The document outlines a plan to develop nanostructured lipid carriers (NLCs) loaded with the antihypertensive drug lacidipine to enhance its oral bioavailability. Key aspects of the plan include: 1) Conducting preformulation studies such as determining lacidipine's solubility, partition coefficient, and developing HPLC and UV methods for analysis. 2) Preparing NLCs using a solvent injection method and studying the effect of formulation variables like lipid type and amount, surfactant concentration, and drug loading. 3) Characterizing the optimized NLCs using techniques like particle size analysis, drug entrapment efficiency tests, and in vitro drug release studies.

1. DEVELOPMENT OF LACIDIPINE LOADED
NANOSTRUCTURED LIPID CARRIERS (NLCs)
FOR BIOAVAILABILITY ENHANCEMENT
By:
Anuradha Kush, Associate Professor
R.K.S.D. College of pharmacy,
Kaithal, Haryana

2. CONTENT
INTRODUCTION
LITERATURE REVIEW
RESEARCH ENVISAGED
PLAN OF WORK
PREFORMULATION STUDIES
PREPARATION, EFFECT OF PROCESS VARIABLES AND
CHARACTERIZATION
IN VITRO AND IN VIVO STUDIES
STABILITY STUDIES
REFERENCES

3. Oral Route
• Most preferred route for administration of drugs
• Offers greatest degree of patient compliance
• Very effective for drugs with high solubility and gastrointestinal
permeability
INTRODUCTION
El-Kattan. et al.2012

4. MECHANISM OF ORALABSORPTION
GIT Anatomy
Following oral dosing, drug molecules can cross the GI
membrane through various mechanisms that involve
• Passive diffusion
• Active transport.
El-Kattan. et al.2012

5. BIOAVAILABILITY
Bioavailability is the rate and extent (fraction or the
percentage of the dose) to which the active drug ingredient
delivered is absorbed from a drug product into the general
circulation.
Poor bioavailability by the oral route can be due to :
 Poor solubility
 Degradation in GI lumen
 Poor membrane permeation
 Pre-systemic elimination
Liang Shen. et al.2008

6. FACTORS AFFECTING BIOAVAILABILITY
Particle size
pKa
Partition coefficient
Stability
Liang Shen. et al.2008

7. BIOAVAILABILITY ENHANCEMENT OF
POORLY WATER SOLUBLE DRUGS
Prodrug approach
Microemulsion formulations
Particle Size Reduction (Micronization, Nanotechnology)
Gupta. et al.2011

8. NANOSTRUCTURED LIPID CARRIERS
Radtke. et al. 2011
Nanostructured lipid carriers (NLCs) are a type of submicron
particulate drug delivery system based on mixture of solid lipids
with liquid lipids.
 The nanostructure lipid carriers (NLCs) are regarded as the
second-generation of lipid nanoparticles.
 Solid lipids and liquid lipids are considerably different in
chemical structure.
 The usual particle diameters of the NLCs are in the range
of approximately 10–1000 nm.
 Molecules of NLCs improve mucosal adhesion due to
small particle size and increases their GIT residence time.
 Accommodate more drug and reduce the drug leakage
during storage.

9. NLCs - The more intelligent system than SLNs
SLN: tendency to form
perfect crystals can not
accommodate larger drug
amount
NLC: inhibit crystallization process
leads to a highly disordered,
imperfect lipid matrix structure
offering space for drug molecules.

10. ADVANTAGES OF CARRIER SYSTEM
Reginald. et al. 2012
 Increases drug loading
 Controlled drug release
 The possibility of production on large industrial scale
 Possess negligible toxicity
 Bioavailability of highly lipophilic molecules can be increased
 Minimum drug leakage during storage

11. GASTROINTESTINALABSORPTION OF NLCs
THROUGH SMALL INTESTINE
Pathak.et al. 2011

12. Solid lipid (Glyceryl monostearate)
 Lipophillic in nature.
 Responsible for more sustained release of the drug.
Liqid lipids (Oleic acid, Linoleic acid)
 Enhances the entrapment efficiency.
 Entropy of the lipid provides space for entrapment of drug
molecules.
Surfactant (Poloxamer 407 )
Supports the absorption of NLCs, due to their effect on intestinal
epithelial permeability.
FORMULATION OF NLCs

13. B.P 2007, page-1205
IUPAC Name Diethyl 4-{2-[(tert butoxycarbonyl)vinyl]phenyl}- 1,4
dihydropyridine -3,5dicarboxylate
Category Antihypertensive(Calcium channel blocker)
Appearance It's a white to pale yellow colored crystalline powder
Molecular Formula C26H33NO6
Molecular Weight 455.54 g/mol
Melting point 183.5-184.5°C
Partition Coefficient 5.49
Dose 2-6mg
Bioavailabity 10%
Solubility Practically insoluble in water, springly soluble in alcohol,
soluble in dichloromethane, freely soluble in acetonitrile
and acetone.
LACIDIPINE PROFILE

14. MECHANISM OF ACTION
TRIPATHI KD., 6th edition. 2010 PAGE- 529,550
 Lacidipine blocks calcium channels.
Act as smooth muscle relaxant.
Thus produces vasodilation.
Lacidipine

15. LITERATURE REVIEW
Author Work Year
Khan et al., Explained the different types of nanocarriers which were
based on solid lipid like solid lipid nanoparticles,
nanostructured lipid carriers, lipid drug conjugates are
with their structural differences.
2012
Reginald et al., Developed and compared solid lipid nanoparticles
(SLNs) and nanostructured lipid carriers (NLCs) of
Clotrimazole and concluded that the tested NLCs
showed better stability than SLNs at 25 °C and shows no
significant difference in drug release profile of NLCs
after 3 months storage.
2012
Bajaj et al., Described various techniques used for the improvement
of the Bioavailability of drugs including size reduction,
solubilising excipients, colloidal drug delivery systems
etc.
2011
Das et al., Reviewed the different formulation techniques, drug
incorporation models, GI absorption and oral
bioavailability enhancement mechanisms, stability and
storage condition of the formulations.
2011

16. Patravale et al., Reviewed various nano-architectures such as nanosuspensions,
lipid and polymeric nanocarriers, inorganic nanostructures and
its advantages and challenges associated with their efficient
delivery of poorly bioavailable drugs.
2011
Pathak et al., Demonstrated the superiority of NLC over SLN for improved
oral delivery and it was deduced that the liquid lipid, oleic acid
was the principal formulation factor responsible for the
improvement in characteristics, pharmacokinetics and
biodistribution of NLC.
2011
Shen et al., Incorporated etoposide into NLCs by an emulsification and low
temperature solidification method and concluded that NLCs is a
promising delivery system to enhance the oral absorption of
poorly water soluble drugs.
2011
Krishnaiah et al., Explained that the prodrug approach is an exciting way of
improving the oral bioavailability of BCS class II drugs with low
solubility and reasonable permeability.
2010
Gupta et al., Reviewed that, for poorly soluble drug candidates, self-
emulsifying drug delivery systems (SEDDS) could provide an
effective and practical solution to the problem of formulating
drugs where low solubility in the fluids of the GIT limits drug
exposure.
2010

17. Gannu R. et al., Developed microemulsion based transdermal therapeutic system
for Lacidipine is a solution for increasing the absorption of drug
into systemic circulation.
2010
Pan et al., Developed an optimized nanostructured lipid carriers (NLC)
of Vinpocetine (VIN) by using high pressur homogenization
method.
2010
Sanad et al., Formulated Oxybenzone into nanostructured lipid carriers
(NLCs) to enhance its sunscreening efficacy and safety.
2010
Fang et al., Investigated that the bioavailability of lovastatin can be
improved by oral delivery of NLCs and concluded that oral
lovastatin bioavailability was enhanced from 4% to 24% when
the drug was administered from NLCs containing Myverol and
soybean phosphatidylcholine.
2009
McCormack et al., Studied the role of Lacidipine in management of hypertension,
its pharmacodynamic and pharmacokinetic properties.
2003

18. Antihypertensive drug Lacidipine is having only 10%
bioavailability so NLC’s are selected to enhance the
bioavailability.
Lipid-based drug delivery systems are expected as promising
oral carriers because of their potential to increase the solubility
and improve oral bioavailability of lipophilic drugs.
The objective of the present study is to prepare and
characterize Nanostructured lipid carriers (NLCs) for delivering
orally the hypertensive drug Lacidipine for enhancement of
bioavailability.
RESEARCH ENVISAGED

19. PLAN OF WORK
Preformulation Studies
• Absorption maxima (λmax)
• FT- IR Spectrum
• Melting Point
• Solubility study
• Partition coefficient
• Estimation of Lacidipine by UV and HPLC
Preparation and Effect of Formulation Variables
• By solvent injection technique
• Effect of Formulation Variables
 Amount of liquid lipid
 Amount of solid lipid
 Concentration of surfactant
 Amount of drug

20. Characterization of Nanostructured Lipid Carriers (NLCs)
• Morphology (SEM & TEM)
• Particle Size and size distribution
• Drug Entrapment Efficiency (%)
• In vitro drug release and kinetics
Stability Studies
• At 4°±1°C and 25±2°C (60±5% Relative Humidity(RH))
• 3 months study
• Effect on size and (%) drug remaining
In vivo study
• Bioavailability Study

21. PREFORMULATION
STUDIES

22. FT-IR SPECTRUM OF LACIDIPINE
743.25
763.68
1098.75
1151.87
1192.74
1307.15
1364.36
1495.12
1625.88
1650.40
1670.83
1703.52
2978.43
3350.28
3411.58
0
10
20
30
40
50
60
70
80
90
100
%T
500
1000
1500
2000
2500
3000
3500
4000
Wavenumbers (cm-1)
Peak (cm-1) Reference peak (cm-1) vibration Functional group
3411.58 3500 – 3100 -NH Due to amino group present
2978.43 3000 – 2850 -CH(C-C) Alkane group present
1670.83 1600 – 1680 -C=O Carbonyl group present
1625.88 1680 – 1600 -C=C- Double bond b/w carbon
1495.12 1500 – 1400 -C-N- Due to –C-N- in pyridine ring
1151.8 1300 – 1000 C-O-C Due to C-O-C in ester linkage
B.P 2007 page-1205

23. U.V. SPECTRUM OF LACIDIPINE
WITH METHANOL
283 nm
Prasanthi N.L. et al. 2010 (reported is 284nm in methanol)

24. STANDARD PLOT FOR LACIDIPINE
Solvent - Methanol
Wavelength - 283nm
Concentration
(µg/ml)
Absorbance
2 0.190
4 0.262
6 0.381
8 0.482
10 0.621
12 0.728
14 0.846
y = 0.056x + 0.052
R² = 0.994
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
0 5 10 15
Absorbance
Concentration (µg/ml)

25. MELTING POINT Confirms purity of drug
DSC OF LACIDIPINE
Melting point of Lacidipine was detected as 183.51 ̊C and reported is 183.5-184.5 ̊C.
(http://pharmacycode.com/Lacidipine.html).

26. PARTITION COEFFICIENT
The partition coefficient of drug was
determined by shake flask method using
n-octanol and water
The logP of the drug was found to be
5.33± 0.2
Reveals the
lipophilic nature
of drug
Reported is
5.49
B.P 2007 page-1205

27. CHROMATOGRAPHIC ANALYSIS OF
LACIDIPINE
Acetonitrile:ammonium acetate buffer
(90:10)
Flow rate 1 ml/min.
Wavelength used 283nm
HPLC Chromatogram
Gannu R. et al. 2009 (Method was modified from the reported one)

28. STANDARD PLOT OF LACIDIPINE BY
HPLC
Acetonitrile:ammonium acetate buffer
(90:10)
Flow rate 1 ml/min.
Wavelength used 283nm
Concentration
(µg/ml)
Area of
peak
50 1029124
100 2084987
150 3339629
200 4178042
250 5247138
y = 21058x + 17059
R² = 0.997
0
1000000
2000000
3000000
4000000
5000000
6000000
0 50 100 150 200 250 300
Concentration (µg/ml)
Area

29. SOLUBILITY STUDIES
B.P 2009, vol1, page 23
Solvents Solubility (mg/ml) Inference
Absolute Ethanol 8.799±0.20 Sparingly soluble
Water 0.007±0.01 Insoluble
Dichloromethane 88.36±0.49 Soluble
Acetonitrile 169.48±0.6 Freely Soluble
Acetone 158.58±0.7 Freely soluble
Methanol 82.20±0.40 Soluble

30. Lipids Inference
Solid
Glycerol Monostearate
Stearic acid
+
+
Liquid
Almond oil
Oleic acid
Olive oil
Linoleic acid
Sunflower oil
Coconut oil
_
+
_
+
_
_
LIPID SOLUBILITY OF LACIDIPINE
( +; Soluble, - ; Insoluble) Pardeike.et al.2011

31. PREPARATION, EFFECT OF VARIABLES
AND CHARACTERIZATION OF
NANOSTRUCTURED LIPID CARRIERS
FORMULATION

32. METHOD OF PREPARATION
Solvent injection technique
Pathak.et al. 2011
2 ml isopropyl alcohol
Stirring at 400 rpm for 30 min
Drug + Glycerol monostearate +
oleic acid/linoleic acid
Heating at melting point
of Glycerol monostearate
(52-54)°C
20 ml of 0.8%
poloxamer solution

33. 4 ml of 0.1 N HCl to decrease the pH
Centrifuged at 1000 rpm for 30 min at 10°C
Resuspended in 10 ml of 4% poloxamer solution
Pathak.et al. 2011
Filtered to solidify and stored at 4°C.

34. DETERMINATION OF DRUG ENTRAPMENT
EFFICIENCY
Accurately weighed 50mg grounded powder of nanostructured lipid carriers were
soaked in 50 ml of acetonitrile and sonicated using bath sonicator for 2 hrs.
After that the whole solution was centrifuged by using a centrifuge to remove the
polymeric debris.
Then the polymeric debris was washed twice with fresh solvent to extract any adhered
drug.
The clear supernatant solution was filtrated through a 0.45μm filter then analyzed for
drug content by HPLC .
%Drug entrapment efficiency =
Amount of drug recovered
Total amount of drug added
× 100
Thatipamula. et al.2011

35. EFFECT OF FORMULATION VARIABLES
Amount of liquid lipids (linoleic acid/oleic acid)
Amount of solid lipid (GMS)
concentration of surfactant (Poloxamer 407)
Amount of Drug (Lacidipine)

36. Formulation
code
Solid lipid
(mg)
Liquid lipid(mg) Surfactent
conc(%)
Amount of
drug(mg)
Oleic acid Linoleic
acid
F1 100 10 o.8 30
F2 100 15 o.8 30
F3 100 20 o.8 30
F4 100 25 o.8 30
F5 100 30 0.8 30
F6 100 10 o.8 30
F7 100 15 o.8 30
F8 100 20 o.8 30
F9 100 25 o.8 30
F10 100 30 0.8 30
F11 100 20 o.8 30
F12 150 20 o.8 30
F13 200 20 0.8 30
DIFFERENT PARAMETERS FOR
FORMULATIONS

37. F14 100 20 o.8 30
F15 150 20 o.8 30
F16 200 20 o.8 30
F17 150 20 o.8 30
F18 150 20 1 30
F19 150 20 1.2 30
F20 150 20 o.8 30
F21 150 20 1 30
F22 150 20 1.2 30
F23 150 20 1 10
F24 150 20 1 20
F25 150 20 1 30
F26 150 20 1 40
F27 150 20 1 50
F28 150 20 1 10
F29 150 20 1 20
F30 150 20 1 30
F31 150 20 1 40
F32 150 20 1 50

38. EFFECT OF AMOUNT OF OLEIC ACID
0
50
100
150
200
250
300
0
10
20
30
40
50
60
70
80
90
100
F1 F2 F3 F4 F5
Entrapment efficiency Particle size
Entrapment
efficiency
(%)
Batch code
Particle
size
(nm)

39. EFFECT OFAMOUNT OF LINOLEIC ACID
0
50
100
150
200
250
300
0
10
20
30
40
50
60
70
80
90
100
F6 F7 F8 F9 F10
Entrapment effeciency Particle size
Entrapment
efficiency
(%)
Particle
size
(nm)
Batch code

40. 0
50
100
150
200
250
300
0
10
20
30
40
50
60
70
80
90
100
F11 F12 F13
Entrapment efficiency (%) Particle size
Particle
size
(nm)
Batch code
Entrapment
efficiency
(%)
EFFECT OF AMOUNT OF SOLID LIPID
WITH OLEIC ACID

41. 0
50
100
150
200
250
300
0
10
20
30
40
50
60
70
80
90
100
F14 F15 F16
Entrapment efficiency (%) Particle size
Entrapment
efficiency
(%)
Batch code
Particle
size
(nm)
EFFECT OF AMOUNT OF SOLID LIPID WITH
LINOLEIC ACID

42. 0
50
100
150
200
250
300
0
10
20
30
40
50
60
70
80
90
100
F17 F18 F19
Entrapment efficiency (%) Particle size (nm)
Entrapment
efficiency
(%)
Particle
size
(nm)
Batch code
EFFECT OF SURFACTANT CONCENTRATION
WITH OLEIC ACID

43. 0
50
100
150
200
250
300
0
10
20
30
40
50
60
70
80
90
100
F20 F21 F22
Entrapment efficiency (%) Particle size (nm)
Entrapment
efficiency
(%)
Batch code
Particle
size
(nm)
EFFECT OF SURFACTANT CONCENTRATION
WITH LINOLEIC ACID

44. 0
50
100
150
200
250
300
0
10
20
30
40
50
60
70
80
90
100
F23 F24 F25 F26 F27
Entrapment efficiency (%) Particle size (nm)
Entrapment
efficiency
(%)
Particle
size
(nm)
Batch code
EFFECT OF DRUG AMOUNT WITH
OLEIC ACID

45. 0
50
100
150
200
250
300
0
10
20
30
40
50
60
70
80
90
100
F28 F29 F30 F31 F32
Entrapment efficiency (%) Particle size (nm)
Entrapment
efficiency
(%)
Particle
size
(nm)
Batch code
EFFECT OF DRUG AMOUNT WITH
LINOLEIC ACID

46. Formulations
Particle size
(nm)±S.D
Polydispersity index
Drug entrapment
efficiency(%)±S.D
Blank formulation 235.32±6.2 0.327±0.54 _
Selected
formulation with
oleic acid
249.52±6.8 0.250±0.78 86.9±0.5
Selected
formulation with
linoleic acid
241.4±8.4 0.246±0.47 92.65±0.01
*(Mean ± S.D.)(n = 3)
SELECTED NANOSTRUCTURED LIPID
CARRIERS (NLCS) FORMULATION

47. PARTICLE SIZE OF NANOSTRUCTURED
LIPID CARRIERS
(Zeta sizer)
Particles size of NLCs obtained was 241.4 nm

48. MORPHOLOGY OF NLCs
NLCs were spherical in shape and had smooth surface
SEM image TEM image

49. IN VITRO DRUG RELEASE STUDY
 In vitro drug release from the nanostructured lipid carriers was tested with dialysis
technique.
 Dialysis bag of cellulose dialysis membrane (MW cut- off 10,000 Da) was soaked in
the distilled water overnight.
 1ml of the preparations was placed in dialysis bag and sealed both ends with threads.
 Initial studies were carried out in 100 ml of 0.1N HCl (pH 1.2) for 2 hours and then in
phosphate-buffered saline (PBS) pH 6.8 at 37°C on magnetic stirring moving at a
speed of 50 rpm for 24hrs.
 Samples were withdrawn at predetermined time intervals and replaced with fresh
media.
 Samples were filtered and then analyzed using HPLC at λmax of 283 nm.
Sanad.et al. 2010

50. IN-VITRO DRUG RELEASE STUDY
0
10
20
30
40
50
60
70
80
90
100
0 10 20 30
oleic acid NLCs linoleic acid NLCs
Time (hr)
Cumulative
%
drug
release

51. DRUG RELEASE KINETICS
Models Regression coefficient (r2)
Linoleic acid Oleic acid
Zero order 0.996 0.769
First order 0.982 0.825
Higuchi 0.983 0.986
Korsmeyer-peppas 0.923 0.950

52. BIOAVAILABILITY STUDY
All the in vivo studies were carried out under the guidelines compiled by
CPCSEA (Committee for the Purpose of Control and Supervision of
Experiments on Animal, Ministry of Culture, Government of India)
Approval No. RITS/IAEC/2012/05/26
All the study protocols were approved by the local institutional Animal Ethics
Committee.

53. Albino male wistar rats (250 gm -300gm) are divided in to
three groups and six rats were assigned each group
First group received saline
solution of PBS (pH 7.4)
by oral route
Second group received
aqueous drug solution
by oral route
Third group received
Lacidipine loaded NLCs
by oral route
Blood samples (1.0 ml) were collected from retro-
orbital plexus after 1, 2, 4, 6, 8, 12, 18, 24 hrs.
Serum separated by centrifuge for 20 min at 3000 rpm
Serum samples were stored at -20 °C till assayed by high pressure
liquid chromatographic (HPLC) analysis.
Pan. et al.2010

54. PLASMA PROFILE CURVE
0
100
200
300
400
500
600
700
800
900
0 5 10 15 20 25 30
Drug Solution Lacidipine Loaded Nanostructured Lipid Carriers
Time (hr)
Lacidipine
concentration
(ng/ml)

55. Formulation AUC
(h*ng/ml)
Cmax
(ng/ml)
Plasma
clearance
(ml/hr)
MRT
(h)
t1/2
(h)
Relative
bioavailability
Drug suspension 2064.7586 571.770 5392.32 2.35 1.32 1
Lacidipine
loaded
nanostructured
lipid carriers
(NLCs)
8225 813 1236.82 10.0606 4.03 3.9
MEAN PHARMACOKINETIC PARAMETERS
OF NANOSTRUCTURED LIPID CARRIERS
FORMULATIONS

56. Effect on particle size in blank NLCs
0
50
100
150
200
250
300
350
4 ± 1 °C 25 ± 2 °C
Initial
15 Days
30 Days
45 days
60 Days
75 Days
90 Days
Temperature (˚C)
Particle
size
(nm)
*
*
*
*
*
**
**
*
**
*
**
Temperature (˚C)
Particle
size
(nm)
*
*
*
*
*
**
**
*
** **
*
**
Values are represented as mean ± S.D. (n=3) and significance was tested using
one way ANOVA and Dunnett post test where * represents P< 0.05 and **
represents P<0.01 using initial as control.
STABILITY STUDIES

57. Effect on particle size in Lacidipine loaded
nanostructured lipid carriers
Values are represented as mean ± S.D. (n=3) and significance was tested
using one way ANOVA and Dunnett post test where * represents P< 0.05 and
** represents P<0.01 using initial as control.
0
50
100
150
200
250
300
350
4 ± 1 °C 25 ± 2 °C
Initial
15 Days
30 Days
45 Days
60 Days
75 Days
90 Days
Temperature (˚C)
Particle
size
(nm)
*
*
*
*
*
**
**
*
*
*
**
**

58. 80
82
84
86
88
90
92
94
96
98
100
102
4 ± 1 °C 25 ± 2 °C
Initial
15 Days
30 Days
45 Days
60 Days
75 Days
90 Days
*
*
**
**
*
*
*
*
*
**
%
Drug
remaining
Temperature (˚C)
**
**
Effect on % drug remaining in Lacidipine
loaded nanostructured lipid carriers
Values are represented as mean ± S.D. (n=3) and significance was tested
using one way ANOVA and Dunnett post test where * represents P< 0.05 and
** represents P<0.01 using initial as control.

59. Preformulation studies includes FT-IR spectrum, UV spectrum, melting point
confirms identity and purity of Lacidipine. Solubility profile was analyzed. Log P
Value of Partition coefficient confirms its lipophilc nature.
Preparation of NLCs using glycerol monostreate and linoleic acid by solvent
injection technique. Effect of different variables was observed on the bases of
amount of liquid lipid, amount of solid lipid and surfactant concentration and amount
of drug. The final optimized parameter values were found to be 20mg, 150mg, 1.0%
and 30mg respectively.
The nanostructured lipid carriers was further evaluated by in-vitro drug release and
in-vivo study.
SUMMARY

60.  NLCs formulated with oleic acid and linoleic acid were studied for in vitro
release study, results obtained from the dissolution profile of NLCs was fitted
into various mathematical models and the data was found to fit best into the
zero release model, which suggests that the mechanism of drug release was
constant or independent on concentration.
Stability studies were performed at 4±1°C and 25±2°C (60±5%RH) for 3
months to access the stability of the NLCs. Results of stability studies have
shown that no significant changes were observed in %drug remaining and
particle size.
In vivo bioavailability study was performed in albino rats. The result
showed that the Lacidipine loaded NLCs formulation shows more than thrice
the relative bioavailability of drug solution.

61. CONCLUSIONS
Results from preformulation studies proved the purity of the drug and
compatibility with the excepients.
 Lacidipine loaded NLCs developed for oral delivery of Lacidipine possessed site
specific targeting ability, better stability and higher entrapment efficiency, easy to
scale up.
Linoleic acid caused numerous crystal defects in solid lipid and provided
imperfections in highly ordered crystal. In this space more drug molecules get
entrapped due to which the increase in entrapment efficiency and reduction in
particle size takes place.

62. REFERENCES
 Thomas, V. H., Bhattachar, S., Hitchingham, L., Zocharski, P., Naath, M., Surendran,
N. Stoner, C. L. & El-Kattan. The road map to oral bioavailability: an industrial
perspective. Expert Opin Drug Metab Toxicol 2006, 2(4): 591-608.
 Hurst, S., Loi, C. M., Brodfuehrer, J. & El-Kattan, A. Impact of physiological,
physicochemical and biopharmaceutical factors in absorption and metabolis mechanisms
on the drug oral bioavailability of rats and humans. Expert Opin Drug Metab Toxicol
2007; 3(4): 469-489.
 Varma, M. V., Ambler, C. M., Ullah, M., Rotter, C. J., Sun, H., Litchfield, J., Fenner,
K. S. & El-Kattan, A. F. Targeting intestinal transporters for optimizing oral drug
absorption. Curr Drug Metab 2013; 11(9): 730-742.
 Varma, M. V., Obach, R..S., Rotter, C., Miller, H. R., Chang, G., Steyn, S. J., El-
Kattan, A. & Troutman, M. D. Physicochemical space for optimum oral bioavailability:
contribution of human intestinal absorption and first-pass elimination. J Med Chem
2010; 53(3): 1108-1098.

63.  Karlsson, J., Ungell, A., Grasjo, J. & Artursson, P. Paracellular drug transport across
intestinal epithelia: influence of charge and induced water flux. Eur. J. Pharm. 1999;
9(1): 47-56.
 Lipinski, C. A. Computational alerts for potential absorption problems: profiles of
clinically tested drugs. J. Pharmacol. Tox. Methods 1995; 44(1): 235-249.
GIT Anatomy(www.edoctoronline.com), accessed on April 27, 2013.
 Lennernas, H. Does fluid flow across the intestinal mucosa affect quantitative oral
drug absorption? Is it time for a reevaluation?, Pharm. Res.1995; 12(11): 1573-1582.
Avdeef, A. Physicochemical profiling (solubility, permeability and charge state). Curr.
Top. Med. Chem. 2001; 1(4): 277-351.
 Gursoy, R.N. and Benita, S. Self-emulsifying drug delivery systems (SEDDS) for
improved oral delivery of lipophilic drugs. Biomed. Pharmacother. 2004; 58: 173-182.
 Amidon, G.L. A theoretical basis for a biopharmaceutic drug classification: the
correlation of in vitro drug product dissolution and in vivo bioavailability. Pharm. Res.
1995; 12: 413–420.

64.  Date, A.A. Self-nanoemulsifying drug delivery systems: formulation insights,
applications and advances. Nanomedicine 2010; 5: 1595-1616.
http://en.wikipedia.org/wiki/Bioavailability accessed on April 29, 2013.
 Lappin, Graham; Rowland, Malcolm; Garner, R Colin. The use of isotopes in the
determination of absolute bioavailability of drugs in humans. Expert Opinion on Drug
Metabolism & Toxicology. 2006; 2 (3): 27-419.
 Lappin, Graham; Stevens, Lloyd. Biomedical accelerator mass spectrometry: Recent
applications in metabolism and pharmacokinetics. Expert Opinion on Drug Metabolism &
Toxicology. 2008; 4(8): 33-1021.
 Hoag, Stephen W.; Hussain, Ajaz S. The Impact of Formulation on Bioavailability:
Summary of Workshop Discussion. The Journal of Nutrition. 2001; 131(4): 1389S–91S.
Rita Wegmuller, Michael B. Zimmermann, Diego Moretti, Myrtha Arnold, Wolfgang
Langhans and Richard F. Hurrell. Particle size reduction and encapsulation affect the
bioavailability of ferric pyrophosphate in rats. Amer Soci Nutri Sci J. Nutr. 2004; 134:
3301-3304.

65. Shargel, L.; Yu, A.B. Applied biopharmaceutics & pharmacokinetics (4th edition)1999;
453-430.
 EI-Laithy HM. Self-nanoemulsifying drug delivery system for enhanced bioavailability
and improved hepatoprotective activity of biphenyl dimethyl dicarboxylate. Curr Drug
Deliv. 2008; 5(3): 6-170.
 Wu JZ, Ho PC. Evaluation of the in vitro activity and in vivo bioavailability of realgar
nanoparticles prepared by cryo-grinding. Eur J Pharm Sci. 2006; 29(1): 44-35.
 Kotyla T, Kuo F, Moolchandani V, Wilson T, Nicolosi R. Increased bioavailability of a
transdermal application of a nano-sized emulsion preparation. Int J Pharm. 2008; 22: 8-
144.
 Hecq J, Deleers M, Fanara D, Vranckx H, Boulanger P, Le Lamer S, Amighi K.
Preparation and in vitro/in vivo evaluation of nano-sized crystals for dissolution rate
enhancement of ucb-35440-3, a highly dosed poorly water-soluble weak base. Eur J Pharm
Biopharm. 2006; 64(3): 8-360.
 Lohitnavy, M., Lohitnavy, O., Thangkeattiyanon, O., Srichai, W. Reduced oral
itraconazole bioavailability by antacid suspension. J. of Clin Pharm. 2005; 30(3): 6-201.

66. Magdalene Radtke, Eliana B. Souto, Rainer H. Muller. Nanostructured Lipid Carriers: A Novel
Generation of Solid Lipid Drug Carriers. Pharmaceutical Technology Europe. 2005; 17(4):45–50.
Gupta MM, Saini TR. Preformulation Parameters Characterization to Design, Development and
Formulation of Vancomycin Hydrochloride Tablets for Psudomembranous Colitis. Int J Pharm
Res Dev. 2009; 1(9):1-7.
Prasanthi NL, Rama Rao N. Formulation evaluation of lacidipine tablets employing lacidipine-
starch phosphate binary systems. Research j. Pharm. and Tech. 2010; 3(2): 458-460.
Pathak kamla, Tiwari Radheshyam, Nanostructured lipid carrier versus solid lipid nanoparticles
of simvastatin: Comparative analysis of characteristics, pharmacokinetics and tissue uptake,
International Journal of Pharmaceutics. 2011; 415: 232– 243.
Tiecheng Zhang, Jianian Chen, Yi Zhang, Qi Shen,Weisan Pan. Characterization and evaluation
of nanostructured lipid carrier as a vehicle for oral delivery of etoposide. European Journal of
Pharmaceutical Sciences. 2011; 43:174–179.
Chun-Yang Zhuang, Ning Li, Mi Wang, Xiao-Ning Zhang, Wei-San Pana, Jun-Jie Penga, Yu-
Sheng Pana, Xin Tang. Preparation and characterization of vinpocetine loaded nanostructured
lipid carriers (NLC) for improved oral bioavailability. International Journal of Pharmaceutics.
2010; 394 : 179–185.

67. ACKNOWLEDGMENT
I wish to express my gratitude to my respected guide Dr. Senthil Kumar M., Professor and
Head Department of Pharmaceutics, Rajendra Institute of Technology and Sciences,Sirsa.
I am profoundly grateful to Dr. Rajendar Singh sra, Chairman, and Mr. Omprakash,
Secretary of Rajendra Institutes, Sirsa, for providing me all necessary facilities required for
completion of my project.
My sincere thanks to Dr. Vikramdeep Monga, Dr. Anupama Setia, Mr. Ashwani Sharma,
Mr. Rampal, Mr. Shammi, Miss Madhu, Mrs. Suman.
I express my thanks to SAIF staff of AIIMS, Delhi, for carrying out SEM and TEM of my
sample. I also would like to thanks staff of Nanotechnology GJUS&T, Hisar for carrying out
Zeta sizer and zeta potential of my sample.

68. PUBLICATIONS
Research Paper
Entitled “Development of Lacidipine Loaded Nanostructured Lipid Carriers
(NLCs) for Bioavailability Enhancement”
Communicated to Journal of Drug Targeting
POSTERS PRESENTATIONS
Title: Nanoparticulate Delivery System for Tuberculosis Treatment
Conference: National seminar on Importance of Cell Lines in Pharmaceutical
Research in India. Held at School of Pharmaceutical Sciences, Shoolini
University, Solan, 14-15 Nov, 2011
Title: Nanostructured Lipid Carriers (NLCs)- A Review
Conference: Pharmaceutical Drug Discovery & Development- Future
Perspectives. Rajendra Institute of Technology and Sciences, Sirsa, 28-29 July,
2012