International Journal of Pharmaceutical Sciences

INT.J.PH.SCI.,MAY-AUG, 2010;2(2):515-521
ISSN 0975-4725
www.ijps.info

Original Research Manuscript

MICROCRYSTALS: FOR IMPROVEMENT OF SOLUBILITY AND DISSOLUTION RATE OF

LAMOTRIGINE

Mali Sandip L*, Nighute Ashok B, Deshmukh Vivek, Gonjari Indrajeet D, Bhise Satish B
Dept. of Biopharmaceutics, Govt. College of Pharmacy, Karad, MS, Pin: 415 124.
Correspondence e-mail address: e-mail: sandip22686@yahoo.co.in

ABSTRACT
The emulsion solvent diffusion method was employed for the preparation of microcrystals of Lamotrigine (LTN). To achieve the
combined effect of surfactant and size reduction on the solubility of LTN, the drug was formulated using constant concentrations of the
surfactants such as PVA, PEG 6000 and PVP K- 30 at stirring speed of 3000 rpm. As it was noted that the speed of stirring is responsible
for micronization of crystals, the speed of stirring was optimized to get uniform sized crystals. The effect of changing the type of
surfactant on the formation of LMT microcrystals was investigated. Microcrystals were dried at room temperature. Saturation solubility
and dissolution characteristics were also investigated. Crystalline state evaluation before and following particle size reduction was also
conducted through differential scanning calorimetry (DSC) and powder X-ray diffraction (PXRD).
KEY WORDS: Lamotrigine, Emulsion solvent diffusion, microcrystals, polymer effect.

INTRODUCTION surface area, increasing surface area by reducing particle sizes to

Solubility and dissolution rate of the drugs are key determinants
of its absorption behavior from gastrointestinal tract. Numbers of
newly synthesized drugs are poorly soluble in water. BCS class
II (i.e. less water soluble) drugs require innovative approaches to
reach a sufficiently high bioavailability when administered by
oral route.[1] Poorly water soluble drugs can exhibit a number of
negative clinical effects including potentially serious issues of
interpatient variability and subsequent erratic absorption
following dosing to individual patients.[2] Reducing the particle
size of an active pharmaceutical ingredient has opened new
formulation opportunities. Noyes-Whitney equation supports the
statement that the linear dependence of the dissolution rate on the
one or more order of magnitude increase in the dissolution rate.[3]
Lamotrigine is an anticonvulsant drug used in the treatment of
epilepsy and bipolar disorders. For epilepsy it is used to treat
partial seizures, primary and secondary tonic-clonic seizures, and
seizures associated with Lennox-Gastaut syndrome.[4]
Lamotrigine is a BCS class II drug, due to which dissolution rate
is the rate limiting step for its absorption.[5] Reduction of particle
size, solubilization in surfactant systems, formation of water
soluble complexes, use of pro-drugs are some of the methods to
increase the dissolution rate of the drug.[6]
Even though the microparticles are prepared it should be given in
a dosage form like tablets, capsules or injectable preparations. If
it is given orally in the form of tablet or capsules flow properties

515 Int.J.Ph.Sci., May-August 2010;2(2):

Mali Sandip L et al: MICROCRYSTALS: FOR IMPROVEMENT OF SOLUBILITY AND DISSOLUTION RATE OF
LAMOTRIGINE
of the drug does matter. In the present work, microcrystals of
Lamotrigine were prepared and evaluated for various parameters
along with flow properties for an optimized sample.
MATERIALS AND METHODS
Materials
Lamotrigine was obtained as gift sample from Alembic
Pharmaceuticals (Baroda, India). Povidone (PVP K-30),
polyethylene glycol (PEG 6000) and polyvinyl alcohol (PVA)
were procured from Glenmark (Mumbai, India). Methanol and
hydrochloric acid were of AR grade (Qualigens, Mumbai, India).
Preparation of Microcrystals
Microcrystals were prepared by emulsion solvent diffusion
method. A weighed amount of drug was dispersed homogenously
in 7 ml of methanol. This organic phase was added at room
temperature, under constant mechanical stirring (3000 rpm, Remi
Stirrer, Mumbai, India) to 100 ml of 0.2 % w/v aqueous solution
of surfactants (PVA, PEG 6000, PVP K30). Stirring was
continued for 30 min. Microcrystals were collected by filtration
and dried at room temperature.
Drug Content
An amount of 25mg of the samples was weighed and dispersed
in 10 ml of 0.1 N HCl. It was sonicated for 10 min; the samples
were centrifuged at 2000 rpm for 10 min. Supernatant was
diluted with suitable quantity of 0.1N HCl and analyzed through
UV-Visible Spectrophotometer (Shimadzu UV-1700, Tokyo,
Japan) at 267nm.
Saturation Solubility
Apparent saturation solubility measurements were assayed
through UV spectrophotometer. To 10 ml of the 0.1 N HCL
excess quantity of the samples (50 mg) were added. Apparent
solubility study was performed by standardized shake flask
0
method at 37 C. After shaking for 48 hrs, the samples were
filtered through 0.2 µm membrane filters (PALL life Sciences,
Mumbai, India) and the filtrate was appropriately diluted with
HPLC mobile phase prior to analysis.
516
In-Vitro Dissolution Studies
In vitro dissolution studies were carried out using eight station
USP type II dissolution apparatus (LABINDIA 2000, Mumbai).
The samples were checked for dissolution rate in 900 ml of 0.1N
HCl. Dissolution medium was maintained at 37 ± 0.50c
temperature. The basket was rotated at 50 rpm. After
predetermined time intervals, 5ml of samples were withdrawn
and 5ml of fresh dissolution media was added to maintain the
sink. The withdrawn samples were analyzed using UV-Visible
Spectrophotometer (Shimadzu UV-1700, Tokyo, Japan).
Scanning Electron Microscopy (SEM)
Morphological evaluation of the collected microcrystals and the
drug was carried out by JSM-6400 scanning electron microscope
(JEOL, Tokyo, Japan). Samples were fixed on aluminum stubs
with conductive double sided adhesive tape and coated with the
gold by sputter coater at 50 mA for 50 s.
X-Ray Powder Diffraction
Crystallinity of the drug and the spherical microcrystals were
determined using the Philips Analytical X-RD (Model: PW 3710,
Holland), with copper target. The conditions were: 40 kV
voltage; 30 mA current; at room temperature. The scanning angle
0 0
ranged from 5 to 60 of 2è, steps were 0.02 of 2è and the
counting rate was 0.4 s/step.
Differential Scanning Calorimetric (DSC)
Thermal properties of the drug and the samples were analyzed by
DSC. The samples were heated in a hermetically sealed
aluminum pans. Heat runs for each sample were set from 30 to
350 0C at a heating rate of 10 0C/min, using nitrogen as blanket
gas.
Flow Properties
Flow properties of the drug and microspheres were studied by
determining the bulk density (ób), tap density (ót), Carr’s Index
and Hausner ratio.
Carr’s Index = [(ót – ób)/ ót] x 100
Hausner ratio = (ót)/ (ób)
Int.J.Ph.Sci., May-August 2010;2(2):
Mali Sandip L et al: MICROCRYSTALS: FOR IMPROVEMENT OF SOLUBILITY AND DISSOLUTION RATE OF
LAMOTRIGINE
Wettability
The powder bed hydrophilicity test was done to confirm the
wettability of the microcrystals by placing the samples on a
sintered glass disk forming the bottom of glass tube on which
methylene blue crystals were placed. The whole device was
brought into contact with water. The time taken for the capillary
rising of water to the surface so as to dissolve methylene blue
crystals was noted. The shortest rising time would correspond to
the most hydrophilic drug leading to good wettability.
Moisture Uptake Study
A weighted quantity of crystals (10 mg) were placed in crucibles
0
at accelerated condition of temperature & humidity, 40 ± 2 C &
75 ± 5 % RH respectively (environmental test chamber, HMG
INDIA, Mumbai). The changes in weight of samples were
determined.
Stability Study
The optimized samples passed through accelerated stability
studies. The samples (each 5mg, n=3) were kept for stability
0
studies at 40 ± 2 c and 75 ± 3% RH for a period of 3 months in
stability chamber. The samples were kept in glass vials sealed
with rubber plugs. After 30 and 60 days, the samples were taken
out and analyzed for drug content.
RESULTS AND DISCUSSION
Preparation of Microcrystals
The emulsion solvent diffusion method was used as a method of
choice for the preparation of the microcrystals of Lamotrigine.
The method was simple and efficient because it does not
consume energy for homogenization. The method includes the
Table 1: The drug content and solubility of the pure drug and microcrystals
Batches Batch ID
LMT L -
LMT/PVA L1
LMT/PEG 6000 L2
LMT/PVP K30 L3
*
mean ± S.D.
517
formation of microcrystals by addition of organic phase
containing drug to aqueous solution of the surfactants by using a
syringe through16# gauge needle. Methanol as selected as the
organic phase. Stirring at 3000 rpm using mechanical stirrer was
optimized and found to be efficient to get smaller particle size
microcrystals. PEG, PVA and PVP K-30 were selected as
stabilizers. Concentration of the surfactants was kept constant.
The dispersion of drug and methanol added gradually in the
aqueous phase containing dissolved polymer, and the added
droplets solidified into the microcrystals. It was found that the
preparations of microcrystals were controlled by two processes,
drug-polymer complexation and solidification. The combined
effect of stirring and stabilizers result in reduction of size and
increased hydrophilic characters of the drug. The solidified
crystals were dried at room temperature.
The manufacturing of a microcrystals implies the creation of
additional surface area and hence interface. As the Gibbs free
energy change, associated with the formation of additional
interface is positive, the microcrystals formed are
thermodynamically unstable and will tend to minimize their total
energy by agglomeration. Kinetically, the process of
agglomeration depends on its activation energy. This activation
energy can be influenced by adding stabilizers to the system. A
first requirement for a stabilizing system is that it provides
wetting of the hydrophobic surfaces of the drug particles.[7,8]
Drug Content
The drug content was found to be good and uniform among the
different batches of the prepared samples and ranged from 98.03
to 99.65 % (Table 2).
% Drug Content* Solubility* (mg/ml)
0.172±0.032
99.45 ± 0.14 0.327 ± 0.063
98.03 ± 0.05 0.346 ± 0.015
99.65 ± 0.07 0.412 ± 0.034
Int.J.Ph.Sci., May-August 2010;2(2):
Mali Sandip L et al: MICROCRYSTALS: FOR IMPROVEMENT OF SOLUBILITY AND DISSOLUTION RATE OF
LAMOTRIGINE

Solubility Studies dissolution.[13] Surfactants applied in the process have been

As water is a universal solvent, apparent solubility studies were
carried out in deionized water. In solubility studies of the
samples, the crystals prepared using PVP K30 have showed
highest solubility of the drug in water (0.412 ± 0.034 mg/ml) as
compared with the untreated drug (0.172 mg/ml) (Table 1).
In-Vitro Dissolution Studies
The dissolution profile of the drug and its microcrystals are
shown in Figure 1. The dissolution studies were carried out in 0.1
N HCL which is mentioned as USP dissolution media. The drug
microcrystals prepared with polymers exhibited better dissolution
rate when compared with plain drug, which indicates the
deposition of polymer on the drug surface.[9,10] The dissolution
profile of the pure drug and the polymeric microcrystals explains
that the particle size reduction was an effective and versatile
[11]
option to enhance the rate of dissolution. The microcrystals
prepare without surfactant shown 39.88% dissolution after
completion of 45 minutes, while that with PVA, PEG 6000 and
PVP K30 shown 85.65, 83.35 and 97.89% dissolution of
microcrystals. PVP K30 was proved more efficient of all used
polymers.
shown to reduce the aggregation tendencies of particles
compared to milling. The mechanism for how this process yields
enhanced dissolution properties is believed to be a
microenvironment surfactant effect whereby surfactant
dissolution creates a local surfactant concentration in the
boundary layer surrounding the drug particles, providing a lower
energy pathway for drug dissolution.[14] This process of
microcrystal preparation is believed to be particularly effective at
enhancing the rate of drug dissolution because the drug particles
are maintained in direct contact with the surfactants particles
during drug dissolution.
SEM, XRD AND DSC
The particle morphologies before and after comminution were
not simply related. Figure 1a and 1b shows the particles before
and after comminution without using surfactants, indicating very
less change in the particle morphology. However Figure 1c-1e
shows the particles after comminution with various surfactants
were morphologically quite different from the starting material.
Therefore, the addition of surfactants aids in breaking and
stabilization of the larger particles.

Figure 1: In-vitro drug release profile of (L) the drug and its
crystals prepared (L1) without surfactant and with (L2) PVA,
(L3) PEG 6000, (L4) PVP K30

According to the Noyes- Whitney and Ostwald-Freundlich

equations, size reduction is an effective tool for dissolution
elevation.[12] Those samples prepared with surfactants showed
the faster dissolution rate, with approximately more than 50% of
the drug being released within 15 min compared to 30 % for the
samples prepared without surfactant. At the end of 50 min, more
than 90 % of the drug was released from all crystals except for
the crystals prepared without surfactants. This effect can be
explained by an increased specific surface area which is
hydrophilized due to the adsorbed hydrophilic polymers. The fact
that the microcrystals prepared by solvent change approach
exhibited a faster dissolution rate than the control shows that the Figure 2: SEM photomicrographs of (a) the drug and its crystals
prepared (b) without surfactant and with (c) PVA, (d) PEG 6000,
solvent change method itself was responsible for increased (e) PVP K30

518 Int.J.Ph.Sci., May-August 2010;2(2):

Mali Sandip L et al: MICROCRYSTALS: FOR IMPROVEMENT OF SOLUBILITY AND DISSOLUTION RATE OF
LAMOTRIGINE

XRD pattern of Lamotrigine (Figure 2a) showed intense and peak towards lower temperature dictates decreased melting point
sharp peaks indicating its crystalline nature. The diffractogram of of the drug in the crystals. This decreased melting point accounts
the samples after treatment attested no modifications occurred for increased solubility of the drugs.[16] From the DSC analysis of
after comminution. The diffraction pattern of microcrystals with the crystals; positive influence of the hydrophilic polymer on the
PEG 6000 and PVP K30 showed no change in the peak intensity solid state of the drug was attested.
of Lamotrigine at the same angle, indicating retention of
crystalline state of Lamotrigine. Both the formulations were
found to contain crystalline material, as shown by their X-ray
diffraction patterns (Figure 2b and 2c) which indicates most
stable form of drug. It means that inclusion of hydrophilic
surfactant enhance solubility and ultimately the dissolution by
retaining crystallanity of the drug.[15] Amorphous transition leads
to unstable form that is difficult to formulate so it is
advantageous fact regarding formulation development.

Figure 4: DSC thermo-grams of (a) the Lamotrigine and its

crystals prepared with (b) PEG 6000 and (c) PVP K30

Powder Flow Properties of Microcrystals

The speed of the stirring is the key determinant of the
size of the crystals. The size of the resultant crystals was
determined by the balance between the interfacial tension of
emulsion droplets and the shearing force applied to the droplets
under stirring. The higher the speed of the stirring smaller the
crystals because of the increase in the shearing force applied.[17]
Figure 3: XRD pattern of (a) the Lamotrigine and with (b) PEG
Effect of various polymers on the bulk density, tap density,
6000 (c) PVP K30
Hausner’s ratio and Carr’s index is shown in the Table 2. Among
Thermal behavior of pure drug and crystals are shown in Figure
the used polymers, PVP was found to be best in all respects.
3. The DSC curve showed that LMT appeared as sharp
Result of the Carr’s index is an indicative of improved
endothermic peak at about 123.04 oC corresponding to its
compaction behavior compared to that of the drug. Compaction
melting. However, the crystals prepared with PVP K30 and PEG
behavior plays a very important role in manufacture, processing
6000 shows shift of endothermic peak towards lower temperature
and packaging techniques.
at 95.75 oC and 103.14 oC respectively. Shift of the endothermic

519 Int.J.Ph.Sci., May-August 2010;2(2):

Mali Sandip L et al: MICROCRYSTALS: FOR IMPROVEMENT OF SOLUBILITY AND DISSOLUTION RATE OF
LAMOTRIGINE

Table 2: Flow properties of prepared microcrystals Shivaji University, Kolhapur for getting facilities to perform

Parameter L1 L2 L3 L4 XRD and DSC.

Bulk Density (g/ml) 0.6110 0.6849 0.7246 0.6944

REFERENCES
Tap Density (g/ml) 0.8475 0.8197 0.8620 0.8197
1) Moneghini M, Kikie I, Oinovich D, Perissutti B, Filipovic-Grcic
Hausner Ratio 1.3647 1.1968 1.1896 1.188
J. Processing of carbamazepine-PEG 4000 solid dispersions
Carr’s Index (%) 27.9056 16.45 15.93 15.28
with supercritical carbon dioxide: preparation, characterization
and in vitro dissolution. Int J Pharm 2001; 222: 129-138.
Wettability, Moisture Uptake and Stability Studies 2) Crisp MT, Tucker CJ, Rogers TL, Williams III, Johnston KP.
The wettability studies were done by using powder bed Turbidimetric measurement and prediction of dissolution rate of
hydrophilicity test. For LMT, LMT/PVA, LMT/PEG 6000, poorly water soluble drug nanocrystals. J Control Rel 2007;
LMT/PVP K30 the methylene blue crystals were dissolved after 117: 351-359

53, 49, 31 and 33 min respectively, whereas even after 53 min
the methylene blue crystals did not get dissolved for the
untreated drug, indicating an augmented wetting property of the
drug in the prepared crystals.
The moisture uptake study is important to check hygroscopic
nature of the prepared crystals. The change in weight was not
seen, indicates no hygroscopic characters.
The accelerated stability studies shown there was no considerable
change in the drug content after study duration. The drug content
3) Lee RW, Mcshane J, Shaw JM, Wood RW. Particle size
reduction. Liu R. Water-Insoluble drug formation. Interpharm
press. 2000: 455-492.
4) Poselli GM, Pennisi EM, Roberti FTR, Garelli FF, Bandinelli
PL. Psychotic disorders after lamotrigine. Ital J Neurol Sci
1998, 19:124-125.
5) Nielsen KA, Dahl M, Tommerup E, Wolf P. Comparative daily
profiles with different preparations of lamotrigine: A pilot
investigation. Epilepsy and Behavior 2008, 13: 127-130.
6) Mutalik S, Parambil A, Krishnan M, Achutha NU. Enhancement

after accelerated stability studies was not less than 98.30%. It is
notification of stable nature of crystals towards accelerated
stability studies.
CONCLUSION
From the above discussion is concluded that the emulsion solvent
diffusion method is efficient as it gives uniform micro sized
crystals with improved solubility and dissolution rate of the
prepared crystals. XRD and DSC results showed the crystalline
of dissolution rate and bioavailability of aceclofenac: A
chitosan-based solvent change approach. Int J Pharm 2008,
350: 279-290.
7) Huang Y, Chung T, Tzeng T. Drug release from PLA/PLG
microparticulates. Int J Pharm 1997; 156: 9-15.
8) Bernard VE, Guy VM, Patrick A. Top-down production of drug
nanocrystals: Nanosuspension stabilization, miniaturization and
transformation into solid products. Int J Pharm 2008; article in
press.
9) Singh B, Ahuja N. Studies on dissolution enhancement and

nature and thermal behavior of prepared microcrystals. Drug
release profile confirms the efficiency of methodology. This
indicated that both the emulsion solvent diffusion and the
inclusion of the hydrophilic surfactant contributed to enhanced
dissolution rates of Lamotrigine.
ACKNOWLEDGMENT
We are thankful to Glenmark Pharma, Mumbai & Alembic
Pharmaceuticals, Baroda, India for gift samples of drug and also
mathematical modeling of drug release of a poorly water
soluble drug using water-soluble carriers. Eur J Pharm
Biopharm 2007; 65: 26-38.
10) Nayak A, Mutalik US, Reddy M, Kustagi P. Preparation and, in
vitro, preclinical and clinical studies of Aceclofenac spherical
agglomerates. Eur J Pharm Biopharm 2008; 70: 674-683.
11) Meziani MJ, Desai T, Sun YP. Nanosizing drug particles in
supercritical fluid processing. J Amer Chem Soc 2004; 126:
10842-10843.
12) Hecq J, Deleers M, Franara D, Vranckx Lamer SL. Preparation
and in vitro/in vivo evaluation of nano-sized crystals for

520 Int.J.Ph.Sci., May-August 2010;2(2):

Mali Sandip L et al: MICROCRYSTALS: FOR IMPROVEMENT OF SOLUBILITY AND DISSOLUTION RATE OF
LAMOTRIGINE
dissolution rate enhancement of ucb-35440-3, a highly dosed
poorly water soluble weak base. Eur J Pharm Biopharm 2006;
64: 360-368.
13) Paradkar AR, Pawar AP, Chordiya JK et al. Spherical
crystallization of Celecoxib. Drug Dev Ind Pharm 2002; 28:
1213-1220.
14) Nielloud F, Marti-Mestres G. Pharmaceutical Emulsions and
Suspensions, Drugs and the pharmaceutical sciences. Marcel
Dekker Inc., New York, 2000, 105: 127-190
15) Wong SM, Kellaway IW, Murdan S. Enhancement of the
dissolution rate and oral absorption of a poorly water soluble
drug by formation of surfactant-containing microparticles. Int J
Pharm 2006; 317: 61-68.
16) Wong J, Brugger A, Khare A. Suspensions for intravenous (IV)
injection: A review of development, preclinical and clinical
aspects. Adv Drug Del Rev 2008; 60: 939-954.
17) Cui F, Yang M, Cun D. Design of sustained-release Nitredipine
microspheres having solid dispersion structure by quasi-
emulsion solvent diffusion method. J Control Rel 2003; 91: 375-
384.

Author Information:

Article History:
Date of Submission: 17-02-2010
Date of Acceptance: 12-06-2010
Conflict of Interest: NIL
Source of support: NONE

521 Int.J.Ph.Sci., May-August 2010;2(2):