Floating microspheres of cephalexin

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RESULTS AND DISCUSSION

1 Drug polymer interaction study

2 Standard curve of Cephalexin

3 Discussion of Preliminary batches

for process optimization

4 Optimization of various parameters

on the best batch of cephalexin

5 Layout of 32 full factorial design

6 Results of optimization batches of

factorial design

7 Characterization of optimized
batches

8 Validation of optimum floating

microspheres formulation

9 Drug Release Kinetics

10 Photomicrographs of microspheres

11 SEM

12 Stability Study

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1.Drug polymer interaction study (FTIR Sectra study)

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Figure-5.1(a) Cephalexin

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IR spectra of cephalexin and physical mixture of drug and polymer showed in figure 5.1 a,b, in ir
spec tra of cephalexin ,peak is observed at and .Two peak cm observed because of
functional group having two peaks in range.IR spectra of physical mixture of drug and polymer
are nearly same to the plain compound.There is no significant change in IR spectra of physical
mixture. all prominent peaks of Cephalexin. The major IR peaks observed in
Cephalexin were 3335.12 (3300‐3500) (N‐H), 1686.44 (1680 ‐ 1760
(C=O), 3054.69 (3300 ‐ 2500 (O‐H), 2525.12 (2590 – 2550 (S‐H),
1196.61 1220 ‐1020 (C‐N) and 1282.43(1000 –1300) (C‐O)

2 Preparation of Standard curve of Cephalexin

Standard curve was prepared according to procedure given in chapter-4.4 the method obeys
Beer’s Law in the concentration range of 10 to 80 mcg/ml.The result of standard curve
preparation are shown in Table-1 and figure 1

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 Floating microspheres of cephalexin

Table 1: Standard curve data of the observed U.V. analysis of

cephalexin
date] in 0.1N HCl.
the
[Pick Conc Absorbance at 257 Average Std. Calculated
( µg/ml nm absorban deviatio absorbance
) S1 S2 S3 ce n

0 0 0 0 0 0 0.007
10 0.109 0.10 0.108 0.109 0.000516 0.203
9
20 0.250 0.25 0.250 0.250 0.001033 0.403
1
30 0.358 0.35 0.356 0.357 0. 000516 0.603
7
40 0.465 0.46 0.465 0.465 0. 000516 0.803
5
50 0.567 0.56 0.568 0.567 0. 000516 1.003
7
60 0.668 0.66 0.668 0.668
8
70 0.790 0.79 0.791 0.790
0
80 0.901 0.90 0.901 0.901
0

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Figure 2 Standard calibration curve of Cephalexin in 0.1 N HCL

3 OPTMIZATION OF PROCESS VARIABLES

The effect of the following was studied for blank as well as drug loaded
microspheres.
A. VOLUME OF AQUEOUS PHASE FOR PRIMARY EMULSION

B. VOLUME OF ORGANIC PHASE

C. ACETONE:DICHLOROMETHANE RATIO

D. STIRRING RATE

E. POLYMER:DRUG RATIO

Table : Optimization of volume of aqueous phase for primary emulsion

Batch Volume of aqueous phase Characteristic

No.
A1 5 ml Thick emulsion(Not easily porable)
A2 10 ml Proper amount to form emulsion
A3 15ml Break down of emulsion
A. VOLUME OF AQUEOUS PHASE FOR PRIMARY EMULSION

Results and discussion:

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 Floating microspheres of cephalexin

• The volume of aqueous phase (0.1NHCL)was found have significant influence on

the formation of microspheres.
date] • It was found difficult to form emulsion by use of 5ml,it was not easily pourable
the
[Pick • Microspheres could not be recovered when more than 15 ml of the aqueous was
used.It was found that emusion was breakdown.
• Relatively large microspheres were obtained when 15 ml of aqueous phase was
used. Afterwards the other process parameters were adjusted to obtain discrete and
spherical microspheres.
• 15ml of the aqueous phase ( A2 ) was selected for further studies.

B. VOLUME OF ORGANIC PHASE

Table : Optimization of volume of the organic phase

Batch Volume of the organic Phase Product characteristic
No.
B1 10 ml Aggregated lump
B2 20 ml Spherical microspheres
B3 30 ml Coarse, not complete spherical

Results and discussion:

• The volume ( 10 ml ) of the organic phase was difficult to handle. The main
difficulties were high viscocity and the formation of opaque polymer dispersion.
• A minimum 20 ml of solvent mixture was required to have necessary probability and
clarity of the solution.
• The rate of addition of organic phase into the aqueous phase was important in the
process of microspheres formation lump formation and coagulation of polymer was observed
when the organic phase was added to a faster rate into the aqueous phase.
• Hence, organic phase was slowly delivered into the aqueous phase with a syringe
fitted with an 18 gauge needle.
• 20 ml of organic phase( B3 ) was selected for further studies.

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C. ACETONE: DICHLOROMETHANE RATIO

Table : Optimization of Acetone:Dichloromethane ratio

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Batch No. ACT:DCM Product characteristic Aggregation

ratio
C1 3:1 Spherical microspheres ++
C2 2:1 Spherical microspheres +
C3 1:1 Irregular microspheres ++
C4 1:2 Spherical microspheres +++
C5 1:3 Spherical microspheres +++
+= no/low aggregation +++= high aggregation
Results and discussion:

• The results shown in the table reveals that, as the proportion of dichloromethane in
solvent blend was increased, the yield of blank microspheres declined. The probable reason may
be the high degree of volatility of dichloromethane from the surface of ethyl cellulose
microspheres.
• The 2:1 ratio of acetone: dichloromethane (2:1 ) ( C2) was selected for further studies.
Note: Residual solvent limits for microspheres.
Dichloromethane class 2 solvent , 600 ppm limit
Acetone Class 3 solvent.
Gas chromatography is used to determine residual solvents.
D .STIRRING RATE
Table Optimization of stirring rate

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Batch No. Stirring rate Product characteristic Aggregation

D1 350 rpm Very large size particles ****
D2 550 rpm Largesize,Nearly spherical ***
D3 750 rpm Small size, spherical **
D4 950 rpm Small size microparticles *
D5 1150rpm very small,braekdown of *
microparticles
* = Low aggregation **** = High aggregation
Results and discussion:
• The higher stirring rate produced irregular sized microspheres.
The probable reason would be the collision of droplets and rupturing of solidified
microspheres.

• Satisfactory microspheres were produced in the range of 550 rpm to 950 rpm.
• 950 rpm ( D4 ) was selected for further studies.

E . POLYMER : DRUG RATIO

Table : Optimization of polymer : drug ratio

Batch No. Polymer: Product Aggregation % yield

drug ratio characteristic
P1 6:1 Spherical +
P2 5:1 Spherical +
P3 4:1 Spherical +
P4 3:1 spherical +
P5 2:1 Spherical + 79
P7 1:1 Spherical ++ 75
P4 1:1.5 Spherical ++ 64
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P5 1:2 Irregular +++ -----

*= low aggregation **= High aggregation
Results and discussion:
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[Pick • Polymer to drug ratio was tried at five levels as shown in the table. Size, shape and
various other morphological characteristics were found to be dependent on the polymer :
drug ratio.
• To a certain point, an increase in polymer ratio resulted in the formation of discrete
spherical microspheres. After that point, Irregular microspheres were formed.( i.e. Polymer to
drug ratio 1:2).
• It could be remarked that with 2:1 ratio of polymer to drug (P1). microspheres with
highest in-vitro availability and an efficient entrapment of the drug were obtained.
• Considering superior yield and morphological characters, batches P1 to P4 selected
for in vitro dissolution studies

Table : Shows the optimized parameters

Parameter Selected batch Specification

Volume of aqueous phase A2 10 ml of aqueous phase

Volume of organic phase B3 20 ml of organic phase

Acetone : DCM ratio C2 2:1 ratio of ACT:DCM

Stirring rate D3 550 rpm

750 rpm
950 rpm

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Table 42: Micromeritic properties of batches S1 to S9

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[Pick repose density density ratio
(º) (g/cm3) (g/cm3)
S1 19.29±0.22 0.361±0. 0.428±0.00 15.65±0.6 1.18±0.3
005 2 96 3
S2 21.00±0.34 0.385±0. 0.464±0.00 17.02±0.4 1.20±0.3
006 8 32 4
S3 22.19±0.29 0.389±0. 0.474±0.01 17.93±1.4 1.21±0.3
007 0 65 3
S4 18.67±0.18 0.422±0. 0.493±0.00 14.40±0.5 1.16±0.3
008 9 37 1
S5 22.58±0.65 0.447±0. 0.518±0.01 13.70±0.4 1.15±0.3
009 3 26 3
S6 24.95±0.22 0.450±0. 0.522±0.00 13.79±1.2 1.16±0.2
006 9 31 8
S7 19.29±0.65 0.473±0. 0.570±0.01 17.01±0.7 1.20±0.4
010 5 42 4
S8 25.17±0.54 0.532±0. 0.631±0.01 15.68±0.8 1.18±0.0
016 6 82 2
S9 27.11±0.27 0.543±0. 0.695±0.02 16.75±0.7 1.27±0.0
018 1 65 4

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4.7 RESULTS AND DISCUSSION

Table 29 : Results of dependent variables

Batch Y1 (Particle Y2 (%encapsulationefficiency) Y3( t80%

code size)
K1 480±1.32 64.10 340
K2 378±1.53 59.46 327
K3 210±0.95 62.00 332
K4 487±0.99 68.55 274
K5 381±1.17 61.82 310
K6 2131±.11 60.00 295
K7 491±1.56 60.68 276
K8 383±0.99 71.40 282
K9 217+1.56 76.23 320
Mean ± S.D., n=3

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