C Basic & Clinical Pharmacology & Toxicology 2004, 95, 263265.

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Copyright C
ISSN 1742-7835

A Two-Way Cross-Over Bioequivalence Study

Comparing Two Products of Diclofenac Sodium
Suppositories in Healthy Human Volunteers
Mazen Hasan1, Sameer Otoom1, Naji Najib2 and El-Sayed Sallam3
Faculty of Medicine and 2Faculty of Pharmacy, Jordan University of Science and Technology, Irbid 22110, Jordan

(Received May 5, 2004; Accepted August 18, 2004)

Abstract: This report presents the results of two treatment cross-over investigations on 20 healthy male volunteers to
assess the bioequivalence of two suppository products of diclofenac sodium. The study was carried out under US Food
and Drug Administration Guidelines. The two products were voltarenA (100 mg) suppository (Ciba-Giegy), as a reference
product, and InflabanA (100 mg) suppository (The Arab Pharmaceutical Manufacturing Company, Ltd. APM), as a
test product. Both products were administered rectally as a single dose (100 mg) separated by a one-week wash-out period.
Following drug administration, blood samples were collected over 12 hr, and serum harvested from the blood was analyzed
for diclofenac sodium using a sensitive and specific high performance liquid chromatographic assay. The results of this
investigation indicated that there were no statistically significant differences between the two products in either the mean
concentration-time profiles or in the obtained pharmacokinetic parameters, including area under the serum concentrationtime curve for 12 hr (AUC012h), lag time between product administration and first appearance of the drug in serum
(Tlag), peak serum concentration (Cmax), and time to reach this peak serum concentration (Tmax). Concerning the relative
extent of absorption, assessed by the AUC ratio (Inflaban/Voltaren) for 12 hr, the average value was found to be 1.000.09
with a 95% confidence limits (C.L.) of 0.821.18. Thus, these findings clearly indicate that the two products are bioequivalent in terms of rate and extent of drug absorption.

banA and VoltarenA). The bioavailability of the test product

(Inflaban) relative to the reference product (Voltaren) will
be assessed by comparing the concentration-time profiles as
well as the obtainable pharmacokinetic parameters; T1ag,
Tmax, Cmax, AUC012h, AUC0inf, Ke, T1/2e, and MRT (mean
residence time) derived from the corresponding concentration-time profiles.

Diclofenac sodium is a non-steroidal anti-inflammatory

drug, widely used in the treatment of rheumatic disorders
and other chronic inflammatory diseases (Brogden et al.
1980). Due to its analgesic properties (Charles & Navil 2002),
it is also used in the management of acute conditions such as
dental pain (Matthews et al. 1984), renal colic (Lundstam et
al. 1982), postoperative and posttraumatic pain (Kantor
1986). Moreover, The drug was found to reduce the incidence
and severity of postoperative pruritus (Colbert et al. 1999).
Diclofenac sodium is characterized by rapid systemic clearance that necessitates repeated daily dosing when a course of
treatment with this drug is required (Charles & Navil 2002).
Therefore, interest has emerged in the production of slow release formulations of diclofenac sodium (Hasan et al. 1992),
to enable the administration of the drug once daily and reduce the incidence of the occurrence of multiple high peaks
(and potential adverse effects) associated with the conventional products of this drug slow release dosage forms, however, have disadvantages, such as the possibility of dose
dumping due to faulty formulation, reduced potential for accurate dose adjustment and possible reduction in systemic
availability due to slow or incomplete release (Welling 1983).
Another method to increase efficacy and reduce the adverse effects of diclofenac sodium is to use the rectal formulations (Sastry et al. 1992). The purpose of this study is to
compare the rectal absorption and disposition kinetics of
two suppository formulations of diclofenac sodium (Infla-

Drug analysis. A rapid and sensitive high-performance liquid chromatographic method has been developed for the determination of
diclofenac sodium in serum using flufenamic acid as an internal
standard. All solvents used were HPLC grade. Serum protein was
precipitated with acetonitrile. The drugs were eluted from a 5 mm
C-8 reversed-phase column at ambient temperature with a mobile
phase consisting of acetonitrile-water (50:50%, V/V) adjusted to pH
3.3 with glacial acetic acid, at a flow rate of 2 ml/min. with UV
detection at 280 nm. The quantitation of the chromatogram was
performed using peak height ratios of the drug to the internal standard. and the relative and absolute recoveries varied from 90 to
98%. A representative standard curve of the diclofenac sodium/flufenamic acid peak height ratio over the diclofenac serum concentrations ranging between 504000 ng/ml, resulted in the following
linear least-squares regression equation: Y0.0013 X0.0004 (r
0.9999). Detection limits for diclofenac sodium in serum was found
to be 20 ng/ml. Intra-day coefficients of variation (CVs) ranged
from 2.5 to 4.6% and inter-day CVs from 3.5 to 7.0% at three different concentrations. Serum samples of diclofenac sodium were
stored in a freezer at 20 until the time of analysis. The results
demonstrated that the drug can be stored frozen in serum for at
least two weeks without degradation.

Author for correspondence: Sameer A. Otoom, Faculty of Medicine, Arabian Gulf University, P.O. Box 22979, Al Manama, Kingdom of Bahrain (fax 962 2 7095010, e-mail sato/just.edu.jo).

Subjects. Twenty healthy adult male volunteers were included in this

study. Their ages ranged from 20 to 36 years (27.31.0 years) and
their weight between 60 and 85 kg (71.81.9 Kg) with body mass
index (BMI) of 25.60.67. On the basis of medical history, clinical

Materials and Methods

264

MAZEN HASAN ET AL.

for infinity (AUMC0inf) by AUC). Ka and t1/2 of absorption was
calculated using Wagner-Nelson method (Pedraz et al. 1988). Other
pharmacokinetic parameters, such as Tlag, Tmax and Cmax were estimated by inspecting the obtained concentration-time curves.
Statistical analysis. All results are expressed as meanS.E.M. The
coefficient of variation (CV) was calculated for the obtained
pharmacokinetic parameters. Statistical analysis was performed by
two-way analysis of the variance for the pharmacokinetic parameters. On the basis of variance homogeneity, the ANOVA of all
the parameters were carried out on the linear data. The t-test for
paired data was employed for serum drug concentrations. Differences between 2 related means were considered statistically significant for P values equal or less than 0.05.

Fig. 1. Mean serum concentration-time profiles (linear scale) of diclofenac sodium for 20 volunteers following rectal administration
(100 mg suppository) of VoltarenA (open circles) versus InflabanA
(filled circles). The vertical bars represent S.E.M.
examination and laboratory investigations (haematology, blood biochemistry and urine analysis), none of the participants has revealed
any medical abnormality. In addition, the included subjects have no
history of hospitalization or involvement in any clinical trials within
12 weeks, and none of them had received any regular course of drug
therapy within 4 weeks prior to this study. Informed written consent
was obtained from the volunteers and the protocol of the study was
approved by the Ethical Committee on Human Research at Jordan
University of Science and Technology. Each subject received the two
products in two treatment days, separated by a one week wash-out
period. The order of product administration was done according to a
randomized cross-over design. Each volunteer reported to the Clinical Trials Laboratory at 7 a.m. after an overnight fast of 10 hr. Three
standard meals were served at 0.5, 5, and 11 hr after dosing. Cigarettes, and food or beverage-containing caffeine were not allowed
over 12 hr. Blood samples (10 ml) were collected in plain glass vacutainers at 0 (pre-dose), 0.25, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 6.0, 8.0,
10.0, and 12.0 hr after drug administration. Following clot retraction,
the samples were centrifuged at 3,000 rpm for 15 min. and the obtained serum was frozen at 20 until assayed.
Pharmacokinetic analysis. The pharmacokinetic parameters were
calculated using the PKCALC Computer Program (Schumacher
1986). The elimination rate constant (Ke) and the half-life of elimination (T1/2e) were calculated by linear regression of the terminal
slope of the serum concentration-time profiles. Areas under the concentration-time curves for 12 hr (AUC012h) were estimated by the
hybrid logarithmic/linear trapezoidal rule, and AUC for infinity
(AUC) was obtained by adding C*/Ke to AUC012h, where C* is the
last detectable concentration. The mean residence time (MRT) was
obtained by calculating the ratio of the area under the first moment

Results
Fig. 1 shows the mean serum concentrations of diclofenac
sodium following rectal administration of the 2 products
(Voltaren A and Inflaban B). No statistically significant
differences were observed between the two products at any
time period over the entire sampling interval. The mean
values of the pharmacokinetic parameters of the two products (Tlag, Tmax, Cmax, AUC012h), are presented in table 1.
No statistically significant differences were observed between the 2 products for any of the derived parameters.
Concerning the relative extent of absorption as assessed by
the AUC ratio (B/A) for 24 hr, the average value was found
to be 1.000.09 with a 95% confidence limits (C.L.) of
0.821.18. No adverse effects were reported by the volunteers after administration of either product.
Discussion
The aim of this study is to compare the relative bioavailability of two suppository products of diclofenac sodium;
namely InflabanA in the form a suppository containing 100
mg of diclofenac sodium produced by The Arab Pharmaceutical Manufacturing Company, Ltd. (APM), Sult, Jordan, and VoltarenA (a suppository containing 100 mg of diclofenac sodium) produced by Ciba-Giegy, Switzerland.
Our results showed that there is no significant differences
between the two products of diclofenac sodium in either
the mean serum concentration-profiles or in the obtained

Table 1.
A statistical comparison of the average values (S.E.M.) of the pharmacokinetic parameters of diclofenac sodium derived from the concentration-time curves of VoltarenA and InflabanA following rectal administration (100 mg suppository) to 20 subjects.
Pharmacokinetic
parameter
.

T max (hr)
Cmax (ng/ml)
AUC012h (ng hr/ml)
AUC (ng hr/ml)
Ka (min.1)
t1/2a (min.)
Ke (hr1)
T1/2e (hr)
MRT (hr)
C*/Ke (ng/ml hr)
.

Product
VoltarenA
0.930.05 (0.831.03)*
2856251 (23643348)
5570472 (46456495)
5742475 (48116673)
0.1250.009 (0.1070.143)
6.50.8 (4.98.1)
0.4300.028 (0.3750.485)
1.770.13 (1.522.02)
2.710.14 (2.442.98)
32.657

InflabanA
0.860.05 (0.760.96)
2753183 (23943112)
5407274 (48705944)
5579277 (50366122)
0.1340.007 (0.1200.148)
5.60.6 (4.46.8)
0.3950.020 (0.3550.435)
1.890.15 (1.602.18)
2.640.11 (2.422.86)
29.25

P-value
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05

Range of Tmax (hr) was 0.751.5 for VoltarenA and 0.51.5 for InflabanA. *Values between brackets represent the 95% confidence limits.

BIOAVAILABILITY OF DICLOFENAC SODIUM SUPPOSITORIES

pharmacokinetic parameters indicating that Voltaren and

Inflaban suppositories are bioequivalent in terms of the rate
and extent of drug absorption.
Rectal administration of diclofenac sodium is preferred
over the oral route to minimize the gastric adverse effects.
This method is useful when patient can not swallow, uncooperative or have severe gastrointestinal ulceration (Sastry
et al. 1992), Recently, the use of rectal diclofenac in clinical
practice is becoming more evident. The drug was found to
significantly improve postoperative analgesia in varicose
vein surgery. This effect was achieved when the drug was
given at induction or 3045 min. before operation (Lyons
et al. 2003). Moreover, the drug was found to reduce the
incidence of pancreatitis if given rectally immediately after
endoscopic retrograde cholangiopancreatography (Murray
et al. 2003). However, preoperative rectal diclofenac sodium
was found to increase intraoperative blood loss when used
in tonsillectomy surgery (Schmidt et al. 2001).
Landsdorp et al. (1990) studied the pharmacokinetics of
rectal diclofenac and its hydroxy metabolites in man. They
found that the apparent half-lives of diclofenac to be 1.30.3
hr and 4.31.0 hr for its hydroxy metabolite. These half-lives
were estimated to be 1.80.9 hr and 3.60.5 hr respectively
when derived from the renal excretion rate-time profile. Diclofenac was excreted for 13.66.5% with renal clearance of
3.231.03 ml/min.. The pharmacokinetic parameters of diclofenac sodium were also studied in rabbits, Cmax (mg/ml)
was 67.304.2, Tmax (hr) was (1.080.15). AUC (mg hr/ml)
was 247.2014.8. Interestingly, the T1/2 and MRT in rabbits
were comparable to our data in human 1.950.1 and
3.20.08 hr respectively (Ramakrishna et al. 1996a).
Many studies have compared the rectal pharmacokinetic
parameters of diclofenac sodium to the oral ones. It was
found that the two formulations had similar pharmacokinetic profiles as evidenced by the insignificant difference
between Cmax, T1/2, MRT and AUC (Ramakrishna et al.
1996a). Another study showed bioequivalence between rectal suppositories and oral tablets with respect to AUC and
Cmax in 12 healthy male human volunteers, but with different Tmax (Ramakrishna et al. 1996b). On the other hand,
one study conducted by Hanses et al. (1996) showed that
rectal and oral diclofenac have similar extent bioavailability
but the rate of drug input was lower for the suppositories.
In conclusion, our results show no significant differences
between the two products of diclofenac sodium in either the
mean serum concentration-profiles or in the obtained
pharmacokinetic parameters reflecting the rate (Tmax, Cmax,
Ka, T1/2a) and extent (AUC010h, AUC) of drug absorption.
The 95% C.L. of the AUC ratios for 12 hr ours and for infinity were found to be within the FDA accepted limits {0.80
1.25) for bioequivalent products. Thus, the data clearly indicate that VoltarenA and InflabanA suppositories are bioequivalent in terms of the rate and extent of drug absorption.
Acknowledgements
This work was supported by a grant from the deanship
of Jordan University of Science and Technology. The

265

authors like to thank Ms Raedah Swaidan from the Department of Pharmacology, College of Medicine for her technical support. The authors have no conflict of interest directly
relevant to the content of the study. No financial support
was obtained from the producer of Inflaban.

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