suppositoria ibuprofen

A comparative bioavailability study of two ibuprofen formulations after
single-dose administration in healthy volunteers
Metta Sinta Sari Wiria, Fransiscus D. Suyatna
Abstrak
Penelitian ini dilakukan untuk mengetahui apakah bioavailabilitas formulasi ibuprofen suppositoria 125 mg yang diproduksi oleh PT
Kalbe Farma,Tbk. (Ibukal®
) bioekivalen dengan produk yang sama dari komparatornya (Proris®
). Parameter farmakokinetik yang dinilai
dalam studi ini ialah luas daerah di bawah kurva kadar -waktu selama 10 jam (AUC0-t
), luas daerah di bawah kurva kadar -waktu sampai
waktu tak terhingga (AUC0-inf), kadar puncak (Cmax), dan waktu untuk mencapai kadar puncak (tmax). Penelitian ini menggunakan
rancangan menyilang acak, tersamar tunggal yang mengikutsertakan 12 sukarelawan dewasa sehat. Sukarelawan dipuasakan semalam
dan keesokan harinya diberi 1 suppositoria obat uji (produk PT.Kalbe-Farma) atau 1 suppositoria obat pembanding (produk
komparatornya). Contoh darah diambil pada jam ke 0 (kontrol), 20 min; 40 min; 1; 1,5; 2; 2,5; 3; 4; 6; 8; dan 10 jam setelah pemberian
obat. Setelah 1 minggu periode washout, prosedur ini diulang dengan memberikan obat pembandingnya. Kadar obat ditentukan dengan
kromatografi cair kinerja tinggi dengan detektor ultraviolet. Pada penelitian bioavailabilitas ini, rerata (SD) AUC0-t
, AUC0-inf,Cmax dantmax
dari obat uji masing-masing adalah 28,59(3,37) µg.jam.mL-1
, 30,47(3,56) µg.jam.mL-1
,8,24(1,44)µg/mL, dan 1,33(0,44) jam. Rerata (SD)
AUC0-t
, AUC0-inf, Cmax dan tmax dari obat pembanding masing-masing adalah 28,13(8,14) µg.jam.mL-1
, 30,56(8,05) µg.jam.mL-1
,
8,27(2,88) µg/mL, dan 1,79(0,33) jam. Rasio rerata geometrik obat uji terhadap obat pembandingnya ialah 104,38% untuk AUC0-t
,
101,97% untuk AUC0-inf, dan 104,02% untuk Cmax, Nilai 90% confidence intervals(CI) nya ialah 90,38-120,54% untuk AUC0-t, 89,51-
116,16% untuk AUC0-inf, dan 85,73-126,16% untuk Cmax. Tidak ada efek samping yang dijumpai dalam penelitian ini. Dari hasil penelitian
ini disimpulkan bahwa Ibuprofen suppositoria 125 mg produksi PT. Kalbe Farma,Tbk. (Ibukal®
) bioekuivalen dengan produk yang sama
dari komparatornya (Proris®
).(Med J Indones 2007; 16:181-6)
Abstract
This study was aimed to investigate the bioequivalence of ibuprofen 125 mg suppository formulation (Ibukal®
, test formulation from PT.
Kalbe Farma, Tbk., Jakarta) and the ibuprofen suppository comparative formulation (Proris®
, from PT. Pharos Indonesia, Jakarta) in 12
healthy volunteers. The pharmacokinetic parameters used in this study were the area under the concentration-time curve from time zero to
hour 10 (AUC0-t
), the area under the concentration-time curve from time zero to infinite (AUC0-inf), the maximum concentration (Cmax), and
the time needed to reach the maximum concentration (tmax). The study was designed as a random cross-over fashion, single-blinded which
included 12 healthy adult volunteers. The volunteers were fasted overnight and in the morning they received a suppository of the test drug
(Ibukal®
) or a suppository of the comparative drug (Proris®
). Blood samples were withdrawn on hour 0 (control), 20 min; 40 min; 1; 1,5;
2; 2,5; 3; 4; 6; 8; and 10 time points after the administration of the drug. Following a wash-out period of 1 week, this procedure was
repeated using the other drug. The serum concentration of the drug was determined by means of high-performance liquid chromatography
with ultraviolet detection. The results of the study showed that, the mean (SD) of AUC0-t
, AUC0-inf, Cmax and tmax of the test drug were,
respectively, 28.59(3.37) µg.h.mL-1
, 30.47(3.56) µg.h.mL-1
, 8.24(1.44) µg/mL, and 1.33(0.44) h. The mean (SD) of AUC0-t
, AUC0-inf,Cmax
andtmaxof the comparative drug were, respectively, 28.13(8.14) µg.h.mL-1
, 30.56(8.05) µg.h.mL-1
, 8.27(2.88) µg/mL, and 1.79(0.33) h. The
geometric means ratio of the test to the comparative drug were 104.38% (CI 90%: 90.38-120.54%) for AUC0-t
, 101.97% (CI 90%: 89.51-
116.16%) for AUC0-inf, and 104.02% (CI 90%: 85.73-126.16%) for Cmax. There was no side effect of the drug detected in this study. From
the results we can conclude that the 125 mg of ibuprofen suppository of PT Kalbe Farma, Tbk. (Ibukal®
) is bioequivalent to that of the
comparative drug (Proris®
).(Med J Indones 2007; 16:181-6)
Keywords: ibuprofen suppository, high-performance liquid chromatography, bioequivalence
Departement of Pharmacology and Therapeutics, Faculty of Medicine
University of Indonesia, Jakarta, Indonesia182 Wiria and Suyatna Med J Indones
Ibuprofen (2-(4-isobutylphenyl)-propionic acid, C13H18O2 )
is a non-steroidal antiinflamatory drug. It is a racemate
of (S)-(+) dan (R)-(-) enantiomer with the ratio 1:1,
but only the S-enantiomer is active. Both enantiomers
posses the same pharmacokinetic (concentration vs.
time) profile. The analgesic and antiinflamatory effect
of the drug have been used clinically to treat post
operative pain in young adults and children with minor
side effect. The main side effect of ibuprofen is gastrointestinal tract irritation e.g., gastritis. The mechanism
of antiinflamation of ibuprofen is through the inhibition
of the prostaglandin and leukotriene biosynthesis.
Ibuprofen is rapidly absorbed after oral administration
in men, and peak plasma concentration is observed
after 1 to 2 hours. The half-life in plasma is about 2
hours. Although the rectal preparation may have some
delay in reaching the peak plasma concentration,
ibuprofen suppositories are absorbed efficiently. The
peak plasma concentration (Cmax) is about 5-20 mg/L
(oral, single dose of 200 mg) or about 12.4-30.1 mg/L
(suppository, doses of 20 mg/kg BB). The bioavailability
of suppository formulation is about 73%.
Ibuprofen is extensively bound to plasma proteins (90-
99%), penetrates slowly from plasma to synovial
spaces and may accumulate there in higher concentration. The drug is metabolized by hydroxylation
and carboxylation to form two inactive metabolites,
which are eliminated after conjugation to glucuronic
acid. The excretion of ibuprofen is rapid and complete;
more than 90% of an ingested dose is excreted in the
urine as metabolites or their conjugates, and no
ibuprofen is found in the urine.1,2,5
The bioavailability of two (or more) formulations of
the same active ingredient could differ from one to the
other; therefore bioequivalence studies become the
important part of registration dossiers. If they are
equivalent, then one may subsequently claim that the
therapeutic efficacy of both formulations is similar. It
may also mean that the beneficial and side effects of
the two drugs are identical, and hence the formulations
are interchangeable.
The aim of this study was to compare, the pharmacokinetic
profiles and to evaluate the bioequivalence of two
formulation of 125 mg ibuprofen suppositories (Ibukal®
from PT.Kalbe Farma Tbk., Jakarta and Proris®
from
PT. Pharos Indonesia, Jakarta), after single dose rectal
administration in healthy volunteers of both sexes.
METHODS
Clinical protocol
Twelve healthy subjects were recruited in this study.
They were of both sexes between 18-55 years of age
and with a normal body weight (Body Mass Index
=18-25). Physical examination and laboratory check
up (hemoblobin, hematocrite, total and differential
white cell count, creatinine, alkaline phosphatase,
Alanin Trans Aminase /ALT, total bilirubin, albumin
and total protein, and routine urinalysis) were
performed.Pregnant or nursingwomen, individualwith a
history of hypersensitive to ibuprofen or other antiinflamatory Non Steroidal Drugs (NSAIDs), and those
who had any gastrointestinal tract problem(s), were
excluded from the study.
All subjects gave written informed consent and the
protocol had been approved by Medical Research
Ethics of the Medical Faculty, University of Indonesia.
The study was conducted in accordance with the Good
Clinical Practice (GCP) standard.
The study was a single-dose, 2-way randomized crossover
design with a one week washout period between the
doses. The subjects were instructed to abstain from
taking any medication for at least 1 week before and
during the study period. The subjects were also not
permitted to consume alcohol, or beverage or food
containing xanthines, such as tea, coffee and cola, or
fruit juice.
The subjects were fasted for 10 hours prior to drug
administration. On the morning of study phase as
scheduled, the volunteers were given a single dose of
either formulation (reference or test) of ibuprofen
suppository. No food was permitted for 4 hours after
dosing. Breakfast, lunch, and dinner were given to all
the volunteers according to the time schedule. The
volunteers were instructed to stay in bed for 4 hours
after drug administration, afterwards they were
allowed to walk but were prohibited from strenuous
activity; they were under direct supervision at the
study site. Blood pressure, pulse, and adverse events,
which might occur, were monitored and recorded in
the Case Report Form (CRF).
Blood samples (approximately 5 mL) from a suitable
antecubital vein were collected into tube before (0 h)
and at 0; 20 min; 40 min; 1; 1,5; 2; 2,5; 3; 4; 6; 8, and
10 hours after dosing. Blood samples were centrifuged
at 3000 rpm for 10 min, serum were separated and
stored frozen at -200 C until assayed. Vol 16, No 3, July – September 2007 Bioavailability study of two ibuprofen formulations 183
After a washout period of 7 days, the study was repeated
in the same manner to complete the crossover design.
Formulations
The following formulations were employed: Ibukal®
125 mg suppository (Test formulation from PT. Kalbe
Farma,Tbk., batch No. 821001A, expiration date January
2008) and Proris®
125 mg suppository (Reference
formulation from PT. Pharos Indonesia, batch No.
B5L44D, expiration date November 2008).
Drug analysis
Serum samples were analyzed for ibuprofen according
to the HPLC method developed at the PT. Pharma
Metric labs laboratory, and validated following
international guideline.
To a tube of 0.5 mL serum containing ibuprofen was
added 40 µL of internal standard (mefenamic acid,
125 µg/mL); the mixture was shaken for 30s by mean
of a vortex mixer. 250 µL of 1.0 M phosphate buffer
pH 2.0 and 4.0 mL diethylether was then added and
the mixture was again vigorously shaken for 1 min.
The mixture was then centrifuged for 10 min at 3000
rpm.The supernatant was transferred to a clean glass
tube and evaporated to dryness under nitrogen stream
in a waterbath 400C. The residue was redissolved in
200 µL methanol, and vortex-mixed for 30s. The solutions
were then transferred to the auto-injector microvials.
An aliquot (20 µL) of each serum extract was injected
into a Waters Symmetry C18 analytical column, (150
mm x 4.6 mm i.d.). The compounds were eluted by
pumping the mobile phase (acetonitrile – 0.01M
potassium dihydrogen phosphate, pH 7.0 (35:65; v/v))
at a flow rate of 1.0 mL/min. The effluent was
monitored using an ultraviolet detector at 225 nm
wavelength. Under this condition, typical retention
times were 3.15 and 5.60 minutes for ibuprofen and
mefenamic acid, respectively.
Accuracy, Precision and Recovery
Accuracy, precision and recovery of quality control
serum samples were measured at the concentration of
1.5, 15.0, and 30.0 µg/mL of ibuprofen.
Stability
Stability of quality control serum samples (1.5, and
30.0 µg/mL) were subjected to 7, 14, 28 days storage,
and 2 freeze/thaw (-20 to 250C) cycles test. Subsequently,
the ibuprofen concentrations were measured and
compared to freshly prepared samples.
Pharmacokinetics and statistical analysis
The bioequivalence between the 2 formulations was assessed
by calculating individual test/reference ratio for:
1. Area under the curve (AUC) of serum concentration
until last concentration (Ct) observed (AUC0-t),
AUCt were calculated by applying the linear
trapezoid rule
2. Area under the curve (AUC) between the first
sample (pre-dosage) and infinity (AUC0-inf), AUC0-
inf were calculated by the following equation:
AUC0-inf = AUCt + C0-t
/ Ke; Ke = 0,693 / t½ ; t½
= 0,693 / (2,303 x slope)
3. Peak serum concentration (Cmax)
4. Time to peak serum concentration (tmax)
The mean maximum serum concentration and the
mean time to peak serum concentration obtain
directly from observed data.
The AUC and Cmax data for the 2 formulations were
analyzed by ANOVA using Equiv Test®
versi 2.0
programme (Statistical Solution Ltd., Saugus. MA,
USA) to establish whether the 90% CI of the ratios
was within 80-125% interval (for AUC) and 70-143%
interval for Cmax, indicating bioequivalence as proposed
by US Food and Drug Administration.
Parametric and non parametric analyses of lntransformed arithmetic means and individual tmax
difference between the test and reference formulations
were also performed.
RESULTS
The volunteer involved in the study had the following
characteristic (expressed as mean ± SD (range)): age:
20.0±5.0 years old, (19-36) years old, height:
164.8±6.6 cm (150-176 cm), body weight: 61.2±7.1
kg (51-75 kg).
The HPLC chromatograms of the sample is shown in
Figure 1, in which the retention times of ibuprofen and
mefenamic acid are 3.15 and 5.60 minutes, respectively.
The lower limit of quantification (LOQ) of our method
was 0.5 µg/mL. Ibuprofen concentrations were
quantified using linear regression of area under the
curve (AUC) ratios (Ibuprofen/IS) versus concentration.
The calibration curves were linear over the concentrations range of 0.5 to 40.0 µg/mL. Table 1 shows the
precision, accuracy, recovery and stability data for QCs.184 Wiria and Suyatna Med J Indones
Figure 1. The HPLC chromatogram of serum sample after suppository administration of ibuprofen 125 mg suppository formulation in
healthy volunteer
Table 1. The precision, accuracy, recovery, and stability data for ibuprofen from the pre-study validation in human serum
Intra-day validation
Nominal concentration 1.5 15.0 30.0
Precision (%) 7.94 6.58 7.95
Accuracy (%) 15.73 9.24 5.52
Inter-day validation
Nominal concentration 1.5 15.0 30.0
Precision (%) 8.06 4.85 6.64
Accuracy (%) 7.78 2.74 3.78
Recovery data
Nominal concentration 1.5 15.0 30.0
Range (%) 114.5-118.4 103.1-104.7 104.3-110.7
Mean (%) 116.3 103.7 107.1
Stability on storage Concentration measured at:
Actual concentration Day-0 Day-7 Day-14 Day-28
1.5 (µg/mL) 1.67(11.33) 1.43 (6.89) 1.59 (5.78) 1.45(6.67)
30.0 (µg/mL) 30.71(2.49) 28.79(5.38) 29.28(2.40) 30.96(11.41)
Freeze/thaw cycle Cycle-1 Cycle-2 Cycle-3
1.5 (µg/mL) 1.67(11.33) 1.43 (6.89) 1.60 (6.44)
30.0 (µg/mL) 30.71(2.49) 28.79(5.38) 31.12(3.74)
Ibuprofen was found to be well tolerated and no
significant adverse reaction was encountered during
the trial.
The ibuprofen serum mean concentration vs. time
profile obtained after the single suppositopies administration
of each 125 mg suppository formulation is shown in
Figure 2.
The mean pharmacokinetic parameters obtained from
12 volunteers after administration of a 125 mg
suppository formulation is shown in Table 2.Vol 16, No 3, July – September 2007 Bioavailability study of two ibuprofen formulations 185
Figure 2.Ibuprofen serum mean concentration vs. time profile obtained after the single suppository administration of 125 mg of
ibuprofen suppository formulation in 12 volunteers
Table 2. Mean pharmacokinetic parameters obtained from 12 volunteers after administration of each of 125 mg ibuprofen formulation
Proris®
(Reference drug) Ibukal® (Test Drug)
Mean SD (±) Min Max Mean SD (±) Min Max
tmax (h) 1.79 0.33 1.50 2.50 1.33 0.44 1.00 2.50
tlast (h) 8.17 1.80 6.00 10.00 8.33 1.44 6.00 10.00
t1/2 (h) 1.90 0.45 1.36 2.66 1.85 0.29 1.50 2.42
Ke (1/h) 0.35 0.10 0.27 0.49 0.38 0.06 0.29 0.46
Cmax (µg/mL) 8.27 2.88 4.18 13.49 8.24 1.44 6.62 11.42
Ct (h) 0.86 0.33 0.56 1.48 0.73 0.28 0.52 1.32
AUC 0-t (µg.h/mL) 28.13 8.14 8.29 45.79 28.59 3.73 23.36 36.43
AUC 0-inf (µg.h/mL) 30.56 8.05 21.46 48.42 30.47 3.56 25.36 38.35
Table 3 shows the geometric mean of the individual
Cmax, AUC0-t
, AUC0-inf, (test/reference formulation),
the respective 90% confidence intervals (CI), and
Intrasubject CV for 12 volunteers. The calculated 90%
CI for mean AUC0-t and AUC0-inf, individual ratios
were within the 80-125%, and for Cmax were within
75-143% interval defined by the US Food and Drug
Administration(US FDA) and Indonesian FDA, thus
establishing the bioequivalence of the two formulations
There is not statistically significant difference of tmax
value between the test and the reference drug.
Table 3. Geometric mean of individual AUC0-t, AUC0-inf, and
Cmax ratios (test/reference formulation), the respective
90% confidence interval (CI) and inter-subject CV
Geometric
Mean
90% CI Intra
Subject CV%
AUC0-t 104.39 90.38 – 120.54 19.44
AUC0-inf 101.97 89.51 – 116.16 17.61
Cmax 104.02 85.73 – 126.16 26.01
0.00
3.00
6.00
9.00
12.00
0 1 2 3 4 5 6 7 8 9 10
Time (hour)
Concentration mean (µg/mL)
Ibuprofen - test
Ibuprofen-reference186 Wiria and Suyatna Med J Indones
DISCUSSION
Ibuprofen has been determined in plasma or serum and
other biological fluids by several methods. The method
used in this study is a modification of that describes
previously by Rustam AM et al.
4
The assay was
performed using C18 analytical column, and mefenamic
acid as the internal standard (IS). The HPLC chromatograms of the sample gave a good resolution and
selectivity between ibuprofen and the internal standard.
From the pre-study validation data (precision, accuracy,
recovery and stability) obtained, it can be shown that
the developed method is sufficient for ibuprofen
pharmacokinetic study.
After rectal administration of the ibuprofen suppositories,
the observed ibuprofen mean peak serum concentrationin (Cmax) value were similar to those reported in
the literature (Kyllonem M. et al.).
CONCLUSION
From this study, it can be concluded that ibuprofen
125 mg suppository produce by PT. Kalbe Farma, Tbk.
(Ibukal®
) is bioequivalence to its comparative drug
(Proris®
) for both rate and the extent of absorption.
Acknowledgement
We thank the volunteers for the participation and PT.
Kalbe Farma, Tbk. for the sponsorship in this study.
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