PARENTERALS

Definition

“ Parenterals are the sterile dosage forms intended for administration other than the enteral route and exert their action by directly entering into the systemic circulation.”

LEAKER TEST PYROGEN TESTPARTECULATE MATTER TESTSTERILITY TEST

Tests for Parenterals

This test is used to detect incompletely sealed ampoules.It is performed on each and every ampoules.

Used to prevent microbial contamination.

3methods are used for this: dye bath liquid loss high voltage conductivity

LEAKER TEST(USP_NF)

REQUIREMENTS:Healthy rabbits of either sex. Weight should greater than 1.5kg.Should be free from antibiotics.Animal should not lose its body weight 1week

before test performance.Mean rise in temperature is >than 1.2degree.EQUIPMENTS:Thermometer,glassware,syringes,needles,retaining boxes.

PYROGEN TESTING(B.P)

PROCEDUREPRELIMINARY TEST MAIN TEST Counter check test about

the authenticit y of product.

Material inserted is 0.9% w/v NaCl i.e pyrogen free.

Conducted 1-3 days before main test.

Dose of inserted substance is 10ml/kg BW.

Solution is at 38.5degree C.

Divide animals into 4 groups each group contain 3rabbit.

Sample inserted is in liquid or solid form.

Dose of inserted material is 0.5_10ml/kg BW.

Inserted in marginal vein in ear

of rabbit in 4sec.

2things noted in it.INITIAL TEMPERATURE: noted before insertion

of the pyrogen free substance. 90mint. before should be noted.

FINAL TEMPERATURE: Noted after 3hoursof

insertion and noted on regular interval and max. reading is final temp.

RESPONSE:=final temp.-initial temp.

2things noted in it.INITIAL TEMPERATURE: Initial temp. is mean of 2

temperature readings recorded for a rabbit with an interval of 30mint. within 40 mint. immediately before start of test.

FINAL TEMPERATUER: Max. temp within

3hours.RESPONSE:=final temp.-initial temp.

OBSERVATIONS

NO.OF RABBITS MATERIALS PASSED IF SUMMED RESPONSE DON’T EXCEED

MATERIAL FAILED IF SUMMED RESPONSE EXCEED

3 1.15 2.65

6 2.80 4.30

9 4.45 5.95

12 6.60 6.60

INTERPRETATION OF RESULTS

In vivo by U.S.P:The test involves measuring the rise in

temperature of rabbits after the I.V injection. In the test rabbit the dose not to exceed 10ml per kg injected intravenously within the period of not more than 10 minutes.

PYROGEN TESTING(USP_NF)

Take 3 rabbitsNot more than 30 mins prior to inj of test

dose note the control temperature.According to the monograph inject the dose

into the ear vein of rabbit.Dose should be 10 ml per kg body weight of

rabbit & complete the dos within 10 mins.Record the temperature at 30 min intervals

between 1 & 3 hrs subsequent to the injection

PROCEDURE

If no rabbit shows an individual rise in temp of 0.5 degree or more above control temp the test is passed.

If any rabbit shows rise in temp of 0.5 or more, take 5 more rabbits. If not more than 3 of 8 rabbits shows individual rise in temp of 0.5 or more, and if the sum of the 8 individual max temp rises does not exceed 3.3, the test is passed otherwise failed.

Test interpretation

Reagent used: LAL reagent is used.The presence of endotoxins are determined by the following methods :

Gel clot techniqueTurbimetric methodChromogenic method

IN VITRO TESTING OF PYROGENS

According to B.P Manual method Automatic methodAccording to U.S.P Optical microscopy Light blockage method

PARTICULATE MATTER TESTING

Articles are tested by membrane filtration. If the membrane filtration is unsuitable ,use the

direct inoculation of the culture medium method.

Aseptic conditions must be provided for the test.

Personnel must be trained. When evidence of microbial contamination in

the article is obtained , failure of the article to meet the requirements of the test for sterility is determined

STERILITY TESTING

MEDIUM USED:Fluid thioglycollate medium: is primarily

intended for the culture of anaerobic bacteria. However it can also detect aerobic bacteria.

Soybean-casein digest medium: is suitable for the culture of both fungi and aerobic bacteria.

PROCEDUREMembrane filtration

Direct inoculation

used for filterable aqueous preparations, alcoholic or oily preparations etc.

Membrane filters having pore size not greater than 0.45 um are used in 50 mm diameter .

transfer the quantity of the preparation to be examined in directly into the medium so that the volume of the product is not more than 10% of the vol. of medium.

If the product to be examined has antimicrobial activity then this activity should be neutralized.

After the incubation period, examine the media for microscopic evidence of microbial growth.

If no evidence of microbial growth is found, the product to be examined complies with the test for sterility.

if evidence of microbial growth is found, the product to be examined does not comply with the test for sterility.

RESULTS