Quality control. Parenteral preparations are sterile preparations intended for administration by injection, infusion or implantation into the human.

Quality control

Parenteral preparations are sterile preparations intended for administration by injection, infusion or implantation into the human or animal body.

Injections are sterile, pyrogen free preparations intended to be administered parenterally (outside alimentary tract).

All parenterals, as well as nasal, ophthalmic solutions, must be sterile, including packaging materials.

Parenteral preparations

Container for parenteral preparations are made as far as possible from materials that are sufficiently transparent to permit the visual inspection of the contents, except for implants.

Parenteral preparations are supplied in glass containers or in other containers such as plastic containers and prefilled syringes.

The tightness of the container is ensured by suitable means. Closures ensure a good seal, prevent the access of micro-organisms and other contaminants and usually permit the withdrawal of a part or the whole of the contents without removal of the closure.

Closures for multidose containers are sufficiently elastic to ensure that the puncture is resealed when the needle withdrawn.

Contamination

Any effect or action that has a negative impact on a product's integrity making it unfit for use

–Chemical composition –pH –Sterility (e.g.microorganism contamination)–Pyrogenicity –Physical appearance –Particulate matter (e.g. dust, glass or precipitation)

Sources of product contamination

● People (most common)– Touch contamination– Generation of particulates from shedding cells or hair● Air Supply– Heating, Ventilation and Air Conditioning (HVAC)● Infiltration– Particles from adjacent spaces (e.g. entrance, hall)● Internal generation– Walls, floors, ceilings, packaging, equipment

SterilityThe complete destruction of all living organisms and their spores or their complete removal from the formulation.

Aseptic TechniqueThe technique for preparation and manipulation of compounded sterile products and parenteral preparations that prevents contamination.

Importance of Aseptic Technique –Parenteral administration bypasses the skin and gastrointestinal

tract, the bodies natural barriers to infection. –Giving a patient a contaminated product can cause serious adverse

effects including DEATH. –Parenteral medications account for >40% of all medications

administered in institutional practice.

Necessities of Parenteral preparations: –Sterility (must)–Free from Pyrogen (must)–Free from particulate matter (must)–Clarity (must)–Stability (must)–Isotonicity (should)–Solvents or vehicles used must meet special purity and other standards.–Restrictions on buffers, stabilizers, antimicrobial preservative. Do not use coloring agents.–Must be prepared under aseptic conditions.–Specific and high quality packaging.

●Parenteral products require additional tests, which include those of sterility, pyrogen, clarity, and particulate analysis, and for glass-sealed ampoules, leaker test.

1.Volume in container

● The total fluid volume that must be filled into a unit parenteral container is typically greater than the volume that would contain the exact labeled dose.

●The fill volume is dependent on the viscosity of the solution and the retention of the solution by the container and stopper.

●The USP provides a procedure for calculating the fill dose that is necessary to ensure the delivery of the stated dose. It also provides a table of excess volumes that are usually sufficient to permit withdrawal and administration of the label volume.

An injection container is filled with a volume in slight excess of the labeled size.

Determination of filled volume: 10 mL or more 1 container3-10 mL 3 containersLess than 3 mL 5 containers

Labeled sizeMobile liquidViscous liquid

0.5 mL0.1 mL0.12 mL

1 mL0.1 mL0.15 mL

2 mL0.15 mL0.25 mL

5 mL0.3 mL0.5 mL

10 mL0.5 mL0.7 mL

20 mL0.6 mL0.9 mL

50 mL or more2%3%

2 .Clarity testing and particulate analysis:a. Clarity examination:

●Clarity is defined as the state or quality of being clear or transparent to the eye.

●Clarity specifications are not given in the USP, other than to state that all injections be subjected to visual inspection.

●Visual method is a simple method, in which the filled containers are examined against strong illuminated screen by holding neck & rotating it slowly or inverted it to keep out the foreign matter.

●Instruments that measure scattered light, such as the Photo-Nephelometer are used to evaluate and set clarity standards for parenteral preparations. Nephlometer readings are insensitive to contamination by large (visible) particulates.

b. Particulate matter:

● It is defined in the USP as extraneous, mobile, undissolved substances, other than gas bubbles, unintentionally present in parenteral solutions.

●Test methods and limits for particulates are stated in the USP for large-volume injections and small-volume injections.

●Parenteral solutions are sorted for foreign particles, such as glass, fibers, precipitate, and floaters.

●All products containing clear solutions should be inspected against a black and sometimes a white background using a special light source. Although manual visual inspection is the most common means of inspection, electronic particle detection equipment and computer-controlled electrooptic systems are replacing manual inspection.

Sources of particulate matter

●Raw materials ●Processing and filling equipment

●The container ●Environmental contamination

●Electronic particulate counters have been applied to parenteral because of the rapidity at which they do particulate analysis.

●Their main disadvantages are the lack of differentiation of various types of particulates, and the fact that particle size is measured differently from microscopic analysis.

●The USP tests for particulate matter in injections utilize both the microscopic and light obstruction methods.

Particulate matter in injections

●Particulate matter in injections and parenteral infusions consists of mobile undissolved particles, other than gas bubbles, unintentionally present in the solutions.

●For the determination of particulate matter, two procedures: Method 1 (Light Obscuration Particle Count Test) and

Method 2 (Microscopic Particle Count Test), are specified hereinafter .

●When examining injections and parenteral infusions for sub-visible particles Method (1) is preferably applied. However, it may be necessary to test some preparations by the light obscuration particle count test followed by the microscopic particle count test to reach a conclusion on conformance to the requirements.

●When Method (1) is not applicable, e.g. in case of preparations having reduced clarity or increased viscosity, the test should be carried out according to Method (2), Emulsions, colloids, and liposomal preparations are examples.

Similarly, products that produce air or gas bubbles when drawn into the sensor may also require microscopic particle count testing.

If the viscosity of the preparation to be tested is sufficiently high so as to

preclude its examination by either test method, a quantitative dilution with appropriate diluents may be made to decrease viscosity, as necessary, to allow the analysis to be performed.

Method (1) Light obscuration particle count test:

●Use a suitable apparatus based on the principle of light blockage which allows an automatic determination of the size of particles and the number of particles according to size.

●The apparatus is calibrated using dispersions of spherical particles of known sizes between 10µM and 25µM. These standard particles are dispersed in particle-free water.

General precautions:

● The test is carried out under conditions limiting particulate matter, preferably in a laminar-flow cabinet.

● Very carefully wash the glassware and filtration equipment used, with a warm detergent solution and rinse with abundant amounts of water to remove all traces of detergent. Immediately before use, rinse the equipment from top to bottom, outside and then inside, with particle-free water.

● Take care not to introduce air bubbles into the preparation to be examined, especially when fractions of the preparation are being transferred to the container in which the determination is to be carried out.

Procedure:

● Mix the contents of the sample by slowly inverting the container 20 times successively. If necessary, cautiously remove the sealing closure. Clean the outer surfaces of the container opening using a jet of particle-free water and remove the closure, avoiding any contamination of the contents. Eliminate gas bubbles by appropriate measures such as allowing to stand for 2 min or sonicating.

● For large-volume parenterals, single units are tested. ● For small-volume parenterals less than 25 ml in volume, the contents of 10 or more units is combined in a cleaned container to obtain a volume of not less than 25 ml; the test solution may be prepared by mixing the contents of a suitable number of vials and diluting to 25 ml with particle-free water or with

an appropriate particle-free solvent when particle-free water is not suitable .

●Small volume parenterals having a volume of 25 ml or more may be tested individually.

● Remove four portions, each of not less than 5 ml, and count the number of particles equal to or greater than 10 μm and 25 μm. Disregard the result obtained for the first portion, and calculate the mean number of particles for the preparation to be examined.

Evaluation:For preparations supplied in containers with a nominal volume of more than 100 ml, apply the criteria of test 1.A.

The preparation complies with the test if the average number of particles present in the units tested does not exceed 25 per ml equal to or greater than 10 μm and does not exceed 3 per ml equal to or greater than 25 μm.

●For preparations supplied in containers with a nominal volume of less than 100 ml, apply the criteria of test 1.B.

● The preparation complies with the test if the average number of particles present in the units tested does not exceed 6000 per container equal to or greater than 10 μm and does not exceed 600 per container equal to or greater than 25 μm.

● If the average number of particles exceeds the limits, test the preparation by the Microscopic Particle Count Test.

Method (2) Microscopic particle count test:

The microscope is adjusted to 100 ± 10 magnifications

and is equipped with an ocular micrometer calibrated with an objective micrometer, a mechanical stage capable of holding and traversing the entire filtration area of the membrane filter, and two suitable illuminators to provide episcopic illumination in addition to oblique illumination.

Test 2.A — Solutions for parenteral infusion or solutions for injection supplied in containers with a nominal content of more than 100 mL.● The preparation complies with the test if the average number of particles present in the units tested does not exceed 12 per mL equal to or greater than 10 μm and does not exceed 2 per mL equal to or greater than 25 μm.

Test 2.B — Solutions for parenteral infusion or solutions for injection supplied in containers with a nominal content of less than 100 ml.● The preparation complies with the test if the average number of particles present in the units tested does not exceed 3000 per container equal to or greater than 10 μm and does not exceed 300 per container equal to or greater than 25 μm.

3 .Pyrogen test:

● Pyrogenic substances are primarily lipid polysaccharide products of the metabolism of microorganisms, they may be soluble, insoluble or colloidal.

● Pyrogen produced by gram-negative bacilli are generally the most potent.Minute amounts of pyrogens produce a wide variety of reactions in both animals and humans, including fever, leukopenia and alteration in blood coagulation.

● Large doses can induce shock and eventually death.

●One pyrogen test is a qualitative biological test based on the fever response of rabbits. The test aims of measuring the rise in body temperature in healthy rabbits by the intravenous injection of a sterile solution of the substance under test.

●If a pyrogenic substance is injected into the vein of a rabbit, a temperature elevation will occur within three hours.

Sources of pyrogen contamination

-Solvent-possibly the most important source.

-The medicament

-The apparatus

-The method of storage between preparation and sterilization

Rabbit pyrogen test ●Rabbits must be healthy and mature

●Either sex may be used

●Must be individually housed between 20 and 23°C

●Weighing not less than 1.5 kg

●Fed a complete and balanced diet not containing antibiotics, and not showing loss of body mass during the week preceding the test.

Rabbit test ● Selection of animals (healthy, adult, not less than1,5kg,…)

● Housing of animals (environmental problems: presence of strangers (unknown place), noise, T,…)

● Equipment and material used in test (glassware, syringes, needles)

● Retaining boxes (comfortable for rabbits as possible)

● Thermometers or electrical device (standardized position in rectum, precision of 0.1°C)

Preliminary test:

- After selection of the animals, one to three days before testing the product to be examined.

- Intravenous injection of 10 ml per kilogram of body mass of a sterile pyrogen-free saline solution warmed to about 38.5°C.

-To exclude any animal showing an unusual response to the trauma (shock) of injection.

- Record the temperatures of the animals, beginning at least 90 min. before injection and continuing for 3 h after the injection of solution.

-Any animal showing a temperature variation greater than 0.6°C is not used in the main test.

-All glass ware, syringes and needles must be pyrogen free by heating at 250°C for not less than 30min.

Main test:

-Group of 3 rabbits

-Preparation and injection of the product:Warming the product to approximately 38.5°C before the injection.

-Dissolving or dilution the product to be examined.

-Injection site: inject the solution slowly into the marginal vein of the ear of each rabbit.

-The injected volume: not less than 0.5 ml per kg and not more than 10 ml per kg of body weight over a period not exceeding 4 min.

-Duration: Record temperature at 30-min intervals for 3 hours

Determination of the initial and maximum temperature:

●The initial temp. of each rabbit is the mean of two temp. readings recorded for that rabbit at an interval 30 min in the 40 min immediately preceding the injection of the product to be examined.

● The maximum temp. of each rabbit is the highest temperature recorded for that rabbit in the 3 h after the injection.

●Record the temp. of each rabbit at intervals of not more than 30 min, beginning at least 90 min before the injection of the product to be examined and continuing 3 hr after the injection.

●The difference between the max. temp. and the initial temp. of each rabbit is taken to be its response.

●Rabbits showing a temp. variation greater than 0.2°C between two successive readings in the determination of the initial temp. are withdrawn from the test.

All rabbits having an initial temp. higher than 39.8°C or less than 38°C are withdrawn from the test.

Interpretation of results:● Test is carried out first on a group of three rabbits, repeat if necessary on further groups of three rabbits to a total of four groups depending on the results obtained.

● If the summed response of the first group does not exceed the figure given in the second column of the table, the substance passes the test.

Number of rabbitsProduct passes if summed response doesn't exceed

Product fails if summed response exceed

31.15°C2.65°C

62.8°C4.3°C

94.45°C5.45°C