CAPECITABINE LOADED LIPOSOMAL GEL: FORMULATION CHARACTERISATION AND OPTIMIZATION

1. CAPECITABINELOADEDLIPOSOMAL GEL: FORMULATIONCHARACTERISATION
ANDOPTIMIZATION
PRESENTED BY,
tanmay panigrahy
m.PHARM, 2nd year
R.I.P.S
UNDER THE ESTEEMED GUIDANCE OF,
dr.goutam ku. jena
ASSo .PROFESSOR,,R.I.P.S
1

2. CONTENT
 Introduction
 Plan of work
 Drug profile
 Methodology
 Evaluation
 conclusion
 Reference

3. INTRODUCTION
 Definition :-
An artificial microscopic vesicle
consisting of an aqueous core enclose
d in one or more phospholipids layers,
used to convey vaccines, drugs, enz
ymes, or othr substances to target c
ells or organs.

5. Composition of liposomes:
 Phospholipids
 The most common natural
phospholipid is the
phospatidylcholine(PC) is the
amphipathic molecule and also known
as lecithin.
 Cholesterol

9. Aim and Objective
 To prepare Capecitabine loaded liposomal gel
for sustaining drug release, decreasing dosing
frequency and increasing efficacy of drug for
the effective treatment of colorectal cancer.
Plan of work
Pre-formulation studies
 Preparation of calibration curve by UV method
 Fourier transform spectroscopy (FT-IR)
 Differential Scanning Calorimetry (DSC)

10. Plan of work
 Preparation of Liposome by Lyophilization
monophase solution method.
 Characterization of Liposomes
 Drug entrapment efficiency
 In vitro diffusion study
 Optimization
 Formulation of optimized Liposome into a
Topical gel.
 Characterization of Liposomal gel
 Physical appearance & Homogeneity
 Viscosity
 In vitro diffusion study

11. Drug profile
 Name : Capecitabine
 Structure
 IUPAC Name : Pentyl[1-(3,4-dihydroxy-
5-methyltetrahydrofuran-2-yl)-5-fluoro-2-
oxo-1H pyrimidin-4-yl]carbamate
 Molecular Formula: C15H22FN3O6
 Molecular Weight: 359.35
 Melting point : 110-121° C

12.  LogP : 0.4
 Solubility : It is soluble in water
(26mg/ml)
 Category : Antineoplastic.
 T½ : Approximately 45-60 minutes.
 BCS : Class Type IV (low solubility, Low
permeability)
 Protein binding: Less then 60%

13. METHODOLOGY
Standard graph of Capecitabine in Phosphate
buffer PH 7.4
 Stock solution was prepared by dissolving 10mg of drug in
10 ml buffer.
 2.5ml solution was taken out into a 25 ml volumetric flask
and fill the volume to get working standard solution.
 From this solution 0.5 to 5 ml were transferred in to 10 ml
vol.flask and fill the volume with help of buffer to get conc.of
5-50 µg/ml.
 Absorbance was analyzed by UV- Visible spectrophotometer
at λmax of 237nm and calibration curve was plotted by taking
concentration (µg/ml) and absorbance at X-axis and Y-axis
respectively.

14. PREFORMULATION STUDY
Differential scanning calorimetry (DSC)
study
 The DSC measurements were performed on a DSC-60
with a thermal analyzer.
 All samples were placed in sealed aluminum pans before
heating under nitrogen flow (20 mL/min) at a scanning
rate of 100C/min from 25 to 2500C.
 DSC study of pure drug along with blank and drug
loaded formulation was carried out

15. FT-IR spectroscopy study
 Interactions of drug with other polymers were
assessed by FT-IR spectroscopy.
 FT-IR spectra of Capecitabine and its blank
formulation and drug loaded formulation were
recorded on IRAffinity-1, using KBr discs.
 The instrument was operated under dry air purge
and the scans were collected at a scanning speed
of 2 mm/s with resolution of 4 cm-1 over the
region 4000–400 cm-1.
 FT-IR study of drug along with blank and drug
loaded freeze dried optimized formulation was
carried out

16. FORMULATION OF
CAPECITABINE LIPOSOME
 lyophilization monophase solution method was used For the
preparation of the liposome.
 Capecitabine,soya lechithine,cholesterol was weighed
accurately .then all the ingredient was dissolved in TBA in
450c.then water soluble lyoprotectant mannitol was dissolved
in distilled water in 450c.
 Then these two solution were mixed to get a third clear
solution volume of 60 ml.
 All the formulations were characterized in terms of
entrapment efficiency (%) and amount of drug released after
24 h of diffusion study (Q24). Each formulation is to be
optimized by Box-Behnken method of central composite
design (CCD) with 2 level variables by Design expert-11
software.

17. LEVEL USED
FACTORS Low (-1) Medium (0) High (+1)
Independent
variables:
X1= Lipoid S100
X2= Cholesterol
X3= TBA
360
60
40%
480
120
50%
600
180
60%
Dependent variables:
Y1= %EE, Y2= Q24,

18. CHARACTERIZATION OF
LIPOSOME
Entrapment efficiency
 It determines the % of drug entrapped into liposome
that in turn affects drug release pattern from the
formulation.
 In 1st step 1ml of formulation was pipetted out.
%EE was determined by separating unencapsulated
fraction of capecitabine from the formulated
liposome after centrifugation in micro-centrifuge at
9000 rpm for 30 min. at a temperature of 150C.
 UV-spectrophotometer was used at 237nm for
quantification of unencapsulated fraction of
capecitabine in liposome formulation.

19. In-vitro Diffusion study
 For this dialysis membrane-60 was used. Dialysis membrane
was cut into appropriate size and soaked in distilled water for
24hr before use.
 From it 5ml was pipette out. Then other end of dialysis
membrane was tied out & hanged to a stand.
 It was immersed in a beaker filled with 100ml PBS (pH-7.4).
 A magnetic bead was put into it & the whole system was kept
on a magnetic stirrer with continuous stirring at room
temperature & 100rpm.
 1ml sample was pipetted out from beaker at 30,1,2,4,6,8,12 &
24hr interval into 10ml volumetric flask and volume was
made of to the mark.
 Each sample was studied for the absorbance then % drug
release was calculated from the data.

20. PREPARATION OF GEL
 First of all an aqueous solution of 0.001% propyl paraben
solution was prepared. A separate aqueous dispersion of
Carbopol 934 was prepared by dispensing it in 20 mL of
water.
 Then the preservative solution was mixed into it. The
dispersion was allowed to hydrate for 7-9 h.
 Then additional distilled water was mixed until gel like
consistency appeared. In the next step, 3% w/v of glycerin
was added to the dispersion of Carbopol 934.
 optimized liposome (E11) was added to the above dispersion.
To the above mixture, sufficient quantity of triethanolamine
was added and stirred for 5 min with a glass rod until the
desired pH (7-8) comes.
 Then the product was allowed to stand overnight for removal
of entrapped air.

21. CHARACTERIZATION OF GEL
Physical appearance & Homogeneity
 Physical appearance and homogeneity of gel
was determined by visual observation.
Measurement of pH
 By the help of pH meter the pH was
determined.
Measurement Viscosity
 Viscosity of the formulation was determined
by the help of Brookfield viscometer DV-I &
spindle number 62 and RPM was set to 50.

22. Diffusion Study
 Dialysis membrane was cut into appropriate size and soaked
in distilled water for 24hr before use.
 Then one end of open ended diffusion glass tube was tied
properly with rubber.
 Glass tube was filled properly upto top with gel. It was fixed
in a burette stand & immersed in a beaker, filled with 200ml
PBS (pH-7.4).
 A magnetic bead was put into it & the whole system was kept
on a magnetic stirrer with continuous stirring at room
temperature & 100rpm.
 1ml sample was pipetted out from beaker at 30,1,2,4,6,8,12 &
24hr interval into 10ml volumetric flask and volume was
made upto the mark by PBS (pH-7.4). Simultaneously 1ml
fresh PBS was put into the beaker
 Each sample was studied for the absorbance then cumulative
% drug release was calculated from the data.

23. Standard curve of
capecitabine in PBS pH-7.4
Standard graph of capecitabine in PBS (pH-7.4)
Solvent medium λmax(nm) Equation R2 value
Phosphate buffer(pH-7.4) 237 y = 0.0239x - 0.0202 R² = 0.9995

24. Preformulation study
DSC study
DSC thermograms A. Capecitabine pure form, B. Blank
liposomes and C. Capecitabine loaded Liposomes
From the DSC studies of Capecitabine, blank liposomes and Capecitabine loaded Liposomes, it
was found that the peak for pure drug ((116.32ºC) and the peak of formulation appeared at
114.42ºC and there is no peak for blank formulation. As the peak of drug and drug loaded
liposomes appeared approximately at same temperature, hence it can be confirmed that there is
no interaction between formulation ingredients and the drug.

25. FT-IR study
FT-IR spectra of pure Capecitabine, Blank formulation,
Formulation and Gel

26. Independent variables
Run Drug
(in mg)
Lipoid S 100 (X1)
[mg]
Cholesterol (X2)
[mg]
TBA (X3)
[in %]
Mannitol(1:2)
(mg)
F1 20 360 180 50 180
F2 20 480 120 50 240
F3 20 600 180 50 300
F4 20 600 120 60 300
F5 20 360 60 50 180
F6 20 600 60 50 300
F7 20 480 180 60 240
F8 20 480 60 40 240
F9 20 480 180 40 240
F10 20 360 120 40 180
F11 20 600 120 60 300
F12 20 360 120 60 180
F13 20 480 60 60 240
Formulation table of Capecitabine loaded liposome
several trial batches were formulated to determine the concentration
range of the excipients. Drug concentration was kept fixed (20mg). A total
volume of 60ml liposomal was prepared.

27. CHARACTERIZATION OF
LIPOSOME
 All the formulations were characterized in terms of entrapment
efficiency (%), and amount of drug released after 24 h of diffusion
study
 It was observed that amount of drug released after 24 hr of E11 was
satisfactory
Different Capecitabine Liposomes formulation as per DX 11
software (CCD)

28. IN VITRO DIFFUSION STUDY

29. EVALUATION OF LIPOSOMAL GEL
Physical appearance & Viscosity and pH of 3 gel formulations
% drug release of liposomal
gel

30. CONCLUSION:
 The Capecitabine loaded liposome was successfully
prepared by Lyophilization Monophase Solution
Method by using different concentration of
soyalecithin, cholesterol and different volume % of
TBA as independent variables followed by
optimization with respect to % EE and Q24 as
dependent variables.
 The optimized formulation F11 was selected on the
basis of highest %EE and better drug release as
compared to other formulations and incorporated with
carbopol 934 to produce a transparent gel for
exhibiting more sustaining effect, minimizing
frequency of administration and side effects for
providing better chemotherapy for colorectal cancer.

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