Exercise in Pharmaceutical Analysis II

EXPERIMENT NO. 3

DISSOLUTION TEST OF AMOXICILLIN CAPSULES USING THE ROTATING

BASKET ASSEMBLY (Apparatus 1)

I- OBJECTIVE: to determine the concentration of the amoxicillin absorbed by unit

dosage form and its compliance with USP/NF requirement by using the
Dissolution Apparatus and the UV-VIS Spectrometer.

II - DISCUSSION:

Dissolution test is a requirement of FDA in testing solid dosage forms,

such as tablets and capsules. The rotating basket assembly, which is also called
as Apparatus 1 is applied to testing hard and soft gelatin capsules.

Dissolution test determine the quantity / amount of the drug which is

absorbed in a certain liquid medium, at a specified temperature of 37⁰C.

The test characterizes the basic bioavailability profile of a drug.

Aside from temperature, there are other parameters that must be

controlled;

a. Speed or rpm of the rotation of the basket

b. Kind and volume of dissolution media.
c. Time of run.
d. Wavelength of the spectrophotometer in absorbance
readings

Apparatus:

A. Dissolution Apparatus using the Rotating Basket.

Speed : 100 rpm

Temperature : 37oC

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Exercise in Pharmaceutical Analysis II

B. UV / VIS Spectrophotometer
Spectrophotometer- is a photometer (a device for measuring light intensity)
that can measure intensity as a function of the color and wavelength of the
light.

Parts:

Radiation source – for ultraviolet measurement, it is usually a hydrogen to

deuterium lamp and a tungsten for visible measurements.

Monochromator – to select a narrow band of wavelength to pass through the

sample cell, an important property of monochromator is it band pass or
bandwidth.

Sample cells or Cuvette

⮚ That contain the sample to be analyzed, should have parallel sides that
are perpendicular to the radiation source\
● UV – silica or quartz
● VIS – glass or plastic
● The cuvette used is usually has a path length of 1cm.
Detector
⮚ Changes the radiation transmitted from the UV / VIS spectrophotometer
into a current or Voltage for the readout device to use.
⮚ Types: phototube, photomultiplier tubes (wavelength region 150 –
1000nm)

Readout device
⮚ Use the current or voltage produced by the detector and amplifies it.
⮚ The readouts device then converts the Current or Voltage into a form that
analyst can read as a print out.

Readings: Absorbance; Transmittance; Concentration.

Principle involved:
⮚ The sample absorbs a portion of the incident radiation; the remainder is
transmitted to a detector, where it is changed into an electrical signal and
displayed usually after amplification, on a meter, chart recorder or some
other type of readout device.

III - MATERIALS: Amoxicillin capsules (any branded or generic product)

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Exercise in Pharmaceutical Analysis II

Amoxicillin reference standard (or innovator brand)

Distilled water
Dissolution apparatus, basket assembly
UV – VIS Spectrophotometer and Glass Cuvettes
Analytical Balance
Beaker (1 Liter)
Volumetric flasks, 1L, 100mL 50mL capacities
Pipets, 10 mL, 5mL 2mL, 1mL calibrations
Laboratory thermometer
Hot plate / stove
Rubber aspirators
Forceps
Watch glass
Wash bottle
Tissue paper, lint free
Glass funnels
Filter paper

IV - ASSAY PROCEDURES:

1. Preparation of Dissolution media

Place 900 ml plain, distilled water in each of the six – 1L glass beakers. Place the
beakers on the hot plate and heat until it reaches 37⁰C (+/-5). Monitor the
temperature with a thermometer. Keep the temperature constant. Set aside.

2. Preparation of the amoxicillin reference standard solution

a. Weigh exactly 100mg of the standard amoxicillin powder in a watch glass.
b. Add about 70 ml of the dissolution media (distilled water) to a 100 ml
volumetric flask add the weighed standard amoxicillin powder. Cover and
shake the flask to dissolve the powder completely.
c. Wash the watch glass with portions of distilled water and add the rinsing to
the volumetric flask fill to the mark with more distilled water to make 100ml
solution. Mix thoroughly. This is the first solution.
d. Using a pipet transfer 1ml of the first dilution to another 100ml volumetric
flask. Dilute with more dissolution medium (distilled water) to make 100ml
solution. This is the final dilution that contains 10ug/ml amoxicillin.

3. Preparation of the dosage unit samples

Weigh no less than 10 capsules and take the average weight. Take six
capsules and weigh individually in the analytical balance for use in the test.

4. Assembly and Operation of the Dissolution apparatus

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Exercise in Pharmaceutical Analysis II

A. Hanson dissolution apparatus

1. Pour in a clear acrylic water bath a suitable amount of top water.
2. Turn on the soft flow heater to maintain the desired temperature for
testing.
3. Place the six 1 ml glass vessels into the large holes of the base, fasten
properly with the clips.
4. Fill the six-1L glass vessels with the 900ml previously heated distilled
water. Cover each vessel.
5. Stabilize the temperature to 37⁰C.
6. Set the height of each basket shaft so that it will be 25 mm (+/-2) from the
bottom of each vessel.
7. Set the basket speed to 100 rpm.
8. Place 1 capsule in each of the six cylindrical baskets.
9. Snap the capsule-filled basket onto the metal shafts, previously placed in
the apparatus.
10. Lower the cylindrical baskets to the desired height and turn the speed
control to 100 rpm.
11. After 90 minutes, withdraw an aliquot portion equivalent to 0.5 mg of
Amoxicillin (based on the labeled claim of the product), passing the portion
through a filter. Dilute to 50ml of the medium in 50ml volumetric flasks.
12. Aliquot sampling should be carried out from the point midway between the
tip of the paddle and the surface of the medium and no less than 1 cm
from the vessel walls.
13. The final dilution should contain about 10 mcg/ml.
14. Determine the absorbance of each of the 6 capsules at a wavelength of
maximum absorbance at 272 nm in comparison with a standard having
10ug/ml in the final dilution.
15. Calculate the amount of the Amoxicillin absorbed in solution per tablet (in
mg/capsule) and the % labeled claim of each capsule.

B. Erweka dissolution Tester DT 700

1. Plug the Erweka dissolution tester DT 700 into a properly installed wall outlet.
2. Fill the water bath with distilled water up to the designated maximum level.
3. Place the testing level inside the water bath then pour the required dissolution
medium.
4. Turn on the instrument by pressing the (On/Off) switch located at the back
potion of the drive head.
5. Set the parameters: temperature, time and speed by pressing the necessary
numerical key.
6. Place the sample to be tested inside the basket, then connect them to the
metal shaft in the drive head.
7. Lower the metal shaft the press [START] key to start the test.

C. Operating Procedures

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Exercise in Pharmaceutical Analysis II

1. Precautions before use:

● When requiring axial base up and down, remove obstacles to prevent
hazards.
● The main water bath has floating ball water level switch. Note that the
water level shall be higher than the floating ball to activate heating system
and circulation system.
● The heating system has a temperature (50⁰C). Thus, the temperature
setting should not exceed 50⁰C.
● For the return rotating axis match the no. and insert into the rotating axis. It
must be lock tight. Use +25m/m calibrates to determine calibration height.
● Use dedicated socket.
● Do not share socket with large amperage machine.
● When using test basket, place the height increasing padding under the
lower stop column (between the 2 support columns at the right sides) so
that when the lowering of the axis base will stop at the correct position.

2. Make sure that the machine is placed on top of flat and stable top. Use level
instrument to make sure the level of the machine.
3. Check if the voltage match and make sure they are connected correctly.
Connect the 2 types of the connectors for upper and lower seats. Make sure
that they are connected correctly and lock tight.
4. Feed water into the water tank to appropriate level. The round bottom tank is
placed inside the water tank to prevent floating switch stuck.
5. Push the beaker at the right rear side to on the start power.
6. Press POWER to start power.
7. Press JACK to move axis base up or down to the fixed position
8. Press HEAT to start heating system and circular system, set up the
temperature to maintain constant temperature.
9. Press STIR to start mixer system. Adjust the speed adjustment knob to the
required speed.
10. When assembling and dismantling the paddle blades and test basket move
the axis base to upper position. Tighten the screw.
11. Cover the beaker and maintain temperature inside the cup.
12. After use, store the accessories properly. Keep the machine clean.

DISSOLUTION SPECIFICATIONS FOR AMOXICILLIN:

Dissolution medium Plain, distilled water at 37⁰C

Time 90 minutes

Apparatus used Apparatus 1 Rotating Basket Assembly

Speed 100 rpm

Wavelength 272 nm

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Exercise in Pharmaceutical Analysis II

USP Specifications Not less than 80% of the labeled claim of amoxicillin
is absorbed in 90 minutes.

CALCULATIONS:

The concentration of the Amoxicillin per capsule and its compliance with the USP
requirement may be calculated as follows:

Au / As x concentration of RS (mg/ml) x DF x ave. wt. per capsule

1. mg/capsule =
Wt. of capsule in mg
Where:
Au = Absorbance reading of the sample
As = absorbance reading of the reference standard
DF = dilution factor

Note: if the capsules used contain 500 mg of Amoxicillin, the aliquot portion
equivalent to 0.5 mg Amoxicillin is 0.9ml, which is calculated as follows:

500mg: 900mL = 0.5mg: x

900ml (0.5mg)
X= = 0.9ml
500mg

900 50
DF = X = 50,000
0.9 1

Note: The DF is 50,000 if the dilution done is as follows:

One 500mg capsule was dissolved in 900ml of dissolution medium, then

0.9ml was withdrawn and diluted to 5oml its final dilution is 10ug/m if the capsule
used is 500mg of Amoxicillin.

Actual amount obtained

2. % of labeled claim = x 100
Labeled claim

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Exercise in Pharmaceutical Analysis II

WORK SHEET

Name : Princess L. Ronsable Date of Analysis: ______________

Group No. : _________________________ Year & Section: ______________
Score:
______________

EXPERIMENT NO. 3

DISSOLUTION TEST OF AMOXICILLIN CAPSULES USING THE ROTATING

BASKET ASSEMBLY (Apparatus 1)

RESULTS OF ANALYSIS:

1. Tabulation of Data and Results:

2.730

Absorbance reading if the RS (As)

10 ug/mL

Concentration of the RS (ug /ml)

50,000

Dilution factor

3.46 g

Weight of the 10 capsules

0.4325 g

Average weight for capsule

Amount % Labeled
obtained claim
Capsule # Actual wt. Au readings

1 420 mg 2.749 518.46 mg 103.69%

2 460 mg 2.732 470.45 mg 94.09%

3 430 mg 2.799 515.62 mg 103.12%

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Exercise in Pharmaceutical Analysis II

4 450 mg 2.700 492.70 mg 98.54%

5 440 mg 2.31 491.66 mg 98.33%

6 400 mg 2.733 541.22 mg 108.24%

2. Calculations

3. Quality Control Disposition

Certified Correct:

_________________________
Signature over Printed Name
Of QC Inspector

Date: __________________

ANSWER THE FOLLOWING QUESTIONS BRIEFLY:

1. In the dissolution test for capsules, why is necessary to use the cylindrical
basket instead of the paddle?

It is because the pores in the mesh allow the dissolving drug to move from the
basket into the holding vessel, creating a uniform solution. The sticky capsules
containing HPMC or similar compounds as a binder are tested using a basket because
tablet swells in water & may stick to the walls of the dissolution vessel.

2. Why is the sampling procedure done at a point midway between the tip of
the basket and the surface of the dissolution medium?

Due to the reason that it is where the concentration is exact. The sample at the
bottom of the vessel is at is concentrated state while at the top is diluted.

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Exercise in Pharmaceutical Analysis II

3. In tabulated form, present the different apparatus numbers and the

description/names of each.

APPARATUS NAME AND DESCRIPTION

1 Rotating Basket
2 Paddle
3 Reciprocating Paddle
4 Flow Cell
5 Paddle over disk
6 Cylinder
7 Reciprocating Disk
8 Non-USP NF
9 Non-USP NF
10 Non-USP NF

4. Sketch and label the parts of the Dissolution Apparatus.

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