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  1. 1. PREFORMULATION PREPARED BY:- PANDYA DEEP R. M.PHARM SEM-1 PHARMACEUTICS Guided By:- Mr. Keyur Patel Pharmaceutics Department K.B.Raval College Of Pharmacy 1
  2. 2. Incompatibility- general aspects ►inactivation of drug through either decomposition or loss of drug by its conversion to a less favorable physical or chemical form. ►It affects safety, therapeutic efficacy, appearance or elegance. ►When we mix two or more API and / or excipient with each other & if they are antagonistic & affect adversely the safety, therapeutic efficacy, appearance or elegance then they are said to be incompatible. 2
  3. 3. Importance of Drug Excipient Compatibility Study:-  Stability of the dosage form can be maximized.  It helps to avoid the surprise problems.  DECS data is essential for IND (investigational new drug ) submission.  Determine a list of excipients that can be used in final dosage form. 3
  4. 4.  Aspects of above tests are- ►Identification of compatible excipients for a formulation ►Identification of stable storage conditions for drug in solid or liquid state. 4
  5. 5. Types Of Incompatibility  Physical incompatibility:- It involves the change in the physical form of the formulation which involves color changes, liquefaction, phase separation or immiscibility.  Chemical incompatibility:- It involves undesirable change in formulation which is due to formation of new chemical comp. with undesirable activity or our formulation undergoes hydrolysis, oxidation, reduction, precipitation, decarboxylation, racemization.  Therapeutic incompatibility:- It is type of in vivo compatibility. 5
  6. 6.  Solid state reactions: much slower and difficult to interpret.  Liquid state reactions: easier to detect - Acc. to Stability Guidelines by FDA following conditions should be evaluated for solutions or suspensions . 6
  7. 7.  FOR SOLID STATE REACTIONS: SampleA: -mixture of drug and excipient SampleB: -SampleA+ 5% moisture SampleC: -Drug itself without excipients 7
  8. 8.  All the samples of drug-excipient blends are kept for 1-3 weeks at specified storage conditions.  Then sample is physically observed .  It is then assayed by TLC or HPLC or DSC.  Whenever feasible, the degradation product are identified by MASS SPECTROSCOPY, NMR or other relevant analytical techniques. 8
  9. 9. FOR LIQUID STATE REACTIONS:  Place the drug in the solution of additives.  Both flint and amber vials are used.  This will provide information about Susceptibility to oxidation. – Susceptibility to light exposure. – Susceptibility to heavy metals.  In case of oral liquids, compatibility with ethanol, glycerin ,sucrose, preservatives and buffers are usually carried out. 9
  10. 10. Technique Name  1) TMA a) DSC b) DTA  2) ASS  3) FT-IR  4) DRS  5) Chromatography  6) Radiolabelled Techniques  7) Vapour Pressure osmometry  8) Flourescence Spectroscopy 10
  11. 11. 11
  12. 12. KNOWN INCOMPATIBILITIES Functional group Incompatibility Type of reaction Primary amine Mono & Di-saccharides Amine-Aldehyde & Amine-Acetal Ester, Lactone Basic component Ring opening, Ester base hydrolysis Aldehyde Amine, Carbohydrate Aldehyde-Amine, Schiff base Or Glycosylamine formation Carboxyl Base Salt formation Alcohol Oxygen Oxidation to Aldehyde & Ketones Sulfhydryl Oxygen Dimerization Phenol Metal Complexation Gelatin- Capsule Shell Cationic Surfactant Denaturation 12
  13. 13. 1) INTRODUCTION Thermal method of analysis are group of techniques in which changes in physical and /or chemical properties of a substance are measured as a function of temperature, while substance is subjected to controlled temp programme. 13
  15. 15.  They are unique methods in the field of polymer analysis & of high value for a solid state analysis.  They finds wide application in  Detection of impurity  Determination of moisture content in any drug substance or any excipient  Study of polymorphism  Characterization of hydrates & solvates  Degree of Crystallinity  Study of phase diagram  Drug excipient compatibility study  Study of complexation 15
  17. 17.  PRINCIPLE: TG is a technique in which a change in the weight of a substance is recorded as a function of temperature or time.  Instrument: Instrument used for thermogravimetry is thermobalance → Major components of a Thermobalance  Sample container ,usually shallow platinum crucible.  Furnace Assembly  Automatic recording Balance(Micro balance 17
  18. 18.  Factors affecting Thermogravimetry analysis are  Heating rate  atmosphere  Sample characteristic 18
  20. 20.  PRINCIPLE: It is a technique in which the energy necessary to establish a zero temp difference between the sample & reference material is measured as a function of temp.  Endothermic reaction. E.g. Melting, boiling, sublimation, vaporization, desolvation.  Exothermic reaction: E.g crystallization, degradation, polymerization. 20
  21. 21. DIFFERENTIAL SCANNING CALORIMETRY (DSC)  DSC is widely used to measure glass transition temp & characterization of polymer.  Glass Transition temp(Tg): Temp at which an amorphous polymer or an amorphous part of crystalline polymer goes from hard ,brittle state to soft, 21
  22. 22. C) DIFFERENTIAL THERMAL ANALYSIS (DTA)  PRINCIPLE: A Technique in which the temperature difference between a substance & a reference material is measured as a function of temperature, while the substance & reference are subjected to a controlled temperature programme.  The Difference in temperature is called as Differential temp(∆t) is plotted against temperature or a function of time.  Physical changes usually result in Endothermic peak ,whereas chemical reactions those of an oxidative nature are exothermic. 22
  23. 23. MAJOR APPLICATION OF THERMAL ANALYSIS IN PREFORMULATION  A) Characterization of hydrates and solvates .  B) study of polymer  C) Detection of impurity.  D) Prediction of stability of drug  E) Degree of crystallinity  F). Compatibility study of drug with excipient.  G) Study of complexes & inclusion compounds. 23
  24. 24. MINOR APPLICATIONS  - DSC is a valuable tool in choice of suppository base.  - In study of polymer composition ,miscibility & individual characterization. - Study of tablet coating - Determinations of melting point. etc - Determination of moisture content in drug. - Checking technological quality grade of disintegrate. - Study of solid drug dispersion. - Determination of drying temp. for different excipients. 24
  25. 25. LIMITATIONS OF THERMAL ANALYSIS  1) low sensitivity for transitions involving small energies.  2) The interpretation of the curves & DSC studies should always Include several conditions & replicate.  3) Impurity consisting of molecules of same size, shape, & character as those of the major component are not detected by DSC, Because these impurity fit into the matrix of the major component without disruption of lattice forming solid solution or inclusion ,such impurity are not detected by DSC.  4) TGA used to studies hydrates & moisture study is not always reliable. 25
  26. 26. X-RAY DIFFRACTION IN PREFORMULATION  METHOD:- Principle:- «- An x-ray irradiation produce a highly specific diffraction pattern from a crystal of material.  An X-ray diffraction pattern from the crystal is formed a series of dots of varying intensity with fixed angular & recorded on photographic film. «-  It is powerful tool for particle size analysis. 26
  27. 27. FTIR TECHNIQUE:  It has been used to quantify binary mixtures of polymorphs.  In identification of polymorphs , only solid samples (as mineral oil mulls & KBr pellets) can be used .  In solutions polymorphs of a compound have identical spectra .  Techniques for solid state stability studies:  Advantages: - Rapid. • TECHNIQUE IS QUALITATIVE & QUANTITATIVE. 27
  28. 28. (1) INTRODUCTION  Impurities in pharmaceuticals are the unwanted chemicals that remain with active pharmaceutical ingredients (APIs),or develop during formulation, or upon aging of APIs and formulated APIs to medicines.  Webster‘s dictionary defines impurity are something that is impure or makes something else impure. .  ►Organic volatile impurities OVIs are generally solvents used in the synthesis or during the formulation of the drug product. 28
  29. 29.  a) impurities associated in with APIs  b) impurities that forms are created during formulation & or with aging or that are related to formulated forms.  According to ICH guidelines impurities associated with APIs are classified in to following categories- (a)Organic impurities (b)Inorganic impurities (c)Residual solvents (d)Other materials 29
  30. 30. Starting materials Intermediates Penultimate intermediate(Final intermediates) By-products Interaction products Related products Degradation products 30
  32. 32. ICH HARMONISED TRIPARTITE GUIDELINE  GENERAL PRINCIPLES Classification of Residual Solvents by Risk Assessment  The term "tolerable daily intake" (TDI) is used by the International Program on Chemical Safety (IPCS) to describe exposure limits of toxic chemicals and "acceptable daily intake"(ADI) is used by the World Health Organization (WHO) and other national and international health authorities and institutes. The new term "permitted daily exposure" (PDE) is defined in the present guideline as a pharmaceutically acceptable intake of residual solvents to avoid confusion of differing values for ADI's of the same substance.  Residual solvents assessed in this guideline are listed in Appendix 1 by common names and structures. They were evaluated for their possible risk to human health and placed into one of three classes as follows: 32
  33. 33. CLASSES EVALUATED ON THE BASES OF THEIR POSSIBLE RISK  Class 1 solvents: Solvents to be avoided carcinogens and environmental e.g.,Benzene, Carben tetrachloride, 1,1-Dichlorobenzene, etc  Class 2 solvents: Solvents to be limited Non-genotoxic animal carcinogens or possible causative agents of other irreversible toxicity such as neurotoxicity or teratogenicity. e.g.,Acetonitrile,Chlorobenzene,Chloroform,Cyclohexane  Class 3 solvents: Solvents with low toxic potential Solvents with low toxic potential to man;Class 3 solvents have PDEs of 50 mg or more per day. e.g.,Acetone, Anisol, 1-Butanol, Cumene, Ethyl ether,etc .  Class4 solvents: Solvents for which no toxicological data have been found Sovents for which no adequate toxicological have been found. e.g.,1,1-Dietoxypropane,Trifluroacetic acid,Ethyl acetate,etc. 33
  34. 34. METHODS FOR ESTABLISHING EXPOSURE LIMITS:-  The Gaylor-Kodell method of risk assessment, is appropriate for Class 1 carcinogenic solvents. Only in cases where reliable carcinogenicity data are available should extrapolation by the use of mathematical models be applied to setting exposure limits. Exposure limits for Class 1 solvents could be determined with the use of a large safety factor  Acceptable exposure levels in this guideline for Class 2 solvents were established by calculation of PDE values. PDE is derived from the no-observed-effect level (NOEL), or the lowest-observed effect level (LOEL) in the most relevant animal study as follows:  PDE = NOEL x Weight Adjustment F1x F2 x F3 x F4 x F5  The PDE is derived preferably from a NOEL. If no NOEL is obtained, the LOEL may be used. 34
  35. 35. METHODS FOR ESTABLISHING EXPOSURE LIMITS:-  F1 = A factor to account for extrapolation between species, F1 takes into account the comparative surface area: body weight ratios for the species concerned and for man.  F2 = A factor of 10 to account for variability between individuals.  F3 = A variable factor to account for toxicity studies of short-term exposure  F4 = A factor that may be applied in cases of severe toxicity, e.g. non-genotoxic carcinogenicity, neurotoxicity or teratogenicity.  F5 = A variable factor that may be applied if the no-effect level was not established 35
  36. 36. LIMITS OF RESIDUAL SOLVENTS :-  Solvent Concentration Limit (ppm)  Benzene 2  Carbon tetrachloride 4  1,2-Dichloroethane 5  1,1-Dichloroethene 8  1,1,1-Trichloroethane 1500 Solvents to Be Avoided:- 36
  37. 37. LIMITS OF RESIDUAL SOLVENTS Class 2 solvents in pharmaceutical products. .  SOLVENT CONCENTRATION LIMIT (PPM)  Acetonitrile 410  Chlorobenzene 360 Chloroform 60  Cyclohexane 3880  1,2-Dichloroethene 1870 Dichloromethane 600  N,N-Dimethylacetamide 1090  N,N-Dimethylformamide 880  Ethylene glycol 620  Formamide 220  Hexane 290  Methanol 3000 37
  38. 38. LIMITS OF RESIDUAL SOLVENTS  Acetic acid, Heptane, Acetone,  Isobutyl acetate, Anisole, Isopropyl acetate,  1-Butanol, Methyl acetate, 2-Butanol,  3-Methyl-1-butanol, Butyl acetate, Methylethyl ketone,  tert-Butylmethyl ether, Methylisobutyl ketone, Cumene,  2-Methyl-1-propanol, Dimethylsulfoxide, Pentane,  Ethanol, 1-Pentanol, Ethyl acetate, 1-Propanol, Ethyl ether, 2-Propanol, Ethyl formate, Propyl acetate Formic acid, Class 3 solvents which should be limited by GMP or other quality based requirements. 38
  39. 39. LIMITS OF RESIDUAL SOLVENTS Solvents for which no adequate toxicological data was found are given below. o 1,1-Diethoxypropane, o Methylisopropyl ketone, o 1,1-Dimethoxymethane, o Methyltetrahydrofuran 2, o 2-Dimethoxypropane, o Petroleum ether, o Isooctane, o Trichloroacetic acid, o Isopropyl ether 39
  40. 40. AS PER U.S. PHARMACOPOEIA REGULATORY LIMITS  Organic volatile USP/NF limits USP/NF 2003 impurities before 2003 (ppm) limits (ppm) Benzene 100 Not specific o Chloroform 50 60 o 1,4-Dioxane 100 380  Methylene chloride 500 600  Trichloroethylene 100 80 40
  41. 41. PREFORMULATION STUDY OF BIOTECHNOLOGICAL PRODUCTS  INTRODUCTION TO BIOTECHNOLOGY  "Biotechnology means any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific use." 41
  42. 42. PHYSICO-CHEMICAL PROPERTIES OF BIOTECHNOLOGICAL PRODUCTS  1. Purity, Impurities and Contaminants  2. Structural Characterization  3. Conformation Study  4. Physical Characterization 42
  43. 43. 4. PHYSICAL CHARACTERIZATION OF BIOTECHNOLOGICAL PRODUCTS  1 SOLID STATE PROPERTIES. 1 Crystallinity 2 Hygroscopicity 3 Molecular weight 4 Thermal Denaturation temperature 5 Extinction coefficient.  2 SOLUTION PHASE PROPERTIES 1 Ionic Strength 2 pH 3 Adsorption 4 Solubility 5 Surface interaction 43
  44. 44. 1. PURITY, IMPURITIES AND CONTAMINANTS  The impurities in the products are:  Process related cell-culture media, host cell proteins, DNA, monoclonal antibodies  Product related product during the manufacture &/or storage 44
  45. 45. PURITY DETERMINATION TECHNIQUE  Protein based product  SDS-PAGE ,HPLC ,Immunological approaches like immunoassay  Amino acid analysis ,N-terminal sequencing ,peptide mapping  Pyrogen detection  Rabbit pyrogen test (exogenous pyrogen) and LAL Test(endotoxins)  DNA  DNA Hybridization studies(dot-blot assay)utilizes radiolabel led DNA probes  Microbial  virus –specific DNA probes , immunoassays. 45
  46. 46. 2.STRUCTURAL CHARACTERIZATION  Protein architecture are typically described in terms of primary ,secondary ,tertiary & quaternary & the order of stability is from primary>>>quaternary.  Protein structure is determined by following techniques, SDS-gel electrophoresis ,HPLC (mainly RP-HPLC) ,Amino acid analysis ,N- terminal sequencing etc 46
  47. 47. 3. CONFORMATION  A protein’s conformation is the most elusive of its physical properties in that it may be the most difficult not only to ascertain but to maintain.  One must establish the other conformational forms present have no undesirable chemical, physical and biological effects.  It can be determined by the following technique ,X-ray diffraction ( to study 3Dprotein conformation ,Circular dichrosim ,Peptide mapping ,N-terminal sequencing or amino acid to determine amino acid sequence. ,Spectroscopic techniques ( UV,IR, Raman, Fluorescence) 47
  48. 48. 4. PHYSICAL CHARACTERIZATION SOLID STATE PROPERTIES  Crystallinity:  Lack of crystallinity is characterized by the low intensity peaks or broad bands in the  POWDER X-ray pattern.  Other methods used are X-ray Diffraction pattern, Solid state NMR (SS-NMR),Differential scanning calorimetry ( DSC), Thermo gravimetric analysis (TGA).  Hygroscopicity:  The changing environmental conditions can affect the stability of the hygroscopic products .The estimation of water content done by the HPLC method 48
  49. 49. 4. PHYSICAL CHARACTERIZATION Molecular weight:  The molecular size of peptides and proteins are 1 to 3 orders of magnitude larger than that of conventional drug molecules, is a major factor limiting their diffusion across the biological membranes.  Biopharmaceuticals molecular weight ranges from 600 to more than 10,000daltons in their non- aggregated state  Larger the size ,lower the permeation across biological membranes g.i.t, ocular epithelium ,stratum corneum considered as impermeable barriers to the permeation to the latter.  Molecular size also restricts diffusion of the biopharmaceutical across the polymeric barriers, used in the fabrication of proteins and peptides delivery systems. 49
  50. 50. 4. PHYSICAL CHARACTERIZATION Extinction coefficient (molar absorptivity):  For quantification purpose, it is desirable to measure at a particular UV-VIS wavelength for the measurement of the protein content. Thermal denaturation temperature:  Most proteins can be reversibly denatured with an increase in temperature and continued increase can result in irreversible denaturation (seen in enzymes).  Also in some case, protein unfolds and refolds into new structure different then original native one.  FREEZE –DRYING of proteins in presence of appropriate buffer excipients can minimize the problem 50
  51. 51. 4. PHYSICAL CHARACTERIZATION SOLUTION PHASE PROPERTIES Adsorption:  In the case of where adsorption is due to ionic interaction of the peptide with the silanol groups on the glass surface, can be prevented by silylating the glass (flasks, syringes,etc) with organosilanes such as Prosil 28. Solubility and charge:  The solubility of a protein is a useful indicator of changes that may occur in the protein conformation  If a protein denatures, more hydrophobic groups may be exposed to the aqueous phase from the interior of the protein. 51
  52. 52. 4. PHYSICAL CHARACTERIZATION PH:  Ph has profound effect on protein stability ,folding and rates of reaction which chemically alter the amino acid residues  In some cases it may be possible to formulate the protein under conditions where chemical modifications and proteolysis are minimized as much as possible while maintaining the protein in misfolded conformation which is readily and rapidly refolded to “native” form upon administration. Ionic strength:  The concentration of counter ions is known to be an important property in mediating electrostatic interactions in molecule, and have large effect on stability of a protein as well as its solubility.  For example: Guanidium HCl is a strong naturant of protein where as Guanidium sulphate at equivalent concentration can stabilize proteins. 52
  53. 53. BIOLOGICAL CHARACTERIZATION  Validated biological assays should be performed at the preformulation stage to assess the biological activity.  For the complex molecules, the physio-chemical information may be extensive but unable to confirm the higher order structures, a biologic activity with a specific quantitative measure.  Radio receptor assays (RIA) may be used to relate binding of a protein hormone to a cellular receptor and may be used to asses activity protein fragments, produced by the enzymatic degradation  Immunogenicity is ability to induce the formation of antibodies when combined with antigenicity is the ability to react with specific antibodies can potentially result in the development of hypersensitivity reactions.  Techniques for minimizing the immunogenicity (considered at the preformulation stage ) 1. Cloned peptides and proteins 2. Conjugation of peptides and proteins to low molecular weight compounds 3. Conjugation of peptides and proteins to various polymers such as dextran, albumin, dl- amino acids, poly vinyl pyrrolidone, polyethylene glycol,etc 53
  54. 54. ORAL DELIVERY OF INSULIN BY CONJUGATION WITH VITAMIN B12  Limitation of oral delivery of insulin: 1. Proteolysis in GIT 2. Absorption at intestinal entrocyte  To overcome this problem, we decided to use the dietary uptake pathway of vitamin B12. Mammals have an active transport mechanism in the GIT for the absorption and the cellular uptake of relatively large vitamin B12. 54
  55. 55. DEGRADATION PROCESSES OF PROTEIN PHARMACEUTICALS  PHYSICAL INSTABILITY 1 Denaturation 2 Surface adsorption 3 Aggregation 4 Precipitation.  CHEMICAL INSTABILITY 1 Hydrolysis 1.1 Deamidation 1.2 Intramolecular aminolysis 1.3 Transpeptidation 2 Oxidation 3 Racemization 4 β-elimination 55
  56. 56. METHODS TO IMPROVE PHYSICAL STABILITY  Additives such as metal ions ,polyalcohol ,surfactants ,reducing agents ,chelating agents and other proteins ,amino acids ,fatty acids and phospholipids  Specific ion binding sites can be introduced on protein molecules in relation to the aggregation and precipitation. eg salts  Both ionic and non-ionic surfactants can stabilize proteins by preventing adsorption of proteins at interfaces  At low concentration in aqueous solutions ,polyhydric alcohol such as glycerol by selective salvation .(excluding the water molecules) 56
  57. 57. METHODS TO IMPROVE CHEMICAL STABILITY:  1. Chemical modification of protein molecules: Synthetic polymers like PEG and Polyoxyethylene can be coupled to protein molecules chemically to enhance the chemical stability. Comparatively, lipids coupled with Insulin can increase the absorption of Insulin and hence can increase the activity.  2 Site directed mutagenesis: This technique creates specific mutations that alter the amino acid sequence of proteins and create new proteins. 57
  58. 58. METHODS TO IMPROVE CHEMICAL STABILITY:  3. Appropriate choice of conditions:  pH,  temperature,  preservative  ionic strength. 58
  59. 59. FORMULATION APPROACHES TO PROTEIN STABILIZATION Protein stabilization in solution using additives:  Concentration of salts has profound effect on protein solution and aggregation.  Non ionic surfactants like, polysorbate 80 inhibits protein aggregation because of greater tendency of surfactant molecule to align themselves at the liquid-air interface, so, excluding the proteins from interface and inhibiting surface denaturation.  Use of cyclodextrin or 2-Hydroxy propyl--cyclodextrin solubilizes ovine growth hormone and prevents shaking induced precipitation. 59
  60. 60. FORMULATION APPROACHES TO PROTEIN STABILIZATION 2. Protein stabilization in the dried solid state: Lyophilization and spray drying used to dehydrate heat sensitive molecules and thereby inhibit the degradative reactions that may be observed when proteins are formulated in solution  Mechanism for Stabilization of Dried Proteins:  Binding of additives to the dried protein to act as a “water substitute” after removal of hydration shell. As water substitutes, sugars such as lactose can serve to partially satisfy Hydrogen bonding requirements of polar groups in dried proteins. 60
  61. 61. FORMULATION APPROACHES TO PROTEIN STABILIZATION  Lyophilization and protein formulation development:  It is characterized by two primary concerns:  1 Excipient selection to optimize long term stability of protein in the dried state.  2. Effect of excipient on processing parameters of Lyophilization cycle such as maximum and minimum drying temperature and cycle length and on the appearance of the lyophilized powder cake.  Freezing causes gradual concentration of solutes surrounding the protein molecule s water is removed by the phase change.  Spray drying of protein pharmaceuticals:  It is alternative to freeze drying proteins as a process capable of drying thermally labile materials.  The particles are dried in the air stream in seconds, owing to high surface area contact with drying gas.  Its major advantage over Lyophilization is particle size and shape of final dried powder. 61
  62. 62. GUIDELINES ON THE STABILITY OF THE BIOTECHNOLOGICAL PRODUCT  1 Preamble  The applicant should develop the proper supporting stability data for biotechnological product and consider many external conditions that can affect the product's potency, purity, and quality.  The purpose of this document is to give guidance to applicants regarding the type of stability studies that should be provided in support of marketing applications. 62
  63. 63. GUIDELINES ON THE STABILITY OF THE BIOTECHNOLOGICAL PRODUCT  2 Scope of the annex  Stability Testing of New Drug Substances and Products'' applies to well-characterized proteins and polypeptides, their derivatives and products of which they are components, and which are isolated from tissues, body fluids, cell cultures, or produced using recombinant deoxyribonucleic acid (r-DNA) technology.  The document does not cover antibiotics, allergenic extracts, heparins, vitamins, whole blood, or cellular blood components. 63
  64. 64. GUIDELINES ON THE STABILITY OF THE BIOTECHNOLOGICAL PRODUCT 3 Selection of Batches  3.1 Drug Substance (Bulk Material) Where bulk material is to be stored after manufacture, but before formulation and final manufacturing, stability data should be provided on at least three batches for which manufacture and storage are representative of the manufacturing scale of production.  3.2 Sample Selection Where one product is distributed in batches differing in fill volume (e.g., 1 milliliter (mL), 2 mL, or 10 mL), unit age (e.g., 10 units, 20 units, or 50 units), or mass (e.g., 1 milligram (mg), 2mg, or 5 mg), samples to be entered into the stability program may be selected on the basis of a matrix system and/or by bracketing. 64
  65. 65. GUIDELINES ON THE STABILITY OF THE BIOTECHNOLOGICAL PRODUCT 4. Stability-Indicating Profile  Consequently, the manufacturer should propose a stability- indicating profile that provides assurance that changes in the identity, purity, and potency of the product will be detected.  At the time of submission, applicants should have validated the methods that comprise the stability-indicating profile, and the data should be available for review.  The determination of which tests should be included will be product-specific.  4.1 Protocol  The protocol should include all necessary information that demonstrates the stability of the biotechnological product throughout the proposed expiration dating period including, for example, well-defined specifications and test intervals. 65
  66. 66. GUIDELINES ON THE STABILITY OF THE BIOTECHNOLOGICAL PRODUCT  4.2 Potency  For the purpose of stability testing of the products described in this guideline, potency is the specific ability or capacity of a product to achieve its intended effect and is determined by a suitable in vivo or in vitro quantitative method.  For that purpose, a reference material calibrated directly or indirectly against the corresponding national or international reference material should be included in the assay. 66
  67. 67. GUIDELINES ON THE STABILITY OF THE BIOTECHNOLOGICAL PRODUCT 5. Storage Conditions Temperature  As most finished biotechnological products need precisely defined storage temperatures, the storage conditions for the real-time/real-temperature stability studies may be confined to the proposed storage temperature. Humidity  Biotechnological products are generally distributed in containers protecting them against humidity. Therefore, where it can be demonstrated that the proposed containers (and conditions of storage) afford sufficient protection against high and low humidity, stability tests at different relative humidity can usually be omitted. 67
  68. 68. GUIDELINES ON THE STABILITY OF THE BIOTECHNOLOGICAL PRODUCT Accelerated and Stress Conditions  The expiration dating should be based on real- time/real-temperature data. However it is strongly suggested that studies be conducted on the drug substance under accelerated and stress conditions.  Also useful in determining whether accidental exposures to conditions other than those proposed (e.g., during transportation) are deleterious to the product and also for evaluating which specific test parameters may be the best indicators of product stability.  Light 68
  69. 69. GUIDELINES ON THE STABILITY OF THE BIOTECHNOLOGICAL PRODUCT Container/Closure  Stability studies should include samples maintained in the inverted or horizontal position (i.e., in contact with the closure), as well as in the upright position, to determine the effects of the closure on product quality.  The applicant should demonstrate that the closure used with a multiple-dose vial is capable of withstanding the conditions of repeated insertions and withdrawals so that the product retains its full potency, purity, and quality for the maximum period specified in the instructions-for-use on containers, packages, and/or package inserts. Stability after Reconstitution of Freeze-Dried Product  The stability of freeze-dried products after their reconstitution should be demonstrated for the conditions and the maximum storage period specified on containers, packages, and/or package inserts. 69
  70. 70. GUIDELINES ON THE STABILITY OF THE BIOTECHNOLOGICAL PRODUCT 6. Testing Frequency  The shelf lives of biotechnological products may vary from days to several years.  With only a few exceptions, however, the shelf lives for existing products and potential future products will be within the range of 0.5 to 5 years.  When shelf lives of 1 year or less are proposed, the real-time stability studies should be conducted monthly for the first 3 months and at 3 month intervals thereafter.  For products with proposed shelf lives of greater than 1 year, the studies should be conducted every 3 months during the first year of storage, every 6 months during the second year, and year of storage, every 6 months during the second year, and annually thereafter. 70
  71. 71. GUIDELINES ON THE STABILITY OF THE BIOTECHNOLOGICAL PRODUCT 7. Specification:  Although biotechnological products may be subject to significant losses of activity, physicochemical changes, or degradation during storage, international and national regulations have provided little guidance with respect to distinct release and end of shelf life specifications.  Recommendations for maximum acceptable losses of activity, limits for physicochemical changes, or degradation during the proposed shelf life have not been developed for individual types or groups of biotechnological products but are considered on a case-by-case basis. 71
  72. 72. GUIDELINES ON THE STABILITY OF THE BIOTECHNOLOGICAL PRODUCT 8. Labeling  For most biotechnological drug substances and drug products, precisely defined storage temperatures are recommended.  Specific recommendations should be stated, particularly for drug substances and drug products that cannot tolerate freezing.  These conditions, and where appropriate, recommendations for protection against light and/or humidity, should appear on containers, packages, and/or package inserts. 72
  73. 73. QUE.  a) A Pharmacist wants to develop a transdermal patch of hormone. ,What test should be carried out in order to ascertain drug-excipient incompatibility? Give outline and significance of each test.  Discuss the considerations of OVIs in preform  Give an applications of Differential scanning calorimetry and FTIR in preformulation studyulation study.  Why OVIs are considered in preformulation study?  How the preformulation studies for biotechnologically derived products are carried out? 73
  74. 74. 74