Nanosuspensions accelerate drug substance dissolution rates by increasing surface area and reducing particle size. The key to nanosuspension development is the identification of a suitable delivery system, such that nano-technology.

1. Formulation and Development of
Garcinia indica Nano-suspension
By :Ahmad Abdulhusiaan Yosef Kinani
Co-guide
Prajakta C. Jagtap
Guide -
Dr. Kiran S. Bhise
M.C.E. Society’s
Allana college of pharmacy
Azam campus
Pune-411001
2011-2013
1

2. 1) Introduction
2) Challenges , Hypothesis , Justifications , Objectives , Plan of work
3) Drug and Excipient profile
4) Experimental
5) Results and Discussion
6) Summary and Conclusion
7) Publication
8) References
Content
2

3. Garcinia indica
 Garcinia species are distributed widely throughout the tropical Asian
and African countries and have tremendous potential, both as spice
and medicinal plants
 Useful products
1. HCA regulate obesity( Anti-hyperlipidemia )
2. Benzophenone Derivatives of dried fruit rind contains 2-3%
Garcinol, a poly benzophenone derivative has some anti-biotic
activities and has been found a potent inhibitor of histone
acetyltransferase so it is considered as a potential anti-cancer agent ,
anti-oxidant and free radical scavenging activity
Introduction
3

4. Nano-suspension can be defined as a biphasic system consisting of pure
drug particles dispersed in a aqueous vehicle in which the diameter of
the suspended particle is less than 1μm in size. Nano-suspension consist
of the poorly water soluble compound without any matrix material
suspended in dispersion
Need for Nano-suspension
Most of the drugs coming from high-screening are poorly water
soluble
 Formulation of poorly water soluble drug is always being a
challenge.
 One of the major problem associated with them is low bioavailability
due to less absorption
 This problem can be overcome by using Nano-suspension
Introduction
4

5. The particle size distribution of the solid particles in nanosuspensions
is usually less than one micron with an average particle size ranging
between 200 and 600 nm
 Nanosuspensions differ from nanoparticles .
Nanoparticles are commonly polymeric colloidal carriers of
drugs whereas solid lipid nanoparticles are lipidic carriers of
drugs. In nanosuspension technology, the drug is maintained
in the required crystalline state with reduced particle size,
leading to an increased dissolution rate and therefore
improved bioavailability
Introduction
5

6. Introduction
Bottom-up
Nanoprecipitation
method
Nano edge
Top-down
technology
Media
milling
High pressure
Homogenization
in water
High pressure
Homogenization
in non-aqueous
Sonication
probe
Methods of preparation
6

7. Application of Nano-suspension
Bioavailability enhancement
Ocular administration
Intravenous administration
Pulmonary administration
Targeted drug deliver
Topical formulations
Introduction
7

8. Pharmaceutical Suspension
A Pharmaceutical suspension is a coarse dispersion in
which internal phase is dispersed uniformly throughout the external
phase.
The internal phase consisting of insoluble solid particles having a
specific range of size which is maintained uniformly throughout the
suspending vehicle with aid of single or combination of suspending
agent.
The external phase (suspending medium) is generally
aqueous in some instance, may be an organic or oily liquid for non
oral use.
Introduction
8

9. Classification
1.Based On General Classes
Oral suspension
Externally applied suspension
Parenteral suspension
2. Based On Proportion Of Solid Particles
Dilute suspension (2 to10%w/v solid)
Concentrated suspension (50%w/v solid)
3. Based On Electrokinetic Nature Of Solid Particles
Flocculated suspension
Deflocculated suspension
4. Based On Size Of Solid Particles
Colloidal suspension (< 1 micron)
Coarse suspension (>1 micron)
Nano suspension (10 ŋg)
Introduction
9

10. Advantages
Suspension can improve chemical stability of certain drug. E.g.
Procaine penicillin G
Drug in suspension exhibits higher rate of bioavailability than other
dosage forms. bioavailability is in following order,
Solution > Suspension > Capsule > Compressed Tablet > Coated tablet
Duration and onset of action can be controlled. E.g. Protamine Zinc-
Insulin suspension
Suspension can mask the unpleasant/ bitter taste of drug. E.g.
Chloramphenicol palmitate
10
Introduction

11. Disadvantages
 Physical stability, sedimentation and compaction can causes
problems.
 It is bulky sufficient care must be taken during handling and
transport.
 It is difficult to formulate
 Uniform and accurate dose can not be achieved unless
suspension are packed in unit dosage form
11
Introduction

12. Features Desired In Pharmaceutical Suspensions
The suspended particles should not settle rapidly and sediment
produced, must be easily re-suspended by the use of moderate
amount of shaking.
It should be easy to pour yet not watery and no grittiness.
It should have pleasing odor, color and palatability.
Good syringeability.
It should be physically, chemically and microbiologically stable.
Parenteral/ophthalmic suspension should be sterilizable.
12
Introduction

13.  C Jacobs 2001 Production and characterization of Nano suspensions for the
formulation of bupravaquone; to overcome the problem of the high elimination rate
caused by diarrhea, bupravaquone was formulated as a Nano suspension. In this
study different polymers/hydro gels were employed to create a prolonged retention
time for the drug in the infected gastrointestinal tract (GIT).
 Min Sun, Yan 2006 Formulation and Development of Nano suspension for oral
delivery of Quercetin to enhance dissolution rate and oral bioavailability of a poorly
water-soluble drug, Quercetin loaded Nano suspension (QT-NS) was fabricated by
Nano-precipitation (NP) and high pressure homogenization (HPH) method. The
formulation of Nano suspension was optimized by screening different stabilizers to
enhance the bioavailability
 P Kocbek 2006 Preparation and evaluation of ibuprofen Nano-suspensions for
enhancing the dissolution of poorly soluble drugs by melt emulsification method
without using of organic solvent
 A Koeberlein2009 , Identification of 5-lipoxygenase and microsomal prostaglandin
E2 synthase-1 as functional targets of the anti-inflammatory and anti-carcinogenic of
Garcinol preparation .
13
Literature survey

14.  Robert Becker 2009 Pharmaceutical Nano suspensions for medicament
administration as systems with increased saturation solubility and rate of solution
 Rockville Pike2001 An aqueous intravenous Nano suspension with reduced adverse
effects.
 Chavhan SS2011 Nano suspensions in drug delivery: recent advances, patent
scenarios, and commercialization aspects.
 Wang Y2013 Effects of Nano-suspension Formulations on Pharmacokinetics, In
Vivo Targeting and Efficacy for Poorly Water-Soluble Drugs.
 Mark I. ,Hongtao Zhang2013 Garcinol Nano-suspension oral control release
capsule induces lysosome-dependent degradation and limits Treg cell functions to
enhance targeting therapy against cancers.
 Bassil,Nicholas,Charles2002Method for the preparation of pharmaceutical Nano
suspensions using supersonic fluid flow.
14
Literature survey

15. 15
Literature survey

16. Challenges
The aim of current research is to enhance the aqueous solubility of
Garcinia indica.
Hypothesis
The aqueous solubility of Garcinia indica powder in water would be
enhanced due to the complexation with β-Cyclodextrine by inclusion
complexation method.
Formulation of Garcinia indica powder Nano-suspension would
improve the bio- acceptability of the drug
16
Challenges & Hypothesis

17. Justification
The literature survey reveals that the limited work has been reported
in improving aqueous solubility of Garcinia indica in other delivery
systems as well as Nano-suspension.
Furthermore there was no significant work reported in the area of
formulation of Garcinia indica powder / β-CD complex as an oral
suspension
Hence, the presence work focuses on the formulation and design of
Garcinia indica Nano-suspension as an oral dosage form.
Objectives
The objectives of the present paper are
Formulation and development of Garcinia indica Nano-suspension
Evaluation the safety of Garcinia indica
17
Justification & Objectives

18. 18

19. 19

20. Garcinia indica
Family: Clusiaceae
The composition of the fresh Kokum rind is found to be as follows
20
Drug Profile
Moisture (%) 87.50
T.S.S. (°Brix) 16.44
Protein (Nx 6.25%) 1.92
Crude fiber (%) 14.28
Total ash (%) 2.57
Tannins (%) 2.85
Pectin (%) 5.71
Starch (%) 1.00
Crude fat (%) 10.00
(Hexane extract) Acid (as hydroxy citric acid) 22.80
Pigment (%) 2.4
Ascorbic acid (%) 0.06
Carbohydrates by difference (%) 35.0

21. 21
Drug Profile

22. Sr.no. API & POLYMERS NAME OF THE INDUSTRY BATCH NO.
1. Garcinia indica ( Kokum ) Hadapsar, Magarpatta, Pune - 411011,
Maharashtra
2486
2. β-Cyclodextrin Research lab fine chem industries, Mumbai. 1633281210
3. HPMC Research lab fine chem industries, Mumbai. 33060305
4. Sodium Alginate Research lab fine chem industries, Mumbai. 13912681
5. DPPH (1, 1-Diphenyl –2-
picrylhydrazyl)
Research lab fine chem industries, Mumbai 43751001
6. Ascorbic acid Research lab fine chem industries, Mumbai 666091145C
7. Aluminium chloride Research lab fine chem industries, Mumbai 110044321AD
8. Potassium acetate Research lab fine chem industries, Mumbai 165439001P
9. Tween 80 Research lab fine chem industries, Mumbai 70043889 T
22
Materials

23. 23
Instruments
Sr.no Name of instrument Manufactured by Model no.
1. Double Beam UV Spectrophotometer Jasco Corporation,Tokyo, Japan V-630
2. FTIR Spectrophotometer Jasco Corporation, Tokyo, Japan 4100
3. Magnetic Stirrer Spectra lab Whirlmatic, India TH 100
4. Dissolution Test Apparatus ELECTROLAB TDT- 08L
5. Melting Point Apparatus Veego VMP-I
6. Electronic Weighing balance METTLER TOLEDO AB54-S Switze
7. Scanning Electron Microscope Jeol, Japan JSM-6360A
8. Stability Chamber Thermo lab Pvt. Ltd, Mumbai TH 90S
9. Differential Scanning Calorimeter METTLER TOLEDO 821 E
10. Soxhlet apparatus Whirlmatic, India TH120
11. Rotary evaporator Equitron MFG.CO 9075.CRP.AJJ.0
34
12. Cooling centrifuge C24 DL Remi
13. Probe sonicator EN-30US Enertech, fast
clean, India

24. 24
Experimental

25. Preformulation studies:
Prior to the development of the dosage forms, it is essential that certain
fundamental physical and chemical of the drug molecule and other
excipients are determined. This information is necessary for subsequent
events and approaches in the formulation development.
Organoleptic Properties:
The colour, odour and appearance of each material were studied
Analytical studies
Including UV spectroscopy , FT-IR ,Electronic microscopy and DSC ,
these studies were required to study the properties of drug and excipient
and the compatibility between them during experimental work
25
Results & Discussion

26. Calibration curve of Garcinia indica
in Methanoal at 253nm
FT-IR Spectrophotometry of Garcinia indica
26
Results & Discussion
Wave no. cm-1 Functional group
683.64 C-O, C-C
823.455 C-C
1060.66 C-C , Ring
1159.01 C-C
1381.75 C=C
1629.55 C=O
2357.66 C≡C
2848.35 C-H
3426.89 O-H , ≡C-H

27.  TLC study:
 DSC :
Differential scanning calorimeters
Is a process of measuring the
heat of chemical reactions or
Physical changes as well as heat
capacity due to the presence of
different functional groups
in a given sample
27
Results & Discussion
Mobile phase chloroform/ethyl acetate/formic acid(10:8:2)
Rf value for extract 0.6
Rf value reported 0.63

28.  FT-IR Drug-Excipients Compatibility Study
28
Results & Discussion
The Drug/β-CD complex and
drug-Excipients compatibility
at the initial time and after
1 week .FT-IR analysis shown
that the wavenumber peaks
in the graph ; there was no
possibility of interaction
and form undesirable reactions
during experimental work.

29. Phytochemical evaluation
The Methanolic extract of Garcinia indica fruit was studied to their
various active constituents found in the Garcinia indica powder
29
Results & Discussion
Sr. No. constituent result
1 Carbohydrate +ve
2 Alkaloids +ve
3 Steroids +ve
4 Glycosides +ve
5 Saponins -ve
6 Favonoids +ve
7 Tannic and phenolic
compound
+ve
8 Proteins and amino acid +ve
9 Fixed oils. +ve
10 Anthraquinone -ve
Sr.no Evaluation
parameter
Value (%)
1 Ash value 20
2 Acid insoluble ash
value
2.5
Concentration
(μg/ml)
Ascorbic acid
Inhibition %
Garcinia indica
Inhibition %
10 15 12
100 70 68
500 95 91
Type of extract Total flavonoids content
Equivalent to Catechin
Conc. (μg/g)
Methanolic extract of
Garcinia indica
134.32
Type of extract Total phenolic Conc. equivalent
to Gallic acid (mg/gm)
Ethanolic extract of
Garcinia indica
0.34

30.  Acute Oral Toxicity (LD 50)
Organization for economic co-operation & development.
Dose preparation: Garcinia indica powder suspension is prepared in honey and
sterile water (1:2:2) to make final concentration of Garcinia indica powder
200mg/ml., and oral dose is given according to adjust 2000m – 5000mg/kg.
Control animals are given honey & water solution accordingly (i.e. 0.2ml and
0.5ml/animal)
 Evaluation of Garcinia indica –(β-Cyclodextrin) complex
Dissolution study revealed that
the dissolution of 1:2 complex
has a high profile than 1:1 complex
as shown in Figure using Dissolution
apparatus which determined the
absorbance by UV spectroscopy
30
Results & Discussion

31. Evaluation of batches (suspension)
The formulation of batches
required to evaluate the samples
for their sedimentation volume,
Viscosity, Re- dispersibility
and % Drug release for the
suspension
31
Results & Discussion
Drug/β-CD
(6gm)of
each F
HPMC
( mg )
Na-
Alginate
( mg )
Tween80
( ml )
PG
( ml )
Ph-
Buffer
( ml )
DW
( ml )
F1 60 600 1 0.5 1 Qs 50
F2 60 400 1 0.5 1 50
F3 60 200 1 0.5 1 50
F4 40 600 1 0.5 1 50
F5 40 400 1 0.5 1 50
F6 40 200 1 0.5 1 50
F7 20 600 1 0.5 1 50
F8 20 400 1 0.5 1 50
F9 20 200 1 0.5 1 50
Formulation F1 F2 F3 F4 F5 F6 F7 F8 F9
Vis. (cP) 60 83 121 152 91 69 85 130 181
SV (24 h) 0.23 0.74 0.76 0.75 0.84 0.95 0.90 0.95 0.92
Re-dispersibility (%) 90 85 85 90 90 95 80 80 85

32. In vitro release kinetics of batches
The study of drug release from the suspension
in vitro by using Dialysis bag to evaluate the
% Drug release from each sample vs. time in
Minutes to obtain the highest % release , F6 was
the highest %Release profile after 600 min.
 Evaluation of Nano-suspensions:
Particle Size Analysis
Photon correlation spectroscopy (PCS) is a technique used to determine the mean
particle size diameter and found that nanometre range should be (<100nm) and the
result was (93.4nm)
Zeta potential Analysis
In colloidal systems according to electric double layer theory, there is net balance of
attractive as well as repulsive forces. Zeta potential imparts positive or negative charge
on surface of particle and should be > +20 mV or > -20 mV ; the result was 23.11 mV.
32
Results & Discussion

33. Particle Size Analysis
Zeta potential Analysis
33
Results & Discussion

34. Regression analysis:
Effect of formulation variables on % Drug release. The factorial equation for percentage
drug where X1 (HPMC) and X2 (Sodium Alginate) As Sodium Alginate increased and
HPMC deceased, it caused more hindrance of drug diffusion from the complex and %
drug release decreased. The combined effect of X1 and X2 has been shown in Figure
Response surface plot. Both in a specific concentration (HPMC40mg factor X1) And
(Sodium Alginate 200mg factor X2) exerted a positive effect on the %Drug Release (Y),
magnitude of these factors increased cumulative drug release. The Counter plot gave an
idea about the effect of formulation variables on % Drug release (Y) which located at
84.41% of the plot(the intercept of 40mg X1 and200mg X2)
34
Results & Discussion

35. Optimized formulation
Using the polynomial equations, the optimized formulation was obtained for the response
parameter. In the trial runs the optimized formulations were arrived Using numerical
optimization in DESIGN EXPERT 8.0.5VERSION. The data for the formulation
variables, the response parameter and the constraints placed on them utilizing this system
to select the best batch which represented by higher % drug release over time
35
Results & Discussion
Dependent variables
Optimized formulation
Experimental
value
Predicted value
(%) Drug release at 11
hrs.
84.41 83.41

36.  Evaluation of factorial batches
The evaluation of factorial batches by determining the sedimentation volume (0.88 for
F4) , viscosity (69 pc for F4) and %re-dispersibility(95% for F4) that confirmed the best
batch among the nine formula (F4 was optimize one)
36
Results & Discussion
Formulation F1 F2 F3 F4 F5 F6 F7 F8 F9
Vis. (cP) 60 83 121 69 91 75 85 130 181
SV (24 h) 0.23 0.74 0.76 0.88 0.84 0.60 0.90 0.95 0.92
Re-dispersibility (%) 90 85 85 95 90 90 80 80 85

37. Cumulative % Drug release of factorial batches
Where F4 was the highest % Drug release by using
Dialysis bag (84.41%) to be selected as an Optimize
Batch. The figure below shown the % release Vs. Time
For the F4 (Optimize batch)
37
Results & Discussion
Batches Cumulative %drug release
F1 77.68±0.91
F2 79.44±0.54
F3 75.92±0.46
F4 84.41±0.53
F5 78.50±0.87
F6 70.84±0.21
F7 81.90±0.42
F8 76.14±0.56
F9 79.41±0.98

38.  The aim of this research is to enhance the solubility of poor aqueous drug
 The use of Nano-suspension technique was the best solution for this
problem as the drug powder was prepared as a complex with β-Cyclodextrin
by inclusion method ;then by using of Sonicator Probe Apparatus to prepare
Nano-suspension
 The suspension was evaluated for (Sedimentation volume, Viscosity , Re-
dispersibilty and in vitro % Drug release)
 The Nano-suspension was evaluated for its particle size and charge (the
results confirmed that both were within the range)
 The various concentration of X1(20,40 and 60mg) and X2(200,400 and
600mg) used to prepare the factorial batches(9 F)
 Evaluation of these batches (Sedimentation volume, Viscosity , Re-
dispersibilty and in vitro % Drug release)
 Selection the best formula that has best % Drug release and good ( viscosity
and Sedimentation Vo) values.
 The optimize batch represented by the highest cumulative % drug release
with time 38
Summary & Conclusion

39.  By selection a specific ratio of combined action ( X1 40mg and X2 200mg)
Hence , the solubility of poor aqueous Garcinia indica powder was enhanced
by two factors as shown previously in Response Surface Plot and Counter
Plot ;First by complexation of the powder with β-Cyclodextrin which is
considered as carrying agent ; Second by formulation of the complex as
Nano-suspension to increase the solubility
 For future work to develop this dosage form to be more global in science
view to enter the clinical trials, and to be used in treatment of different
disorders which are difficult to be cured by classic treatment . This step was
the beginning of a new formula that includes Herbal Drug using Nano-
suspension Technique , so work can be progressed to establish the
therapeutic utility of these systems by pharmacokinetic and
pharmacodynamics studies.
 The different Novel Technology can be adopted in the formulation which
requires a study in different areas such as (Pre-clinical and Clinical Trials) to
confirm that its action with lowest adverse reactions and high Efficacy.
39
Summary & Conclusion

40. Publication
Communicate to Inventi Journals 2013
Full title: Formulation and Development of Garcinia indica Nano-
suspension
Authors: Ahmad AB kinani ,Prajakta C.Jagtap, Kiran S.Bhise
40
Publication

41. I would like to express my obligation to Shri P.A.Inamdar, President
of M.C.E. Society , Mr. Irfan Shaikh, Joint Secretary of M.C.E.
Society for providing excellent research facilities.
Words seem insufficient to express my deep sense of gratitude, to
beloved and respected Principal and my guide Prof. Dr. Kiran S.
Bhise mam for her excellent guidance, critical supervision, keen
interest and continuous encouragement throughout this study and co-
guide Prajakta Jagtap mam.
I am thankful to the teaching staff and the non teaching staff.
I sincerely thank to Baader Schulz Laboratories- Pharma Division
for providing me the help in the Analysis tests and Evaluation
technique.
I extend a deep expression of gratitude to my Parents.
Last but not the least, I am indebted to him for the love and blessings,
he has showered on me. Thank You GOD.
Besides this, I am thankful to all those who have knowingly and
unknowingly helped me in the successful completion of this
project……. 41
ACKNOWLEDGEMENT

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