Ribavarin (RBN) is categorized as an antihepaciviral, nucleoside
(Anti-HCV); it inhibits replication of RNA and DNA viruses. Chemically it is 1-b-D-ribofuranosyl-1H -1,2,4-triazole-3-carboxamide
(Figure 1) . It is a broad spectrum antiviral agent . It is a white, crystalline powder, freely soluble in
water and slightly soluble in anhydrous alcohol. The empirical formula is C8H12N4O5 and the molecular weight is 244.21. It
is used to stop viral RNA synthesis and viral mRNA capping, thus, it is
a nucleoside inhibitor . It is used to treat chronic hepatitis C virus
infection [4-7], Crimean-congo hemorrhagic fever
[8-9] and lassa fever . It is also used for
treatment of respiratory syncytial virus infection in children , also used
as an anti cancer therapy .
Ribavirin nasal and oral inhalation used for treatment of severe lower
respiratory tract infections (i.e., bronchiolitis, pneumonia) caused by RSV in
hospitalized infants and young children. The primary toxicity of ribavirin is
hemolytic anemia [13-15] Reductions in hemoglobin levels occurred within the
first 1-2 weeks of oral therapy. It Absorbed rapidly from GI tract; peak
plasma concentrations achieved within 1–3 hours, bioavailability is 64%. Ribavirin and/or its metabolites accumulate in
The manufacturing of Pharmaceutical
dosage form, from formulation development to finished product, is very
difficult process. This includes interactions of various types between process
conditions and raw materials. These interactions are of great importance for
the process ability and quality of the finished product, so these interactions
should be taken into account earlier, such that to save later loss of time and
In this present study we developed a new formulation of
Ribavirin and all of the physical and chemical parameters were analyzed i.e.
average weight, weight variation, disintegration, dissolution and assay as per
1 Structure of Ribavirin
Materials and reagents
The API and excipients used in the formulation were
Ribavirin, Microcrystalline cellulose PH 102, Lactose monohydrate,
Crocarmellose sodium and Povidone K 30. All of them were of pharmaceutical
grade. For quantitative analysis, the standards used were Ribavirin (Potency
Manufacturing of New Formulations
Accurately weigh all the ingredients of capsules. Pass the
Ribavirin, Microcrystalline cellulose PH 102, Di-calcium phosphate and
Crospovidone through mesh 0.5 mm. add the PVP K-30 in purified water and make a
clear solution. Add this solution to the above sieved materials and made a
granules. Dry the wet mass at 60 °C
and check the moisture which should be not more than 1.00 %. Pass these dried
granules through 1.00 mm mesh and then add Aerosil 200 and Magnesium stearate
after passing through mesh 0.5 mm, then transferred them into suitable food
grade polyethylene bag. Ingredients were mixed by tumbling action in a large
size poly bag for 5 minutes. Finally adjust the capsules shell into the
capsules filling machine carefully so the body adjust in the lower hole. The
head of capsules were unlocked by adjusting the lever. Fill the blended powder
in the body of capsule. Place the upper plate carrying head of the capsules and
locked them on the body with the help of the lever.
After manufacturing of a new formulation of capsules all of
the described parameters were analyzed i.e. average weight, weight variation,
disintegration, dissolution and assay .
Average Weight and Weight Variation
Weight Variation is the in-process test which ensures that
the content of each dosage units is uniform during encapsulation process. 20
capsules of new formulation were accurately weighed on Electronic Balance
Metler AL-204. Calculate the weight of each capsule that must be within
official limits. USP/ BP states that the capsules containing 130-324 mg of the
total weight may be outside ±7.5 % of the average (NMT two capsules out of the
sample) and all must be within 20%.
Disintegration of the new formulation of capsules was tested
by using apparatus Pharmatest Disintegration Tester Series Type PTZ Auto. Place
one capsule in each of the six tubes of the basket and add a disc. Using water
as the immersion fluid, operate the apparatus by maintaining its temperature at
35- 39 °C. Finally at the specified time, pick up the basket from the fluid and
check whether all of the capsules have disintegrated completely. The test is
repeated on 12 extra capsules if one or two capsules fail to disintegrate. The
requirements are met when out of 18 capsules 16 are disintegrated. According to
USP, capsule should disintegrate in not more than thirty minutes.
Dissolution test was carried out on Dissolution Tester AT7
Smart dissolution apparatus. Assemble the equipment, pour 900ml of DI water in
each of the vessel and maintain the temperature of vessels at 37±0.5°C. Place
one capsule of new formulation in an each vessel before rotation. The rotation
of the basket was set at 100 RPM. A sample of 10 ml is withdrawn after 30
minutes. The sample must be taken from a zone midway between the surface of the
dissolution medium and the top of the rotating basket, at least 10 mm from the
vessel wall and not less than 10 mm below the surface. The quantity of
Ribavirin dissolved was determined as specified in USP .
Assay was performed on High Performance Liquid
Chromatography (HPLC) Shimadzu, with Auto sampler. Sample and standard were
prepared as specified in the USP and tested was carried out at UV 207 nm,
Column: 7.8-mm × 15-mm; 7-µm packing L17, Column temperature: 65°C, Flow rate: 1 mLmin-1, and
Injection volume: 10 µL as per USP .
Average Weight and Weight Variation
weight and Wt. variation test of new formulated capsules proved statistically
that all the capsules were in accordance to the BP/USP requirements. (Table 1,
Table 1. Statistical Weight variation
No. of Capsules
Table 2. Weight variation test
Deviation from BP/USP Specification
Table 3. Disintegration test
Deviation from USP Specification
3 min: 18
Table 4. Dissolution Test
in 30 min
Table 5. Assay Test
Disintegration test was conducted on
newly developed formulation and our results were in accordance to BP/USP (Table
In vitro dissolution studies
The percentages of the labeled amount
of Ribavirin dissolved in 30 minutes are shown in Table-4. As per USP official
limits for Ribavirin capsules, dissolved amount of Ribavirin should NLT 80% (Q)
of the labeled amount in 30 minutes and new formulation is under the specified
The assays described are the official
methods upon which the standard of the pharmacopoeia depend. By using HPLC
assay was carried out on the newly developed formulation of Ribavirin Capsules
contain an amount of Ribavirin equivalent to NLT 90.0% and NMT 110.0% each of
the labeled amount of Ribavirin during the study. All capsules have potencies
in accordance of required specification (Table 5).
In the present study new formulation of Ribavarin capsule
was manufactured. All parameters of (average weight, weight variation,
dissolution, disintegration time and assay) of new formulation were tested and
results showed that they are in accordance with the BP/USP limits. In our
trials Disintegration time was found to be 3 minutes 18 seconds which is within
specified BP\USP limit. And the dissolved amount of Ribavirin should NLT 80%
(Q) of the labeled amount in 30 min. Potency of the Ribavirin equivalent to NLT
90.0% and NMT 110.0% each of the labeled amount of Ribavirin. Our research
group has done these types of formulation studies.
All parameters (average weight, weight variation,
dissolution, disintegration time, and assay) of new formulations were tested
and results showed that average weight, weight variation, disintegration time,
dissolution and assay are in accordance with BP/USP limits. The advantage of
this formulation is that this formulation is a quite simple; less time
consuming, economical and achieved all qualitative and quantitative
characteristics which were required by pharmaceutical industry for a dosage
1 USP 37
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