Insitu - Gel Formulation - B.Pharm Project. For Further Details , Contact me @ varshithgopalakrishna@gmail.com

1. RATNAM INSTITUTE OF PHARMACY
APRIL-2019
STUDY OF EFFECT OF SODIUM ALGINATE AND
CALCIUM CARBONATE COMPOSITION ON
LEVOFLOXACIN INSITU GEL FORMULATION
By
R.GOPALA KRISHNA VARSHITH - 15DM1R0014
P.V.SAI GURU CHARAN - 15DM1R0071
CH.ABHISHEK - 15DM1R0002
K.MALLIKARJUNA - 15DM1R0026
Under the Guidance of
Dr. B.KUMAR., M.Pharm. Ph.D.,
Head of the Department,
Department of Pharmacy Practice
1

2. AIM
OBJECTIVEs
The objectives of the study are :
•To formulate a stable Insitu gel.
•To make the gastro retentive Gel float for longer periods.
•To Increase absorption of drug by Targeted Drug Delivery.
•To increase patient compliance.
•To reduce Adverse Effects.
•To reduce dosing frequency.
“To study the effect of Sodium Alginate and Calcium Carbonate
composition difference on Levofloxacin in-situ Gel Formulation”.
2

3. PLAN OF WORK
 Drug literature Survey.
 Procurement of Drug, Chemicals and other
Excipients.
 Preformulation studies of Active Pharmaceutical
Ingredient.
 Formulation development of Floating Gastro-
Retentive Insitu Gel OF Levofloxacin.
 Evaluation of Levofloxacin Floating In-Situ gel.
 Selection of Best Formulation. 3

4. LITERATURE REVIEW
• O’Morain et al. (2018) reviewed on Treatment of Helicobacter In 2018 and
reported that Whether three antibiotics, or bismuth and two antibiotics are used,
excellent eradication rates are achieved, albeit with increased side effects.
Levofloxacin is mainly used as the second-line therapy for treating H.Pylori
induced peptic ulcers.
• Abrham Temesgen et al. (2015) formulated an evaluated sodium alginate, HPMC
based insitu gel of salbutamol sulfate to prolong gastric residence time to
improve bioavailability CaCo3 use as a cross-linker & floating agent where as
sodium alginate used as gel forming agent and HPMC as viscosity enhancer. The
formulation is evaluated based on gelling capacity, floating behavior, invitro drug
release study etc. The study demonstrated that the insitu gelling formulations
can be used as novel formulations with sustained drug release, improved
bioavailability and thereby improve patient compliance.
4

5. • Shika jain et al. (2014) prepared and evaluated gastro retentive drug delivery systems containing
amoxicillin trihydrate and ranitidine HCL to minimize the side effects, for prolongation of action, to
reduce the frequency of drug administration. The formulation prepared by using HPMCK 15M and
HPMC K4M as floating agents, sodium bicarbonate and citric acid as gas-generating agent. Cross
povidone was used as super disintegrant. The result showed Ranitidine HCL showed immediate
release in 30 minutes and Amoxicillin trihydrate showed sustained release in 12 hours and the best
fitted model for amoxicillin trihydrate is zero order and for Ranitidine hydrochloride is first order
release model.
• Vora Vipu et al. (2013) prepared and evaluated the novel floating insitu gelling system for
controlled delivery of ramipril. Ramipril formulation prepared by using sodium alginate , gellan gum
as polymers and calcium carbonate as a gas forming agent. The formulation was evaluated for FTIR.
DSC, viscosity, invitro buoyancy time, invitro drug release. The results suggested that concentrations
of sodium alginate, gellan gum and calcium chloride significantly affected on the dependent
variables like viscosity, invitro buoyancy and showed invitro sustained drug release up to 12hrs.
• R.Rajalakshmi et al. (2011) formulated and evaluated levofloxacin hemihydrate oral floating in-situ
gelling system for controlled release of drug for the eradication of Helicobacter pylori. Polymer
based floating Insitu gelling system prepared by using gellan gum, sodium alginate and caco3 as a
gas forming agent. Calcium ions from calcium carbonate interacted with the polymer help in
gelation. They found that levofloxacin hemihydrate as a insitu floating gelling system exhibited
increase in the residence time of the drug in stomach and thereby increase the absorption.
5

6. DRUG PROFILEDRUG PROFILE
Name : Levofloxacin (Discovered in 1987)
Chemical formula : C18H20FN3O4
Molecular mass : 361.373 g/mol
Category: : Anti Bacterial Agent
Physical Properties : Light yellowish-white to yellow-white crystal
or crystalline powder.
Melting point : 225-227 °C
Protein Binding :24-38% (to plasma proteins)
Mechanism of action:
Levofloxacin inhibits bacterial type-II topoisomerases, topoisomerase-IV, DNA
gyrase-A subunits and two subunits encoded by gyrA gene which results in the breakage of
bacterial chromosome, supercoiling, and resealing, which leads to inhibition of DNA replication
and transcription which causes death of the organism. 6

7. Absorption Rapidly absorbed from GI tract. Peak plasma concentrations usually
attained 1–2 hours after an oral dose; steady-state plasma concentrations
attained within 48 hours with once-daily regimens.
Distribution Widely distributed into body tissues and fluids, including skin, blister fluid,
and a into CSF. Following IV administration of 400 or 500 mg twice daily,
CSF concentrations have been reported to be up to 47% of concurrent
plasma concentrations.
Metabolism Limited Metabolism, drug is mostly (87%) eliminated in unchanged form
Elimination Drug is eliminated unchanged in Urine.
Half life 6-8 hours.
Bio availability Bioavailability is Approximately 99%. 7

8. METHODOLOGY
Sodium Alginate was dissolved in D.I water containing CaCl2
and Sodium Citrate with stirring
Heat to 700
C and then cool to below 300
C
Add CaCO3
and Drug solution to the above solution with stirring
Add Citric Acid & Saccharin sodium with Stirring
Stored in Amber coloured bottle
8

9. FORMULATION DEVELOPMENT
INGREDIENTS F1 F2 F3 F4 F5 F6 F7 F8
Levofloxacin 5% 5% 5% 5% 5% 5% 5% 5%
Calcium
Chloride
0.075% 0.075% 0.075% 0.075% 0.075% 0.075% 0.075% 0.075%
Sodium Citrate 0.25% 0.25% 0.25% 0.25% 0.25% 0.25% 0.25% 0.25%
Sodium Alginate 0.5% 1.5% 2.5% 3.5% 1.5% 1.5% 1.5% 1.5%
Calcium
Carbonate
1% 1% 1% 1 % 0.5 % 1% 1.5% 2 %
Citric Acid 0.2% 0.2% 0.2% 0.2% 0.2% 0.2% 0.2% 0.2%
Saccharin
Sodium
0.05% 0.05% 0.05% 0.05% 0.05% 0.05% 0.05% 0.05%
DI Water
Upto
100ml
Upto
100ml
Upto
100ml
Upto
100ml
Upto
100ml
Upto
100ml
Upto
100ml
Upto
100ml
Table 1:Formulation Table
9

10. Fig 1: Formulation of Levofloxacin Floating In-Situ Gel (F1-F4)
In-situ gel containing In-situ gel containing
0.5% SA & 1.0% CaCO3 (F1) 1.5% SA & 1.0%CaCO3(F2)
In-situ gel containing In-situ gel containing
2.5% SA & 1.0% CaCO3 (F3) 3.5% SA & 1.0% CaCO3 (F4) 10

11. Fig 2: Formulation of Levofloxacin Floating In-Situ Gel (F5-F8)
In-situ gel containing In-situ gel containing
1.5% SA & 0.5% CaCO3 (F5) 1.5% SA & 1.0% CaCO3 (F6)
In-situ gel containing In-situ gel containing
1.5% SA & 1.5% CaCO3 (F7) 1.5% SA & 2.0% CaCO3 (F8)
11

12. RESULTS AND DISCUSSION
Table 2: Preformulation Study of Active Pharmaceutical Ingredient
Characteristics Results
Physical appearance
Light yellowish-white to yellow-white crystal
or crystalline powder.
Solubility
Freely soluble in glacial-acetic acid,
chloroform
Practically insoluble in water.
Bulk density 0.72gm/ml
Tap density 0.84gm/ml
Melting point 225-2270
c
Molecular weight 367.373 g/ml
12

13. Concentration
(mcg/ml)
Absorbance
5 0.152
10 0.308
15 0.453
20 0.624
25 0.760
CALIBRATION CURVE OF LEVOFLOXACIN
Table 3: Standard graph of
Levofloxacin by using 0.1N
HCl
Fig 3:Standard curve of Levofloxacin 0.1N
HCl at λmax = 290nm
13

14. 1. Physical Appearance and Clarity
All the formulations had off white to pale yellow coloured
solution.
Clarity of the all formulations was found to be satisfactory.
2. pH of the Formulations
Formulation Code pH
F1 7.72
F2 7.79
F3 7.91
F4 7.93
F5 8.03
F6 8.1
F7 8.27
F8 8.41
Table No.4: pH of the In-Situ Gelling Formulations
14

15. 3 . Viscosity of In-situ Gelling solutions
Table 5: Viscosity of the In-Situ Gelling
Formulations
Formulation
Code
Viscosity (cps)
F1 172
F2 289
F3 383
F4 461
F5 224
F6 258
F7 294
F8 311
Fig 4: Measurement of viscosity of Levofloxacin
Oral Floating In-Situ Gel(F1-F4)
Fig 5: Measurement of viscosity of Levofloxacin
Oral Floating In-Situ Gel (F5-F8)
15

16. 4. Floating
BehaviourTable 6: Floating lag time (sec) and Floating
Duration (hr) of the In-Situ Gelling
Formulations
Formulation
Code
Floating Lag
Time
(sec)
Floating
Duration
(hr)
F1 122 9
F2 38 10
F3 31 10
F4 28 11
F5 108 8
F6 35 10
F7 25 12
F8 19 12
Figure 6: Floating Behaviour of
Levofloxacin In-Situ Gel
16

17. 5. Gelling Capacity of the Formulations
Formulation Code Gelling Capacity
F1 +
F2 ++
F3 ++
F4 +++
F5 +
F6 ++
F7 +++
F8 +++
Table 7: Gelling Capacity of the In-Situ Gelling Formulations
+ = gelation immediate (< 10 s), weak gels dissolve after few hours (6 hr)
++ = gelation immediate (< 10 s), stiff gels remaining for 10 hr
+++ = gelation immediate (< 10 s), more stiff gels remaining for 12 hr
17

18. 6. Drug Content
Formulation Code Drug Content
(%)
F1 97.72
F2 98.35
F3 99.12
F4 99.16
F5 98.39
F6 99.25
F7 97.56
F8 97.72
Table No.8: Drug content of the In-Situ Gelling
Formulations
Figure No.7: Drug content of Levofloxacin Oral
Floating In-Situ Gel (F1-F8)
18

19. 7. Water uptake
Formulation Code Water Uptake (%)
F1 18.63
F2 33.29
F3 50.36
F4 62.65
F5 30.78
F6 39.63
F7 47.31
F8 58.52
Table 9: Water Uptake of the In-Situ Gelling
Formulations
Figure No.9: Measurement of water uptake by
Levofloxacin Oral Floating In-Situ Gel (F5-F8)
Figure No.8: Measurement of water uptake by
Levofloxacin Oral Floating In-Situ Gel (F5-F8)
19

20. 8. Invitro Drug Release
StudyTable 10: In-vitro Drug release of the In-Situ Gelling Formulations
TIME
(Hr)
FORMULATION CODE
F1 F2 F3 F4 F5 F6 F7 F8
0 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
1 44.11 45.87 43.11 41.98 49.34 47.13 42.76 41.07
2 66.24 64. 16 61.43 60.76 67.43 63.59 60.44 61.23
3 79.54 71.06 66.54 64.98 78.56 72.67 66.65 63.11
4 90.57 78.02 73.32 69.44 86.89 78.56 71.32 70.16
5 98.23 84.12 79.43 75.81 94.99 83.23 76.54 77.54
6 98.52 91.10 84.65 79.09 97.14 89.34 82.82 83.12
7 96.07 88.32 83.10 98.33 95.41 87.81 88.65
8 98.08 94.47 87.12 97.23 91.23 92.45
9 98.41 97.57 92.69 95.19 96.42
10 97.86 96.40 96.74 97.23
11 96.56 96.96 97.51
12 98.28
20

21. Figure No.10 In-vitro Drug release of the In-Situ Gelling of F1-F4 Formulations
21

22. Figure No.11 In-vitro Drug release of the In-Situ Gelling of F5-F8 Formulations
22

23. CONCLUSION
 The aqueous solution of sodium alginate based in-situ gelling formulations showed different
properties based on their difference in composition.
 Expect for the formulation with lowest level of sodium alginate, all systems preserved their
integrity for the study period. The formulations showed satisfactory content uniformity and PH
ensuring their safe use.
 The rheological studies showed that the formulations possessed optimal viscosity which can
facilitate easy administration of the required dose.
 In-vitro drug release study proves that release rate of amoxicillin was depends on the amount of
sodium alginate and Caco3used.
 In comparison to all the other formulations Formulation-F8 is found to effective in different
aspects. However, its drug release was at 97.72%. Therefore, the perfect composition of Sodium
Alginate and Calcium Carbonate for best Insitu gelling formulation (F8) was concluded to be 1.5%
and 2.0% respectively.
23

24. BIBLIOGRAPHY
1. Prasad Garrepally et al, Design, Development and Evaluation of
Stomach Specific Insitu Gel for Antibiotics: Cefdinir, Research
Article Pharmaceutical Sciences, 2014, 4(1):128-137.
2. Lena Murad Thomas, Formulation And Evaluation Of Floating Oral
In-Situ Gel of Metronidazole, International Journal of Pharmacy
and Pharmaceutical Sciences, 2014, 6(10):265-269.
3. Avinash Sharma et al, Development and Characterization of Insitu
Oral Gel of Spiramycin, BioMed Research International, 2014:7.
4. Linda Nassour et al, Floating in-situ-gelling gellan formulations of
metformin hydrochloride, Journal of Chemical and Pharmaceutical
Research, 2014, 6(7):1509-1517.
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 V. Hari Baskar Sir, Head of the Department, Department of
Pharmaceutical Analysis , Ratnam Institute of Pharmacy for his
help in getting the Drug.
 SAI MIRRA INNOPHARM PVT LIMITED , Chennai for providing
Drug as Gift Sample

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