International Journal of Pharmaceutics 304 (2005) 178–184

Development of sustained release gastroretentive drug delivery

system for ofloxacin: In vitro and in vivo evaluation
Mahesh Chavanpatil ∗ , Paras Jain, Sachin Chaudhari, Rajesh Shear, Pradeep Vavia
Mumbai University Institute of Chemical Technology, Pharmaceutical Sciences Division,
Near Nathalal Parikh Marg, Matunga, Mumbai 400019, India

Received 5 June 2005; received in revised form 18 August 2005; accepted 22 August 2005

Abstract

Sustained release (SR)-gastroretentive dosage forms (GRDF) enable prolonged and continuous input of the drug to the upper
parts of the gastrointestinal (GI) tract and improve the bioavailability of medications that are characterized by a narrow absorption
window. A new strategy is proposed for the development of gastroretentive dosage forms for ofloxacin preferably once daily.
The design of the delivery system was based on the sustained release formulation, with floating and swelling features in order
to prolong the gastric retention time of the drug delivery systems. Different polymers, such as psyllium husk, HPMC K100M,
crospovidone and its combinations were tried in order to get the desired sustained release profile over a period of 24 h. Various
formulations were evaluated for buoyancy lag time, duration of buoyancy, dimensional stability, drug content and in vitro
drug release profile. It was found that dimensional stability of the formulation increases with the increasing psyllium husk
concentration. It was also found that in vitro drug release rate increased with increasing amount of crospovidone due to the
increased water uptake, and hence increased driving force for drug release. The optimized formulation was subjected to stability
studies at different temperature and humidity conditions as per ICH guidelines. In vivo studies were carried out for the optimized
formulation in 24 healthy human volunteers and the pharmacokinetic parameters of developed formulations were compared
with the marketed once daily (Zanocin) formulation. Based on the in vivo performance in a parallel study design in healthy
subjects, the developed formulation shows promise to be bioequivalent to the marketed product (Zanocin). The percent relative
bioavailability of developed formulation was found to be 97.55%.
© 2005 Elsevier B.V. All rights reserved.

Keywords: Ofloxacin; Sustained release; Psyllium husk; Gastroretentive

1. Introduction
∗ Corresponding author. Present address: Eugene Applebaum Col-
Oral sustained release (SR)-dosage forms (DFs)
lege of Pharmacy, 259, Mack Avenue, #3250, Detroit, MI 48201,
USA. Tel.: +1 313 577 8892.
have been developed for the past three decades due
E-mail address: mcp 112000@rediffmail.com to their considerable therapeutic advantages (Hoffman,
(M. Chavanpatil). 1998). However, this approach has not been suitable for

0378-5173/$ – see front matter © 2005 Elsevier B.V. All rights reserved.
doi:10.1016/j.ijpharm.2005.08.009
 M. Chavanpatil et al. / International Journal of Pharmaceutics 304 (2005) 178–184 179

a variety of important drugs, characterized by a narrow line pH conditions prevail; however, precipitation of
absorption window in the upper part of the gastroin- the active compound occurs, which adversely affects
testinal tract, i.e. stomach and small intestine. This is absorption in the lower sections of the intestine. There
due to the relatively short transit time of the DF in these is a need for systems that reside in the stomach over a
anatomical segments. Thus, after only a short period relatively long time and release the active compound
of less than 6 h, the SR-DF has already left the upper there in a sustained manner (Sen and Kshirsagar, 2002).
gastrointestinal tract and the drug is released in non- This necessitated the design and development of sus-
absorbing distal segments of the gastrointestinal tract. tained release gastroretentive drug delivery system for
This results in a short absorption phase that is often ofloxacin using suitable polymers.
accompanied by lesser bioavailability.
It was suggested that compounding narrow absorp-
tion window drugs in a unique pharmaceutical DF with 2. Materials and methods
gastroretentive properties would enable an extended
absorption phase of these drugs. After oral adminis- 2.1. Materials
tration, such a DF would be retained in the stomach
and release the drug there in a sustained manner, so Ofloxacin and psyllium husk were gifted by
that the drug could be supplied continuously to its Macleoid Pharmaceuticals, India. HPMC K100M,
absorption sites in the upper gastrointestinal tract. This PVP K30 and crospovidone were obtained as gift sam-
mode of administration would best achieve the known ples from M/s Rohm Pharma, Germany. Talc and mag-
pharmacokinetic and pharmacodynamic advantages of nesium stearate were gifted by M/s Bayer India Ltd.,
SR-DFs for these drugs (Hwang et al., 1998; Hoffman India. All other solvents and reagents were purchased
and Stepensky, 1999). from Ranbaxy chemicals, India, and were of analytical
The need for gastroretentive dosage forms (GRDFs) grade.
has led to extensive efforts in both academia and indus-
try towards the development of such drug delivery sys- 2.2. Methods
tems (Deshpande et al., 1996). These efforts resulted
in GRDFs that were designed in large part based on 2.2.1. General description of the manufacturing
the following approaches: (a) low density form of the process for sustained release formulation of
DF that causes buoyancy above gastric fluid (Singh ofloxacin
and Kim, 2000); (b) high density DF that is retained Typical sustained release formulations of ofloxacin
in the bottom of the stomach; (c) bioadhesion to the are listed in Tables 1 and 2. Tablets were made by
stomach mucosa (Moes, 1993); (d) slowed motility of using psyllium husk (gelling agent), HPMC K100M
the gastrointestinal tract by concomitant administration (hydrophilic polymer), crospovidone (swelling agent),
of drugs or pharmaceutical excipients (Rubinstein and sodium bicarbonate (gas-generating agent) and beta-
Friend, 1994); (e) expansion by swelling or unfolding cyclodextrin (channeling agent). Tablets were made by
to a large size which limits emptying of the DF through using wet granulation process with PVP K30 (5%, w/v,
the pyloric sphincter (Mamjek and Moyer, 1980). isopropyl alcohol). Compression was done on a Cad-
The objective of present work was to develop gas-
troretentive formulation, which releases drug in the
Table 1
stomach and upper gastrointestinal (GI) tract, and form Formulation composition to study the effect of psyllium husk and
an enhanced opportunity of absorption in the stomach HPMC K100M on in vitro release of ofloxacin
and upper GI tract rather than the lower portions of Composition (mg/tablet) F1 F2 F3 F4 F5
the GI tract. Example of substance whose bioavailabil-
Ofloxacin 400 400 400 400 400
ity is strongly dependent on the local physiology in Psyllium husk 75 100 125 100 100
the GI tract and which preferably is absorbed in the HPMC K100M 40 40 40 30 50
higher sections of the intestine is ofloxacin. Ofloxacin Sodium bicarbonate 70 70 70 70 70
is readily soluble in the acidic environment of the Crospovidone 200 200 200 200 200
PVP K30 (5% in IPA) 20 20 20 20 20
stomach. In the intestine, where neutral to slightly alka-
180 M. Chavanpatil et al. / International Journal of Pharmaceutics 304 (2005) 178–184

Table 2 The rotation speed was 50 rpm. The dimensional

Formulation composition to study the effect of crospovidone, sodium stability of ofloxacin formulations was observed
bicarbonate and betacyclodextrin on in vitro release of ofloxacin
visually.
Composition F6 F7 F8 F9 F10 F11
(mg/tablet)
2.2.5. Bioequivalence studies
Ofloxacin 400 400 400 400 400 400
A total of 24 volunteers participated in the study. The
Psyllium husk 100 100 100 100 100 100
HPMC K100M 40 40 40 40 40 40 volunteers were fasted overnight at least 10 h prior to
Sodium 70 70 60 80 70 70 dosing with water ad lib. Twelve volunteers were given
bicarbonate the test formulation (F11) and 12 the marketed formu-
Crospovidone 0 100 200 200 200 200 lation (Zanocin) with water (parallel study design). At
PVP K30 (5% in 20 20 20 20 20 20
predetermined time interval, 5 ml of blood was with-
IPA)
Betacyclodextrin – – – – 50 100 drawn at 0, 2, 4, 6, 8, 10, 12, 18 and 24 h. Blood
samples were collected from forearm vein using sterile
disposable needle and syringe into sterile glass cen-
mach single station tablet press using caplet shaped trifuge tubes containing heparin. The samples were
punches. centrifuged immediately at 4000 rpm and the plasma
was stored in light protected container at −20 ◦ C till
2.2.2. In vitro release study the time of analysis.
The release of ofloxacin from the tablets was stud- The concentration of ofloxacin from plasma was
ied using USP dissolution Apparatus I. The dissolution measured by validated reversed phase HPLC. HPLC
medium was phosphate buffer pH 1.2 for first 2 h, phos- method was validated for linearity, accuracy, precision
phate buffer pH 4.5 for next 2 h and pH 7.4 for remain- and extraction efficiency. To 0.5 ml of plasma, 2 ml of
ing hours (Wang, 1978), the volume being 900 ml. acetonitrile was added and vortexed for 5 min. This
The temperature was maintained at 37 ± 0.5 ◦ C. The was centrifuged at 4000 rpm for 10 min. The super-
rotation speed was 100 rpm. Five milliliters of aliquot natant was filtered through 0.45 ␮m filter and this solu-
were withdrawn at predetermined time intervals of 1, tion was stored at −20 ◦ C till HPLC analysis. There
2, 3, 4, 6, 8, 10, 12, 14, 16 and 24 h. The medium was no drug loss due to the adsorption of drug on
was replenished with 5 ml of fresh buffer each time. 0.45 ␮m filter during filtration process. The filtered
Sample was analyzed by using UV spectrophotometry solution was injected onto the HPLC column (Waters
at 291 nm. C8 spherisorb, 5 ␮m, 250 mm); the signal was mon-
itored at 293 nm. Mobile phase was prepared with
2.2.3. Buoyancy lag time and the duration of methanol:acetonitrile:water containing 1.5 g phospho-
buoyancy ric acid/l (5:5:80, v/v/v). Tetrabutyl ammonium hydro-
The buoyancy lag time and the duration of buoyancy gen sulphate (0.2%) was used as an ion-pairing reagent
were determined in the USP dissolution Apparatus II in the mobile phase. The flow rate was maintained at
in an acid environment. The time interval between the 1.5 ml/min. The parameters employed to evaluate were
introduction of the tablet into the dissolution medium Cmax , tmax , Kel and t1/2 values. Cmax and tmax were read
and its buoyancy to the top of dissolution medium was directly from the observed mean drug plasma concen-
taken as buoyancy lag time and the duration of buoy- tration against time profile. Kel and t1/2 were computed
ancy was observed visually (Yang et al., 1999). from observed mean plasma concentration against time
profile. The extent of absorption from the test formu-
2.2.4. Dimensional stability lation relative to the marketed one was calculated as
The dimensional stability of the formulations was the relative bioavailability by using the formula given
studied using USP dissolution Apparatus II. The dis- below:
solution medium was phosphate buffer pH 1.2 for first
AUC(0–∞) of test
2 h, phosphate buffer pH 4.5 for next 2 h and pH 7.4 relative bioavailability =
for remaining hours, and the volume being 900 ml. AUC(0–∞) of marketed
The temperature was maintained at 37 ± 0.5 ◦ C. ×100
 M. Chavanpatil et al. / International Journal of Pharmaceutics 304 (2005) 178–184 181

3. Results and discussion lative drug release at the end of 24 h was found to
be 53.60 ± 1.86 and 47.20 ± 1.74%, respectively. This
3.1. Effect of psyllium husk on in vitro drug might be due to the gelling properties of psyllium husk.
release and integrity of formulations Psyllium husk forms thick gel at higher concentra-
tions, which could have contributed to the decrease in
Effect of different concentrations of psyllium husk drug release. Drug associated with the surface of tablet
on in vitro release of ofloxacin was studied. Initially, matrix could have also contributed to the initial burst
tablet containing 75 mg of psyllium husk (F1) could not release. As the surface associated drug was released,
retain its physical integrity for desired period (24 h) of psyllium husk matrix could have contributed to the
time. As the concentration of psyllium husk increases slower drug release over a period of 24 h. Formulation
(F2 and F3), it retains integrity up to desired period (F2) containing 100 mg of psyllium husk maintained its
of time (24 h). It is important to maintain the physical physical integrity for 24 h and showed similar pattern of
integrity of the tablet in case of once daily formulations. drug release as compared to the marketed formulation,
If it does not maintain its physical integrity, tablet could hence, selected for the further studies.
be broken down into smaller fragments and escape Ofloxacin is soluble in aqueous solution with pH
from the upper part of the GI tract. Hence, an attempt between 2 and 5. It is sparingly to slightly soluble in
was made in order to increase the physical integrity aqueous solution with pH 7. Hence, developed for-
of the tablets using psyllium husk. Further, formula- mulations as well as marketed formulation could not
tion (F1) provided higher burst drug release as com- release total amount of the drug into the dissolution
pared to the marketed formulation as shown in Fig. 1. medium by pH change method.
In case of F1 and marketed formulations, burst drug
release after 2 h was 34.06 ± 0.52 and 31.08 ± 0.89%, 3.2. Effect of HPMC K100M on in vitro release of
respectively. Therefore, amount of psyllium husk was ofloxacin
increased to 100 (F2) and 125 mg/tablet (F3). As the
concentration of psyllium husk increases, initial burst Initially, HPMC K100M was tried in concentration
drug release as well as drug release in the latter hours of 30 mg/tablet (F4). The formulation provided higher
decreases as compared to the marketed formulation. In burst drug release as compared to marketed formula-
case of F2 and F3 formulations, burst drug release after tion as shown in Fig. 2. In case of F4 and marketed
2 h was found to be 28.22 ± 0.96 and 24.22 ± 1.28%, formulations, burst drug release after 2 h was found
respectively. In case of formulation F1 and F3, cumu- to be 34.08 ± 0.92 and 31.08 ± 0.89%, respectively.

Fig. 1. Effect of psyllium husk concentration on in vitro drug release Fig. 2. Effect of HPMC K100M concentration on in vitro drug
of ofloxacin (n = 6). Standard deviation was found to be less than 2% release of ofloxacin (n = 6). Standard deviation was found to be less
in all the in vitro drug release profiles. than 2% in all the in vitro drug release profiles.
182 M. Chavanpatil et al. / International Journal of Pharmaceutics 304 (2005) 178–184

Therefore, amount of HPMC K100M was increased to

40 mg/tablet (F2), the formulation provided burst drug
release comparable to the marketed formulation. Fur-
ther increase in amount of HPMC K100M to 50 mg (F5)
provided low burst drug release as compared to the mar-
keted tablets. Formulation (F5) showed 24.01 ± 1.23%
of burst drug release at the end of 2 h. High HPMC
K100M content results in a greater amount of gel being
formed. This gel increases diffusion path length of the
drug. Its viscous nature also affects diffusion coeffi-
cient of the drug. As a result, reduction in drug release
is obtained. Thus, 40 mg (F2) HPMC K100M was
selected for further studies.

3.3. Effect of crospovidone on in vitro release of

Fig. 3. Effect of crospovidone concentration on in vitro drug release
ofloxacin of ofloxacin (n = 6). Standard deviation was found to be less than 2%
in all the in vitro drug release profiles.
In order to improve the release profile of the formu-
lations, concentration of crospovidone was increased. decreases as shown in Fig. 4. This might be due to
Crospovidone acts as a swelling agent, which is capa- the alkaline nature of sodium bicarbonate. Sodium
ble of swelling greater than its original volume and bicarbonate creates an alkaline environment around
preferably to at least twice its original volume when the tablet. Ofloxacin is less soluble in alkaline envi-
coming into contact with an aqueous fluid, such as gas- ronment that decreases the drug release from the
trointestinal fluid. The swelling agent, which normally formulation. Formulation containing 60 mg sodium
swells to several times its original volume in water, bicarbonate (F8) showed 54.91 ± 1.30% cumulative
exhibits a controlled swelling in the presence of water- drug release whereas formulation containing 80 mg
soluble hydrophilic polymers (Talwar and Staniforth, of sodium bicarbonate (F9) showed 47.20 ± 1.35%
2001, WO 01/64183). Three concentrations 0 mg (F6), cumulative drug release at the end of 24 h. Formula-
100 mg (F7) and 200 mg (F2) of crospovidone were tion containing 70 mg sodium bicarbonate (F2) showed
tried. As the concentrations of crospovidone increases
from 0 to 200 mg/tablet, drug release has increased
as shown in Fig. 3. Formulation (F6), which does
not contain crospovidine, showed 38.71 ± 1.64% of
cumulative drug release at the end of 24 h. Formu-
lation containing 100 mg (F7) showed 49.64 ± 1.32%
cumulative drug release whereas formulation contain-
ing 200 mg (F2) showed 52.55 ± 1.02% cumulative
release at the end of 24 h. As the concentration of
crospovidone increases, water uptake capacity of the
formulation increases (data not shown). This increases
the porosity of the matrix, which results in increased
drug release from the matrix system.

3.4. Effect of gas-generating agent on in vitro

release of ofloxacin
Fig. 4. Effect of sodium bicarbonate concentration on in vitro drug
As the concentration of sodium bicarbonate release of ofloxacin (n = 6). Standard deviation was found to be less
increases from 60 to 80 mg/tablet, drug release than 2% in all the in vitro drug release profiles.
 M. Chavanpatil et al. / International Journal of Pharmaceutics 304 (2005) 178–184 183

Table 3
Effect of sodium bicarbonate concentration on onset and duration of
floating
Amount of sodium Onset of Duration of
bicarbonate (mg) floating (s) floating (h)
60 40 ± 3 18
70 30 ± 4 24
80 25 ± 3 24

52.55 ± 1.02% cumulative drug release at the end of

24 h.
In such systems, sodium bicarbonate acts as a gas-
generating agent. It generates gas when it comes into
contact with an acidic environment of the stomach. This
gas entraps into the matrix of water-soluble polymers
Fig. 5. Effect of betacyclodextrin concentration on in vitro drug
and the formulation floats in an acidic environment of release of ofloxacin (n = 6). Standard deviation was found to be less
the stomach. than 2% in all the in vitro drug release profiles.
As the concentration of sodium bicarbonate
increases from 60 (F8) to 80 mg/tablet (F9), floating
lag time has reduced to 25 s. As the concentration of marketed formulations released the total amount of the
sodium bicarbonate increases from 60 to 80 mg/tablet, drug in pH 1.2 phosphate buffer for 24 h (data not
duration of floating has also been increased from 18 to shown).
24 h as shown in Table 3. This might be due to the gas
generated by 60 mg sodium bicarbonate might not be 3.6. Pharmacokinetic studies in healthy human
sufficient to keep formulation floating for prolonged volunteers
period of time whereas in case of gas generated by
80 mg sodium bicarbonate was sufficient to keep for- Pharmacokinetic studies were carried out in healthy
mulation floating for 24 h. human volunteers for test formulation (F11) and mar-
keted once daily (Zanocin) formulation. Cmax value
3.5. Effect of betacyclodextrin on in vitro release for Zanocin was found to be 4.13 ± 0.487 ␮g/ml
of ofloxacin

In order to improve the drug release profile from

the formulation, betacyclodextrin was added in to the
formulation. As the concentration of betacyclodextrin
increases from 0 to 100 mg/tablet, initial burst releases
as well as drug release in the latter hours have been
increased as shown in Fig. 5. In case of formulation con-
taining 0 mg (F2) and 50 mg betacyclodextrin (F10),
cumulative drug release after 24 h was 52.55 ± 1.02 and
57.90 ± 1.32%, respectively. The formulation contain-
ing 100 mg betacyclodextrin/tablet (F11) showed sim-
ilar drug release as that of marketed formulation. Beta-
cyclodextin dissolves rapidly from the tablet matrix
into the medium and it creates porosity to the matrix,
which results in increase in drug release from the tablet
matrix. In vitro drug release studies were also carried Fig. 6. Pharmacokinetic studies of marketed and developed formu-
out in pH 1.2 phosphate buffer, developed as well as lation (n = 12).
184 M. Chavanpatil et al. / International Journal of Pharmaceutics 304 (2005) 178–184

whereas Cmax value for F11 was found to be Acknowledgements

3.87 ± 0.467 ␮g/ml as shown in Fig. 6. Tmax values
for both Zanocin and F11 formulation were found We are thankful to University Grant Commission
to be 6 h. AUC values for Zanocin and F11 were and NAFETEC, India, for providing financial assis-
64.48 ± 6.60 and 62.90 ± 6.65 ␮g/h ml, respectively. tance.
Kel values for Zanocin and F11 were 0.0737 ± 0.0056
and 0.00704 ± 0.0057 h−1 . T1/2 value for Zanocin was
found to be 9.45 ± 0.718 whereas T1/2 value for F11 References
was found to be 9.89 ± 0.899 h. The difference between
the AUC values of F11 and marketed formulation were Deshpande, A.A., Shah, N.H., Rhodes, C.T., Malick, W., 1996.
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found to be insignificant (P > 0.05). The percent rela- residence: an overview. Drug Dev. Ind. Pharm. 22, 531–539.
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