.1 N sodium hydroxide at 37°C for 3 h and
excess alkali solution was removed by washing the slurry several times with distilled water.
The final volume of the suspension was filtered using a suction pump (ABM Greiffenberger,
GmbH, Germany) aided with a porcelain funnel and filter paper (Whatman #1). The residue
(free from lipids and proteins) was placed in a hot air oven (Memmert 854, Schwabach,
Germany) at 40°C to dry for 3 h. The dry mass was size-reduced for 30 min in a grinder
(MFC Janke and Kunkel, GmbH IKA-Werk 7813, Germany) and further pulverized with a
pestle and mortar for another 30 min. The final MAS obtained was passed through a 1.6 mm
sieve, weighed, and stored for experimentation. The percentage yield of the starch polymer
was calculated as shown in Equation 1 below:
Percentage (%) yield of MAS = MSR/MMS x 100 (1)
www.mnf-journal.com Page 8 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
Where:
MSR = mass of starch recovered and
MMS = mass of Musa acuminate slurry
2.2.1.2. Retrogradation extraction (RTG) method (Phase II extraction)
A fraction of the extracted MAS (50% w/w) from phase I was weighed out and made into a
slurry with water in a beaker to obtain the concentration of 5 g/mL. This was followed by the
addition of boiling water in small quantities (1-5 mL) while stirring until a gel was obtained
[13]. The gel was boiled at 110°C for 30 min, transferred to a metallic container, and
refrigerated at a range of 0 to -8°C for 24 h. The ice-cold formed mass was grated into
pellets of varying sizes (lengths 1-10 mm), transferred into an aluminum pan, and then dried
in an oven at 60°C for 24 h. Dried flakes were formed, and these were milled into powder
using the MFC Janke and Kunkel grinder (GmbH Staufen, Germany) and weighed. The
percentage yield of the retrograded starch was calculated as shown in Equation 2 below [6]:
% Yield of retrograded Musa acuminate starch (RMAS) = MRS/IMS x 100 (2)
Where:
RMAS = the retrograded starch polymer of Musa acuminata
MRS = mass of the retrograded starch
IMS = initial mass of the starch used
www.mnf-journal.com Page 9 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
2.2.1.3. Mixing of the extracted products from phase I and phase II
The retrograded product obtained from phase II extraction was mixed with the remaining
extracted product (50% w/w) that was left from phase I. The blended mix gave a unique
polymer named Musa acuminata advanced starch polymer (MAP) having surface-striated
particles that are similar in structural morphology to MCC. MCC is the best direct
compression excipient due to its unique surface-striated morphology that provides good
lubricity and compressibility during tablet formulation.
2.2.2. Microstructure studies by Scanning Electron Microscopy (SEM)
Analysis of samples of ACM, MAP, and predetermined ratios of MAP/ACM blends was done
using SEM, conducted with a Carl Zeiss Field Emission scanning electron microscope (NTS
GmBH AURIGA, Germany). Q150T sputter coater from Quorum Technologies Ltd was used
for sample preparation. A film piece was mounted on aluminum stubs using carbon tape and
subsequently coated with an Au-Pd alloy layer (15-20 nm), allowing surface and cross-
section visualization. Samples were examined using an accelerated voltage of 20 kV at 500
X, 1.00 KX, and 5.00 KX magnifications.
2.2.3. Fourier Transform Infrared spectroscopy
The FTIR spectra were recorded for ACM, MAP, and predetermined ratios of MAP/ACM
solid systems. A sample was placed on the crystal of an FTIR Spectrometer (Spectrum 400,
PerkinElmer, USA). The pressure was applied gradually using the pressure gauge and
www.mnf-journal.com Page 10 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
ranged between 50 and 60%. A resolution of 4 cm-1 and a scanning range of 500 to 4000
cm-1 was used.
2.2.4. Thermogravimetric Analysis and Differential thermal analysis
A thermogravimetric analyzer (TGA 4000, PerkinElmer, USA) was used. A small sample
(2.5-3.0 mg) of ACM, MAP, and a predetermined ratio of MAP/ACM blend was heated at
10°C/min from 25°C to 450°C. Nitrogen at a flow rate of 20 mL/min was used as the purge
gas.
2.2.5. Differential Scanning Calorimetry
Thermal properties of ACM, MAP, and MAP/ACM solid systems were conducted using a
DSC 8000 PerkinElmer calorimeter (Waltham, USA). Nitrogen was used as the purge gas at
a rate of 20 mL/min. A quantity (2.7 mg) of each of the above-mentioned samples was
sealed in an aluminum pan and heated from about 30 to 400°C at a rate of 10°C/min,
followed by a cooling back cycle to 30°C at the same rate.
2.2.6. X-ray Diffraction and Relative Crystallinity
www.mnf-journal.com Page 11 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
Evaluation of the crystallinity patterns of the different MAP/ACM binary mixtures was
achieved from collected diffractograms recorded on a Siemens® X-ray diffractometer (Model
D5000; Germany), using a nickel-filtered Cu Kα radiation (λ = 1.5406 Å) (tube operating at
40 kV and 30 mA). The scanning regions were collected from 5 to 70° (2θ) in a step size of
0.05° (2θ).
2.2.7. Analysis of moisture
The oven-dry method of moisture content determination [14] was employed to determine the
moisture content in a sample of ACM, MAP, or MAP/ACM blend. The percentage moisture
content was calculated as shown in Equation 3 below:
MC = W1 - W2/W2 x 100% (3)
Where:
MC = Moisture content of a sample (ACM, MAP, or MAP/ACM ratio)
W1 = Initial weight of a sample (ACM, MAP, or MAP/ACM ratio)
W2 = Oven-dry weight of a sample (ACM, MAP, or MAP/ACM ratio)
2.2.8. Micromeritic evaluation of powder/granule mixtures
Tablet formulas for direct compression and wet granulation were constructed for all batches
of tablets as shown in Table 2. Micromeritic properties of the powder/granule mixtures (flow
www.mnf-journal.com Page 12 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
rate, angle of repose, bulk density, tapped density, true density, and moisture content) were
characterized as outlined in previous studies [3, 15], and tabulated (Table 1).
2.2.9. Tablet preparation
Different batches of ACM 500 mg tablets of six binary blends were formulated with the first
five batches containing MAP (10, 20, 30, 40, and 50% w/w) and AR (1% w/w), and the
second five batches containing MS, PVP, TC, MTS, and water. Compacts were obtained
using the Manesty Type F3 (Liver Poole, England) single punch tableting machine with a
punch diameter of 0.75 cm set at 32.5 kN compression pressure. The die volume was
adjusted to obtain 500 50% mg ACM.
2.2.10. Mechanical characterization of tablets
The tablet’s resistance to crush test (hardness) [11, 15] was done using the Pharma Test
Type PTB 301, (Germany), the friability test [15] using an Erweka GmbH Type TAD tester
(Germany), and the disintegration test [15] using the Electrolab disintegration tester (model
ED-2AL, India). The RCFR:DT (resistance to crush-friability disintegration time ratio) of
tablets [16] was calculated for all batches and results were obtained in triplicate and plotted.
The RCFR:DT identified as β is a mathematical model used to determine the quality of
tablets based on the type and characteristics of excipients [16]. This was calculated as
shown in Equation (3).
www.mnf-journal.com Page 13 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
β = (RC x FR)/DT (3)
Where:
β = resistance to crush-friability/disintegration time ratio
RC = resistance to crush
FR = friability
DT = disintegration time
2.2.11. In vitro drug release and quantification by HPLC
The in vitro drug release study was performed using the USP type I dissolution rate
apparatus (model 65-1100, NJ08820, USA), fitted with a basket rotating at 100 rpm,
performed in 900 mL of simulated gastric fluid (0.1 N HCl). At predetermined time intervals of
5, 10, 15, 20, 25, 30, 35, 40, and 45 min, a 5 mL sample was withdrawn, and the withdrawn
volume replaced with fresh medium to maintain sink conditions. The drug concentrations
were quantified using an Agilent® High-Performance Liquid Chromatography (HPLC) with
similar fitted accessories and procedures described previously [8]. Run conditions included
UV detection at 243 nm.
2.2.12. Statistical analysis
www.mnf-journal.com Page 14 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
Statistical analysis was performed by GraphPad Prism version 7.04, using Bonferroni's
multiple comparisons test (after two-way ANOVA) to evaluate the excipient activity of the
starch polymer. At a 95% confidence interval, p-values above 0.05 were considered not
significant. All data obtained were in triplicate.
3. Results and discussion
3.1. The two-phase ASR extraction
3.1.1. The MASS method (Phase I extraction)
The calculated percentage yield of the extracted M. acuminata starch polymer was found to
be as high as 81% w/w. Although, Kamali et al [3] had reported a higher yield of 84% w/w, it
proved to be difficult to obtain such a high yield in this study, although the difference is
insignificant.
3.1.2. The RTG method (Phase II extraction)
The percentage yield of the retrograded M. acuminata starch polymer was 67% w/w. While
this was a good yield in comparison to other studies, the gelatinization process promoted a
significant reduction in quantity because the process destroys the molecular order of the
starch granule, promoting easy digestion as explained by Recife et al [9].
www.mnf-journal.com Page 15 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
3.1.3. Microstructure studies by SEM
Fig 1. Presents the SEM analysis of (a) Formulation 1, (the novel formula ingredients) by
direct compression, (b) Formulation 2, (the old traditional formula ingredients) by wet
granulation, and (c) a side-by-side comparison of MCC and RMAS.
The ACM particles in Fig. 1 (a) and Fig. 1 (b) were the same and irregular in shape,
suggesting poor flowability during tablet formulation in line with a previous report [17]. A
glidant with spherical particles was needed to serve as wheels to roll the ACM particles
down the hopper, imparting good fluidity, lubricity, and compressibility.
In Fig. 1 (a), a small quantity of AR powder particles (which are spherical in shape) was
combined with the disc-like shaped MAP particles to provide the excellent flow needed. MS
was selected as a reference starch polymer because of its wide pharmaceutical application
[18].
A portion of the MAS (50% w/w) was retrograded as explained earlier, to produce
biopolymer particles (RMAS) with striated surface appearances and peel-like coverings
similar to those of MCC as compared in Fig 1 (c). The striations are the features that render
MCC as the best direct compression excipient. A dry binder, or MCC, is always necessary
for direct compression [17], but to reduce the number of excipients in this study, RMAS was
considered in the place of MCC. RMAS influences the mechanical and release
characteristics of the ACM formulation in the same way that MCC would do. However, MAP
(MAS/RMAS 50-50% polymer blend) was actually used instead of RMAS alone to further
www.mnf-journal.com Page 16 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
improve the compressibility, lubricity, and release characteristics of the formulation. The
amorphous nature of MAS in the RMAS/MAS 50-50% blend allows it to retain some minimal
quantity of moisture which served to provide the solvent needed. Thus, making MAP
(MAS/RMAS 50-50% polymer blend) a 4-in-1 excipient (serving the functions of a filler,
binder, disintegrant, and solvent).
3.1.4. Characterization of samples by FTIR
The FTIR result of the effect of different percentage concentrations of the MAP on the ACM
is presented in Fig. 2. Bands observed at 3200 to 3400 cm-1 regions traceable to the N-H
group vibration in the ACM were still very evident at the low concentrations of MAP (10 to
50% w/w). At higher MAP concentrations, the ACM bands diminished as shown with 70%
w/w. At 70% w/w MAP, a broad absorption band between 3000 and 3600 per cm-1 emerged.
This can be ascribed to the presence of considerable O-H bonding of water due to the semi-
hygroscopic nature of MAP. Moisture negatively affects the stability of tablets [12], but in this
case, due to the low concentration of MAP used, the minimal moisture served as an
advantage of being a solvent binder to impact good compressibility of the dosage form.
The results show that MAP at low concentrations (less than 50% w/w) did not significantly (P
> 0.05) impact compatibility changes on the ACM, but at higher concentrations of above
50% w/w, it did [Fig. 3 (b)], agreeing with previous studies [19].
3.1.5. Thermogravimetric Analysis (TGA) and Differential thermal
analysis (DTA) of samples
www.mnf-journal.com Page 17 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
In the thermograms overlay in Fig. 3 (a) the mass loss traces were not clearly
distinguishable and it was rather difficult to accurately ascertain the situation. Fig. 3 (b) was
thus created from Fig. 3 (a) by re-scaling the annotated portion of the latter. Thermograms in
Fig. 3 (b) clearly show an increase in percentage mass loss due to moisture with an
increasing concentration of MAP (Supporting Information Table 1). The thermograms’ first
mass loss stage occurring between 30 and 115°C was as result of the moisture evaporation
during heating [19]. The second stage (70-80 % mass loss) occurred between 250 and
350°C as a result of the elimination of polyhydroxy groups, followed by depolymerization and
decomposition [9]. Similar results were obtained with DTA thermograms (Supporting
Information Fig. 1) suggesting no incompatibilities at concentrations lower than 50% w/w.
The thermograms of Fig. 3 (a) show that there is an increase in temperature
of melting with increasing concentration of MAP. This agrees with the work of Gorain et al
[19], proving that small quantities of total excipients found in drugs (5-15% by weight) did not
significantly influence the thermal behavior of the active component, whereas excipients with
concentrations above 65% had significant influence.
3.1.6. Characterization of samples by the DSC
The DSC analysis of samples are shown in Fig 4 (a), while Fig. 4 (b) shows a zoomed-in
portion of the graph indicating the effect of moisture carried by the MAP. Fig. 4 (c) shows
another zoomed-in portion indicating the effect of the MAP on the melting point range of the
ACM.
www.mnf-journal.com Page 18 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
In Fig 4 (b), zero and lower concentrations of the MAP [(i) 0%, (ii) 10% and (iii) 30%] carried
an insignificant (P > 0.05) amount of moisture, while higher concentrations of the MAP [Fig 4
(b) (iv) 50%, (v) 70%, and (vi) 100%] carried a significant amount of moisture, affecting the
quality of the ACM. The thermogram of the MAP did not exhibit any endothermic peaks [Fig
6 (c)], suggesting that the starch polymer is amorphous. All other thermograms exhibited
endothermic peaks at about the same temperature ranges of 169-172°C coinciding with the
official melting point range of ACM and indicating a high compatibility index between the
ACM and the MAP. However, the surface area of transition for all the thermograms vary in
sizes and ranked in the following order: ACM > MAP 30% w/w > 50% w/w > 70% w/w >
100% w/w (Fig 6 (c). This indicated better compatibility with the smaller concentrations of the
polymer. The reason behind this could be the amorphous nature of the MAP making it retain
a small amount of water [20]. The larger the polymer concentration, the less the compatibility
[19].
3.1.7. Characterization of samples by X-ray diffraction
The X-ray diffractograms of samples are shown in Fig. 5. The ACM diffractogram exhibited
pronounced peaks at 15.2, 18, 23, 24, and 26.2° (2θ), which are characteristic of the API.
Samples with lower amounts of the MAP (≤ 50% w/w) still retained the pronounced peaks,
indicating the retention of the ACM activity, however, samples with large amounts of the
MAP (> 50% w/w) produced no significant (P > 0.05) ACM peaks, indicating the loss of
activity of the API. This suggests that the smaller the quantity of MAP used as an excipient,
the better the quality of the formulation. However, Agarwal et al [21], showed that greater
degrees of changes were also associated with lower excipient concentrations. Therefore, the
www.mnf-journal.com Page 19 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
moisture content in the MAP due to its semi-hygroscopic nature might have largely caused
this change in XRD patterns with increasing MAP concentrations [20].
3.2. Micromeritic properties
The powder characterization of the ACM/MAP binary mixture (bulk and tapped densities,
moisture content, angle of repose, and Carr’s compressibility index) was completed (Table
1). Based on the specifications of the British Pharmacopoeia [22], all values obtained during
the powder analyses fall within official acceptable ranges.
Table 2 shows a comparison of two formulas for the ACM formulation; one by direct
compression using the MAP as a 4-in-1 excipient (serving as filler, binder, solvent, and
disintegrant) in comparison to the traditional formulation by wet granulation using the MS.
Five excipients were used for the wet granulation process, whereas, only three were used
for the direct compression, indicating the advantage of this new study [21], and especially in
high-dose tablet formulations where only minimal quantities of excipients are required due to
the already increased dose [22].
3.3. Resistance to crush-friability disintegration time ratio (RCFR:DT)
The characterization of tablets was done following the specifications of the British
Pharmacopoeia [22], and all batches tested were found to pass the resistance to crush,
friability, and disintegration tests as expected of standard uncoated tablets. RCFR:DT, a
www.mnf-journal.com Page 20 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
higher parameter for tablet quality, was also used. The RCFR:DT is good for measuring
tablet quality because, in addition to measuring tablet strength (resistance to crush) and
weakness (friability), it simultaneously evaluates all negative effects of these parameters on
disintegration time [16]. The higher the RCFR:DT values, the better the quality of tablets
[16]. From this study, the order of increasing RCFR:DT based on the percentage
concentration of the MAP was found to rank as follows: 10% > 20% > 30% > 50% w/w
(Supporting Information Fig. 2).
3.4. In vitro release study of tablets
The in vitro release study is shown in Fig. 6. The US-FDA guideline specifies 85% (T85%) of
the labeled amount of drug release within 30 min [21], therefore, all the batches tested in this
study exhibited good drug release properties. Burst release [22] of the ACM was observed
with the MS 30% w/w but normal release for a conventional tablet with the MAP 10, 20, 30,
40, and 50% w/w (Fig. 6). The rank order of observed increasing drug release was as
follows: MAP 10%. < MAP 20% < MAP 30% < MAP 50% < MAP 40% < MS 30%. By
increasing the concentration of the MAP, drug release was increased up to a maximum MAP
concentration of 40% w/w. When proceeded to MAP 50% w/w, a decreased release occurs.
This agrees with the work of Gorain et al [19] which states that for solid binary mixtures with
low excipient content of 80:20 w/w, good compatibility was usually observed when tested,
although increasing the excipient content increases the chances of incompatibilities. A study
of the release mechanism indicated a diffusion-controlled release mechanism with all
batches tested exhibiting good drug release properties (Supporting Information Fig. 3).
4. Conclusion
www.mnf-journal.com Page 21 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
The application of the newly developed ASR method in the extraction of M. acuminata starch
produced MAS and RMAS which were blended in a 50-50% proportion to obtain the unique
biopolymer (MAP). MAP was used as a 4-in-1 excipient (serving as filler, binder,
disintegrant, and solvent) in ACM high-dose tablet formulation by direct compression. The
processes involved the application of selected physicochemical and analytical
characterization to test the compatibility, viability, and quality of the formulation. Lower MAP
concentrations (between 10% and 40% w/w) were found to be more compatible with ACM in
the formulation processes.
Acknowledgments
The authors wish to express gratitude to the staff of the School of Pharmacy at the
University of the Western Cape for their technical support.
Funding: This work was supported by ‘The World Academy of Science’ (TWAS) in
partnership with the ‘National Research Foundation’ (NRF) of South Africa. Award reference:
PD-TWAS150925143012).
Conflict of interest: None.
References
www.mnf-journal.com Page 22 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
1. O. Odeku, Starch - Starke, 2012, doi.org/10./1002/star.201200076
2. R. Matsushima, Morphological variations of starch grains, in Starch, (Eds: Y.
Nakamura), Springer, Tokyo, Japan 2015, Ch. 13.
3. H. Kamali, E. Khodaverdi, F. Hadizadeh, R. Yazdian-Robati, A. Haghbin, G. Zohuri,
J. Drug Del. Sci. Tech. doi.org/10.1016/j.jddst.2018.06.027
4. M. Ahmad, A. Gani. I. Hassan, Q. Huang, H. Shabbir, Sci. Rep. 2020,
doi.org/10.1038/s41598-020-60380-0
5. P.K. Kumar, H.S. Joyner, J. Tang, B. A. Rasco, S.S. Sablan, Food Bioprocess
Technol. 2020, doi.org/10.1007/s11947-020-02488-9
6. A.H. Khan, N. M. Minhas, M. J. Asad, A. Iqbal, M. Ilyas, R. T. Mahmood, Eur. Acad.
Res. 2014, http://www.euacademic.org/UploadArticle/749.pdf.
7. G.L.P. Silva, J.A.C. Bento, L.A.M. Bataus, M.S.S. Júnior, M. Caliari, Starch-Starke,
2021, doi.org/10.1002/star.202000210.
8. S.P. Lizarazo, G.G. Hurtado, L.F. Rodríguez, Agron. Colomb. 2015,
doi.10.15446/agron.colomb.v33n2.47239
9. A.C.D. Recife, A.B. Meneguin, B.S.F. Cury, R.C. Evangelista, J. Drug Del. Sci. Tech.
2017, doi.org/10.1016/j.jddst.2017.06.003.
10. H. Montaseri, P.B.C. Forbes, TrAC, Trends Anal. Chem. 2018,
doi.org/10.1016/j.trac.2018.08.023.
11. United States Pharmacopeia (USP) 34 (NF 29), Chapter 621. Edition; 2011.
12. Q.I. Sun, Z. Han, L. Wang, L. Xiong, Food Chem. 2014,
doi.org/10.1016/j.foodchem.2013.08.129.
www.mnf-journal.com Page 23 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
13. M. Autamashih, A.B. Isah, T.S. Allagh, M.A. Ibrahim, Int. J. Green Pharm. 2011,
doi:10.4103/0973-8258.82095.
14. A.M.C. Nirmaan, B.D. Rohitha Prasantha, B.L. Peiris, Chem. Biol. Technol.
Agric. 2020, doi.org/10.1186/s40538-019-0164-1.
15. K.S. Tumwesigye, E. O'Brien, J.C. Oliveira, A. Crean, M.J. Sousa-Gallagher, Future
Foods, 2020, doi.org/10.1016/j.fufo.2020.100003.
16. M.V. Lawal, M.A. Odeniyi, O.A. Itiola, Asian Pac. J. Trop. Biomed. 2015,
doi.org/10.1016/j.apjtb.2015.05.003.
17. N. Ghazi, Z. Liu, C. Bhatt, S. Kiang, A. Cuitino, AAPS Pharm. Sci. Tech. 2019,
doi:10.1208/s12249-019-1369-0.
18. S. K. Gunatilake, S. S. Samaratunga, F. A. Adekola, Der. Pharma. Chemica. 2016,
8(4): 237-242, www.researchgate.net/publication/301630370
19. B. Gorain, H. Choudhury, M. Pandey, T. Madheswaran, P. Kesharwani, R. K.
Tekade, in Dosage Form Design Parameters. Vol. II (Eds: R.K. Tekade), Academic
Press, 2018, Ch. 11.
20. J.R. Witono, I. Noordergraaf, H.J. Heeres, L.P.B.M. Janssen, H. Heeres, Carbohyd.
Polym. 2014, doi.org/10.1016/j.carbpol.2013.12.056.
21. U. Agarwal, S.A. Ralph, C. Baez, R.S. Reiner, S.P. Verrill, Cellulose. 2017,
doi.10.1007/s10570-017-1259-0
22. Bp, C, British Pharmacopoeia. Stationery Office, London, England. 2012.
23. D. A. Diaz, S. T. Colgan, C. S. Langer, N. T. Bandi, M. D. Likar, L. V. Alstine. AAPS
J. 2016, doi: 10.1208/s12248-015-9830-9
24. C. Jackson, M. Emeje, S. Ofoefule, Bri. J. Pharm. Res. 2015,
doi.org/10.9734/BJPR/2015/14412
www.mnf-journal.com Page 24 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
Figure Legends
Fig 1: Analysis of samples by scanning electron microscopy. (a) Formulation 1 indicates the
ingredients for this novel formulation of ACM by direct compression. (b) Formulation 2
indicates the ingredients for the old traditional formulation of the ACM by wet granulation,
and (c) indicates a side-by-side comparison of the structural morphologies of MCC with
RMAS particles showing morphological similarity.
www.mnf-journal.com Page 25 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
Fig. 2: FTIR spectral analysis of the effect of MAP concentrations on ACM. (a) Indicates
profiles for ACM with added low concentrations of MAP (10 to 50% w/w) while (b) indicates
profiles for ACM with added higher concentrations of MAP (50 to 100% w/w). The annotated
regions indicate the wide hydroxyl group regions as caused by moisture.
www.mnf-journal.com Page 26 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
Fig. 3: TGA analysis of the effect of MAP concentrations on ACM. (a) indicates the moisture
effect of MAP on ACM on full TGA and (b) indicates the same moisture effect rescaled and
zoomed-in for a better view as shown by the annotated regions.
www.mnf-journal.com Page 27 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
Fig 4: DSC analysis of the effect of MAP concentrations on ACM. (a) indicates the effect of
MAP on ACM on full DSC thermograms, (b) indicates the moisture effect of MAP on ACM
(shown by the first annotated region), and (c) indicates the effect of MAP on the melting
point range of ACM (shown by the second annotated region).
www.mnf-journal.com Page 28 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
Fig 5: X-ray diffractograms indicating the effect of MAP concentrations on ACM.
Fig 6: HPLC dissolution of ACM/SP tablets. MAP 10%, 20%, 30%, 40%, and 50% indicates
ACM tablets with MAP concentrations of 10%, 20%, 30%, 40%, and 50% w/w respectively
while MS 30% indicates ACM tablets with MS concentration of 30% w/w.
www.mnf-journal.com Page 29 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
Table 1: Evaluation of ACM/MAP binary solid mix for direct compression showing values of
powder analyses of MAP 10% to 70%, and MS 30%, versus the British Pharmacopoeia
specifications
Table 1: Evaluation of ACM/MAP binary mix for direct compression
Parameters
Granule analysis and BP 2011 specification
MAP10%
MAP30%
MAP50%
MAP70%
MS30%
BP
specification
pH
5,9 ± 0,2
6,2 ± 0,3
6,5 ± 0,1
6,7 ± 0,1
6,36 ±
0,2
≤ 6,5 for ACM
Moisture content (%)
10,2 ± 0,5
11,8 ± 0,9
12,5 ± 0,3
15,9 ± 0,5
11,9 ±
0,8
≤ 15%
True density (g/cm3)
1,19 ± 0,1
1,2 ± 0,5
1,49 ± 0,6
1,18 ± 0,5
1,1 ± 0,2
NA
Bulk density g/cm3
0,35 ±
0,01
0,39 ±
0,06
0,41 ±
0,04
0,52 ±
0,02
0,34 ±
0,01
NA
Tapped density g/cm3
0,41 ±
0,08
0,45 ±
0,03
0,46 ±
0,01
0,58 ±
0,04
0,43 ±
0,04
NA
Angle of repose
30.0 ± 0,7
29,1 ± 0,5
27,97 ±
0,2
25,5 ± 0,5
20,9 ±
0,2
25.0-40.0
Hausner's ratio
1,17 ± 0,3
1,17 ± 0,1
1,12 ± 0,1
0,76 ± 0,4
1,26 ±
0,3
≤1.25
Carr's compressibility
14,63 ±
0,9
13,33 ±
0,4
10,87 ±
0,7
10,17 ±
0,3
20,93 ±
0,1
5.00-16.00 %
www.mnf-journal.com Page 30 Molecular Nutrition & Food Research
This article is protected by copyright. All rights reserved.
Table 2: Tablet formulation of ACM using various MAP concentrations as multi-functional
excipients with AR as a glidant by direct compression versus the formulation of ACM using
MS, TC, and MTS by wet granulation
Table 2: Comparative composition of tablet formulations of ACM: Direct compression
using MAP versus wet granulation using MS
Ingredients
MAP
10%
(mg)
MAP
20%
(mg)
MAP
30%
(mg)
MAP
40%
(mg)
MAP
50%
(mg)
MS
10%
(mg)
MS
20%
(mg)
MS
30%
(mg)
MS
40%
(mg)
MS
50%
(mg)
ACM (active ingredient)
500
500
500
500
500
500
500
500
500
500
MAP (multi-excipient)
45,5
94
142,5
193
341,5
AR (glidant) (1% w/w)
5,5
6
6,5
7
8,5
MS (diluent/disintegrant)
7
25
148
144
189.95
PVP (binder) (2.5%w/w)
13,75
15
16,25
17,5
18,75
TC (glidant) (5% w/w)
27,50
30
32,5
35
37,5
MTS (lubricant) (0.5% w/w)
2,75
3
3,25
3,5
3,75
Water (solvent) qs
0
0
0
0
0
Tablet theoretical weight
550
600
650
700
750
550
600
650
700
750
ACM = Acetaminophen, MAP = Musa acuminata advanced starch polymer, AR = Aerosil® 200, MS = modified maize starch,
TC = talc, MST = magnesium stearate, PVP = Polyvinylpyrrolidone, and qs = quantity sufficient
