6.1 characterization of tablets -...
Post on 06-Feb-2018
218 Views
Preview:
TRANSCRIPT
105
6.1 CHARACTERIZATION OF TABLETS
All the prepared formulations were subjected for following
evaluation parameters and obtained results were showed in Tables 6.3 &
6.4.
6.1.1 Description (Size, Shape, Color)
The tablets were looking good and non sticky. The color, shape of
tablets was analyzed with naked eye. The diameter and thickness of
tablets was performed on 20 tablets from each formulation. Digital
vernier caliper was used for the study, which permits accurate
measurements and provides information of the variation between tablets.
6.2 WEIGHT VARIATION 99,100,102
Weight variation was carried out to ensure that, each of tablets
contains the proper amount of drug. The test was carried out by
weighing the 20 tablets individually using analytical balance, then
calculating the average weight, and comparing the individual tablet
weights to the average.
The percentage of weight variation is calculated by using the
following formula.
Table 6.1 Weight Variation Limits for Tablets (I.P)
S. No. Average weight of tablets (mg) Maximum percentage
difference allowed
1 130 or less ±10.0
2 130-324 ±7.50
3 More than 324 ±5.0
106
6.3 HARDNESS 99,100,102
The resistance of tablets to capping, abrasion or breakage under
conditions of storage, transportation and handling before usage depends
on its hardness. Tablet hardness is defined as the load required crushing
or fracture a tablet placed on its edge. Sometime it is also termed as
tablet crushing strength. The hardness test was performed using
Monsant type (Make: Singhla) hardness tester. The instrument measures
the force required to break the tablet when the force generated by anvils
to the tablet. The tablet was placed between two anvils; force applied to
the anvils, and the crushing strength that just causes the tablet to break
was recorded. The crushing strength test was performed on 20 tablets
from each formulation.
6.4 FRIABILITY 99,100,102
For each formulation, the friability of 20 tablets was determined
using Roche type friabilator. 20 tablets from each formulation were
weighed and tested at a speed of 25 rpm for 4 min. After removing of
dusts, tablets were re-weighed and friability percentage was calculated
using the following equation.
6.5 DISINTEGRATION 99,100,118
The disintegration apparatus, described in I.P was used for the
study. It contains 2 basket rack assembly. Each basket rack assembly
consists of 6 glass tubes that are 3 inches long, open at the top and held
107
against 10 mesh screen at the bottom. Each tablet was placed in each
tube, and the basket rack was positioned in 1-L beaker of distilled water.
The 37±2°C temperature was maintained throughout the study.
6.6 DISSOLUTION 99,100,102
The In-vitro dissolution study of tablets for best formulation was
determined by using USP Type 2 (paddle type) dissolution apparatus. The
test was performed in 200 ml of distilled water at 50 rpm maintained at
37± 0.5°C. The specific amount of samples were withdrawn at
predetermined time intervals for period of 2 hours (15, 30, 45, 60, 90,
120 min.) and replaced with the equal volume of the same dissolution
medium. The samples were filtered through 0.2μm membrane filter.
Amount of catechins, sodium selenite released from the tablets were
analyzed by HPTLC and LC ICP MS methods respectively.
6.6.1 Methodology of Analysis of Sodium Selenite Compound103 LC-ICP-MS technique is used to qualitate and quantitates sodium
selenite. The coupling of LC to ICP-MS is a relatively straight forward
task, and gives an accurate result. LC column is simply connected to the
nebulizer of the ICP-MS by a piece of PEEK™ tubing. When using the
Varian 820-MS equipped with the collisional reaction interface (CRI), it is
possible to measure the sodium selenate at m/z 80.
The LC-ICP-MS system used for this study consisted of a Varian
ProStar 230 Tertiary Solvent Delivery Module, a Varian ProStar 410
biocompatible auto sampler with a 200 µL sample loop, and a Hamilton
108
PRP-X100 (4.1 mm x 250 mm, 10 µm) anion exchange column and the
Varian 820-MS. The Varian 820-MS setup, inducing the CRI gas flow,
was optimized using the ICP-MS Expert software auto optimization
routine. The Varian 820-MS was operated in high sensitivity mode, and
was connected to the LC column using a 20 cm length of PEEK tubing
(0.25 mm ID). Table 6.2 provides detailed listings of the ICP-MS and LC
conditions.
6.6.2 Construction of Standard Plots For Sodium Selenite Marker
Compound
Preparation of Stock solution: Ten mg of marker sodium selenite
was weighed carefully and transferred into 100 ml standard flask. This is
diluted up to the mark with de ionized water. 10 ml of this solution was
diluted to 100 ml with de ionized water, by using standard flask. This
solution was considered as stock solution and its concentration is
10µg/ml.
Preparation of serial dilutions: One ml of stock solution is diluted
up to 1 L with water to give concentration 10µg/l. In the same passion, a
series of solutions, having 20, 30, 40, 50, 60, 70, 80, 90 & 100 µg
sodium selenite per L, were prepared. All the solutions were injected into
LC, and obtained curves were recorded. A 10 point calibration curve of
the purchased sodium selenite was constructed, by taking
concentrations on X axis, and peak areas on Y axis. The prepared
calibration curve was used to determine the amount of sodium selenite
109
present in the dissolution samples. Table 6.5 gives the data of Sodium
Selenite calibration curve. Fig. 6.1 shows calibration curve of Sodium
Selenite.
Table 6.2 Details of LC-ICP-MS Conditions
S. No. Parameter Value
Flow Parameters (L/min)
1. Plasma flow 18.0
2. Auxiliary flow 1.65
3. Sheath gas 0.24
4. Nebulizer flow 0.98
Other
5. RF power (kW) 1.40
6. Pump rate (rpm) 25
Ion optics (volts)
7. First extraction lens -500
8. Second extraction lens -879
9. Third extraction lens -643
10. Corner lens -763
11. Mirror lens left 40
12. Mirror lens right 37
13. Mirror lens bottom 40
14. Entrance lens 1
15. Fringe bias -3.9
16. Entrance plate -30
CRI (mL/min)
17. Skimmer gas source H2
18. Skimmer flow 70
LC Operating Conditions
19. Parameter Settings
20. Mobile Phase 60 m M Ammonium nitrate
21. Flow rate 1.0 mL/min
22. Run time 7 min
Column Anion exchange, Hamilton PRP-X 100,
4.1 mm ID x 250 mm, 10 µm
23. Column Temperature Ambient
24. Sample injection volume 200 µL
25. Detection Varian 820-MS
110
6.6.3 Determination of Amount of Catechins Released
From Tablet 96, 97, 104, 105
The 2 ml sample of dissolution media was withdrawn and replaced
with the equal volume of the fresh dissolution medium. The collected
samples were filtered through 0.2μm membrane filter. 5µl of this filtered
solution was applied per each spot. This test was done in triplicate.
The amount of catechins released from tablet was analyzed with
help of standard plots. The following formula, generated from calibration
curve was used to calculate the amount of catechins. Fig. 6.2 to 6.7
shows HPTLC chromatograms of tablet dissolution samples. The amount
of all the catechins released from tablet was given in Table 6.6 to 6.12
111
6.6.4 Determination of Amount of Sodium Selenite Released From
Tablet
The amount of Sodium Selenite released at corresponding time
intervals was calculated by using the following formula.
Where,
CCC = Concentration obtained from Calibration Curve
DF = Dilution Factor
VDM = Volume of Dissolution Medium
CF = Conversion Factor
The amount of Sodium Selenite released from tablet during
dissolution is showed in Table 6.13. Fig. 6.8 to 6.13 shows LC ICP MS
chromatograms of tablet dissolution samples.
112
6.7 Results and Discussion
The results obtained from above tests were showed and described
below.
Table 6.3 Results of Description of Tablets
S. No. Parameter Result
1 Color Greenish Brown
2 Shape Round, Biconvex
3 Odor Characteristic
4 Size in mm
i. Thickness 5.15±0.12 mm
ii. Diameter 8.65±0.23 mm
Table 6.4 Results of Physico-Chemical Characterization of Tablets
Formulation
Code
Average
Weight
(mg)
Hardness
(kg/cm2)
Friability
(%)
Disintegration Time
GST-1 400±0.20 3.10±0.08 0.69±0.05 30 min & 20 sec
GST-2 400±0.12 4.03±0.14 0.50±0.05 28 min & 24 sec
GST-3 400±0.11 4.21±0.09 0.53±0.03 25 min & 40 sec
GST-4 400±0.09 4.06±0.11 0.84±0.01 20 min & 12 sec
GST-5 400±0.12 4.13±0.12 0.98±0.00 16 min & 14 sec
GST-6 400±0.11 4.03±0.17 0.44±0.05 13 min & 35 sec
GST-7 400±0.07 3.17±0.04 0.84±0.07 110 min & 10 sec
GST-8 400±0.06 3.34±0.33 0.72±0.08 104 min & 12 sec
GST-9 400±0.06 4.13±0.12 0.98±0.11 100 min & 54 sec
GST-10 400±0.09 3.20±0.24 0.63±0.04 092 min & 33 sec
GST-11 400±0.12 3.17±0.07 0.84±0.05 082 min & 23 sec
GST-12 400±0.10 3.14±0.03 0.70±0.09 079 min & 43 sec
113
The maximum weight variation obtained was ± 0.20%, which falls
within the acceptable weight variation range of ± 5%. Hence all the
tablets passed the weight variation test. Hardness for tablets was in the
range of 4.0 to 4.2 kg/cm2, which falls above the limit of not less than
3.0 kg/cm2. None of the tablets showed friability value more than 0.87%
which is less than ideal limit 1%. Out of all the formulations, formulation
GST-6 has passed the disintegration test by showing Thirteen minutes
and Thirty five seconds, which is less than ideal limit Fifteen minutes.
The acceptance criteria of dissolution rates of herbal medicinal product
are as follows.106, 107
1) Europian Pharmacopoeia Limits: 50-85% release within 30-60 min.,
and/or, Mean 75% release within 45 min.
2) F.I.P Limits: For rapid release = >80% within 20-30 min., for non-rapid
release = 20-30% within 60-120 min.
The best formulation has showed release of maximum amount
(around 100%) of catechins and sodium selenite within one hour. Hence
the formulation was passed the dissolution test.
114
Table 6.5 Data of Sodium Selenite Calibration Curve
S. No. Concentration of Sodium Selenite (µg/L) Peak Area
1 0 0
2 10 1600
3 20 3298
4 30 4952
5 40 6621
6 50 8300
7 60 9950
8 70 11712
9 80 13343
10 90 14950
11 100 16519
Calibration Equation y = 166.0x + 0.0
Regression Coefficient R² = 0.999
Fig. 6.1 Calibration Curve of Sodium Selenite.
115
Fig. 6.2 HPTLC Chromatogram of 15 min. Dissolution Sample for
Catechins
116
Fig. 6.3 HPTLC Chromatogram of 30 min. Dissolution Sample for
Catechins
117
Fig. 6.4 HPTLC Chromatogram of 45 min. Dissolution Sample for
Catechins
118
Fig. 6.5 HPTLC Chromatogram of 60 min. Dissolution Sample for
Catechins
119
Fig.6.6 HPTLC Chromatogram of 90 min. Dissolution Sample for
Catechins
120
Fig. 6.7 HPTLC Chromatogram of 120 min. Dissolution Sample for
Catechins
121
Fig. 6.8 LC ICP MS Chromatogram of 15 min. Dissolution Sample for
Sodium Selenite
Fig. 6.9 LC ICP MS Chromatogram of 30 min. Dissolution Sample for
Sodium Selenite
122
Fig. 6.10 LC ICP MS Chromatogram of 45 min. Dissolution Sample for
Sodium Selenite
Fig. 6.11 LC ICP MS Chromatogram of 60 min. Dissolution Sample for
Sodium Selenite
123
Fig. 6.12 LC ICP MS Chromatogram of 90 min. Dissolution Sample for
Sodium Selenite
Fig. 6.13 LC ICP MS Chromatogram of 120 min. Dissolution Sample for
Sodium Selenite
124
Table 6.6 Amount of Catechin Released from Tablet during Dissolution of tablet.
S. No. Sampling
Time
(min)
Peak
Area
Concentration
obtained from
Calibration Curve
Amount of Catechin
Released in 200ml
of dissolution media
(mg)
% of
Catechin
Released
1 15 285.6 15.60 0.624 48.00
2 30 399.8 21.25 0.850 65.40
3 45 500.3 26.22 1.049 80.72
4 60 601.3 31.22 1.249 96.09
5 90 611.0 31.70 1.268 97.56
6 120 619.1 32.10 1.284 98.79
Table 6.7 Amount of EC Released from Tablet during Dissolution of tablet.
S. No. Sampling
Time
(min)
Peak
Area
Concentration
obtained from
Calibration Curve
Amount of EC
Released in 200ml
of dissolution
media (mg)
% of EC
Released
1 15 1977.4 097.25 3.89 46.72
2 30 2679.7 132.00 5.28 63.54
3 45 3649.8 180.00 7.20 86.49
4 60 4109.6 202.75 8.11 97.43
5 90 4129.8 203.75 8.15 97.99
6 120 4215.7 208.00 8.32 100.00
Table 6.8 Amount of GC Released from Tablet during Dissolution of tablet.
S.
No.
Sampling
Time
(min)
Peak
Area
Concentration
obtained from
Calibration Curve
Amount of GC
Released in 200ml
of dissolution
media (mg)
% of GC
Released
1 15 0544.8 027.00 1.80 43.28
2 30 1310.9 065.75 2.63 63.19
3 45 1765.6 088.75 3.55 85.33
4 60 2027.5 102.00 4.08 98.05
5 90 2042.4 102.75 4.11 98.73
6 120 2047.3 103.00 4.12 98.99
125
Table 6.9 Amount of EGC Released from Tablet during Dissolution of tablet.
S. No. Sampling
Time
(min)
Peak
Area
Concentration
obtained from
Calibration Curve
Amount of EGC
Released in 200ml
of dissolution
media (mg)
% of
EGC
Released
1 15 4701.4 238.50 09.54 44.08
2 30 6346.2 322.25 12.89 62.78
3 45 8575.4 435.75 17.43 84.90
4 60 9552.4 485.50 19.42 94.54
5 90 9925.6 504.50 20.18 98.28
6 120 10102.4 513.50 20.54 100.00
Table 6.10 Amount of ECG Released from Tablet during Dissolution of tablet.
S. No. Sampling
Time
(min)
Peak
Area
Concentration
obtained from
Calibration Curve
Amount of ECG
Released in 200ml
of dissolution
media (mg)
% of
ECG
Released
1 15 4580.9 227.70 09.10 47.98
2 30 5715.8 284.90 11.40 60.04
3 45 7831.9 391.56 15.66 82.52
4 60 8662.1 433.40 17.34 91.34
5 90 9477.5 474.50 18.98 100.0
6 120 9477.5 474.50 18.98 100.0
Table 6.11 Amount of EGCG Released from Tablet during Dissolution of tablet.
S. No. Sampling
Time
(min)
Peak
Area
Concentration
obtained from
Calibration Curve
Amount of EGCG
Released in 200ml
of dissolution
media (mg)
% of
EGCG
Released
1 15 19917.7 992.50 39.70 51.76
2 30 25834.8 1287.00 51.48 67.12
3 45 30692.5 1528.82 61.15 79.73
4 60 36413.1 1813.57 72.54 94.58
5 90 38501.5 1917.50 76.70 100.0
6 120 38501.5 1917.50 76.70 100.0
126
Table 6.12 Dissolution Profile Data of all the Catechins Released from
Tablet
S. No. Sampling Time
(min)
% of catechins released
Cat EC GC EGC ECG EGCG
1 15 48.00 46.72 43.28 44.08 47.98 51.76
2 30 65.40 63.54 63.19 62.78 60.04 67.12
3 45 80.72 86.49 85.33 84.90 82.52 79.73
4 60 96.09 97.43 98.05 94.54 91.34 94.58
5 90 97.56 97.99 98.73 98.28 100.0 100.0
6 120 98.79 100.00 98.99 100.00 100.0 100.0
Table 6.13 Amount of Sodium Selenite Released from Tablet during
Dissolution of Tablet.
S.
No.
Sampling
Time (min)
Peak
Area
Concentration
obtained from
Calibration Curve
(µg/L)
Amount of Sodium
Selenite Released
in 200ml of
dissolution media
(mg)
% Amount of
Sodium
Selenite
Released
1 15 4067 24.5 0.49 49
2 30 5810 35.0 0.70 70
3 45 7387 44.5 0.89 89
4 60 8300 50.0 1.00 100
5 90 8300 50.0 1.00 100
6 120 8300 50.0 1.00 100
top related