Varaprasad Bobbarala et al.

/ Drug Invention Today 2009, 1(1),13-15

Available online through
www.ditonline.info
Research Article

Bioequivalence study of formulation Rabiplus-XT with reference Rabium plus in healthy

volunteers
Somasekhar Penumajji 1 and Varaprasad Bobbarala2*
1Vivimed labs Limited, Veeranag towers, Habsiguda, Hyderabad, A.P.
2For U Biosciences, A/4A, Park lane Residency, East point colony, Vizag-17, A.P, India.

Received on: 09-08-2009; Revised on: 23-09- 2009; Accepted on:05-10-2009

ABSTRACT
A randomized, two-way, crossover bioequivalence study in 12 healthy volunteers was conducted to compare the pharmacokinetic param-
eters of RABIPLUS-XT versus Reference Rabium Plus (Rabeprazole Sodium 20mg+Itopride HCl 150 mg SR Capsules). The blood samples
were collected during the 48hour period after drug administration. Rabeprazole and Itopride levels in plasma were determined by high-
performance liquid chromatography. The pharmacokinetic variables of area under the plasma concentration time curve, maximum concentra-
tion, time to maximum concentration, elimination half-life, and mean residence time were computed. The results of this study suggest that the
two formulations are bioequivalent. After the test formulation of RABIPLUS-XT is found to be bioequivalent with the reference formulation
Reference of Rabium Plus.

Keywords: Chronic gastritis, Bioequivalence, High performance liquid chromatography (HPLC), Healthy volunteers.

INTRODUCTION

Rabeprazole is chemically designated as 2-[(4-(3- Itopride hydrochloride is chemically designated as N-[[4-(2-

methoxypropoxy) - 3- ethyl-pyridin-2-yl) methyl sulfinyl] - 1H-
benzoimidazole. Rabeprazole belongs to a class of anti secretory com-
pounds (substituted benzimidazole proton-pump inhibitors) that do
not exhibit anticholinergic or histamine H2-receptor antagonist prop-
erties, but suppress gastric acid secretion by inhibiting the gastric
H+ /K+ ATPase at the secretary surface of the gastric parietal cell 1, 2.
Because this enzyme is regarded as the acid (proton) pump within the
parietal cell, rabeprazole has been characterized as a gastric proton
pump inhibitor. Rabeprazole blocks the final step of gastric acid se-
cretion. In gastric parietal cells, rabeprazole is protonated, accumu-
lates, and is transformed to an active sulfenamide.
Figure 1: Structure of Rabeprazole
*Corresponding author.
Dr. Varaprasad Bobbarala
Scientist In-Charge,
For U Biosciences/IMMA Labs,
A/4A, Park lane Residency, East point colony,
Visakhapatnam, A.P-530017, India.
Tel.: + 91-9949129539
E-mail:varaprasadphd@rediffmail.com
Drug Invention Today Vol.1.Issue 1.November 2009
dimethyl aminoethoxy) phenyl] methyl]-4-dimethoxy-benzamide.
Itopride is a gastrointestinal prokinetic with dopamine D2 receptor
antagonistic and acetyl cholinesterase inhibitory actions 3, 4, 5, 6.
Figure 2: Structure of Itopride
Rabiplus-XT is indicated for relief from symptoms of
dysmotility associated with acid peptic disorders, GERD (gastroe-
sophageal reflux disease), Dyspepsia, chronic gastritis. Pharmacody-
namics indicate that Rabeprazole belongs to a class of anti secretory
compounds (substituted benzimidazole proton-pump inhibitors) that
do not exhibit anticholinergic or histamine H2-receptor antagonist
properties, but suppress gastric acid secretion by inhibiting the gas-
tric H+ , K+ ATPase at the secretory surface of the gastric parietal cell.
Because this enzyme is regarded as the acid (proton) pump within the
parietal cell, rabeprazole has been characterized as a gastric proton-
pump inhibitor. Rabeprazole blocks the final step of gastric acid se-
cretion. In gastric parietal cells, rabeprazole is protonated, accumu-
lates, and is transformed to an active sulfenamide. The anti-secretory
effect begins within one hour after oral administration of 20mg
rabeprazole. The median inhibitory effect of rabeprazole on 24 hour
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 Varaprasad Bobbarala et al. / Drug Invention Today 2009, 1(1),13-15
gastric acidity is 88% of maximal after the first dose. Rabeprazole
20mg inhibits basal and peptone meal-stimulated acid secretion ver-
sus placebo by 86% and 95%, respectively, and increases the percent
of a 24-hour period that the gastric pH>3 from 10% to 65%. This
relatively prolonged pharmacodynamic action compared to the short
pharmacokinetic half-life (1–2 hours) reflects the sustained inactiva-
tion of the H+ , K+ ATPase. Itopride activates the gastrointestinal mo-
tility through synergism of its dopamine D 2 -receptor antagonistic
action and its acetylcholine esterase-inhibitory action. In addition to
these actions, Itopride has an antiemetic action, which is based on its
dopamine D 2 -receptor antagonistic action at CTZ.
The objective of the single dose randomized bioequivalence
study was to compare the pharmacokinetic parameters of RABIPLUS-
XT versus Reference Rabium Plus.
MATERIALS AND METHODS
The Drug controller General of India approved the study to
do at LTM Medical College, Sion, Mumbai. The study was conducted
at the LTM medical college for Creative Health care Pvt Ltd, a division
of Vivimed Labs Limited. Twelve volunteers participated in a random-
ized, two treatment, two way, two period, single dose crossover study.
Their ages range from 21 to 35 years. To be included in this study, the
subjects should be healthy volunteers, 18 to 50 years of age and
within 15 percent of ideal body weight for their height and build. One
week prior to enrolment, a complete physical exam and biochemical
screening was performed. Volunteers were asked to abstain from tak-
ing any drug for at least one-week prior and during the study, and to
fast for at least 10 hours prior to the study.
The study was divided in to two periods where the two
commercial trades RabiPlus-XT (Rabeprazole 20mg & Itopride HClmg
SR Capsules) which is the test sample and a reference sample
Rabiumplus (Rabeprazole 20mg & Itopride HClmg SR Capsules) were
orally taken in a randomized, double-blind, crossover order on each
sunday with a washout period of 28 days. At 8:00 AM, a blood sample
was taken and each subject received a single oral dose of either brand
with 240 ml of water. Volunteers were remained fasting 2 hours after
the administration of the tablet. This was followed by a light break-
fast, standardized lunch and dinner 2, 6 and 12 hours after drug ad-
ministration, respectively. All subjects abstained from smoking and
exercise during the day of the study. Blood samples (5ml) were ob-
tained form the cubital and fore arm veins through an indwelling hep-
arinized plastic tube, put into coded plain tubes at 0.00 (predose),
1.00, 2.00, 3.00, 4.00, 5.00, 6.00, 7.00 8.00, 9.00, 10.00, 12.00, 24.00 and
48.00 hours post dosing. After the 48 hours sampling the volunteers
were released. The withdrawn blood (5 ml) was placed in labelled
tubes each according to the volunteers’ Identification Number, time,
and drug code. Thereafter, these tubes were sent to the lab, and the
plasma was separated by centrifugation (within 30 minutes) at a rate
of 5000 rpm for 10 minutes. Plasma separated was stored at -70°C until
the assay. The stability of these samples at -70°C is three years. All
samples were analyzed in duplicates.
Drug Invention Today Vol.1.Issue 1.November 2009
The plasma samples were analysed by reverse phase HPLC
for the content of Rabeprazole and Itopride, the pharmacokinetic pa-
rameters were computed and subjected to statistical analysis.
Rabeprazole is extracted by taking 0.5mL Plasma adding 5mL
dichloromethane, vortexing for 4 minutes and centrifuging at 2400rpm
for 5 minutes. Filter through sodium sulphate and evaporate the or-
ganic layer to dryness under a stream of nitrogen at room tempera-
ture. Reconstitute the residue with 150µl of mobile phase [1% tri
ethylamine, pH adjusted to 7.4 with ortho phosphoric acid: Methanol
(50:50)] and inject 50 µl into the HPLC C18 column and at a flow of
1.0mL/min and at a wavelength of 290nm. Itopride is extracted by
taking 0.5ml plasma containing Itopride, adding 8mL of tetra butyl
methyl ether and dichloromethane (7:3) and vortexing for 5 minutes,
centrifuging at 3250g for 10minutes. Remove the organic layer and
evaporate to dryness at 40ºC under stream of nitrogen. Reconstitute
the residue in 150 µl of mobile phase [0.05M Potassium Dihydrogen
Phosphate, pH adjusted to 7.5 with Triethylamine : Acetonitrile (80:20)]
and vortex for 1 minute and inject 100 µl supernatant liquid into the
HPLC C18 column and at a flow of 1.0mL/min and at a wavelength of
290nm
For data analysis, the maximum Rabeprazole and Itopride
serum concentrations Cmax and Tmax determine the rate at which the
drug is absorbed and was directly computed from the mean plasma
concentration curve. AUC0-t: Area under the plasma concentration
time curve from time 0 to t was obtained by the trapexoidal rule. Kel is
the elimination rate constant determined by linear regression of the
last three sampling points, T1/2 is determined by the formula T1/2 =
0.693/ Kel. The pharmacokinetic parameters were compared using paired
– test and analysis of variance.
RESULTS
The mean plasma levels and pharmacokinetic parameters were evalu-
ated by S-INV and paired-t programs on a PC. The comparison of
plasma profiles and pharmacokinetic parameters of a single dose of
Rabiplus-XT and single dose formulation of Rabium Plus. The Mean
peak plasma levels of the above formulations are shown in Figure 3
and 4.
Figure 3: Bioequivalence of Rabeprazole
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 Varaprasad Bobbarala et al. / Drug Invention Today 2009, 1(1),13-15
Figure 4: Bioequivalence of Itopride
DISCUSSION
Rabeprazole:
The Pharmacokinetic study was performed upto 48 hours.
The mean peak plasma concentration for Rabeprazole in Rabeprazole
sodium 20mg + Itopride HCl 150mg SR capsules was found to be at
Cmax 615.26±23.34 ng/mL and 636.24±14.77 ng/mL for the test and
reference formulations respectively and these concentrations were
achieved at Tmax 4.08±0.15 hrs and 4.17±0.11 hrs for test and refer-
ence formulations respectively. AUC0-t was 4760.91±152.59ng. hr/mL
and 4819.37±116.92ng. hr/mL for the test and reference formulations
respectively. The plasma elimination half-life (T1/2) for the test prepa-
ration was found to be 3.48±0.21 hrs and for the reference prepara-
tion was observed to be 2.93±0.27 hrs. The Kel for the test prepara-
tion was0.2084±0.0141hr-1, while that for the reference preparation
was 0.2527±0.0165 hr-1. There were no statistical significance differ-
ences in Cmax , Tmax and AUC of the test and reference formulations
for Rabeprazole. The bioavailability of the test formulation as judged
from AUC0-t was 98.97% as compared to the reference formulation of
Rabeprazole.
Source of support: Nil, Conflict of interest: None Declared
Drug Invention Today Vol.1.Issue 1.November 2009
Itopride:
The Pharmacokinetic study was performed upto 48 hours.
The mean peak plasma concentration for Itopride in Rabeprazole
sodium 20mg + Itopride HCl 150mg SR capsules was found to be at
Cmax 763.42±20.94 ng/mL and 731.49±17.42 ng/mL for the test and
reference formulations respectively and these concentrations were
achieved at Tmax 7.25±0.13 hrs and 7.17±0.16 hrs for test and refer-
ence formulations respectively. AUC0-t was 10463.51±332.56 ng.hr/
mL and 10508.87±345.22ng. hr/mL for the test and reference formula-
tions respectively. The plasma elimination half-life (T1/2) for the test
preparation was found to be 3.50±0.52 hrs and for the reference prepa-
ration was observed to be 4.48±1.09 hrs. The Kel for the test prepara-
tion was0.2322±0.02241hr-1, while that for the reference preparation
was 0.2138±0.0268 hr-1. There were no statistical significance differ-
ences in Cmax , Tmax and AUC of the test and reference formulations
for Itopride. The bioavailability of the test formulation as judged
from AUC0-t was 99.57% as compared to the reference formulation of
Itopride.
The test formulation of RABIPLUS-XT (Rabeprazole sodium
20mg + Itopride HCl 150 mg SR Capsules), is bioequivalent with the
reference formulation Reference Rabium Plus (Rabeprazole Sodium
20mg + Itopride HCl 150 mg SR Capsules).
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