Bioequivalence Study - in Vitro Test

PHARMACEUTICAL DOSSIER
GENERIC SEAL Intramammary Suspension

BIOEQUIVALENCE STUDY – IN VITRO TEST
Issue date: Page 1 from 19

PART III.B
BIOEQUIVALENCE STUDY
CONTENT
III.B.1. Product identification ...................................................................................................3

III.B.1.1. Formulation used in bioequivalence study – in vitro tests .......................................3
III.B.1.1.1. Name of the product: GENERIC SEAL ............................................................3
III.B.1.1.2. Indications for use, specifying the target species ...............................................3
III.B.1.1.3. Amounts to be administered and administration route ......................................3
III.B.1.1.4. Qualitative and Quantitative composition ..........................................................4
III.B.1.1.5. Summary description of the active substance ....................................................4
III.B.2. Bioequivalence study - in vitro tests .............................................................................5
III.B.2.1. Objective of bioequivalence study ............................................................................5
III.B.2.2. Composition of the finished product ........................................................................5
III.B.2.3. Selection of batches ..................................................................................................6
III.B.2.4. Dissolution test..........................................................................................................6
III.B.2.4.1. Apparatus ...........................................................................................................6
III.B.2.4.2. Dissolution medium ...........................................................................................7
III.B.2.4.3. Procedure ............................................................................................................8
III.B.2.4.4. Dissolution profile ..............................................................................................8
III.B.2.4.5. Dissolution test .................................................................................................11
III.B.2.4.6. Acceptance limits .............................................................................................14
III.B.2.4.7. Interpretation ....................................................................................................14
III.B.2.5. Particle size distribution ..........................................................................................14
III.B.2.6. Viscosity of the products ........................................................................................15
III.B.2.7. Pharmaceutical form ...............................................................................................16
III.B.2.8. Results/ Comparative profile ..................................................................................16
III.B.3. CONCLUSIONS: ........................................................................................................18
III.B.4. References: ...................................................................................................................19
III.B.1. Product identification
III.B.1.1. Formulation used in bioequivalence study – in vitro tests
III.B.1.1.1. Name of the product: GENERIC SEAL
GENERIC SEAL Intramammary suspension is a generic product and its development

were conducted against the reference product REFERENCE SEAL Intramammary suspension.
The generic finished product GENERIC SEAL is a suspension for intramammary

administration, containing 2.6 g bismuth subnitrate (BSN), indicated for prevention of new
intramammary infections in dairy cows during the dry period. The product is provided in single-
dosage unit (4 g syringe) and should be administered at drying off at a dose rate of one syringe
per udder quarter.
The intramammary suspension based on BSN, prevent mastitis by providing a physical
barrier (non-pharmacological) against the entry of potential pathogens into the teats and udder
during non-milk producing periods.
III.B.1.1.2. Indications for use, specifying the target species
Prevention of new intramammary infections throughout the dry period.
In cows considered likely to be free of sub-clinical mastitis, GENERIC SEAL can be

used on its own in dry cow management and mastitis control.
Selection of cows for treatment with GENERIC SEAL should be based on veterinary
clinical judgement. Selection criteria may be based on the mastitis and cell count history of
individual cows or recognized tests for the detection of subclinical mastitis or bacteriology
sampling.
As a guide, where individual cell counts are available, cows with an average cell count
less than 200,000 cells/ml may be given GENERIC SEAL. A minor increase in cell count during
the last 4 weeks before drying off is normal and may be ignored.
III.B.1.1.3. Amounts to be administered and administration route
For intramammary infusion only.

Infuse the contents of one syringe of GENERIC SEAL into each udder quarter
immediately after the last milking of the lactation (at drying off.). Do not massage the teat or
udder after infusion of the product.
Care must be taken not to introduce pathogens into the teat in order to reduce the risk of
post-infusion mastitis.
III.B.1.1.4. Qualitative and Quantitative composition
Qualitative and Quantitative composition of GENERIC SEAL in terms of active

substance and excipients per dosage form unit (4000 mg) is presented below:
Table 3.b.1.
QUANTITY FUNCTION
COMPOUND NAME
PER UNIT
Active Substance
Sealing agent
BISMUTH SUBNITRATE, HEAVY 2.6 g
(Non-antibiotic)
Excipients
Liquid paraffin Xg Ointment base
Aluminium di tri stearate Yg Thickening agent
Silica, colloidal anhydrous Zg Anticaking agent
III.B.1.1.5. Summary description of the active substance
Bismuth subnitrate is an inorganic salt of bismuth that does not have an associated
antimicrobial activity, its role being producing a physical barrier against the entry of bacteria
thereby reducing the incidence of new intramammary infections during the dry period.
In general, the composition of bismuth salts is variable. For bismuth subnitrate most often
(Bi2O3)6(N2O5)5(H2O)9 is encountered, and both light and heavy forms exist: the light form is a
mixture of bismuth subnitrate and bismuth subcarbonate, whereas the heavy form consists of
the subnitrate only and contain 72% bismuth.
Properties:
IUPAC Name: Bismuth subnitrate
Chemical Name: Bismuth hydroxide nitrate oxide
Structure: 4[BiNO3(OH)2],BiO(OH) (acc. to Ph. Eur)
Molecular formula: Bi5O22H9N4
Molecular weight: 1462
Classification: Sealing agent (Non-antibiotic)
Pharmacotherapeutic group: Various products for teats and udder
ATCvet code: QG52X
Molecular formula: 4[BiNO3(OH)2],BiO(OH)
CAS Number: 1304-85-4
General properties:
Solubility: Practically insoluble in water and in all common solvents (i.e. ethanol,
ether, chloroform). It dissolves in strong mineral acids with decomposition.
Physical characteristics: White, microcrystalline powder, odourless and tasteless.
Bismuth content: 71.0 – 74.0 %
III.B.2. Bioequivalence study - in vitro tests
III.B.2.1. Objective of bioequivalence study

Bioequivalence studies are often part of applications for generic veterinary medicinal
products to allow bridging of safety and efficacy data associated with a reference veterinary
medicinal product.
Efficacy and tolerance studies may also be waived if the following conditions are
fulfilled: the generic product has the same pharmaceutical form and contains qualitatively and
quantitatively the same active substance(s) (same salts), the excipients of the generic are
qualitatively and quantitatively very similar compared to the reference product, and the
physicochemical properties (e.g. crystalline form, particle size distribution, viscosity, relative
density, dissolution profile) of the generic product are similar to those of the reference product.
In-vitro tests aim to demonstrate, by investigational analytical studies, that the generic
GENERIC SEAL and referenced product (REFERENCE SEAL) are very similar, in order to
provide a biowaiver.
This study in-vitro was performed in accordance with “Guideline on the conduct of
efficacy for intramammary products for use in cattle” (EMA/CVMP/344/1999-Rev.2).
III.B.2.2. Composition of the finished product
Qualitative and Quantitative compositions of both reference and generic products, in

terms of active substance and excipients per dosage form unit (4000 mg) are presented below:
Table 3.b.2.
REFERENCE SEAL GENERIC SEAL
QUANTITY QUANTITY
COMPOUND NAME COMPOUND NAME
PER UNIT PER UNIT
Active Substance
BISMUTH SUBNITRATE, BISMUTH SUBNITRATE,
2.6 g 2.6 g
HEAVY HEAVY
Excipients
Liquid paraffin ~Xg Liquid paraffin Xg
Aluminium di tri stearate ~Yg Aluminium di tri stearate Yg
Silica, colloidal anhydrous ~Zg Silica, colloidal anhydrous Zg
Quantitative composition data of reference product REFERENCE SEAL are taken from
„Codex committee on residues of veterinary drugs in foods”
III.B.2.3. Selection of batches
The batches used in this study are identified in table 3.b.3
Table 3.b.3
REFERENCE PRODUCT -
REFERENCE SEAL
BATCH No. Manufacturing date
ref 1 02.2016
ref 2 02.2016
ref 3 02.2016
GENERIC PRODUCT - GENERIC
SEAL
exp 1 07.2015
exp 2 02.2015
exp 3 04.2015
III.B.2.4. Dissolution test
In vitro dissolution studies are used to provide evidence of the similarity of the quality of
the generic and reference products.
The similarity acceptance limits should not be greater than a 10% difference. In addition,
the dissolution variability of the test and reference product data should also be similar, however,
a lower variability of the test product may be acceptable.
III.B.2.4.1. Apparatus
For dissolution test performing, a Rotating Paddle type stirrer dissolution apparatus was
used, shown in Figure 1.
The assembly consists of the following:

 a vessel made of inert, transparent material;
 a motor
 a drive shaft
 a rotating paddle, formed from a blade and a shaft, used as a stirring element.
The shaft is positioned so that its axis is not more than 2 mm from the vertical axis of the
vessel, at any point, and rotate smoothly without significant wobble. The distance of 25 ± 2 mm
between the bottom of the blade and the inside bottom of the vessel is maintained during the
test.
Figure 1
III.B.2.4.2. Dissolution medium
The products intended to be tested are intramammary seals, of which therapeutic purpose
is based on their physical properties not to disintegrate at the site of administration throughout
the treatment, the removal being a physical one (by hand during milking or by the calf ingestion).
Therefore, the dissolution medium will be chosen to encourage their dissolution; the only
purpose of this study it is creating ability to compare the generic and the reference products from
this point of view
Also, the dissolution medium is chosen thus not influence the results obtained, taking into
account the method of analysis.
Considering the information above, it was used a solution containing 200 ml water and
tert-butyl methyl ether (10 ml), to facilitate the dissolution process. Dissolution profiles are
compared using dissolution temperature of 37°C, that represent normal physiologically body
temperature being considered relevant for experimental.
Working conditions
 Stirrer dissolution apparatus: Rotating Paddle;
 Rotational speed: 200 rpm;

 Temperature of dissolution medium: 37 ± 1°C;
 Amount of the product undergoing the dissolution test: 4000 mg (a unit dose);
 Volume of dissolution medium: 210 ml (mixture of 200 ml water and 10 ml tert-
butyl methyl ether).
Mentions:
Replace the aliquots withdrawn for analysis, at any sampling times, with equal volumes
of fresh dissolution medium at 37 ± 1°C.
Keep the vessel covered for the duration of the test and verify the temperature of the
medium at suitable times.
III.B.2.4.3. Procedure
 Place the stated volume of the dissolution medium in the vessel of the apparatus;
 Assemble the apparatus, equilibrate the dissolution medium to 37 ± 1°C;
 Discard the total content from the individual container in the dissolution medium;
 Operate the apparatus at the specified rate (200 rpm);
 At each of the time stated, withdraw a sample from a zone midway between the
surface of the dissolution medium and the top of the rotating blade, not less than
1 cm from the vessel wall.
III.B.2.4.4. Dissolution profile
The dissolution profile aims to establish the time required to release the active substance
from the oily matrix to a level above 85%.
Where more than 85% of the drug is dissolved within 15 minutes, dissolution profiles
may be accepted as similar based on a single point.
In case more than 85% is not dissolved at 15 minutes, at least three-time points are
required. Thus, the following sampling time-points were chosen for both reference and generic
products: 15 min.; 30 min; 40 min and 60 min.
Recoveries will be calculated for each sampling time and the release trend of the active
substance over time will be observed as well as the similarity between the generic product and
the reference product.
Results of time dissolution profile for GENERIC SEAL are given in table 3.b.4.
Table 3.b.4.
GENERIC SEAL
Sampling Volume of Sampled Titration
Factor Recovery
time dissolution volume volume Cp Ct
%
medium (ml) (Vsample) ml (ml)
15 min 200 20 1.2 1.01934 357.66 2600 13.76
30 min 200 20 5.9 1.01934 1758.525 2600 67.636
40 min 200 20 7.4 1.01934 2205.607 2600 84.83
60 min 200 20 8.1 1.01934 2414.246 2600 92.86
Results of time dissolution profile for REFERENCE SEAL are given in table 3.b.5.
Table 3.b.5.
REFERENCE SEAL
Sampling Volume of Sampled Titration
time dissolution volume volume Factor Cp Ct Recovery %
medium (ml) (Vsample) ml (ml)
15 min 200 20 1.1 1.01235 325.61 2600 12.52
30 min 200 20 6.2 1.01235 1835.27 2600 70.59
40 min 200 20 7.5 1.01235 2220.08 2600 85.39
60 min 200 20 8.2 1.01235 2427.29 2600 93.36
Where:
Vsample = volume of sample withdrawn used in titration;
CP = amount of dissolved active substance, expressed as content of active substance per
unit dose (concentration obtained);
Ct = amount of the labelled content (theoretical concentration).
The results presented above, show that the active substance from both Generic seal (generic)
and Reference seal (reference) is released quite hardly in the dissolution medium. Within 40
minutes of contact with the medium and under continuous stirring, approximately 85% of the
bismuth subnitrate of both products is released from the matrix of excipients and will be present
as a suspension in the aqueous dissolution medium.
The similarity of dissolution profiles for both products can be seen from the graphical
representation below:
Graph 1
Dissolution time for REFERENCE SEAL
100
90
80
70
RECOVERY %
60
50
40
30
20
10 Series1
0
15 30 40 60
TIME (minutes)
Graph 2
Dissolution time for Generic seal
100
GENERIC SEAL
90
80
70
60
50
40
30
20
10 Series1
0
15 30 40 60
TIME (minutes)
To support statistically, the results obtained, a larger number of tests will be performed
for each batch of the product, using three lots of both products studied.
The sampling time point is 40 minutes, this being the time at which the active substance
is released more than 85% from oily matrix in the dissolution medium.
III.B.2.4.5. Dissolution test
Before starting the actual tests, it was performed the assay for each batch of product.
The results obtained for assay of the active substance in finished product for each batch
subjected to the dissolution test are shown in table 3.b.6.
Table 3.b.6.
GENERIC REFERENC
LOT mg BSN/ 4000 mg

E SEAL
ref 1 2623
ref 2 2579
ref 3 2613
LOT mg BSN/ 4000 mg
SEAL
exp 01 2582
exp 02 2619
exp 03 2608
A number of 12 tests for each sample of the three batches of the two products tested is
considered sufficient to show statistically that the time and mode of dissolution of the active
substances is robust.
Working conditions
 Stirrer dissolution apparatus: Rotating Paddle;
 Rotational speed: 200 rpm;
 Temperature of dissolution medium: 37 ± 1°C;
 Amount of the product undergoing the dissolution test: 4000 mg (a unit dose);
 Volume of dissolution medium: 210 ml (mixture of 200 ml water and 10 ml tert-
butyl methyl ether).
 Number of samples: 12 per each batch
 Sampling time-point: 40 minutes
Results of dissolution test for GENERIC SEAL are given in table 3.b.7.
Table 3.b.7.
GENERIC SEAL Bach exp 1 Bach exp 2 Bach exp 3
BSN/ Nr. BSN/unit dose BSN/unit dose BSN/unit dose
Recovery Recovery Recovery
API unit dose sam obtained obtained obtained
% % %
(ct) ple (cp) (cp) (cp)
2600 1 2184.74 84.03 2148.98 82.65 2253.30 86.67
BSMUTH SUBNITRATE, HEAVY
2600 2 2241.37 86.21 2211.57 85.06 2202.63 84.72

2600 3 2217.53 85.29 2178.78 83.80 2268.20 87.24
2600 4 2280.12 87.70 2250.32 86.55 2172.82 83.57
2600 5 2303.97 88.61 2274.16 87.47 2214.55 85.17
2600 6 2238.39 86.09 2238.39 86.09 2199.65 84.60
2600 7 2232.43 85.86 2172.82 83.57 2262.24 87.01
2600 8 2199.65 84.60 2190.70 84.26 2265.22 87.12
2600 9 2256.28 86.78 2193.68 84.37 2160.90 83.11
2600 10 2274.16 87.47 2184.74 84.03 2196.67 84.49
2600 11 2265.22 87.12 2265.22 87.12 2271.18 87.35
2600 12 2235.41 85.98 2262.24 87.01 2169.84 82.77
Average 86.31 Average 85.165 Average 85.32
SD 1.314 SD 1.618 SD 1.6997
RSD 1.522 RSD 0.8997 RSD 1.992
Results of dissolution test for GENERIC SEAL are given in table 3.b.8.
Table 3.b.8.
REFERENCE SEAL Bach ref 1 Bach ref 2 Bach ref 3
BSN/ Nr. BSN/unit dose BSN/unit dose BSN/unit dose
Recovery Recovery Recovery
API unit dose sam obtained
%
obtained
%
obtained
%
(ct) ple (cp) (cp) (cp)
2600 1 2140.16 82.31 2175.68 83.68 2169.76 83.45
BSMUTH SUBNITRATE, HEAVY
2600 2 2228.96 85.73 2223.04 85.50 2193.44 84.36

2600 3 2116.48 81.40 2157.92 83.00 2154.96 82.88
2600 4 2163.84 83.22 2246.72 86.41 2223.04 85.50
2600 5 2178.64 83.79 2211.20 85.05 2255.60 86.75
2600 6 2152.00 82.77 2163.84 83.22 2187.52 84.14
2600 7 2169.76 83.45 2143.12 82.43 2175.68 83.68
2600 8 2205.28 84.82 2228.96 85.73 2178.64 83.79
2600 9 2202.32 84.70 2249.68 86.53 2154.96 82.88
2600 10 2226.00 85.62 2258.56 86.87 2169.76 83.45
2600 11 2255.60 86.75 2279.29 87.66 2246.72 86.41
2600 12 2175.68 83.68 2220.08 85.39 2205.28 84.82
Average 84.02 Average 85.12 Average 84.34
SD 1.552 SD 1.684 SD 1.2897
RSD 1.847 RSD 1.9784 RSD 1.529
As can be observed, the rates of release of the active substance under identical
dissolution conditions (temperature, time) are similar between reference and generic products.
Thus, an average rate of 85.598% of REFERENCE SEAL and 84.495% of GENERIC
SEAL product were obtained. The coefficient of variation (RSD%) of the values with which
dissolution rates were calculated is satisfactory (RSD <2%), so we can state that the test is
accurate and repeatable, and it is also comparable between the two products which are the
subject of the study.
The way in which the release of the active substance from the two finished products
(generic vs. reference) was achieved during the 40 minutes of contact with the dissolution
medium is represented by graph 3, where the similarity between them can be observed again.
Graph 3
Reference Seal Generic Seal
90
DISOLUTION TEST Reference Seal vs Generic Seal
88.61
88 87.7 87.47 87.47 87.66

87.12 87.12 87.24 87.12 87.35
86.78 86.75 86.87 87.01
86.67 86.75
87.01
86.55
86.41 86.53 86.41
86.21 86.09 85.98 86.09
85.73 85.86 85.73
86 85.62 85.5 85.39 85.5
85.29 85.06 85.17
84.8284.7 85.05 84.82
84.6 84.72 84.6 84.49
RECOVERY %
84.37
84.26 84.36 84.14
84.03 84.03
84
83.79
83.45 83.68 83.8
83.68 83.57 83.45 83.57 83.79
83.68 83.45
83.22 83 83.22 83.11
82.77 82.65 82.88 82.88 82.77
82.31 82.43
82 81.4
80
78
76
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36
SAMPLE NUMBER
III.B.2.4.6. Acceptance limits

The similarity acceptance limits should not be greater than a 10% difference. In addition,
the dissolution variability of the test and reference product data should also be similar, however,
a lower variability of the test product may be acceptable.
III.B.2.4.7. Interpretation
Following the dissolution test described herein, it has been found that:
 the difference between the results obtained for both the generic product and the reference
product is not more than 10%;
 it is released into the dissolution medium ~ 85% of the amount of active substance
contained in the finished product, under identical work conditions, within 40 minutes,
both for the generic product and for the reference product;
 the difference between the results obtained for the reference and the generic products is
not greater than the variation of the values obtained between the batches of the reference
product.
III.B.2.5. Particle size distribution
The rate of dissolution of the active substance is proportional to the surface area of the active
substance particles exposed to the dissolution medium. According to the guideline for
intramammary preparations "EMA/CVMP/344/1999-Rev.2-Chapter 6," the particle size
distribution of the active substance in the generic product should be similar to the particle size
distribution of the reference product.
Test method: Particle size analysis by Laser Light Diffraction
Equipment: Mastersizer 2000
Measuring interval: 0.02 μm – 2000 μm
table 2. b.9- Particle size distribution test results

Bach Particle size distribution
Product tested d (0.1) d (0.5) d (0.9)
REFERENCE ref 1 0.255 µm 0.889 µm 5.657 µm
SEAL ref 2 0.556 µm 1.959 µm 4.094 µm
-reference ref 3 0.391 µm 2.135 µm 4.151 µm
product- mean 0.4 µm 1.661 µm 4.634 µm
GENERIC exp 1 0.316 µm 1.259 µm 3.374 µm
SEAL exp 2 0.385 µm 2.332 µm 4.454 µm
-generic exp 3 0.257 µm 0.879 µm 5.757 µm
product- mean 0.319 µm 1.49 µm 4.528 µm
D-values can be thought of as the diameter of the sphere which divides the sample's
mass into a specified percentage when the particles are arranged on an ascending mass basis.
d (v, 0.5), d (v, 0.1) and D(d, 0.9) are standard “percentile” readings from the
analysis.
d (0.1)- is the size of particle for which 10% of the sample is below this size.
d (0.5) is the size of particle at which 50% of the sample is smaller and 50% is larger
than this size. This value is also known as the Mass median diameter (MMD).
d (0.9)- gives a size of particle for which 90% of the sample is below this size.
As can be seen in table 2. b.9, 90% of the sample mass contains particles smaller than
16 μm (15.657 μm for REFERENCE SEAL and 5.757 μm for GENERIC SEAL respectively);
50% of the sample mass is composed of particles whose dimensions are less than 3 μm (2.135
μm for REFERENCE SEAL and 2.331 μm for GENERIC SEAL); 10% of the sample mass
has particle diameter <1 μm (0.556 μm for REFERENCE SEAL and 0.385 μm for GENERIC
SEAL);
III.B.2.6. Viscosity of the products
According to the guideline for intramammary preparations "EMA/CVMP/344/1999-Rev.2-

Chapter 6," the viscosity of the generic product (GENERIC SEAL) should be similar to the
viscosity of the reference product (REFERENCE SEAL).
Viscosities of products are compared using a measurement temperature of 37°C, that
represent normal physiologically body temperature being considered relevant for experimental.
Viscosity is measured using a spindle viscometer, accordingly to the method described
in EurPh., monograph 20210 (SPINDLE VISCOMETER). The principle of the method is to
measure the force acting on a rotor (torque) when it rotates at a constant angular velocity
(rotational speed) in a liquid
Equipment’s and materials
- Viscometer Fungilab
- Berzelius beaker
Procedure
- the measurement is performed at 37°C temperature;
- fill the beaker with a sufficient quantity of the suspension to be examined, ensuring the
level that the spindle is immersed to the level indicated on it;
- set the rotational speed of the spindle;
- monitor 1-2 minutes until stabilization of the viscosity value (Pa·s).
table 2. b.10- Results for viscosity

Vâscozitate (Pa·s)
Product LOT testat (37°C)
REFERENCE ref 1 63
SEAL ref 2 65
-reference ref 3 55
product- mean 61
GENERIC exp 1 67
SEAL exp 2 61
-generic exp 3 58
product- mean 62
Conclusions: The rheological profile of the generic GENERIC SEAL product is almost identical
to that of the reference product, REFERENCE SEAL.
III.B.2.7. Pharmaceutical form
Pharmaceutical form is the physical characteristics of the combination of active substance

and excipients (non-active ingredients) forming a medicinal product.
In accordance with, Guideline on the conduct of efficacy studies for intramammary products for
use in cattle”, EMA/CVMP/344/1999-Rev.2, the pharmaceutical form should be the same,
and the appearance of the generic and the reference products should be similar.
Both, generic product Generic seal and reference product Reference seal are
intramammary suspensions as greyish white, smooth, unctuous cream used as intramammary
infusion only, for prevention of new intramammary infections throughout the dry period in
cattle.
Conclusions:
Generic product Generic seal and reference product Reference seal have the same
pharmaceutical form.
III.B.2.8. Results/ Comparative profile

TEST REFERENCE SEAL - reference product GENERIC SEAL - generic product-

BATCH BATCH
ref 1 ref 2 ref 3 exp 1 exp 2 exp 3
pharmaceutical form intramammary intramammary intramammary intramammary intramammary intramammary
suspension suspension suspension suspension suspension suspension
Appearance smooth, smooth, smooth, smooth, smooth, smooth,
unctuous cream unctuous cream unctuous cream unctuous cream unctuous cream unctuous cream
Colour greyish white greyish white greyish white greyish white greyish white greyish white
bismuth subnitrate content mg/4000 mg 2614 2591 2625 2625 2613 2589
Dissolution rate bismuth subnitrate % 84.02 85.12 84.82 86.31 85.165 85.32
Particle size dv (0.1): max. 2 μm 0.255 0.556 0.391 0.316 0.385 0.257
distribution dv (0.5): max. 10 μm 0.889 1.959 2.135 1.259 2.332 0.879
dv (0.9): max. 50 μm 5.657 4.094 4.151 3.374 4.454 5.757
viscosity Pa·s 37±1°C 63 65 55 67 61 58
composition active substance bismuth subnitrate heavy: 2600 mg bismuth subnitrate heavy :2600 mg
mg compound/ excipients Liquid paraffin: ~x mg Liquid paraffin: x mg
4000 mg product Aluminium di tri stearate: ~ y mg Aluminium di tri stearate: y mg
Silica, colloidal anhydrous: ~ z mg Silica, colloidal anhydrous: z mg
III.B.3. CONCLUSIONS:
Following a study to demonstrate the in vitro bioequivalence of the GENERIC

SEAL generic product as compared to the reference product REFERENCE SEAL, as set
out in the Annex to the Guidance on the conduct of efficacy studies for intramammary
products for use in bovine animals, Guideline on the conduct of efficacy studies for
intramammary products for use in cattle-EMA / CVMP / 344/1999-Rev.2-Chapter 6 "It
was noted that:
- Generic product Generic seal and reference product Reference seal, based on present
study showed to have the same qualitative composition and approximately the same
quantitative composition.
- the pharmaceutical form is identical for both
- their rheological properties are very similar with values of very close viscosities
- their physical properties are identical (appearance, colour, density),
- the dissolution rate of each active substance in the finished product is very similar
between the two products studied,
- the particle size distribution of the active substance in the GENERIC SEAL generic
product is similar to the particle size distribution of the active substance in the reference
product, REFERENCE SEAL.
As the admissibility conditions are met, it can be said that the generic product
GENERIC SEAL is equivalent to the reference product REFERENCE SEAL
III.B.4. References:
1. Guideline on the conduct of efficacy studies for intramammary products for use in
cattle-EMA/CVMP/344/1999-Rev.2-capitolul 6
2. https://www.drugs.com/international/bismuth-subnitrate.html
3. https://pubchem.ncbi.nlm.nih.gov/compound/16717608#section=Top
4. EMEA/MRL/201/97-FINAL
5. EMEA/MRL/705/99-FINAL
6. JOINT FAO/WHO FOOD STANDARDS PROGRAMME CODEX COMMITTEE ON
RESIDUES OF VETERINARY DRUGS IN FOODS, Twenty-third Session, Houston,
Texas, United States of America, 17 – 21 October 2016-PRIORITY LIST
NOMINATION- Submitted by New Zealand,758,
7. JOINT FAO/WHO FOOD STANDARDS PROGRAMME CODEX COMMITTEE ON
RESIDUES OF VETERINARY DRUGS IN FOODS, Twenty-third Session, Houston,
Texas, United States of America, 17 – 21 October 2016-PRIORITY LIST
NOMINATION- Submitted by New Zealand,758,
8. Complexometric Titration -James P. Leyda- Lederle Laboratories Pearl River, New
York
9. The Veterinary Formulary -sixth edition- Yolande Bishop
10. European Pharmacopoeia, Edition IX

Bioequivalence Study - in Vitro Test

Încărcat de

Informații document

Titlu original

Drepturi de autor

Formate disponibile

Partajați acest document

Partajați sau inserați document

Opțiuni de partajare

Vi se pare util acest document?

Este necorespunzător acest conținut?

Drepturi de autor:

Formate disponibile

Bioequivalence Study - in Vitro Test

Încărcat de

Drepturi de autor:

Formate disponibile

PHARMACEUTICAL DOSSIER

GENERIC SEAL Intramammary Suspension

Issue date: Page 1 from 19

BIOEQUIVALENCE STUDY – IN VITRO TEST

Issue date: Page 2 from 19

III.B.1. Product identification ...................................................................................................3

Issue date: Page 3 from 19

III.B.1. Product identification

III.B.1.1. Formulation used in bioequivalence study – in vitro tests

III.B.1.1.1. Name of the product: GENERIC SEAL

GENERIC SEAL Intramammary suspension is a generic product and its development

The generic finished product GENERIC SEAL is a suspension for intramammary

III.B.1.1.2. Indications for use, specifying the target species

Prevention of new intramammary infections throughout the dry period.

In cows considered likely to be free of sub-clinical mastitis, GENERIC SEAL can be

III.B.1.1.3. Amounts to be administered and administration route

For intramammary infusion only.

Issue date: Page 4 from 19

III.B.1.1.4. Qualitative and Quantitative composition

Qualitative and Quantitative composition of GENERIC SEAL in terms of active

III.B.1.1.5. Summary description of the active substance

Issue date: Page 5 from 19

III.B.2. Bioequivalence study - in vitro tests

III.B.2.1. Objective of bioequivalence study

III.B.2.2. Composition of the finished product

Qualitative and Quantitative compositions of both reference and generic products, in

Issue date: Page 6 from 19

III.B.2.3. Selection of batches

The batches used in this study are identified in table 3.b.3

III.B.2.4. Dissolution test

The assembly consists of the following:

Issue date: Page 7 from 19

III.B.2.4.2. Dissolution medium

Issue date: Page 8 from 19

 Rotational speed: 200 rpm;

III.B.2.4.4. Dissolution profile

Issue date: Page 9 from 19

Issue date: Page 10 from 19

Dissolution time for REFERENCE SEAL

Dissolution time for Generic seal

Issue date: Page 11 from 19

III.B.2.4.5. Dissolution test

LOT mg BSN/ 4000 mg

Issue date: Page 12 from 19

2600 2 2241.37 86.21 2211.57 85.06 2202.63 84.72

2600 2 2228.96 85.73 2223.04 85.50 2193.44 84.36

Issue date: Page 13 from 19

88 87.7 87.47 87.47 87.66

Issue date: Page 14 from 19

III.B.2.4.6. Acceptance limits

table 2. b.9- Particle size distribution test results

Issue date: Page 15 from 19

III.B.2.6. Viscosity of the products

According to the guideline for intramammary preparations "EMA/CVMP/344/1999-Rev.2-

Issue date: Page 16 from 19

table 2. b.10- Results for viscosity

III.B.2.7. Pharmaceutical form

Pharmaceutical form is the physical characteristics of the combination of active substance

III.B.2.8. Results/ Comparative profile

Issue date: Page 17 from 19

TEST REFERENCE SEAL - reference product GENERIC SEAL - generic product-

Issue date: Page 18 from 19

Following a study to demonstrate the in vitro bioequivalence of the GENERIC