RESEARCH ARTICLE

QbD Based Formulation Development of Delayed-Release Beads for Better

Management of Nocturnal Asthma
Kumar V V1, Ismail Y1*, Uthumansha U1, Pradeep K2
1
Crescent School of Pharmacy, B.S. Abdur Rahman Crescent Institute of Science and Technology, GST Road, Vandalur,
Chennai-600048, Tamil Nadu, India.
2
Faculty of Health Sciences, Villa college, Male, Maldives

Received: 23rd Sep 2024; Revised: 8th Oct, 2024; Accepted: 22nd Nov, 2024; Available Online: XX

ABSTRACT
The present study discusses about the quality by design (QbD) based formulation development and characterization of
delayed release beads for better management of Nocturnal Asthma. Ionotropic gelation technique was used to create
BUD-loaded pectin-alginate beads, and calcium chloride was added as a crosslinking agent. The central composite design
model was used to optimize the beads. The response of beads was 91.9861 % for loading efficiency, 0.998 mm for bead
size, and 360 min for time required for 90% drug release. The optimal formulation variables for a formulation were found
to be 4.14 mg of pectin, 1.82 mg of alginate, 14.36% of Cacl2, and 5.99 hours of cross-linking time. PEC-ALG beads
showed no release in 0.1 N HCl (pH 1.2). In contrast, increased Ca2+ and Na+ ion exchange as well as solvent
penetration into the pectin-alginate network have been linked to the quick release from cross-linked beads in phosphate
buffer pH 7.4. FTIR results showed that there was no interaction between the medication and the polymers, as evidenced
by the significant peaks of BUD detected in the beads. An in vitro study of the toxicity of beads on A549 cell lines
revealed significantly higher cell viability than the group of cells treated with pure BUD.BUD loaded PEC-ALG beads
could have potential for chrono modulated delivery system for targeting nocturnal asthma.
Keywords: Chronotherapy; Delayed Release Beads; Quality by design; Nocturnal Asthma.
How to cite this article: Kumar V V, Ismail Y, Uthumansha U, Pradeep K. QbD Based Formulation Development of
Delayed-Release Beads for Better Management of Nocturnal Asthma. International Journal of Drug Delivery
Technology. 2024;14(4):XX. doi: 10.25258/ijddt.14.4.X
Source of support: Nil.
Conflict of interest: None

INTRODUCTION absorption, distribution, metabolism, and elimination, are

The use of targeted and controlled drug delivery has regulated by the circadian rhythm.4 With chrono-
surpassed conventional dosage forms in recent years. In modulated drug delivery systems, drugs are released under
order to target the precise site, this system has focused on controlled conditions at predetermined intervals to
a continuous, variable, and sustained drug delivery correspond with the body's natural rhythm. Nocturnal
system.1 The term "chrono modulated drug delivery asthma, that affects two-thirds of asthma patients, is
system" refers to such a system.2 It is also referred to as a typified by an increase in symptoms like wheezing,
pulsatile drug delivery system or a chrono modulated drug tightness in the chest, increased airway reactivity, and a
delivery system. Two-thirds of asthma patients have decline in lung function at night. In particular, at 4:00 am,
nocturnal asthma, which is characterized by an increase in and between midnight and 8:00 am, these symptoms
symptoms such as wheezing, tightness in the chest, manifest. Several strategies have been explored for
increased responsiveness of the airways, and worsening of designing chrono-modulated drug delivery systems, such
lung function during the night Between midnight and 8:00 as implantable devices, transdermal patches, and oral
am, and particularly around 4:00 am, these symptoms formulations with specialized release profiles. These
manifest. Therefore, since the patient is asleep, taking systems incorporate various mechanisms, including pH-
medication at midnight is inconvenient2. For the best sensitive coatings, osmotic pumps, and programmable
course of treatment, maintaining a steady drug level is not electronics, to achieve the desired temporal release
always necessary. A medication should only be pattern.5-7 The benefits of chrono-modulated drug delivery
administered in the bare minimum amount necessary in systems are extensive. By targeting drug administration
terms of BCS classification, it is in class 3. The intention during specific periods of disease activity or maximum
was to have a 6-hour lag time, meaning that the need, therapeutic concentration levels can be optimized,
medication is taken at bedtime and released into the leading to improved patient outcomes. Furthermore, by
system at 4:00 am, or after 6 hours.3 By coordinating drug minimizing drug exposure during inactive disease phases,
delivery with disease patterns and circadian rhythms, side effects and drug resistance can potentially be
chrono-modulated drug delivery systems have become a reduced.8 This study focused on developing chrono-
viable strategy for improving therapeutic outcomes. modulated drug delivery systems for targeting nocturnal
Numerous physiological processes, such as drug asthma.9

*Author for Correspondence: [email protected].

 QbD Based Formulation Development

The quick in vitro release of the entrapped material is a Since alginate aqueous solutions have extremely high
significant drawback of alginate, one of the most widely viscosities even at low concentrations, the matrix material
utilized natural biopolymers as a delivery method for low for encapsulation can be employed at a maximum
molecular weight phytochemicals. concentration of roughly 2-4%, for example, in the case of

Figure 1. Effect of the Independent Variable on Bead Size (mm)

Figure 2. Effect of the process parameters on the Loading Efficiency of the beads

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 QbD Based Formulation Development

Ca-alginate. Consequently, the resultant gel network lacks (Stat-Ease, Minneapolis, MN, USA)
the required barrier effect and has a low density. As a (https://www.statease.com/docs/v11/ viewed on April 28,
result, a strategy for developing novel techniques to 2024). Pectin (X1), sodium alginate (X2), calcium
produce water-insoluble microparticles with decreased chloride (X3), and cross-linking [x4] were the four
porosity and delayed release of entrapped materials is formulation components (independent variables) that were
needed.10,11 Alginate can be used in conjunction with put to use. Three responses (dependent variables) were
complementing plant-derived biopolymers, like Based on studied in order to determine the best formulation: the time
the literature, the present work aims on design and required for drug 90% release (Y3), loading efficiency
development of chrono-modulated drug delivery systems (Y2), and bead size (Y1). The experimental design
using alginate-pectin combination polymers with consisted of a randomized sequence, six replication cases
ionotropic gelation technique.12 with central points, six axial points, and eight designated
factorial points. To test the method's repeatability, the
MATERIALS AND METHODS focal point was iterated five times.14 The response surface
Optimization of Delayed Released Beads (Design of the regression technique was employed to assess the
Experiment [DOE]) information. A polynomial model was chosen by taking
As a possible colon administration technique, bud-beads into consideration the significant terms (p < 0.05),
have been developed and designed with the best possible coefficient of variance, least significant lack of fit, and
formulation using the response surface methodology.13 multiple correlation coefficients that were obtained from
The Central Composite Design approach was used to the Design Expert tool. According to the independent
optimize the parameters that make up the BUD-Beads variables, Table 1 shows the highest and lowest levels.
formulation using Design Expert Software version 11 Preparation of Ca++ ion Cross-linked Pectin-Alginate

Figure 3. Effect of the process parameters on the Time required for drug 90% release

Figure 4. Pertubation Graph a) Bead Size b) Loading efficiency c) Time required 90% drug release

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 QbD Based Formulation Development

Beads gently stirred at room temperature and added to a 250 ml

In double-distilled water, sodium alginate and pectin were solution of CaCl2 (at concentrations of 0.05 M and 0.20
dissolved at concentrations of 1-3.0% w/v and 2–6% w/v, M) using a syringe (no. 23) with a drop-in grate set at 1
respectively. Then, while stirring, precisely weighed BUD ml/min. The calcium pec-alginate beads that had formed
(drug: polymer, 1:1) was added. The dispersions were were left to stand in the gelling solution for two to ten

Figure 5. Observed values of optimal condition

Figure 6. Formulation of Budesonide-PEC-ALG Beads.

Figure 7. The combined graphical image of FT-IR. The line which indicatesred color is pure API,the line sky bluecolor
indicates pectin, theline green colorindicates alginate and blue color indicate beads. Overall prepared beads show the
combined graph.

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 QbD Based Formulation Development

hours. After that, they were filtered out and given two water. The beads were then baked for 24 hours at 50°C to
successive washes in 100 milliliters of double-distilled dry them out.15
90.00
80.00
70.00

Swelling Index
60.00 PEC_ALG Beads
50.00
40.00
30.00
20.00
10.00
0.00
0 1 2 3 4 5 6
Time in Hours
Figure 8. SEM image of Delayed release beads Figure 9. Swelling index bar graph

Figure 10. Release kinetics of Budesonide from PEC-ALG Beads.

Figure 11.In vitro Cytotoxicity Assay of optimized beads a) Budesonide DRB (A549 cell lines) and b) Plain Budesonide
drug (A549 cell lines)

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 QbD Based Formulation Development

Practical Yield moisture remained on the dish's surface in order to

After the last beads were gathered, the practical yield was determine the weight. The bead was then
calculated using the equation 𝑆𝑡 − 𝑆𝑖
𝑆𝐼 = 𝑥100
Practical yield = [(practical weight of beads)/(theoretical 𝑆𝑖
weight of beads)] × 100. Where Si = initial PEC-ALG dry bead weight, and St =
Encapsulation Efficiency wet PEC-ALG bead weight at time t (hours). (Figure-9).
Every batch of designed beads was individually weighed In vitro Drug Release Studies
at 50 mg and added to 100 mL of pH 6.8 phosphate buffer For the dissolution investigation, a USP Type II (paddle
containing 0.5% w/v Tween 80. After that, BUD was type) dissolving apparatus was used, with a paddle speed
released when the beads were broken down by of 75 rpm and a temperature of 37±0.5 °C. In order to
ultrasonication. Whatman's filter paper (diameter 0.45 μm) simulate gastrointestinal conditions, the core cross-linked
was used to filter the samples, and a UV beads were first tested for dissolution in 500 mL of 0.1N
spectrophotometer was used to examine them at 240 nm. HCl (pH 1.2) with 0.5% w/v tween 80, and then for further
Encapsulation efficiency was determined by the below hours in 900 mL of phosphate buffer pH 6.8 with 0.5%
given formula15, w/v tween 80. The amount of medication was taken into
Encapsulation efficiency = (Actual drug loading/ account when placing beads into size 1 hard gelatin
Theoretical drug loading) × 100 capsules.15-17 At predetermined intervals, five milliliters of
Measurement of Beads Size the sample were removed, replaced with fresh matching
A ten-piece string BUD-Beads were chosen at random. media, and examined at two distinct wavelengths using a
Each type of bead was measured for size and form using a UV-Visible spectrophotometer at 240 nm in 0.1N HCl and
computerized vernier caliper (Mitutoyo, Kawasaki, Japan). 240 nm in phosphate buffer.
Ten beads were chosen at random to establish the mean In vitro drug release kinetics analysis
diameter of the beads. Each bead's length and width were To identify which mathematical model best matched the
measured, and its diameter was used to compute the mean. drug release profile, the received release data were fixed
Scanning Electron Microscopy (SEM) into numerous models, including the (i) Zero Order, (ii)
Double-sided adhesive was used to adhere the beads on First Order, (iii) Higuchi, Korsemeyer-Peppas, and (iv)
the plate, and gold spray was used afterwards. Using an Hixson Crowell models.18 The release from systems where
accelerating voltage of 10 kV, the morphological features the particle diameter and surface area change is described
of the samples (Supra 55, Zeiss, Oberkochen, Germany) by the equation. Data from in vitro drug release studies
were examined and captured on camera.16 were plotted as the cube root of the drug percentage
Fourier Transform Infrared Spectroscopy (FT-IR) remaining in the matrix versus time to study the release
The drug's physical interaction and the surface chemistry kinetics. The result is a straight line.
of the polymers were determined by FT IR spectra. The In vitro cell line study
samples were combined with KBr and compressed into Preparation of test solutions
transparent tablets. A Jasco 6700 FT-IR spectrometer was For MTT assay, serial two-fold dilutions (6.25 – 100 µg)
used to analyze the samples for FT-IR spectrometry across were prepared from this assay.
the 4000-400 cm-1 range17(Figure 7). Cell lines and culture medium
Swelling Index Penicillin (100 IU/mL), streptomycin (100 μg / mL), and
Using a digital vernier caliper, 10 BUD's length and 10% inactivated fetal bovine serum (FBS) were added to
width-The weights of the beads were calculated after they DMEM media containing the A549 cell line until it
were measured. After being placed in 20 milliliters of pH reached confluency. The culture was kept in a humidified
1.2 buffers, each type of bead was stirred for two hours at environment with 5% CO2 at 37 °C.
50 rpm and 37 °C.to mimic stomach conditions. Beads Procedure
were transferred to a pH 7.4medium after two hours.14 For The monolayer cell culture was trypsin zed and the cell
six hours, the beads were examined in the medium for count was adjusted to 1.0 x 105 cells/mL using the proper
signs of swelling, erosion, and water absorption. Each wet media containing 10% FBS. In each well of the 96-well
bead's moisture content was removed, and then the bead microtiter plate, 100 μL of the diluted cell suspension (1 x
was carefully wiped over a dry Petri dish until no visible 104 cells/well) was added19-21. After 24 hours, when a

Table 1. Formulation factors for preparation of bud-beads and corresponding level

Factor Name of High Medium Low Alpha(+) Alpha(-)
Variable Parameters
A Pectin 6 4 2 8 0
B Sodium Alginate 3 2 1 4 0
C Calcium Chloride 20 15 10 25 5
d Cross linking time 10 6 2 14 -2
RESPONSE Response Units
Y1 Beads size Mm
Y2 Loading efficiency %
Y3 Time required 90 % drug release minutes

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 QbD Based Formulation Development

partial monolayer had formed, 100 μL of different test Beads. The research study looked to determine whether
sample concentrations were added to the partial monolayer various amounts of cholesterol, Chitosan, and surfactants
in microtiter plates, the supernatant was disposed of, and affected the previously mentioned responses. In
the monolayer was once more washed with medium. The accordance with the main compound layout, twenty
plate was then incubated for a full day at 37°C with 5% equations in a three-factor, three-response format with six
CO2 in the air. After the test solutions were removed, 20 in central locations were created. Furthermore, the
μL of MTT (2 mg/1 mL of MTT in PBS) was added to responses were assessed using analysis of variance
each well. (ANOVA), as well as the matching response. A uniform
and regulated spherical bead size is crucial for
RESULTS AND DISCUSSIONS understanding the behaviour of drug release in the GIT
The BUD-Beads for this study were developed using Stat- area. As a result, it was thought that the goal of optimizing
Ease Design Expert software version 11 and the response size should be circular naturally and have a minimal value.
surface methodology. The main compound model was After considering dual straight regression equations, the
used to investigate the primary formulation variables for Y1 result can be written as follows:
the experimental design that influence the responses of the 𝑩𝒆𝒂𝒅𝑺𝒊𝒛𝒆 (𝒀𝟏 ) = +𝟖𝟓𝟎. 𝟎𝟎 − 𝟑𝟓𝑨 − 𝟒𝟖. 𝟕𝟓𝑩
Bead size (Y1), loading efficiency (Y2), and time required +𝟏. 𝟐𝟓𝑪 + 𝟒𝟐. 𝟓𝟎𝑨𝑩 + 𝟐𝟕. 𝟓𝟎𝑨𝑪
for 90% drug release (Y3) of the Pectin, Alginate, and

Table2. Optimization using CCD Design

Factor 1 Factor 2 Factor 3 Factor 4 Response 1 Response 2 Response 3
D: Cross Time required
Std Run A: PEC B: ALG C: CaCl2 Bead Size LE
Linking Time 90% drug release
% % % hr mm % minutes
7 1 2 3 20 2 0.86 66 180
5 2 2 1 20 2 0.79 60 120
10 3 6 1 10 10 1.21 72 240
1 4 2 1 10 2 0.6 61 90
24 5 4 2 15 14 1.02 92 240
13 6 2 1 20 10 0.69 70 270
16 7 6 3 20 10 1.08 58 480
15 8 2 3 20 10 0.76 42 180
11 9 2 3 10 10 0.99 40 180
28 10 4 2 15 6 0.89 90 390
14 11 6 1 20 10 0.97 60 240
8 12 6 3 20 2 1.42 49 480
2 13 6 1 10 2 1.21 65 420
26 14 4 2 15 6 1.02 92 360
22 15 4 2 25 6 2.04 86 300
12 16 6 3 10 10 1.38 67 480
25 17 4 2 15 6 1.02 92 360
23 18 4 2 15 -2 0.84 87 120
6 19 6 1 20 2 0.976 51 420
4 20 6 3 10 2 0.923 49 480
27 21 4 2 15 6 1.02 92 360
17 22 0 2 15 6 0.258 10 0
18 23 8 2 15 6 1.58 9 540
9 24 2 1 10 10 0.856 44 120
21 25 4 2 5 6 1.85 87 270
20 26 4 4 15 6 1.01 90 300
30 27 4 2 15 6 1.02 92 360
3 28 2 3 10 2 0.798 79 240
29 29 4 2 15 6 1.02 92 360
19 30 4 0 15 6 0.368 98 0

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 QbD Based Formulation Development

Table 3: Optimized formulation based on desirability

Factor Name Level Low Level High Level Coding
A PEC 4.14 2.00 6.00 Actual
B ALG 1.82 1.0000 3.00 Actual
C CaCl2 14.36 10.00 20.00 Actual
D Cross Linking Time 5.99 2.00 10.00 Actual

Table 4: Experimental and predicted values under optimal conditions for Oil Entrapped Beads
Solution 1 of 99 Predicted Predicted Observed Std Dev SE Mean
Response Mean Median
Bead Size 0.998432 0.998432 1.02 0.178968 0.0725966
LE 91.9861 91.9861 90.56 9.69966 3.93457
Time required 90% drug 360 360 360 68 28
release

−𝟏𝟎. 𝟎𝟎𝑩𝑪 + 𝟒𝟓𝑨𝟐 + 𝟐𝟕. 𝟓𝑩𝟐 lessen BUD's gastrointestinal discomfort. Following this
+ 𝟓𝟐. 𝟓𝑪 lag, a pulse with full drug release in phosphate buffer
In contrast to beads with more spherical morphology and occurred in 360 minutes. The PEC-ALG beads exhibited a
medium and high polymer concentrations (+ level), low notable delay in drug release at an acidic pH, which could
concentration beads (-level) have rough and uneven shapes potentially be attributed to the drug's insolubility with
because of inadequate molecular packing and cross- pectin. Calcium pectinate may protonate into an insoluble
linking. As the PEC and SA concentrations were increased form with decreased swelling at an acidic ph. It is possible
this was the case, as demonstrated by the response surface to relate the second pulsed release stage in pH 7.4
plots relating bead size (Figure1). The variations in the phosphate buffer to the calcium pectinate gel's fast
size and morphology of the beads with different crystalline expansion and gel relaxing at an alkaline ph. There was a
levels of PEC and SA were caused by changes in the notable impact of matrix beads on the time needed for
availability of reacting/binding sites for cross-linking 90% drug release22-23 (Figure 3).
cations (Ca2+). When the cross-linking agent content was Optimization of Formulation Factors
increased with SA, the result was smooth, spherical, well- The second-order polynomial equations explain the model
packed beads with a distinct structure; however, similar in terms of the coded parameters, and the central
material in this instance of CS resulted 21. composite design chose A four-dimensional equation. To
Effect of the process parameters on the Loading link the answers and the Separated factors. The multiple
Efficiency of the beads (Y2) correlation coefficient (R2) and confidence interval (P) of
Using polynomial equation, the impact of Examined were the revised model were employed for assessment. The
the primary and compound Impact of separate variables on equations for (LE), (size), and (DR) have R2 values of
EE. An ANOVA revealed that the effect was statistically 0.9707, 0.9690, and 0.9718, respectively, as shown in
significant (P < 0.0001). Table 2. An R2 level above 0.80. Indicates the method of
𝐸𝐸 (𝑌1 ) = +82.00 + 7.25𝐴 + 1.63𝐵 regression used is suitable and is considered a good fit
+1.13𝐶 + 2.00𝐴𝐵 − 2.50𝐴𝐶 − model. This suggests that the model was unable to account
0.25𝐵𝐶 − 3.88𝐴2 − 5.63𝐵2 − 2.62𝐶 2 for only 2% of the response variation and that the process
The model's predictive power was demonstrated by the parameters under investigation account for more than 98%
model fitting criterion of R2 coefficient calculation. The of the diversity in the features of pectin, alginate, and
Y2 (LE) score for each trial batch displayed a strong R- beads (Figure 4).
squared of 0.9976. The equation shows that the ANOVA
independent variables A, B, C, and D have "p" < 0.05, After the responses were collected, they were adjusted,
which significantly affects the LE. The findings and an ANOVA was performed in order to estimate the R2
unequivocally demonstrate that the interaction between the based on the Central Composite Design. Table provides
Ca2+ ion and the COO groups in both polymers may have the adjusted and predicted R2, precision, percentage CV,
led to the development of a "egg-box" structure in PEC and P values.24
within alginate beads (Figure2). This can, as predicted, Predicted and observed values
lessen the issue of drug leaking during bead After obtaining and tabulating the experimental and
manufacturing. Particle size variations in response to predicted values, the percentage error of the responses was
simultaneous changes in two independent variables were also obtained (Figure 5).
demonstrated by the interaction terms (AB, AC, and BC). Optimization of PEC-ALG beads factors
The coefficient values in the given equation.21 A quadratic model that connects the independent
Time required for drug 90% release parameters and the DRB (response) was chosen by the
Regardless of time, every bead delivered 3-4% of the Central composite design. The second-order polynomial
medication in an acidic media. Another benefit of the equation, which is shown in Table 3&4, describes the
modest drug release in the acidic medium is that it helps quadratic model in terms of the coded parameters. The

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 QbD Based Formulation Development

multiple correlation coefficient (R2) and confidence Ionotropic Ally gelled PEC-ALG beads release BUD in
interval (P) of the revised model were employed for vitro in a pulsatile manner (Figure 10). After two hours in
assessment. The R2 values for Equation (Size), (LE), and the alkaline dissolving medium (phosphate buffer; pH
TR were 0.9707, 0.9690, and 0.9718, in that order. An R2 7.4), there was a faster release of BUD in comparison to
value of higher than 0.09 indicates that the applied the acidic dissolution media (pH 1.2; 0.1 N HCl). This
coefficient regression model is suitable and is considered a could be because the medication released in the alkaline
good fit model. This suggests that the model was unable to media was relatively higher due to the beads' tendency to
account for all but 2% of the response variation, and that expand faster in the alkaline medium than in the acidic
the process factors under investigation account for more one. It is likely that the ionized carboxylic acid groups of
than 98% of the diversity in DRB features. the pectin backbone repelled each other electrostatically
Preparation of Ca++ ion Cross-linked Pectin-Alginate produced a significant swelling force in an alkaline
Beads medium. The PEC-ALG beads in an alkaline environment
The ideal formulation variables for a formulation that is were expected to grow following liquid uptake during the
optimized following to the CCD model and it was found first drug, which then diffused through the hydrated
that 4.14 mg of Pectin, 1.82 mg of Alginate, and 14.36 % viscous layer. The dissolution profiles revealed that the
of Cacl2 and 5.99 hours of cross-linking time using beads released the medication slowly at first, but then
predicted values that are 91.9861 % for Loading more quickly (Figure 10). The overall findings indicate
Efficiency, 0.998 mm for Bead Size and 360 min for time that the dried beads did not swell in the stomach. When
needed for 90% BUD release(Figure 6).The observed the beads were transported to the colon, they began to
values were strikingly comparable to the projected values, expand and served as a matrix for chrono-modulated drug
confirming the validation and dependability of the CCD release.27
model used for DRB formulation optimized. This is why In vitro Cytotoxicity Assay of optimized beads
the % error was so low. The MTT method is accurate, user-friendly, and yields
The loading efficiency (EE%) of the PEC-SA beads was repeatable results. 3- [4, 5-dimethylthiazol-2-yl]-2, 5-
calculated, and the result was observed to be 91%. The diphenyl tetrazolium bromide, or MTT, is the primary
practical yield of the beads was discovered to be 95%. component. It is a tetrazolium salt that dissolves in water
According to the SEM photomicrograph of the complete and, when mixed with salt solutions or other media
bead structure and the cross-sectional view of the without phenol red, yields a yellowish solution. After
optimized formulation displayed in the figures, MTT is dissolved, it becomes an insoluble purple
respectively, the BUD-entrapped beads had a spherical formazan (Figure 11).
shape and a rough, porous exterior surface that helped the
PEC-Alginate beads release the drug under control.25 A CONCLUSION
web of tiny fissures and cracks covers the exterior Beads of BUD loaded pectin-alginate beads were made by
surfaces; these may have formed as a result of the ionotropic gelation, in which calcium chloride was added
polymeric gel's drying process and water molecules as a crosslinking agent. FT-IR analysis revealed
moving around shrinking as a result. It is evident that the significant peaks of BUD was found in beads, indicates
polymers' successful crosslinking with Ca2+ resulted in good compatibility between drug and polymers. Sodium
the spheres' uniform creation. These beads' surface did not alginate was used to prepare the pectin alginate beads,
exhibit any drug crystals, suggesting that the pectin- which were then optimized using the central composite
alginate matrix contained a finely distributed drug. The design model. 4.14 mg of pectin, 1.82 mg of alginate,
scanning electron microscopy study reveals that optimized 14.36% of cacl2, and 5.99 hours of cross-linking time
beads were found to be spherical in shape. The size of the were found to be the appropriate formulation variables.
beads was ranged from 0.798 – 1.30 mm and their mean The response of the beads was 91.9861 % for loading
value was found 1.02 mm of PEC-ALG matrix delayed efficiency, 0.998 mm for bead size, and 360 minutes for
released beads (Figure.8). the amount of time needed for 90% drug release. SEM
Swelling index images revealed that the polymers' successful crosslinking
PEC-ALG bead swelling behavior was compared at two with Ca2+ resulted in the sphere-shaped of beads with
distinct pH values (1.2 and 7.4) to simulate how the beads uniform distribution. A maximum degree of swelling was
in the GIT change. Two distinct solutions were used to observed in the bead swelling characteristics at pH 7.4,
study the characteristics of bead swelling: one at pH 1.2 and the results revealed that a significant higher in the
for two hours, and the other at pH 7.4 at 37°C.The impact PEC ratio was accompanied by a significant increase in
of PEC on the optimized beads' degree of swelling is the swelling index. PEC-ALG beads released in vitro in an
depicted in (Figure 8). After two hours of incubation in 0.1 alkaline medium may swell upon liquid absorption as the
M HCl, the percentage weight gain was measured to first medication pass through the diffusion mechanism into
estimate the swelling index. Because of the ionized the high degree hydrated and leads to viscous layer. The
carboxylic groups, which are highly hydrophilic, beads dissolution of PEC-ALG beads profiles revealed beads
showed a maximum degree of swelling at pH 7.4, and as first released the medication more slowly and then
the PEC ratio rose, the swelling index dramatically rose as released it more quickly. An In vitro study of the toxicity
well.26 of beads on A549 cell lines revealed significantly higher
In vitro release study cell viability than the group of cells treated with pure

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 QbD Based Formulation Development

BUD.BUD loaded PEC-ALG beads could have potential Therapy. Journal of Pharmaceutical Research
for chronomodulated delivery system for targeting International, 34(31B), 28-45.
nocturnal asthma. Further in-vivo animal studies are 13. Murata, Y., Miyamoto, E., & Kawashima, S. (1996).
needed to prove the novel concept. Additive effect of chondroitin sulfate and chitosan on
Acknowledgements drug release from calcium-induced alginate gel
The authors are gratefully acknowledged the facilities beads. Journal of controlled release, 38(2-3), 101-108.
provided at the Crescent School of Pharmacy, B.S. Abdur 14. N. Njoku C, K. Otisi S. Application of Central
Rahman Crescent Institute of Science and Technology. Composite Design with Design Expert v13 in Process
Optimization [Internet]. Response Surface
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Chronotherapeutic Approach for Optimum

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chronosystem. Future Journal of Pharmaceutical and evaluation of brain targeting efficiency in

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