1. ENVIRONMENTAL CONTROL:-
  • Traffic control: A carefully designed arrangement to control and minimize the traffic.

Personnel should be permitted to enter aseptic areas only after following rigidly prescribed procedures.

  • Surface disinfection

Personnel:

  1. must be inherently neat, orderly, reliable and alert
  2. should be in good health.
  • Air control:- HEPA (High Efficiency Particulate Air):- It is composed of glass fibers and filters. It is 99.97% efficient. Removes particles of 0.3µm size & larger. Velocity is 100±20 ft/min.  

2. pH MEASUREMENT:- 2 different types of methods used in the measurement of pH.

  • Dip a piece of pH paper into the sample.
  • pH meter
  1. VISCOSITY:- Viscosity and consistency directly relates with stability of solutions. Viscosity is measured by viscometer.
  2. CONTROL ON VOLUME FILLED:- The personnel working in the filling area should have complete control on filling equipments.
  3. OSMOLALITY (Occasionally):- Osmolality is a count of the number of particles in a fluid sample. The osmolality of a solution can be measured using an osmometer. The most commonly used instrument in modern laboratories is a freezing point depression osmometer freezing point depression osmometer.
  4. CONDUCTIVITY MEASUREMENT:- Measured by using conductometer. It is also necessary measure the conductivity of the vehicle used in sterile preparations. The conductivity of the pure water is 0.055 µS/cm (micro-Siemens/cm).
  5. TEMPERATURE FOR HEAT STERILIZED PRODUCTS:- It is important to maintain the constant temperature during heat sterilization of products. The temperature changes may cause some undesirable changes like change in potency, change in isotonicity etc. The temperature can be determined normal thermometer, digital thermometer.
  6. LEAKAGE TEST:- Leakage test is employed to test the package integrity by three methods:-
  • VISUAL INSPECTION
  • BUBBLE TEST
  • DYE TEST

 

9.CLARITY TESTING:- Clarity testing is carried out to check the particulate matter in the sample. It is practically impossible that every unit of lot is perfectly free from visible particulate matter, that is, from particles that are 30 to 40 µm and larger in size. USP limits for large volume infusions:

 

Particle Size Particles limit
10µm and larger/ml 50
25µm and larger/ml 5

  • VISUAL INSPECTION BY NAKED EYE:– Each injectable is inspected visually against white and black backgrounds. The white background aids in detection of dark colored particles. The light or reflective particles will appear against the black back ground.
  • INSTRUMENTAL METHOD:– Also called as the particles count method particles counting may be based on any one of the following principles:
  1. change in electrical resistance
  2. light absorption
  3. light scattering
  4. STERILITY TEST:- The test method for sterility of the product:
  5. Membrane filtration
  6. Direct inoculation of the culture medium
  7. Membrane filtration:- Solutions to be examined must be introduced and filtered under aseptic conditions All steps of this procedure are performed aseptically in a Class 100 Laminar Flow Hood
  • pore size of 0.45 mm
  • effectiveness established in the retention of micro-organisms.
  • the size of filter discs is about 50 mm in diameter.

 

11.  PYROGEN TEST:-

  • Group of 3 rabbits are taken
  • preparation and injection of the product:
  1. warming the product
  2. dissolving or dilution
  3. duration of injection: not more than 4 min
  4. injected volume: not less than 0.5 ml per 1 kg and not more than 10 ml per kg of body mass.
  • determination of the initial and maximum temperature
  1. all rabbits should have initial T: from 38.0 to 39.8 ° C

The Result of Pyrogen Test:-

 

No. Of rabbit

Individual temp. Rise

OC

Temp. Rise in group OC

Test

3 rabbits

0.6

1.4

Pass

If above not passes

3+5 = 8 rabbits

0.6

3.7

Pass

If above test not passes perform the test again. Again on failure the sample said to be pyrogenic.

  1. LAL TEST:- Limulus amebocyte lysate (LAL) test to detect or quantify endotoxins of gram-negative bacterial origin reagent amoebocyte lysate from horseshoe crab ( Limulus polyphemus ). LAL reacts with bacterial endotoxin. It involves 3 methods:
  • Gel clot method
  • Turbidimetric method
  • Chromogenic method