Ipqc tests for parentrals

1. IPQC TESTS FOR PARENTRALS MOSES.M M.PHARMACY 256212886050 GUIDED BY: Dr. NIRMALA Dept. of Pharmaceutics Presented By:

2. Introduction to Parentrals : What are parentrals ? Greek para = "beside" and enteron = "intestine", because it bypasses the intestines .

3. Parentrals are the dosage form for conveying drug by means of injection through the skin or mucous membrane. Parenteral drugs are administered directly into the veins, muscles or under the skin, or more specialized tissues such as the spinal cord. Circumvents the highly efficient first line body defense that is skin and mucous membrane. Thus they should be free from microbial contamination and should have high purity .

4. Definition: Parenterals are Sterile Pyrogen -free & Free from particulate matter which are injected into the internal body compartment.

5. Quality Assurance: It is the sum total of the organized arrangements with the objective of ensuring that the products will be of the quality required for their intended use.

6. GMP: Is that part of Quality Assurance aimed at ensuring that the products are consistently manufactured to a quality appropriate to their intended use.

7. Quality Control: Is that part of GMP concerned with sampling, specification & testing, documentation & release procedures which ensure that the necessary & relevant tests are performed & the product is released for use only after a.scertaining it’s quality.

8. In Process Quality Control IPQC means controlling the procedures involved in manufacturing of the dosage forms starting from raw material purchase to dispatch of the quality product in ideal packaging. It monitors all the features of the product that may affect its quality and prevent errors during processing. It is the activity performed between QA and QC.

9. Importance: To minimize human errors. Provides accurate, specific and definite description of the procedure to be employed. It is a planned system to identify materials, equipments processed and operations. Is to detect the errors if and when it does occurs. Is to enforce the flow of manufacturing and packing operations according to established routes and practice.

10. IPQC For Parenterals : Each filled vials and ampoules are subjected to inspection for particles, volume, cap or seal conditions. Assay: Drug content Clarity test: For particulate matter by Visual method Coulter counter method Filtration method/Microscopic count method Light blockage method

11. Continued.. Conductivity test Fill volume Leak test: Dye bath test Liquid loss test High voltage test Spark test

12. Continued.. Sterility test: Membrane filtration method Direct inoculation method Pyrogen test: Testing on Rabbits Limulus Amoebocyte Lysate (LAL) test pH Label check Pass ability of Parenteral preparation through needle(Viscosity)

13. Drug Content: Assay: Assay is performed according to the method given in the monograph of that parenteral preparation in the pharmacopoeia. Assay is done to check the quantity of medicament present in the parenteral preparation.

14. Clarity test: VISUAL INSPECTION BY NAKED EYE : Each injectable is inspected visually against White and Black Backgrounds. The White backgroud aids in detection of dark colored particles . The light reflective particles will appear against the Black background .

15. Coulter Counter method The sample solution is added to an electrolyte solution which is drawn through a small orifice. As particle passes through the orifice it displaces its own volume of electrolyte. Particle detected by the increase in electrical resistance. Voltage pulses are proportional to the particle size. Particles below 0.2µm can also be detected.

16. Particle size SVP LVP ≥10µm 3000/container 12/ml ≥25µm 300/container 2/ml Limits for detection of subvisible Particulate matter as prescribed in USP

17. Microscopic count method Membrane filter and microscopes are used.

18. Light blockage/Obstruction method This method uses an electronic counter that produces a light beam of high intensity. The solution is allowed to pass under this bright light. A shadow is formed if a particle is present. The particles are counted by the no. of shadows. Limits for subvisible particulate matter as prescribed in USP Particle size SVP LVP ≥10µm 6000/container 25/ml ≥20µm 600/container 3/ml

19. Leakage Test Ampoules are subjected to this test. Leakage test not done for vials and bottles. 1. using methylene blue solution/Dye bath test 2. spark test

20. Leakage test (with methylene blue solution) The ampoules are immersed in vacuum chamber consisting of 1% methylene blue solution A vacuum of about 27” (inch) Hg is created for about 15 to 30 min This causes the solution to enter the ampoules with defective sealing. The vacuum is released and ampoules are observed. If a leakage is present, the solution in the ampoules appear blue color.

21. Spark test The machine uses high precision electrodes to inspect the full circumference of the containers, including the closure zone. All containers are presented inividually to the electrodes. Any moisture that has penetrated through capillary forces in a crack, pinhole or just weak glass is registered as a change in resistance. All products with a measured voltage higher than a defined maximum value are separated from the good products.

22. High Voltage Leak Dectection

23. Advantages of HVLD test Vials and ampoules also can be tested. High accuracy of inspection. High speed of processing.

24. Sterility Test Sterility testing attempts to reveal the presence or absence of viable micro-organism in a sample number of containers taken from batch of product. Based on results obtained from testing the sample a decision is made as to the sterility of the batch. Sterility testing is made after the product exposition to the one of the possible sterilization procedures.

25. Principle Sterility test is based on the principle that when micro-organisms are supplied with nutrient medium and water, and incubated at favorable temperatures, they multiply. The presence of micro organisms can be identified by turbidity in the clear medium.

26. Culture Media The culture media used for sterility test must be capable of promoting the growth of a wide range of micro organisms. Types of media- 1. Fluid thioglycollate medium (for anaerobic bacteria). 2. Soyabean -casein digest medium (for aerobic bacteria and fungi). Incubation of media: 14 days at 30-35 o C

27. Fluid Thioglycolate Medium Ingredients Quantity for 1000 ml L- cysteine 0.5 g Sodium chloride 2.5 g Dextrose 5.5 g Agar 0.75 g Yeast extract 5.0 g Pancreatic digest of casein 15.0 g Sodium thioglycollate 0.5 g Resazurin (0.1% fresh solution) 1.0 ml Distilled water Upto 1000 ml

28. Soyabean -casein Digest Medium Ingredients Quantit y for 1000 ml Pancreatic Digest of casein 17 g Peptic digest of soyabean meal 3 g Sodium chloride 5 g Dibasic potassium phosphate 2.5 g Dextrose 2.5 g Distilled water Upto 1000 ml

29. Minimum sample size related to Batch size No. of items in the batch Min. No. of items recommended to be tested Injectable preparations a) upto 100 containers 10% or 4 containers whichever is greater b) 101-500 containers 10 containers c) >500 containers 2% or 20 containers whichever is less Ophthalmic and other non – Injectable preparations a) upto 20 containers 5% or 2 containers whichever is greater b) >200 containers 10 containers

30. No. of items in the batch Min. No. of items recommended to be tested Solids a) <4 containers All containers b) 4 - 50 containers 20% or 4 containers whichever is greater c) > 50 containers 2% or 20 containers whichever is greater

31. For injectable preparations Quality of each container Min. Quality to be used for each culture medium For liquids a) <1 ml Total contents of a container b) 1 ml – 4 ml Half the contents of a container c) 4 ml – 20 ml 2 ml d) 20 – 100 ml 10% of contents of a container e) > 100 ml NLT half the containers For solids a) <50 mg Total contents of a container b) 50 mg – 200 mg Half the contents of a container c) > 200 mg 100 mg

32. Test methods The test methods for the sterility of the products are: 1. Membrane filtration method 2. Direct inoculation of the culture medium

33. Membrane Filtration method Appropriate for: (advantage) Filterable aqueous preparations Alcoholic preparations Oily preparations Preparations miscible with or soluble in aqueous or oily (solvents with no antimicrobial effect) Solutions to be examined must be introduced and filtered under aseptic conditions

34. Selection of the filters Pore size of 0.45µm Effectiveness established in the retention of micro organisms Appropriate composition The size of filter discs is about 50 mm in diameter

35. Procedure Sterilization of filtration system and membrane filtration of examined solution under aseptic conditions (suitable volume, dissolution of solid particles with suitable solvents, dilution if necessary) The membrane is removed, aseptically transferred to container of appropriate culture medium

36. Scheme for sterility test by membrane filtration

37. Direct inoculation of the culture medium Suitable quantity of the preparation to be examined is transferred directly into the appropriate culture medium Volume of product is not more than 10% of the volume of the medium Suitable method for aqueous solutions, oily liquids, ointments and creams

38. Scheme for sterility test by direct inoculation method

39. Interpretation of the results The culture media is examined during and after incubation period to detect the possible microbial growth. The sample passes the test if microbial growth is not found If microbial growth is present, a retest is performed. If growth is absent. Then sample passes the test If microbial growth is present in the retest also, identify the organisms If same organisms are found as in the first test, then the sample fails the test If different organisms are found, retest is performed using twice the number of samples. Passes if microbial growth is not found.

40. Pyrogen Test Pyrogens are the metabolic products of gram negative bacteria Endotoxin – complex of pyrogenic lipopolysaccharide , a protein and inert lipid; Lipid part of the lipopolysaccharide is the main pyrogenic agent; polysaccharide part increases solubility

41. Sources of pyrogen contamination Solvent – possibly the most important source The medicament The apparatus The method of storage between preparation and sterilization

42. Animal and Equipment Selection of animals (healthy, adult, NLT 1.5 kg) Housing of animals Equipment and material used in test (glassware, syringes, needles) Retaining boxes (comfortable for rabbits as possible) Thermometers (standardized position in rectum, precision of 0.1 o C)

43. Housing of Rabbits

44. Preliminary Test Intravenous injection of sterile pyrogen -free saline solution To exclude any animal showing an unusual response to the trauma (shock) of injection Any animal showing a temperature variation greater than 0.6 o C is not used in the main test

45. Main Test Group of 3 Rabbits Preparation and injection of the product Warming the product Dissolving or dilution Duration of injection: NMT 4 min The injected volume: NLT 0.5 ml per 1 kg and NMT 10 ml per kg of body mass Determination of the initial and maximum temperature All rabbits should have initial T: from 38.0 to 39.8 o C The differences in initial T should not differ from one another by more than 1 oC

46. The result of pyrogen test: No. of Rabbits Individual Temp. rise ( o C ) Rise in group ( o C ) Inference 3 0.6 1.4 Pass 3 + 5 = 8 0.6 3.7 Pass

47. Limulus Amoebocyte Lysate Test To detect or quantify endotoxins of gram negative bacterial origin Reagent: Amoebocyte lysate from horseshoe crab ( limulus polyphemu or Tachypleus tridentatus ) Bacterial Endotoxin Test (BET) Monocyte Activation Test (MAT)

48. The endotoxin characteristics Thermostable Water soluble Unaffected by the common bactericides Non volatile These are the reasons why pyrogens are difficult to destroy once produced in a product

49. Horse shoe crab

50. Mechanism of LAL Test The test is based on the primitive blood-clotting mechanism of the horseshoe crab

51. Commercially derived LAL reagent Bleeding adult crabs blood into an anticlotting solution Washing and centrifuging to collect the amoebocyte Lysing in 3% NaCl Lysate is washed and lyophilized for storage

52. Procedure Test: Equal volume of LAL reagent and test solution (usually 0.1 ml of each) are mixed in a depyrogenated test-tube Incubation at 37 o C, for1 hour Remove the tube – invert in one smooth motion (180 o ) Observe the result

53. Different Techniques Three different techniques: The gel-clot technique – gel formation The turbidimetric technique – the development of turbidity after cleavage of an endogenous substrate The chromogenic technique – the development of color after cleavage of a synthetic peptide – chromogen complex

54. Gel Clot Technique A solid gel is formed in the presence of endotoxins This technique requires positive and negative controls Positive controls – a known concentration of endotoxin added to the lysate solution Negative controls – water, free from endotoxins , added to the lysate solution

55. Turbidimetric Technique The test is based on the measurement of opacity change due to the formation of insoluble coagulin Opacity is directly proportional to the endotoxin concentration This technique is used for water systems and simple pharmaceutical products

56. Chromogenic Technique This is based on the measurement of color change which is caused by the release of the chromogenic chemical p - nitroanilide The quantity of the p - nitroanilide produced is directly proportional to the endotoxin concentration

57. Advantages of LAL Test Fast Greater Sensitivity Less Variability Much Less Expensive Alternative to Animal Model More accurate than other Is performed in the pharmaceutical laboratory Specific for endotoxins of gram-negative origin Particularly useful for: Radiopharmaceuticals and cytotoxic agents Products with marked pharmacological or toxicological activity in the rabbit ( e.g. Insulin) Blood products which sometime give misleading results in the rabbit Water for injection where LAL test is potentially more stringent and readily applied

58. pH measurement pH meter is used to measure the pH of parenteral preparations. The pH of injectables should be w.r.t pH of blood (i.e., 7.35 to 7.45) The pH of opthalamics should be w.r.t pH of lacrymal fluid (i.e., 7.2 ± 0.2)

59. Parenterals Problems Remedy Leakage of ampoules Discard the ampoules Use new ampoules with proper sealing Perforation in filter leading to defective filtration Changing the filter

60. Foreign particles/ dust Membrane filtration Fibres Optical and visual inspection pH Use of buffers Leaching Internal coating of glass container

61. References USP, Appendix 788, 56 IP p 659 – 660 Remington, The science and Practice of Pharmacy, 21 st edition p 1367 – 1374 Michael J. Akers and Daniel S. Larrimore , Parenteral Quality Control http://apps.who.int/phint/en/p/docf/

62. Thank you