[PRELIMS] PANA2: Intro, Quality Control

Powder fineness/Micromeritics

Control tests

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→ To check if powder blend can be compacted/compressed

Storage condition and stability test

Control tests

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→ To make sure it will reach expiry date/long shelf life

Tablet thickness

→ Important in ***reproducing tablets identical*** in appearance, but also to ensure that every production lot will be usable with selected packaging components

→ Also used as ***counting mechanism*** using filling equipment

→ Determined with a caliper or thickness gauge

5

Requirement: ⩽ _% of standard thickness (tablet)

Diameter

Factors affecting tablet thickness

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_____ of the die

Fill

Factors affecting tablet thickness

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Amount of _____ permitted to enter the die

Compaction

The _____ characteristics of the fill material

Stokes Hardness Tester

* compressible spring between 2 plungers

Strong-cobb apparatus

* pressure operated air pump

Pfizer hardness tester

* ordinary pliers

Schleuniger apparatus

* most widely used instrument for tablet hardness test

Chewable tablet

Acceptable Hardness:

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Buccal tablet

Acceptable Hardness:

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10 kg

Acceptable choice

Acceptable Hardness:

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3\.5 - 7 kg

Application of Content Uniformity (CU) and Weight Variation (WV) Tests for Dosage Forms

Content Uniformity

→ Developed to ensure the content consistency of active pharmaceutical ingredients (API) within a narrow range around the label claim

→ Tablets of 30 are selected from the batch, 10 are assayed

→ Met if the content of each of the 10 tablets is within the limits of 85 and 115% of the average of the limits specified in the potency definition in the individual monograph

10

\[Compendial Requirement for Weight Variation Tolerances for Uncoated Tablets\]

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What’s the % Difference?

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7\.5

\[Compendial Requirement for Weight Variation Tolerances for Uncoated Tablets\]

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What’s the % Difference?

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130-324

5

\[Compendial Requirement for Weight Variation Tolerances for Uncoated Tablets\]

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What’s the % Difference?

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> 324

Disintegration testing

→ Temperature required: 37±2°C

→ Equipment: Basket rack assembly composed of 6 cylindrical tubes w/ 10-mesh wire cloth at bottom portion & disks

Dissolution testing

a physical test to predict drug delivery to a target area in the proper amount at the right time

Dissolution Rate

amount of active ingredient in a solid dosage form dissolved in a unit time under standardized condition of liquid or solid interface temperature and media composition

Rotating Basket

* USP apparatus 1 - _____
* Standard:
* 40 mesh
* 900 mL water

Rotating Paddle

* USP apparatus 2 - _____
* Standard: 
* Teflon or stainless
* 900 mL water
* Wire sinker for floaters

Tablets

**USP Apparatus 1**

* ***_____*** (immediate release)

Useful at higher rotation frequencies fort modified-release oral dosage forms ***(50 rpm)***

Capsule

**USP Apparatus 2**

* ***_____*** (immediate release)
* More useful at higher rotation frequencies for modified-release oral dosage forms ***(100 rpm)***
* Immediate release dosage forms that tend to flat or disintegrate slowly

Type 1 - Basket apparatus

* Rot. speed: 50-120 rpm
* Dosage form: Conventional tablets, chewable tablets, CR

Type 2 - Paddle apparatus

* Rot. speed: 25-50 rpm
* Dosage form: Orally disintegrating tablets, chewable tablets, CR, suspensions

Type 3 - Reciprocating cylinder

* Rot. speed: 6-35 rpm
* Dosage form: CR, chewable tablets

Type 4 - Flow through cell apparatus

* Rot. speed: N/A
* Dosage forms: ER, poorly soluble API, powder, granules, microparticles, implants

Type 5 - Paddle over disk

* Rot. speed: 25-50 rpm
* Dosage form: Transdermal

Type 6 - Cylinder (Rotating cylinder)

* Rot. speed: N/A
* Dosage form: Transdermal

Type 7 - Reciprocating holder

* Rot. speed: 30 rpm
* Dosage form: CR (non disintegrating oral and transdermal)

Media selection

→ Distilled water is the preferred medium

→ No enzymes should be incorporated

Control test

1\.Organoleptic evaluation

2\.Assay for the active ingredient(s)

3\.Thickness

4\.Hardness/Tensile Strength

5\.Friability

6\.Uniformity of Dosage Units (Weight Variation or Content Uniformity)

7\.Disintegration test

8\.Dissolution test

Organoleptic

color, odor, texture, taste, shape (surfaces,scoring)

* Color (no mottling=irregular color)
* Odor (film coated tablets)
* Taste (chewable tablets)

Controlling variables

→ Common Operating Speeds: 100 rpm for basket type / 50 rpm for paddle type

→ Temperature: 37+/-0.5°C

→ Usual volume of medium: 500 to 1000 mL, greater volumes (up to 200 mL allowed for drugs with limited solubility)

**Control Tests - Capsules**

1. Organoleptic evaluation
2. Stability tests
3. Uniformity of Dose
4. Disintegration test
5. Dissolution test
6. Moisture permeation test

Stability tests

\[Control test - Capsule\]

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* For capsules are performed to know the integrity of gelatin capsule shell and for determining the shelf-life of capsules. The test helps  in improving the quality of contents of capsule shell and for choosing the appropriate retail package.

Shelf-life Determination

\[Control test: Stability test - Capsule\]

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* shelf-life or the expiry date of packed capsules is determined under normal storage conditions
* Usually stored until 25C

**Shell Integrity Test**

* Performed to find out the integrity of capsule shell
* Once the shell is exposed to moisture, they shrink. Silica gel is put so the shell will stay dry
* Conditions: 40°C and 80% RH
* Standard capsule shells  kept at the room temperature 40°C and 80% RH becomes more soft, sticky and swollen.

 Disintegration test

\[Control test - Capsule\]

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Should be less than 30 mins.

Dissolution test

\[Control test - Capsule\]

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0\.1N HCl resembles the stomach; UV spectrophotometer

**Moisture Permeation Test**

*  Assure the suitability of containers for packaging of capsules
* Foil/Blister packs: Laminated and air is vacuumed
* This test is carried out to assure the suitability of containers for packaging of capsules. The moisture permeating feature of capsules packaged in
* Single unit containers-blister pack or strip pack.
* Unit dose containers glass or plastic bottle is to be determined

Control Tests - **Granules and Powders**

▹Particle size analysis

▹Angle of repose

▹Bulk density

▹Tapped density

▹Hausner’s ratio

▹Flowability

▹Stability tests

Stability tests

\[Control Tests - **Granules and Powders]**

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spray water/ethanol

Control Tests - Ointments and Creams

1. Organoleptic evaluation
2. Assay for active ingredients
\*to know assay: USP
3. Test of rate of absorption
4. Test for non-irritancy
\*rabbit (skin of rabbit is more sensitive than rats)
5. Test of rate of penetration
6. Test of rate of drug release
7. Test of rheological properties
\*viscometer
8. Test of content uniformity
9. ID test for the active ingredients and possible contaminants
10. Loss of water/water content
\*packaging and of drug
11. pH
\*pH 4-7
12. Sterility test
13. Storage condition
\*ex: suppositories in the ref
14. Stability test

Test of rate of absorption

\[Control Tests - Ointments and Creams**]**

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done in vivo  only. The ointment should be  applied  over a definite area of the skin by  rubbing. At regular intervals of time, serum and urine samples should be analyzed for the quantity of drug absorbed.

Test for non-irritancy 

\[Control Tests - Ointments and Creams**]**

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* the bases used  in the formulation of ointment may cause irritation or allergic reactions.
* Non-irritancy is evaluated by patch test. 24 human volunteers  are selected. Definite quantity of ointment is applied under occlusion daily on the back or volar fore  arm  for 21 days. Daily  type of pharmacological action observed is noted. No visible reactions or erythema or intense erythema with edema and vesicular  erosions should occur.
* A good ointment base shows  no visible reactions

Test of rate of penetration

\[Control Tests - Ointments and Creams**]**

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* crucial in the onset  and duration of action of drug.
* Weighed quantity of the preparation should be applied over selected area of the skin for a definite period of time. Then the preparation left over is collected and  weighed. (the amount of preparation penetrated divided by  the area and period of application gives the rate of penetration of the preparation). 
* Performed in triplicates

Test of rate of drug release

\[Control Tests - Ointments and Creams**]**

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ZOI. If bactericidal the agar plate  is previously seed with suitable organism such as S. aureus. ZOI  is measured and correlated with the  rate of release.

**Control Tests - Suppositories**

1. Organoleptic evaluation
2. Physical strength
3. Melting range
\*melting point apparatus
4. Uniformity of drug content
5. Softening time
\*the time that the suppository will melt down

Appearance

**[Control Tests - Suppositories]**

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All the suppositories should be uniform size and shape and should have elegant appearance. Suppositories should be examined for cracks and pits on the surface of suppositories.

Physical strength test

**[Control Tests - Suppositories]**

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* The strength of the suppositories should be considered to assess their ability during normal handling.
* The apparatus used for this is called as **breaking test apparatus**, which contains a double walled chamber in which water is pumped to maintain 37°C temperature in between the two walls of the chamber.
* The inner chamber contains a disc for holding the suppositories. To this disc a rod is attached. The other end of the rod contains a disc for holding the weights.
* When the weights are added (upto 200gms) at one minute time interval until the suppositories crumbles.
* All the weights used are added which gives the tensile strength.
* Tensile strength is the maximum force which the suppository can withstand during production packing and handling. Higher the tensile strength indicates less will be tendency to fracture.

Melting Range Test

Macro melting range

Micro melting range

Macro melting range

\[Melting Range Test\]

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* Conducted using the tablet disintegration test apparatus
* It gives the measure of thermal stability of the suppository. It is the time taken by the entire suppository to melt in a constant temperature water bath.
* The test is conducted using the tablet disintegration test apparatus. The suppository is immersed in a constant water bath, finally the melting range is recorded.

Micro melting range

\[Melting Range Test\]

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* Measured using capillary  tubes.
* ▸Measured using capillary  tubes.

Test for Softening Time

* This test measures the softening or liquefaction time of suppository which indicates the hardness of the base.
* The apparatus consists of cellophane tube tied at the two ends of condenser.
* The two ends of the cellophane tube are opened. Water is circulated through the condenser at a definite rate.
* As a result after sometime the upper half of the tube opens wide and lower half collapses. The time period in which the suppository melts completely is considered as softening time10.

Control Tests - Liquid Dosage Form

1. Organoleptic evaluation
2. Assay of active ingredients and degradation products
3. Pourability
4. Viscosity
5. Isotonicity
\*RBC
6. Particle size agglomeration and particle size distribution
\*Coulter counter
7. Clarity
8. Physical stability
9. Relative density
10. pH
11. Surface tension
12. Microbial limit tests
13. Stability of the active ingredients and identification tests
14. Light stability
\*stability cabinet
15. Container and closure compatibility
16. Redispersibility
17. Suspendibility
18. Storage conditions

Visual

\[Control Tests - Liquid Dosage Form\]

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* physical appearance of products for patient adherence and compliance is critical so it should be good looking and elegant in appearance.

 Viscosity

\[Control Tests - Liquid Dosage Form\]

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* property of liquids that is directly related to the resistance to flow;
* viscosity and consistency directly relates with stability of solutions; if viscosity increases; then there  is a chance of increase in stability.
* Controlled in order to ensure accurate measurement of the volume to be dispensed.
* Increasing viscosity of some formulations may increase palatability.
* There is a viscosity range that the formulations should exhibit

Isotonicity

\[Control Tests - Liquid Dosage Form\]

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parenterals

Crystallization and precipitation

\[Control Tests - Liquid Dosage Form\]

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* concentration  of sucrose is determined. Very high – crystallize the syrup; less sucrose conc.- favors microbial  growth.

Particle size agglomeration and particle size distribution

\[Control Tests - Liquid Dosage Form\]

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* for suspensions and emulsions; shows stability of a disperse system.
* Measured using coulter counter and microscopy, Sedimentation by __**Andreason apparatus.**__

Clarity

\[Control Tests - Liquid Dosage Form\]

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* visual examination of the formulations under light alternatively against white and black backgrounds.

Container and closure compatibility

\[Control Tests - Liquid Dosage Form\]

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 torque test

Light-resistant container

\[Control Tests - Liquid Dosage Form\]

 

*  a container intended to provide protection from light.


* meets the requirements for Light Transmission 

pH control is performed to:

* **Maintain solubility** of  therapeutic agent in the formulated product.
* **Enhance the stability** of products in which the chemical stability of the active agent is  pH dependent.

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* The solubility of a vast number of currently available drugs is pH-dependent and therefore, the solubility of the therapeutic agent in the formulation may be compromised by small changes in  pH. Unless there are issues re: solubility or stability of the therapeutic agent, the usual  pH of oral solutions is 7.0 (5.0 – 8.0).

Container and Closure control

* Torque test - is a measure of circular force required to loosen the cap; and application torque is the amount of force used to tighten the cap.
* Sample size = 10

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* The integrity of the seal between closure and container depends on the geometry of the 2, materials used in construction, the composition of cap liner, and tightness applied to the cap.


* Geometry-controlled by the mold. Mold number is assigned which is used as its identification number.


* Closures should fit the tread of the container. It should sit on a container without tilting, produce no leaks, should not rotate continuously, be reasonably tight.

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1. Minimum titghtness is essential to avoid evap or leakage of product
2. Excessive torque may break the moulded closures
3. Caps applied too tightly may be difficult to remove.

Type: I

Type of test: Powdered Glass

\[Liquid Dosage Form - Glass Types\]

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Highly resistant borosilicate glass

Type: II

Type of test: Water attack

\[Liquid Dosage Form - Glass Types\]

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Treated soda-lime glass

Type: III

Type of test: Powdered Glass

\[Liquid Dosage Form - Glass Types\]

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Soda-lime glass

Type: NP

Type of test: Powdered Glass

\[Liquid Dosage Form - Glass Types\]

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General purpose soda-lime glass

Additional Control Tests - Parenteral/Sterile Liquid Dosage Forms

1. Leaker Test
\*if may leak; bottle is submerged sa blue dye
2. Pyrogen Test
3. Sterility Test
4. Particulate Matter Evaluation
5. Uniformity of Content
6. Ocular Toxicity and Eye Irritation (for eye drops)
7. Test for preservative efficacy

Container and closure compatibility

\[Additional Control Tests - Parenteral/Sterile Liquid Dosage Forms\]

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torque test

**Leaker Test**

* intended to detect incompletely sealed ampoules or vials so that they can be discarded in order to maintain the sterile conditions of the medicines.
* Uses 0.5 – 1.0% methylene blue

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* Leakage occur when a discontinuity exists in the wall of a package that can allow the passage of gas under the action of a pressure or concentration differential
* existing across the wall. Presence of capillary pores or tiny cracks can cause microbes or other dangerous contaminants to enter the ampoules or may lead to the leakage of contents to outside. This may lead to contamination of the sterile contents and also spoilage of appearance of the package.
* Changes in temperature during storage can cause expansion and contraction of the ampoule and its contents, there by accentuating interchange if an opening exists. Leaker test for ampoules is intended to detect incompletely sealed ampoules so that they can e discarded in order to maintain the sterile conditions of the medicines.
* Tip seal are more likely to be incompletely closed than pull seals.
* Open capillaries or cracks at the point of seal result in leakers.

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* Leakers are detected by this process in a visible manner. Ampoules are placed in a vacuum chamber. Completely submerged in a deeply colored dye solution of about 0.5-1% methylene blue. A negative pressure is applied within the ampoule. Subsequent atmospheric pressure causes the dye to penetrate on opening thus making it visible after the ampoule has been washed. The vacuum, about 27 inches Hg, should be sharply released after 30 minutes.  Detection of leakers is prominent when ampoules are immersed in a bath of dye during autoclaving cycle as this has the advantage of accomplishing both leaker detection and sterilization in one operation.
* Result: the color from the dye will be visible within a leaker.

Pyrogen Test

* Pyrogen test is designed to limit to an acceptable level the risks of febrile reaction in the patient to the administration, by injection, of the product concerned.
* Test animals:  healthy, mature rabbits.
* Rabbits – have similar pyrogen tolerance to humans
* Detect non-bacterial endotoxin and bacterial endotoxin pyrogens.
* Qualitative only
* 3 rabbits
* Should be acclimatized first (7 days)
* Procedure: NMT 30 mins prior to injection of test dose, determine “control temp” of each rabbit. Use only rabbits that do not vary by more than 1 deg.C from each other. Do not use any rabbit having temp exceeding 39.8 deg.C. Record tenoerature at 30 minutes interval between 1 and 3 hours subsequent to the injection.
* Requirement: NMT 0.5 deg. Individual rise. If did not meet, add 5 more. NMT 3 of 8 rabbits shows  individual rise of 0.5 deg and total temp rise NMT 3.3 deg.

Disadvantages of Pyrogen Test

* Biological variation
* expensive
* Laborious
* Dose dependent
* Not for antipyretic drug

LAL (Limulus Amebocyte Lysate) Assay

In vitro assay used  to detect the presence and concentration of bacterial endotoxins in drugs and biological products.

LAL tube: 1

Result: Negative

\[Pyrogen Test - LAL\]

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Negative Control (Pyrogen free saline)

LAL tube: 2

Result: Positive

\[Pyrogen Test - LAL\]

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Positive Control (Pyrogen)

LAL tube: 3

Result: Positive

\[Pyrogen Test - LAL\]

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Positive internal control (test sample containing exotoxins)

LAL tube: 4

Result: Negative

\[Pyrogen Test - LAL\]

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Test samples

Sterility Test

* Reveal the presence or absence of viable microorganisms in a product.

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Methods:


1. Membrane Filtration Method
2. Direct Inoculation Method

Particulate Matter Evaluation

* The USP specifies that good manufacturing practice(GMP) requires that each final container of an injection be subjected individually to a visual inspection and that containers in which visible particles can be seen should be discarded.

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Methods:


1. Light obscuration Test
2. Microscopic count test

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* Therefore, all of the product units from a production line currently are being inspected individually by human inspectors under a good light, baffled against reflection into the eye and against a black-and-white back ground.
* The USP has identified two test methods.
* The first test to be used is the light obscuration test, which uses and electronic instrument designed to count and measure the size of the particles by means of a shadow cast by the particle as it passes through a high-intensity light beam.
* If the injection formulation is not a clear, colorless solution, it exceeds the limits specified for the light obscuration test, it is to be subjected to the microscopic count test.

Test for ocular toxicity and irritation

* Test animals: 5 albino rabbits
* Requirement: No change in the eye into which the preparation is instilled.
* Five albino rabbits are selected, the iridal vessels of whom can be easily observed for toxicity and irritation. Based on the type of dosage form, the medicament is extracted using cotton seed oil or saline. Small quantities of the extract are instilled into one eye of all the rabbits, while sterile saline solution is instilled into the other eye. After one hour all the rabbits are observed for irritation, swelling or shrinkage of the eye.
* Result: No change in the eye into which the preparation is instilled indicates that the preparation under test is safe for use.

Test for Preservative Efficacy

* Product are tested using cultures of microorganisms like *Aspergillus niger, Candida albicans, Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa.*
* a. Cultures of microorganisms like Aspergillusniger, Candida albicans, Escherchia coli and Pseudomonas aeruginosa, each containing about 10,000 – 10,00,000 organisms per ml are selected.
* b. Three to four samples of each preparation are taken in sterile test tubes and inoculated with few ml of each culture separately.
* c. They are incubated at 20 - 25°C for a period of 28 days and are observed weekly for the appearance of turbidity.
* d. No growth of microorganisms indicates that the preservative is totally effective.

Safety Test - Biological

* Is intended to detect in an article any unexpected, unacceptable biological reactivity.


* Test animals: select 5 healthy mice weighing between 17 and 23 g.
* Refer to OECD Guidelines

albino mice

\[Biological Reactivity Test (In Vivo)\]

\n Systemic Injection Test

* Test animal:

albino rabbits

\[Biological Reactivity Test (In Vivo)\]

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Intracutaneous Test 

* Test animal:

rabbit

\[Biological Reactivity Test (In Vivo)\]

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Implantation Test

* Test animal:

albino rabbits

\[Biological Reactivity Test (In Vivo)\]

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Eye Irritation Test

* Test animal:

mice

\[Biological Reactivity Test (In Vivo)\]

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Safety Test

* Test animal:

Systemic injection

Designed to evaluate systemic responses to the extracts of materials

Intracutaneous

* designed to evaluate local response to extracts of materials

Implantation

* designed for evaluation of plastic materials and other polymeric materials in direct contact with the living tissues.

Rabbit

\[Partial list of official quantitative biologic & microbiologic tests\]

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Drug: Insulin

Test animal:

Pigeon

\[Partial list of official quantitative biologic & microbiologic tests\]

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Drug: Digitalis & related cardiac glycosides

Test animal:

Dog

\[Partial list of official quantitative biologic & microbiologic tests\]

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Drug: Parathyroid hormone

Test animal:

Rat

\[Partial list of official quantitative biologic & microbiologic tests\]

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Drug: Posterior Pituitary

Test animal: