RESEARCH ARTICLE

Development of Biodegradable In Situ Implant and Microparticle

Injectable Formulations for Sustained Delivery of Haloperidol
TAREK A. AHMED,1 HANY M. IBRAHIM,2 FATHY IBRAHIM,2 AHMED M. SAMY,2 ALAA KASEEM,2 MOHAMMAD T. H. NUTAN,3
MUHAMMAD DELWAR HUSSAIN3
1
Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, King Abdulaziz University, Jeddah,
Kingdom of Saudi Arabia
2
Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt
3
Department of Pharmaceutical Sciences, Irma Lerma Rangel College of Pharmacy, Texas A&M University Health Science Center,
Kingsville, Texas 78363

Received 7 December 2011; revised 29 May 2012; accepted 11 June 2012

Published online in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/jps.23250

ABSTRACT: The objective of this study is to formulate injectable, biodegradable sustained

release in situ implant (ISI), and in situ microparticle (ISM) formulations of haloperidol. Factors
affecting the in vitro drug release, pharmacokinetics, and stability of the formulations were
investigated. The concentration of the polymer, poly(lactide-co-glycolide) acid (PLGA), and the
type of solvents showed a pronounced effect on the in vitro drug release from the ISI and
ISM formulations. The ISM formulation [20% PLGA in N-methyl-2-pyrrolidone (NMP)–peanut
oil, 1:4] showed reduced maximum plasma concentration (60 versus 44 ng/mL) and longer
release (30 days, plasma concentration of 8 ng/mL versus 20 days, plasma concentration of
6 ng/mL) compared with the ISI formulation (20% PLGA in NMP) after intramuscular injection
in rats. The delivery of haloperidol can be extended further by changing the concentration,
molecular weight, and lactide-to-glycolide ratio of the PLGA. These formulations can be easily
administered by both intramuscular and subcutaneous injections. The shelf lives of both systems
were found to be 2 years when stored at 4◦ C. Haloperidol can be formulated as an injectable ISI
or ISM systems suitable for 1 month or longer release. © 2012 Wiley Periodicals, Inc. and the
American Pharmacists Association J Pharm Sci
Keywords: in situ implants; in situ microparticles; haloperidol; PLGA; biodegradable
polymers; injectables; sustained release; pharmacokinetics; stability

INTRODUCTION microspheres,4 as polymer/drug pellet,5 and as surgi-

cal implants with PLGA (75:25) and PLGA (85:15).6
Haloperidol is an antipsychotic drug used in the
Various intramuscular or subcutaneous-controlled
treatment of various psychoses including schizophre-
drug delivery systems in the form of implants
nia, mania, polar disorder, and in behavior dis-
or microparticles containing biodegradable poly-
turbances. Haloperidol is usually given orally as
mers have been developed in the last decades. Al-
small daily doses (2–10 mg every 4–8 h) or long
though some pharmaceutical long-acting parenteral
acting intramuscular injection (50–100 mg every 4
systems such as emulsions,7 liposomes,8 micelles,9
weeks).1–3 The commercially available haloperidol de-
microspheres10,11 have demonstrated some success
canoate injection (administered every 28 days) is sus-
in certain applications but they have several inher-
pended in sesame oil with 1.2% (w/v) benzyl alcohol
ent disadvantages. These include a relatively compli-
as a preservative. Haloperidol was also formulated
cated manufacturing procedure to produce a stable,12
with poly(lactide-co-glycolide) acid (PLGA) (50:50) as
satisfactory entrapment efficiency,13 and the pos-
sibility of microsphere migration from the site of
Correspondence to: Muhammad Delwar Hussain (Tele- injection.14,15 In situ implant (ISI) and in situ mi-
phone: +361-221-0739; Fax: +361-593-4303; E-mail: hussain@
pharmacy.tamhsc.edu) croparticle (ISM) are being developed as alternative
Journal of Pharmaceutical Sciences delivery systems. These systems, made of biodegrad-
© 2012 Wiley Periodicals, Inc. and the American Pharmacists Association able polymers can be injected via a syringe into the

JOURNAL OF PHARMACEUTICAL SCIENCES 1

2 AHMED ET AL.
body. Once injected, they solidify to form semisolid
depots.16–18 One or more combined stimuli may lead
to the in situ gelling formation.19 The stimuli may
be physiological stimuli, for example, temperature
and pH20 ; physical changes in biomaterials, for ex-
ample, swelling, solvent exchange (precipitation),
and self-assembly21–23 ; and chemical reactions, for
example, enzymatic, chemical, and photo-initiated
polymerization.24,25
The ISI consists of liquid drug–polymer formula-
tions, which form implants in situ upon injection
and contact with body fluid through one or more of
the above-mentioned mechanisms. The chosen poly-
mer dissolves in water-miscible solvents such as
N-methyl-2-pyrrolidone (NMP) or dimethyl sulfoxide
(DMSO) and solidifies upon injection.26 ISM consists
of an internal phase containing drug in polymer–sol-
vent phase (polymer phase) emulsified into an exter-
nal phase (oil phase). Upon injection of this emulsion,
the internal polymer phase precipitates and forms mi-
croparticles. ISM has reduced myotoxicity compared
with the ISI.27 Additionally, the preparation process
of the ISM is simpler compared with the classical tech-
niques used for the preparation of microspheres.28
This system had shown further advantage in reducing
the initial drug release (burst effect) when compared
with the ISI.26
The objective of this study was to develop haloperi-
dol ISI and ISM formulations for 1 month or longer
drug release. These formulations can be administered
both intramuscularly and subcutaneously. They are
also easier to prepare compared with the classical mi-
crospheres. The implants are formed in situ after in-
jection in contrast to the conventional implants, which
have to be surgically placed.
EXPERIMENTAL
MATERIALS
The following materials were purchased from Fisher
Scientific (Fair Lawn, New Jersey): haloperidol,
imipramine, NMP, triacetin, benzyl benzoate, DMSO,
ethyl acetate, ethanol, acetone, dichloromethane, hex-
ane, chloroform, tetrahydrofuran, acetonitrile, or-
thophosphoric acid, monobasic potassium phosphate,
and methanol. PLGA (50:50, intrinsic viscosity 0.5 dL/
g, molecular weight (MW) 60,000–70,000 Da), phos-
phate buffer saline (PBS, pH 7.4), peanut oil,
Pluronic R
F 68 NF, perchloric acid (48%–50%),
aluminum monostearate, and rat plasma (Sprague—
Dawley rat) were purchased from Lactel Pharmaceu-
ticals (Pelham, Alabama), Sigma–Aldrich (Gaithers-
burg, Maryland), Spectrum Chemical (Gardena,
California), BASF Corporation (Mount olive, New
Jersey), Alfa Aesar, (Ward Hill, Massachusetts), VWR
(West Chester, Pennsylvania), and Innovative Re-
JOURNAL OF PHARMACEUTICAL SCIENCES
search, Inc. (Novi, Michigan), respectively. All these
materials were of analytical grade or better and were
used without further purification.
Preparation of ISI Formulations
Twelve formulations were prepared by adding 20%,
30%, and 40% (w/v) of PLGA (50:50) to the four
studied solvents (NMP, triacetin, ethyl acetate, and
DMSO). Briefly, the specified amounts of PLGA were
added to the solvent in a scintillation vial at room
temperature and kept in an environmental shaker
until it dissolved completely. Haloperidol (10%, w/v)
was added into the polymer solution and mixed until
the drug is completely dissolved or suspended in the
polymer–solvent solution.
Preparation of ISM Formulations
Solutions of PLGA (50:50) at a concentration of
20% (w/v) in NMP and DMSO were prepared, and
haloperidol (10%, w/v) was dissolved in the resulting
clear solution (polymer phase) as described before.
Pluronic R
F 68 (1%, w/w, based on the amount of
the total formulation) was dissolved in the polymer
phase. Aluminum monostearate (2%, w/w, based on
oil) was added to the peanut oil. Pluronic R
and alu-
minum monostearate were used to increase the stabil-
ity of the emulsions. The ISM systems were prepared
by emulsifying the polymer phase into the oil phase
(peanut oil) at three different ratios 1:1, 1:2, and 1:4
to produce six haloperidol ISM formulations. Emulsi-
fication was carried out using probe sonication using
a Branson Sonifier 250 (Branson Ultrasonic Corpo-
ration, Danbury, Connecticut), at output 250 W and
frequency 20 kHz for 30 s under ice cooling.26
In Vitro Release Study
A quantity of formulation equivalent to 25 mg of
haloperidol was added to 900 mL of PBS at pH 7.4 in
a closed-top 1000 mL bottle. The bottles were stored
in the environmental shaker at 37◦ C, with moderate
shaking at 50 rpm. Each sample was run in tripli-
cate. Aliquots of 1 mL were taken from each bottle at
fixed time intervals and analyzed for the drug. The
dissolution medium was replaced with fresh medium
to maintain a sink condition. The concentration of
the drug in the collected sample was analyzed spec-
trophotometrically at 247 nm where polymer or poly-
mer degradation products did not interfere with the
analysis of the drug.5
Scanning Electron Microscopy Study
Scanning electron microscopy was used for morpho-
logical studies. ISI and ISM formulations were in-
jected into PBS and stored in an environmental
shaker at 37◦ C for 24 h. The samples were collected
and freeze dried. The dried samples were sputter
coated with gold before investigation using scanning
DOI 10.1002/jps
 IN SITU IMPLANT AND MICROPARTICLE FORMULATIONS OF HALOPERIDOL
electron microscope (Philips XL30, Eindhoven,
Netherlands) operated at 4–25 kV. The inner struc-
ture of the prepared ISI formulations was also stud-
ied by scanning the longitudinal cross-section of the
samples.
Quantification of Haloperidol in Plasma by HPLC
Previously described high-performance liquid chro-
matography (HPLC)0 method29,30 with modifications
was used to quantify haloperidol in the plasma sam-
ples. Briefly, imipramine was used as internal stan-
dard (I.S.). Stock solutions of haloperidol and I.S. were
prepared at the concentrations of 2 and 50 :g/mL, re-
spectively in methanol and stored at −20◦ C. Plasma
standards containing known amounts of drugs were
prepared by spiking plasma to get concentrations of
5, 10, 20, 50, 100, 250, and 500 ng/mL. Haloperidol
and the I.S. were extracted from the plasma using a
protein precipitation procedure. Five hundred micro-
liters of plasma sample was transferred to a borosil-
icate glass tube. To the tube 500 :L of acetonitrile–
water solution (1:1, v/v), 500 :L of I.S. (50 :g/mL
solution), and 3 mL of acetonitrile were added. The
mixture was vortexed for 2 min, then centrifuged at
1800 g for 10 min at 4◦ C. The supernatant was sepa-
rated and evaporated under vacuum at 45◦ C to dry-
ness. The dried residue was dissolved in 1 mL of a mix-
ture of phosphate buffer (0.02 M, pH 4.6), perchloric
acid, and acetonitrile (89:1:10, v/v) and injected onto
a Symmetry C18, 150 × 4.6 mm2 , 5 :m, analytical
column. The mobile phase was delivered through the
column at a flow rate of 0.6 mL/min. The sample injec-
tion volume was 50 :L. The samples were analyzed
at 245 nm, which was determined to be the wave-
length of maximum absorption, λmax 1of the drug in
the mobile phase.
Pharmacokinetic Studies
Male Sprague–Dawley rats with average weight of
200–224 g were used for the in vivo study. Animals
were maintained in a 12:12 light–dark cycle with free
access to food with all testing and procedures per-
formed during the light cycle. The rats have been kept
for 7 days with water and food before the study. The
experimental procedures described in this study re-
ceived prior approval from the Institutional Animal
Care and Use Committee and were in compliance
with the guide for the Care and Use of Laboratory
Animals published by the US National Institutes of
Health (NIH Publication No. 85-23, revised 1996).
Animals were divided into three groups and ad-
ministered the drug as follows: Group I, 20% PLGA
in NMP–ISI formulation; Group II, 20% PLGA in NM-
P–oil (1:4) ISM formulation; Group III, control (saline,
0.1 mL). The formulations were administered intra-
muscularly into the right musculus rectus of the rats
(n = 6 per group). The ISI and the ISM formulations
DOI 10.1002/jps
3
equivalent to 14 mg of haloperidol were administered
to each rat. Blood samples were collected using tail-
vein bleeding.31 Each rat was placed in a whole body
restrainer and the tail was warmed using an electric
lamp directed toward the tail. The tail was swabbed
with alcohol and a sharp surgical blade was then used
to make an incision over the tail artery or vein lo-
cated near the tip of the tail. The tail was “milked”
by applying even pressure from the base to the end
so that about 0.5 mL of blood was collected and im-
mediately centrifuged at 4300 g at 8◦ C for 15 min.
The separated plasma samples were frozen at −80◦ C
until analysis. Following the initial blood-collection
procedure, direct pressure was applied to the incision
site with a clean cotton swab for about 10 s to stop the
bleeding and allow for the blood to clot. Blood samples
(0.5 mL) were collected at 6, 12, 15, 18, 21, and 24 h
then at 3, 5, 7, 10, 14, 18, 21, 24, 28, and 31 days. The
plasma concentration of the drug in each sample was
calculated after analyzing the samples by the HPLC
method mentioned previously.
The following pharmacokinetic parameters,
namely maximum plasma concentration (Cmax ), time
point of maximum plasma concentration, area under
the plasma concentration–time curve (AUC), mean
residence time (MRT), and total clearance (Cl) were
computed for haloperidol using KineticaTM (Version
4, Thermo Electron Corporation, Waltham, MA)2.
Examination of the injection site has been per-
formed for both the control and treated groups after
the animals were sacrificed by suffocation with carbon
dioxide and their intact skin along with intramus-
cular tissue was excised. Sections were made using
sharp blade. The skin tissue of the study groups were
compared with those of the control group. The tissues
were visually inspected for any type of swelling, red-
ness, or inflammation. The procedure was followed as
mentioned by Sharma et al.32
Statistical Analysis
The pharmacokinetic parameters obtained from the
two groups of rats were analyzed by Student’s t-test.
The p value was set at 0.05.
Stability Studies
Stability studies for the two formulations used in
pharmacokinetic studies were carried out to deter-
mine the effect of temperature on stability of haloperi-
dol over a defined period of time. The formulations
were placed in 3-mL glass vials, which were inserted
into 50-mL screw-capped bottles. The bottles were
stored at 4◦ C, 25◦ C, and 40◦ C in the dark.33 At prede-
termined time, drug content and pH were measured.
Stability studies were also carried out at 4◦ C for 12
months to determine the kinetic order and shelf life
(time needed for 10% loss, t90 ) of the drug in the se-
lected formulations.
JOURNAL OF PHARMACEUTICAL SCIENCES
4 AHMED ET AL.
Drug content was measured by taking aliquot from
each sample. The aliquot was mixed with a known
volume of acetonitrile, vortex-mixed, centrifuged at
1100 g, filtered with a 0.45 :m filter and then an-
alyzed by a spectrophotometer at 247 nm.34 For the
ISM system, the same procedure was utilized for drug
content analysis except that the peanut oil was sepa-
rated in the bottom and only the top acetonitrile part
was used for the analysis.
The pH was measured in each formulation using
an Accumet AR 60 pH meter (Fisher Scientific). The
electrode of the pH meter was immersed directly into
the formula. The color, apparent physical stability,
and polymer precipitation upon injection into buffer
were also evaluated during and after the study.
RESULTS AND DISCUSSION
Preparation of ISI and ISM Formulations
In this study, ISI and ISM formulations were pre-
pared by using solvents with different water misci-
bility. In case of ISI formulations, different polymer
concentrations were investigated, whereas the drug
concentration was kept constant (10%, w/v). NMP
and DMSO were selected as water-miscible solvents,
whereas triacetin and ethyl acetate were used as par-
tially miscible ones. The lowest polymer concentra-
tion was chosen to be 20% in all the studied solvents
because a polymer concentration less than 20% (w/v)
usually gives faster release and high initial burst be-
cause of the lower viscosity of the formulations.16
One way of controlling the drug release from
these systems is to select different grades of PLGA.
Low MW, low lactide-to-glycolide (L–G) ratio, and
uncapped polymer end groups result in a less hy-
drophobic polymer with increased rates of water ab-
sorption, hydrolysis, and erosion of polymer3.35 The
duration of drug release is highly dependent on these
properties.36,37 Thus, the choice of PLGA is perhaps
the most important tool in drug release modifica-
tion for ISI systems. PLGA is available in L–G ra-
tios of 50:50, 65:35, 75:25, and 85:15. Lactic acid
is more hydrophobic than glycolic acid and hence
lactide-rich PLGA copolymers are less hydrophilic,
absorb less water, and subsequently degrade more
slowly.38 In vitro risperidone release was decreased
with increased L–G ratio.39 PLGA 50:50 with viscos-
ity 0.5 g/dL was chosen for the study as it has medium
hydrophobicity.
Polymer solutions in all the solvents used were
clear before addition of the drug. Haloperidol was dis-
solved in NMP, DMSO, and ethyl acetate polymeric
solutions and suspended in triacetin polymeric so-
lution. Because haloperidol did not dissolve in tri-
acetin polymeric solution, triacetin was not used in
ISM preparations. Peanut oil was chosen as the bio-
JOURNAL OF PHARMACEUTICAL SCIENCES
compatible external oil phase in ISM formulations.
The polymer concentration used for ISM formulations
were 20% (w/v) in the polymer phase. Polymer phase
to peanut oil ratios were 1:1, 1:2, and 1:4.
In Vitro Release from ISI Formulations
The in vitro release of haloperidol from ISI formula-
tions are shown in Figures 1 and 2. Upon injection
of the formulations into the release medium, the sol-
vent dissipates into the buffer medium and the poly-
mer solidifies. Thus, a solid implant is formed. The
in vitro drug release from the formulations followed a
triphasic pattern. A fast initial release phase (burst)
is followed by a second slow release phase lasting
days or weeks and a third rapid release phase. Be-
cause ISI systems are administered as liquid, there
is a lag time between the injection and the forma-
tion of the solid implant. During this period, solvent
along with the dissolved drug moves out of the formu-
lation quickly causing the initial burst release. After
that, a diffusion-controlled slower release phase fol-
lows. Finally, when the MW of PLGA approaches a
certain lower threshold, the weight of the ISI system
decreases rapidly and an erosion-controlled rapid re-
lease phase occurs.40 Drug release from ISI formula-
tions is mostly triphasic but biphasic release was also
found.35 The initial burst of drug may cause tissue ir-
ritation and toxicity. The concentration and MW of the
polymer, the type of solvent used, and the presence of
a surfactant influence the rate of precipitation of the
polymer and were used to control the burst effect.41
Figure 1 shows that 20%, 30%, and 40% PLGA in
NMP–ISI formulations showed an initial drug burst
of 18%, 15%, and 13%, respectively within the first
24 h. The three concentrations of polymers sustained
the release of haloperidol for 28, 35, and 47 days, re-
spectively. The in vitro release for the drug from ISI
formulations prepared with DMSO exhibited higher
release rate than the ISI–NMP systems made from
the same polymer concentration. Figure 1 also reveals
that 20%, 30%, and 40% PLGA in DMSO showed an
initial burst of 20%, 17%, and 15%, respectively. The
release was extended over 24, 31, and 45 days, respec-
tively for these three formulations. The initial burst
of the ISI with both solvents (NMP and DMSO) de-
creased with increasing the polymer concentration.
In situ implant formulations with 20%, 30%, and
40% PLGA in ethyl acetate showed release of 9%,
6%, and 5% haloperidol, respectively within the first
24 h (Fig. 2). After 60 days, 92%, 87%, and 79% drug,
respectively was released from these three formu-
lations. Triacetin gave the slowest in vitro release
among all the studied solvents. Within the first 24 h,
7%, 5%, and 3% of drug was released from 20%,
30%, and 40% PLGA in triacetin ISI formulations,
respectively. Also, 85%, 77%, and 71% of haloperidol,
DOI 10.1002/jps
 IN SITU IMPLANT AND MICROPARTICLE FORMULATIONS OF HALOPERIDOL 5

Figure 1. In vitro drug release from in situ implant systems containing different concentra-
tions of PLGA in NMP and DMSO (Mean ± SD, n = 3).

Figure 2. In vitro drug release from in situ implant systems containing different concentra-
tions of PLGA in ethyl acetate and triacetin (Mean ± SD, n = 3).

DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES

6 AHMED ET AL.
respectively was released from these formulations af-
ter 60 days.
It is found that the type of solvents and the polymer
concentrations had pronounced effects on the in vitro
release of haloperidol from the ISI systems. An ex-
tended release profile and significantly reduced burst
effect result when the aqueous affinity of the solvent
was reduced. That slows the phase inversion rate,
which in turn produces depot morphologies favorable
to prolonged release.42 That explains the slower drug
release from triacetin and ethyl acetate formulations.
The solvents that are relatively soluble in aqueous
medium (e.g., NMP and DMSO) promote rapid migra-
tion of water into the polymer composition and result
in a burst effect.
Haloperidol, a practically insoluble drug (water sol-
ubility: 1.4 mL/100 mL),43 may inhibit water diffusion
into the matrix, thereby slowing surface erosion of the
implant and causing further decrease in drug release.
The diffusion (swelling) rate and degradation rate
are higher for hydrophilic than hydrophobic drugs.5
The same results were also was obtained by Jeong
et al.44 who studied the in vitro release of two different
drugs from 4polyethylene glycol (PEG)–PLGA–PEG
triblock and concluded that ketoprofen, which is a
hydrophilic drug, was released over 2 weeks, whereas
spironolactone, a hydrophobic drug, was released over
2 months.
In Vitro Release from ISM Formulations
The in vitro drug release data for the ISM system
were different from the corresponding ISI system in
two aspects; lower initial drug release by 20%–28%
and extended drug release profile as indicated by time
for complete drug release. The initial burst of 20%,
30%, and 40% PLGA in NMP–ISI formulations were
18%, 15%, and 13%, respectively, whereas those of the
corresponding ISM formulations were 13%, 12%, and
10%, respectively. The value for corresponding DMSO
systems were 20%, 17%, and 15% (ISI) and 15%, 13%,
and 12% (ISM), respectively (Figs. 1 and 3). The ISI
formulation, 20% PLGA in NMP showed in vitro re-
lease for 28 days. Its corresponding ISM formulations,
20% PLGA in NMP–oil (1:1, 1:2, and 1:4) released
the drug for 35, 38, and 45 days, respectively. Similar
trends were found with DMSO formulations, where
the drug release from ISI with 20% PLGA was for 24
days, whereas the drug release was observed for 31,
35, and 42 days, respectively from its corresponding
ISM systems.
The release of haloperidol decreased with decreas-
ing polymer to oil phase ratio (Fig. 3). These find-
ings can be explained by the fact that the external
oil phase acts as a barrier for the solvent and hence
reduced solvent diffusion was observed. Haloperidol
release was investigated for the ISM system from two
solvents (20% PLGA in DMSO and NMP). It is impor-
JOURNAL OF PHARMACEUTICAL SCIENCES
Figure 3. In vitro drug release from in situ microparticle
systems containing 20% PLGA solution in NMP and DMSO
(Mean ± SD, n = 3).
tant to mention that the ISM systems also maintained
triphasic release as found with the ISI system.
Haloperidol release from ISM–DMSO system was
faster than that from the corresponding ISM–NMP
system. The solubility of DMSO in peanut oil is 44.93
mg/mL and NMP is completely miscible in peanut oil.
Complete miscibility of NMP with peanut oil facili-
tate the rapid formation of ISM upon contact with
the buffer, whereas the ISM prepared with DMSO is
formed slowly because of the lower solubility of DMSO
in the oil compared with the NMP. This can explain
the faster release of DMSO from the ISM system, com-
pared with the NMP systems. Similar findings for the
release of bupivacaine hydrochloride from ISM sys-
tems prepared with NMP and DMSO were reported.45
Scanning Electron Microscopy Study
The implants had porous surface and the microparti-
cles showed a smooth nonporous surface after 24 h
of exposure to PBS. Figure 4 shows the scanning
electron microscopy of 20% PLGA in NMP–ISI for-
mulation and the corresponding ISM, 20% PLGA in
NMP–peanut oil (1:4). The figure also shows the in-
ternal structure of the prepared ISI after longitudinal
cut has been made (Fig. 4b). Water diffuses inside the
ISI formulation immediately upon injection in PBS
or after in vivo administration and leads to pores
formation.46 These pores are too small for drug trans-
port during the early stage of this process, but as the
number and size of water-filled pores in the polymer
increase, a porous connected network is formed allow-
ing the drug to release from inside the implant. Also,
the space left vacant after drug release will probably
constitute pores, facilitating further drug release.47
A porous surface for implants and a smooth, non-
porous surface for microparticles were reported.26 The
oil phase created a partial barrier around the polymer
DOI 10.1002/jps
 IN SITU IMPLANT AND MICROPARTICLE FORMULATIONS OF HALOPERIDOL
Figure 4. Scanning electron micrographs of (a) in situ im-
plant (20% PLGA in NMP), (b) a longitudinal cross-section
of implant (20% PLGA in NMP), and (c) in situ microparti-
cles (20% PLGA in NMP–peanut oil, 1:4).
phase, causing slow release of the solvent in the aque-
ous medium and probably resulting in the observed
smooth surface of the formed microparticles.
Quantification of Haloperidol in Plasma by HPLC
The chromatograms of blank plasma and plasma
spiked with the drug and the I.S. showed no inter-
fering peak at the retention times of interest. Under
the described chromatographic conditions, retention
times for the drug and I.S. were 11.4 and 13.5, re-
spectively. The standard curve had an average slope
DOI 10.1002/jps
7
of 0.0102 and correlation coefficient of 0.993 (n = 3).
The method was validated with interday and intra-
day variation of less than 10% for the concentration
range 5–500 ng/mL. The lower limit of quantitation
was 5 ng/mL.
Pharmacokinetic Study
On the basis of the in vitro results, in vivo studies
were performed on selected formulations. The plasma
concentration of haloperidol was determined follow-
ing the intramuscular injection of an ISI formula-
tion, 20% PLGA in NMP and an ISM formulation,
20% PLGA in NMP–peanut oil (1:4). Both NMP and
DMSO have been used in commercial injectable prod-
ucts for human use.48 ISI formulations with NMP
showed longer release in the in vitro release stud-
ies compared with the formulations prepared with
DMSO. ISM–NMP formulations showed 1 month or
longer drug release. Among them, 20% PLGA in NMP
was selected for its better syringeability and ease
of administration. The ISM formulation correspond-
ing to this ISI formulation, that is, 20% PLGA in
NMP–peanut oil (1:4) showed lowest initial burst and
longest release among the ISM formulations.
A dose of 2 mg/kg per day was selected for the phar-
macokinetic studies in rats.49,50 Formulations con-
taining 14 mg haloperidol were used for a 28-day
study as the average rat weight was 250 g.
The plasma concentration–time curve following in-
tramuscular administration of haloperidol formula-
tions are illustrated in Figure 5. The mean Cmax ±
SD value for the ISI formulation was 60.3 ± 0.9 ng/
mL and reached after 18 h. The mean Cmax ± SD was
43.9 ± 1.0 ng/mL for the ISM system of the same
drug and reached after 21 h. These results indicate
that the ISM formulations resulted in a reduced ini-
tial drug release relative to the ISI formulation. The
overall low level of haloperidol in the plasma may be
attributed to the high lipophilicity of haloperidol and
a large volume of distribution.51 Plasma drug concen-
tration of haloperidol does not provide direct informa-
tion on CNS level. The concentration of haloperidol af-
ter chronic administration of the drug in rats showed
18.4 ± 4.9 ng/mL in plasma and 334.7 ± 63.0 ng/
g haloperidol in brain tissue at a brain-to-plasma
ratio of 18:1.52 In postmortem human brain tissue,
haloperidol concentrations were reported to be 10–30
times higher than the plasma concentrations.51 From
our pharmacokinetic studies, the level of haloperidol
in the plasma after first 24 h was 46.5 ± 1.1 and
38.5 ± 1.0 ng/mL for ISI and ISM systems, respec-
tively. These levels are within the therapeutic range
in rats (12–59 ng/mL).53 Additionally, the Cmax for
both the ISI and ISM systems can be tolerated as they
are below the minimum toxic concentration. The esti-
mated human therapeutic plasma range is almost the
same as in rats.54 Plasma concentration of haloperidol
JOURNAL OF PHARMACEUTICAL SCIENCES
8 AHMED ET AL.
Figure 5. Plasma concentration versus time profile of haloperidol following the intramuscular
injection of ISI system containing 20% PLGA in NMP and its corresponding ISM system, 20%
PLGA in NMP–oil, 1:4 (Mean ± SD, n = 6).
for both systems remained within the therapeutic
window for about 20 days after single intramuscu-
lar administration. Both systems can be modified by
increasing concentration, MW, and L–G ratio of PLGA
to obtain even longer release of haloperidol.
The ISM system continued to release the drug be-
yond 30 days in our studies. Compared with the ISI
system, the ISM system has lower initial burst re-
lease and longer drug release and therefore, the ISM
system would be advantageous over ISI system.
The mean AUC0–∝ , which reflects the total amount
of drug reaching the systemic circulation, was found
to be 13.1 and 13.9 :g h/mL for the ISI and ISM
formulations, respectively. The AUMC0–∝ was found
to be 3289 and 5138 :g h2 /mL,5 respectively, whereas
the MRT was calculated to be 251.5 and 369.6 h for
them, respectively. The Cl was 17.8 and 17.0 mL/min
for the ISI and ISM formulations, respectively.
Examination of the site of injection of both the con-
trol and treated groups confirmed biocompatibility of
the prepared formulations. Compared with the skin
tissue of control group, the tissues in treated groups
showed no marked difference in appearance. Thus,
the prepared ISI and ISM formulations of haloperidol
were biocompatible.
Stability Studies
The purpose of this study was to establish recom-
mended storage conditions. The highest temperature
JOURNAL OF PHARMACEUTICAL SCIENCES
used in the stability study was 40◦ C. The use of fairly
moderate temperature is suggested to avoid errors of
interpretation that may occur as a result of the fact
that the decomposition of active drugs or the poly-
mer at high temperatures often differs from the de-
composition at ordinary temperatures as mentioned
earlier.55
The drug content determined for the ISI system
under three different temperatures 4◦ C, 25◦ C, and
40◦ C after 90 days was 99.51%, 98.09%, and 95.65%,
respectively, whereas that for the ISM was 99.93%,
98.55%, and 94.27%, respectively (n = 3, SD is less
than 5% of the mean). A slight increase in the rate
of degradation can be observed with an increase of
the storage temperature from 4◦ C to 25◦ C. However,
increasing the temperature from 25◦ C to 40◦ C led
to more drug degradation in most cases. This evi-
dence was confirmed also by the change of the pH
values of the formulations. The mean pH values for
the ISI changed from 9.65 at the beginning of the
study to 9.60, 8.61, and 7.13 at 4◦ C, 25◦ C and 40◦ C,
respectively, whereas the mean pH values for the ISM
changed from 9.12 to 9.08, 8.00, and 6.42, respectively
at the same temperatures (n = 3, SD is less than 5%
of the mean).
After 90 days of study at 4◦ C or even 25◦ C, haloperi-
dol content was more than 98%. Similar findings were
observed by Morlock et al.56 who studied the acceler-
ated stability of recombinant human erythropoietin
DOI 10.1002/jps
 IN SITU IMPLANT AND MICROPARTICLE FORMULATIONS OF HALOPERIDOL 9

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