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Pre-Formulation Studies


What does it convey?

How important is it?

When is it required?

The Wheels Product Development







Preformulation begins or shall be updated immediately after the synthesis and initial toxicity screening of a new drug when a newly synthesised drug shows pharmacological evidence that requires further evaluation in man when formulation and dosage form changes are required Characterization of all Preformulation bulk lots is necessary to avoid misleading predictions of stability or solubility, which depend on particular crystalline form.

These studies should focus on: Physicochemical properties that affect drug performance and development of efficacious dosage form. These properties may provide: Support the need for Molecular modifications A rationale for Formulation Design

Preformulation investigations are designed to deliver all necessary data (especially physicochemical, Physiomechanical and biopharmaceutical properties of drug substances, excipients and packaging materials) which may Influence
formulation design method of manufacture of drug substance and drug product pharmacokinetic/biopharmaceutic properties of the resulting product packaging of the product

Direct Benefit
Gives directions for development of formulation in choice of dosage form, excipients, composition, physical structure... Helps in adjustment of pharmacokinetic and biopharmaceutical properties Support for process development of drug substance (yield, filtration...) Support for PAT (Process Analytical Technology) (critical process parameters...) Produce necessary and useful data for development of analytical methods.

Where to begin?
Desk work. Active substance Additives Process conditions Analytical means



To Generate the physical and chemical properties information that will assist the formulator in developing:

*Stable *Safe *Effective Dosage Forms that can be mass produced with: Maximum Bioavailability (Support Dosage Form Design)

II- To provide an initial working model of behaviour of dosage form in vitro and in vivo III- To establish the compatibility of the drug with common excipients. IV- To establish the kinetic rate profile of the drug. V- To Provide a scientific data that support the dosage form design and evaluation of the product efficacy, stability and bioavailability

Evaluation Phase
Calculation/Prediction in silico of many of the important physicochemical characteristics of NCE solubility polymorphism morphology of the particles acute toxicity intestinal permeation

Evaluation Phase
Literature search of similar or related compounds to provide an understanding of the degradation process any adverse conditions relevant to the drug bioavailability pharmacokinetics and formulation of similar or related compounds

Dosage form of choice

Dosage Form Considerations IV vs. oral formulations High dose vs. low dose Excipient compatibility Interaction with other actives in potential combination formulations Salts and Other Solubilization Techniques Effect of Salts on Complexation Binding Constants Effect of Salts on Solublization by Surfactants Solubility of Salts in Non-aqueous Solvents Toxicological Considerations

Choosing the right route

Solid dosage forms Effect of micronization and processing such as granulation on solid state properties and chemical stability Effect of excipients on crystallization/nucleation Powder flow properties: bulk density, compression properties and particle size and shapes

Parenteral Dosage Forms Injection site precipitation Pain upon injection Toxicity of new excipients Effect of excipients on crystallization/nucleation
Suspensions Effect of processing and formulation on the physical and chemical stability Effect of excipients on crystallization/nucleation

Study what?



Index of studies

Stress Testing of API Impact of Impurities on API Specifications Pre-Formulation Investigations Solid State Degradation & Stability Assessment Role of Excipients in API Instability Hydrolysis Oxidation Photolysis API Solubility/Solution-state Stability Assessment Selection of API & Drug Product Processing Methods Degradation Issues for Combination Products Role of API Processing in Product Instability

Analytical Preformulation
Know what you start with..
Identity NMR, UV, TLC, DSC Purity Moisture Content, Impurity Profile, DSC, Heavy element, Inorganic elements. Assay UV, HPLC

Appearance, Odour, pH of Slurry, Melting Point


Potentially critical attributes of API

Cross reference to stress testing (forced degradation): 1. Sensitivity to temperature (wet granulation, sterilization) 2. Sensitivity to moisture (wet granulation, hygroscopicity) 3. Sensitivity to light (packing materials) 4. Sensitivity to oxidation (inert gas atmosphere in ampoules) 5. Sensitivity to pH (FDC with HCL salts of weak bases) 6. Sensitivity to metal ions (internal peroxide bond) Expected degradants, manufacturing conditions, etc. This information is partially available from the Open Part of the DMF


Potentially critical attributes of API

Key physicochemical characteristics: 1. 2. 3. 4. 5. 6. 7. Polymorphic or solid state form (amorphous, hydrate, solvate) Solubility at 37 oC over the physiological pH range (e.g., BCS, dissolution testing, cleaning validation) Permeability (octanol-water partition) (BCS) Crystal habit, particle shape and size (pharmaceutical and bioequivalence, processability) Bulk density, untapped and tapped (processability) Flowability (processability) Color, odor, taste, consistency (choice of dosage form)should be discussed and supported by experimental data.


Stress Testing of API

Deliberate forced degradation of API - serves several purposes:

To facilitate development of a stability indicating analytical method, e.g. HPLC To aid in development of the first API specification To understand the degradation pathways of the API to facilitate rational product development To screen for possible formation of potential genotoxins

Initially performed over a short period of time (28days) using accelerated or stress conditions (so that reactions proceed more rapidly); target ~10% degradation.


Stress Testing of API

Deliberate forced degradation of API

Typical conditions for API in solid-state might be:

80/75%RH, 60C/ambient RH, 40/75%RH, Light irradiation

Typical conditions for API in solution state might be:

pH 1-9 in buffered media with peroxide (and/or free radical initiator) Light irradiation


Impurities: Impact on API Specification

The allowable level of any given impurity or impurities that are permitted in API/drug product, without explicit non-clinical safety testing, are defined by ICH Q3A/B.

The amounts of impurities that are allowable are based on the total daily intake of the drug product.
There are separate limits (or thresholds) for reporting, identification and qualification of API impurities.

The reporting threshold is defined as the level that must be reported to regulatory agencies to alert them to the presence of a specified impurity.


Impurities: Impact on API Specification

The identification threshold is defined as the level that requires analytical identification of a specified impurity. Finally, the qualification threshold is defined as the level where the specified impurity must be subjected to non-clinical toxicological testing to demonstrate safety. Threshold limits are defined as a percentage of the total daily intake (TDI) of the drug product, or in absolute terms as the total allowable amount, whichever is lower.


Impurities: Impact on API Specification

Threshold Maximum Daily Dose of API in Drug Product
1g >1g

Threshold Limit Based on TDI


0.1%TDI 0.05%TDI


<1mg 1mg-10mg 10mg-2g >2g

1.0%TDI or 5g 0.5%TDI or 20 g 0.2%TDI or 2mg 0.1%TDI


<10mg 10mg-100mg 100mg-2g >2g

1.0%TDI or 50g 0.5%TDI or 200g 0.2%TDI or 3mg 0.1%TDI


API solid-state stability study

An early indication of stability challenges for product development:
Accelerated stability challenge but not unrealistically severe and so allows confident extrapolation to provide an indication of API shelf-life Conditions less extreme than API stress testing:
40C/75%RH open vial 50C closed vial At least l month storage data; typically 1w, 2w, 4w, 3m (potentially supporting 12m shelf-life at RT) Light stability (ICH conditions); typically 1week HPLC analysis Monitor solid-state form (crystallinity) - DSC, TGA, pXRD


API solid-state stability study

An early indication of stability challenges for product development:
Allows comparison with other versions & forms of same API Provides a control baseline for excipient compatibility studies Important to bear in mind that API development is ongoing so API batch used in this early stability study may become unrepresentative of final selected API version & form.

API degradation pathways

Hydrolysis and Oxidation are the most common pathways for API degradation in the solid-state and in solution

Photolysis and trace metal catalysis are secondary processes of degradation


API degradation pathways

Temperature affects each of the above chemical degradation pathways; the rate of degradation increases with temperature. Extrapolation of accelerated temperature data to different temperatures, e.g. proposed storage conditions, is common practice (e.g. using Arrhenius plots) but we must be mindful of the pit-falls the influence of the various degradation pathways and mechanisms can change with temperature.


API degradation pathways

It is well understood that pH, particularly extremes, can encourage hydrolysis of API when ionised in aqueous solution. This necessitates buffer control if such a dosage form is required. pH within the micro-environment of a solid oral dosage form can also impact on the stability of the formulation where the API degradation is pH sensitive; through understanding the aqueous pH imparted by typical excipients, a prudent choice can overcome this issue.



What is it?


Classifications Of Solid State Crystal Properties

Solid Substances Chemical compound Crystal Habit Other appearance

Internal Structure
Molecular Ordering


Partly Amorphous crystalline

Crystal Form Crystal Defects Polymorphs Solvates

Lowering of Crystallinity 32

Polymorphism is a solid state property of organic
molecule which can be obtained in more than one distinct crystal form. Polymorphism may also include solvation or hydration products and amorphous forms enantiotropic one polymorph can be reversibly changed
into another one by varying the temperature or pressure monotropic the change between the two forms is irreversible

Importance of Pharmaceutical Polymorphism

Polymorphic forms of a drug substance can have different chemical and physical properties, including melting point, chemical reactivity, apparent solubility, dissolution rate, optical and mechanical properties, vapor pressure, and density.

These properties can have a direct effect on the ability to process and/or manufacture the drug substance and the drug product, as well as on drug product stability, dissolution, and bioavailability.
Thus, polymorphism can affect the quality, safety, and efficacy of the drug product.

Characterization of polymorphs
X- Ray Diffraction Microscopy, Thermal analysis Differential scanning calorimetry, thermal gravimetric analysis, and hot-stage microscopy Spectroscopy Infrared [IR], Raman, solid-state nuclear magnetic resonance [ssNMR]

Why this is Important

Most important question that likely arises during the registration process is What assurance can be provided that no other crystalline forms of this compound exist?. The manufacturer of a new drug substance is obliged to show that due diligence has been employed to isolate and characterize the various solid-state forms of a new chemical entity


Decision Tree for Acceptance of Polymorphism


Trees 1. and 2. consider whether polymorphism is exhibited by the drug substance, and whether the different polymorphic forms can affect performance of the drug product.


Trees 1. and 2. consider whether polymorphism is exhibited by the drug substance, and whether the different polymorphic forms can affect performance of the drug product.


Why This IS Important

Preformulation studies gives answers for critical phenomena so that: the desired forms can be consistently
manufactured; the effects of pharmaceutical manipulations are understood, e.g., granulation, milling and compression; and the effect of storage conditions on the dosage form can be evaluated and predicted

Influence of Polymorphism on Drug Substances and Drug products

Solubility and Dissolution - polymorphic forms differ in their internal solid-state structure, a drug substance that exists in various polymorphic forms can have different aqueous solubilities and dissolution rates. BA/BE

Influence of Polymorphism on Drug Substances and Drug products

Influence on manufacturing of Drug products Drug substance polymorphic forms can also exhibit different physical and mechanical properties, including hygroscopicity, particle shape, density, flowability and compactibility, which in turn may affect processing of the drug substance and/or manufacturing of the drug product.

Ritonavir: HIV protease inhibitor



Case history:
ABT-538 discovered Launch of semi-solid capsule/polymorph I Polymorph II appears, <50% solubility Product pulled from the market 1998 - 1999 Massive effort to reformulate the product 1999 Reformulated softgel capsule launched 1992 1996 1998


Summary of Ritonavir Crystal Forms


mp 122 C

mp 125 C

mp 80 C

mp 97 C

mp 116 C

Launch in 1996 Launch in 1996 Summer of 1998 Summer of 1998 Morissette et al. PNAS 100, (2003). TransForm 2002 6 week effort 2002 5 forms found


API Processing


Role of API Processing in Product Instability

High energy processes (milling, lyophilisation, granulating, roller-compaction, drying) can introduce a degree of amorphicity into otherwise highly crystalline material. This can lead to increased local levels of moisture and increased chemical reactivity in these areas.

With some materials, ball milling causes irregularity, surface faults and imperfections in crystals. The degree of crystal damage can be directly correlated with the energy of the milling process.

Selection of Product Processing

Understanding of degradation pathways of API will help to decide on most appropriate process:
For APIs showing severe moisture mediated degradation pathways, choose direct compression or dry granulation


Selection of Product Processing

Understanding of physical properties of API will help to decide on most appropriate process:
For APIs showing flow issues, choose a granulation approach (wet or dry granulation) For APIs showing reduced crystallinity after processing e.g. milling, micronisation, etc., choose wet granulation (presence of water will anneal (crystallise) amorphous API) For APIs with low melting point, choose an encapsulation approach (high speed rotary presses will generate significant frictional forces that could melt API)

Degradation Issues For Combination Products

Objective is to minimise incompatibilities. Degradation pathways of the two APIs could well be different, so a stabilisation strategy for API #1 could destabilise API #2. In this situation, first intent strategy could be to prepare separate compression blends of each individual API and compress as a bi-layer tablet Disadvantages: adds complexity and bi-layer rotary presses are expensive


Degradation Issues For Combination Products

Objective is to minimise incompatibilities. Alternatively, could compress one of the APIs and overencapsulate this into a capsule product, along with the powder blend from the second API Disadvantage are that capsule size could be large, it requires specialised encapsulation equipment to fill tablets and blend process is more complex and expensive If however, simplicity and cost are significant issues, look to produce a common blend (particle size of APIs should be similar), and by understanding of degradation pathways stabilise the blend and compress or encapsulate.

API related..
Preformulation studies are an important foundation tool early in the development of both API and drug products. They influence.

Selection of the drug candidate itself Selection of formulation components API & drug product manufacturing processes Determination of the most appropriate container closure system Development of analytical methods Assignment of API retest periods The synthetic route of the API Toxicological strategy


Studies designed for Insight



Steps in Preformulation Process Pharmaceutical Research

1. Stability a. Solid State (1) Temperature (2) Light (3) Humidity b. Solution (1) Solvent (2) pH (3) Light 2, Solid State Compatibility a. TLC Analysis b. DRS Analysis 3. Physico-chemical Properties a. Molecular Structure and Weight b. Color c. Odor d. Particle size, Shape, and Crystallinity e. Melting Point f. Thermal Analysis Profile (1) DTA (2) DSC (3) TGA g. Hygroscopicity Potential h. Absorbance Spectra (1) UV (2) IR i. Solubility (1) Water and Other Solvents (2) pH-Solubility Profile (3) Salt Forms (4) Cosolvents (5) Complexation (6) Prodrug j. Effect of pH on UV Spectra k. Ionization Constant l. Volatility m. Optical Activity n. Polymorphism Potential o. Solvate Formation 4. Physico-mechanical Properties a. Bulk and Tapped Density b. Compressibility c. Photomicrograph 5. In Vitro Availability Properties a. Dissolution of Drug Crystal Per se b. Dissolution of Pure Drug Pellet c. Dissolution Analysis of Pure Drug d. Rat Everted Gut Technique 6. Other Studies a. Plasma Protein Binding b. Effect of Compatible Excipients on Dissolution c. Kinetic Studies of Solution 53 Degradation

Preformulation Steps
Solubility (aqueous, pKa, log P, log D) Molecular optimization (salt, hydrate, solvate, new analogs,) Crystal Engineering (polymorph, habit, size, surface characteristics) Crystal structure determination Biopharmaceutical classification (BCS) (solubility, dissolution, absorption)


Preformulation Steps
Complete Physical and Chemical Characterization Drug stability evaluation (physical, chemical, solution phase, solid phase, ) Compatibility analyses (drug substance, excipient, packaging materials) Technical characterization (Flowability, Compactability, ...


Solubility may be defined as the maximum concentration of a substance that may be completely dissolved in a given solvent at a given temperature and pressure. Drug candidates are becoming more lipophilic and poorly soluble A Market Survey of 257 Marketed Drug and their liphophilicity

The USP/NF generally expresses the solubility in terms of the volume of solvent required to dissolve 1 gram of the drug at a specified temperature (eg. 1 g ASA in 300 ml H2O, 5 ml ethanol at 25C).


SATURATION SOLUBILITY is understood as a maximum amount of solute that dissolves in a solvent at equilibrium. Equilibrium is a state where reactants and products reach a balance - no more solute can be dissolved in the solvent in the set conditions (temerature, pressure). Such a solution is called a saturated solution.

Solubility parameters
Descriptive term Very Soluble Freely Soluble Soluble Sparingly soluble Slightly soluble Parts of solvent needed for 1 part of solute <1 1 to 10

10 to 30
30 to 100 100 to 1000 1000 to 10,000

Very Slightly Soluble

Practically insoluble or insoluble

> 10,000

Formulation challenge in Poorly Soluble Drugs

Poor dissolution rate Low and variable bioavailability More potential for food effect Inability to deliver high doses for tox studies Difficulty in developing parenteral formulations


Drug to be in Solution To Be Absorbed


Biopharmaceutical Classification System

Classification Class I - High Permeability, High Solubility Class II - High Permeability, Low Solubility Class III - Low Permeability, High Solubility Class IV - Low Permeability, Low Solubility Class Boundaries Highly soluble: the highest dose is soluble in <250 ml water over a pH range of 1 to 7.5 Highly permeable: >90% dose absorbed in humans Rapidly dissolving: >85% of labeled amount of drug substance dissolves within 30 minutes

Solvent for Solubility Studies

For developability assessment: Simulated gastric fluid (SGF) Simulated intestinal fluid (SIF) pH 7.4 buffer Intrinsic solubility to estimate pH-solubility profile For Formulation Development: pH solubility profile


Factor Causing poor solubility

High Crystallinity/high MP Zwitterion formation Insoluble salts H-bonding networks Hydrophobicity/High LogP Lack of ionizable groups High molecular weight

Dissolution rate for poorly soluble compounds may often be the rate limiting step to absorption Examples of drugs with dissolution rate limited absorption: - Digoxin - Penicillin V - Phenytoin
- Quinidine - Tetracyclines

Approaches to improving solubility


Co-solvents (GRAS approved) Additives/complexes -surfactants, polymers, cyclodextrins Lipid-based systems (solutions, emulsions) Solid-state modification (particle size reduction, salt formation, solid-state stabilisation of the amorphous state)


Particle Size


Micromeritics is the science of particle sizes, particle shapes, size distributions, porosity, density and surface areas.
Methods Microscopy Sieving Laser diffraction Others: Sedimentation, Coulter counter etc.

Advantages For Small sample Direct technique Particle shape can be determined Image analysis system available It can be caliberated Disadvantages Statistically poor Problem with broad size distribution Not very fast Sample preparation Two dimensional analysis

Sieve Analysis
Advantages Determining samples having broad size distribution. Easy to operate Calibrated sieves Disadvantages Inaccuracy increases in the size of < 75 um. Large sample size.

Sieve Analysis


Laser Diffraction
Advantages: Precise Fast Analysis. Small sample amount Well automated and statistically good method Accepted by various regulatory authorities. Disadvantages: Needs expertise. Expensive. Sample is destroyed.

Laser Diffraction


Volume Particle size distribution




Effect of Particle Size on Dissolution

Dissolution profiles in USP apparatus 2 at 50 rpm and pH 4.5 for product produced with different particle size of the API

Dissolution Profile of Viramune and Generic Nevirapine Tablets on the Indian Market
USP Type II / 0.01N HCl 50 RPM / 900 ml 120

% Drug Dissolved

100 80 60 40 20 0
20 73

Time (Min) Viramune B.No.992633B Brand C B.No.C00139 Ranbaxy B.No.(1024)17


0 0.0 0.0 0.0

10 83.3 34.3 88.9

20 96.6 51.3 97.6

30 97.7 61.2 99.2

45 99.7 70.8 99.7


How important?


Primary forces at work: Friction Particle size and shape Cohesion Moisture, fats, oils Molecular forces, electrostatic forces Environmental Temperature, pressure, time Physical Bin angle, aperture diameter Wall conditions

Flowability methods
Flowing time


Flowability Tester Model BEP-Auto



Flowability Tester Model BEP-Auto

The BEP Series can be used in 4 modes: Determination of the flow time of a predetermined sample weight Determination of the flow time of a predetermined sample volume Determination of the weight of sample in a predetermined time Plot of time against sample weight (Weight/Time)

Flowability Tester Model BEP-Auto

According to EP, the flowability is expressed in seconds and tenths of seconds, related to 100 grams of sample. The results can be expressed as: (1) the mean of the three results, if none of the individual values
deviate from the mean value by more than 10% (2) as a range, if the individual values deviate from the mean value by more than 10% (3) as a plot of the mass against the flow time or (4) as an infinite time, if the entire sample fails to flow through.


Angle of Repose
= 2H tan 1 D H = Height of cone [4.1 cm] D = Base of cone [9.8 cm] = Angle of repose 2 x 4.1 tan 1 = 39.93 o 9.8

Angle of repose
Flow property Excellent Good Fair-aid not needed Passable- may hang up/ block Poor- must agitate vibrate Very Poor Very, very poor Angle of repose {degrees} 25 - 30 31 35 36 - 40 41 - 45 46 - 55 56 - 65 > 66

Carr Index
Carr Index =
Carr Index (%) 5-15 12-16 18-21 23-35 33-38 > 40
Tapped Density- Bulk Density X 100 Tapped Density

Type of Flow Excellent Good Fair to Passable* Poor* Very poor Extremely Poor

Environment control


Slightly hygroscopic: Increase in mass is less than 2 percent m/m and equal to or greater than 0.2 percent m/m. Hygroscopic: Increase in mass is less than 15 percent m/m and equal to or greater than 0.2 percent m/m. Very hygroscopic: Increase in mass is equal to or greater than 15 percent m/m. Deliquescent: Sufficient water is absorbed to form a liquid.

Rate of water absorption as a function of RH

0,45 0,40
Lg RH, %

35% 0,35 0,30 0,25 0,20

10 13 16 19 22 25

55% 75% 100%

Lg time, t (3 min. units)



Dynamic Vapour Sorption (DVS)

The relative humidity is generated by bubbling nitrogen through a water reservoir where it is saturated with moisture. Using a mixing chamber, the moist nitrogen is mixed with dry nitrogen in a fixed ratio, thus producing the required relative humidity. The moist nitrogen is then passed over the sample, and the instrument is programmed such that the increase in weight due to moisture is monitored with time using an ultrasensitive microbalance. The compound takes up moisture and reaches equilibrium, at which point the next relative humidity stage is programmed to start. The adsorption and desorption of moisture can be studied using this instrument, and the effect of temperature can be investigated as well. Using this technique, a quantity as small as 1 mg can be assessed. 90

Corrosive potential
Affects which stage?


Personnel safety Equipment Pack Scale of operations Cleaning process Handling procedures


What is covered ?
Bulk properties crystallinity and polymorphism Hygroscopicity (water adsorption) Fine Particle Characterization (particle size, shape, and surface area) powder flow Properties (Bulk Density, angle of repose, compressibility)

Ionization Constant
The unionized species are more lipid-soluble and hence more readily absorbed. The gi absorption of weakly acidic or basic drugs is related to the fraction of unionized drug in solution. Factors affecting absorption: - pH at the site of absorption - Ionization constant - Lipid solubility of unionized species pH-partition theory 94

What is covered?
solubility analysis Intrinsic Solubility ionization Constant pH solubility profile partition coefficients Salts Solvents Dissolution

Dissolution testing
Dissolution testing is used for the selection of the formulation and comparison of the dissolution profiles with that of the innovator product and clinical batches. This should be a basic strategy in pharmaceutical development to maximize the chances of bioequivalence. Limits should be set for each API in fixed-dose FPs. The dissolution method should be incorporated into the stability and quality control programs. Multipoint dissolution profiles of both the test and the reference FPs should be compared.

Dissolution profile testing

Three media - 900 ml or less - all at 37C Buffer pH 1.2, SGF without enzymes or 0.1M HCl Buffer pH 4.5 Buffer pH 6.8 or SIF without enzymes Water may be used additionally (not instead of) Paddle at 50 or basket at 100 rpm Twelve units of each product in all 3 media Dissolution samples collected at short intervals, e.g. 10, 15, 20, 30, 45 and 60 minutes Analyse samples for all APIs, when applicable

2. 3. 4.

Stability Analysis..
Serves two purposes: To evaluate the specificity of the stability indicating method, e.g. LC To understand the degradation pathways of the API to facilitate rational product development i.e.Hydrolysis, Oxidation, Photolysis and the role of pH
Wherever possible, commercial pharmaceutical products should have shelf-life of Three years. The potency should not fall below 90 or 95% under the recommended storage conditions and the product should still look and perform as it did when first manufactured.

Drug degradation processes

Drug degradation occurs by four main processes: Hydrolysis due to H2O, H3O+, OH-, pH Oxidation O2 Photolysis UV and visible Trace metal ion catalysis Fe2+, Fe3+, Cu2+, Co2+, etc

Hydrolysis and oxidation are the most common pathways!


Typical Stress Condition for FP/s in Solid State

Storage conditions
40C, 75 storage** % RH; open

Testing period*
3 months 3 months according to ICH

50-60 C, ambient RH; open storage Photostability; ICH according to

* 3 months or 5-15% degradation, whatever comes first ** For API1-API2, or API-excipient, or FPP (finished pharmaceutical product) without packing material, typically a thin layer of material is spread in a Petri dish. Open storage is recommended, if possible.


Stress Testing of API in Solution

Storage conditions
pH 2, room temperature pH 7, room temperature pH 10-12, room temperature H2O2, 0.1-2% at neutral pH, room temperature

Testing period*
2 weeks 2 weeks 2 weeks 24 hours

* Storage times given or 5-15% degradation, whatever comes first


Arrhenius Equation
Thermal effects are superimposed on all four chemical processes mentioned above. Typically a 10C increase produces a two- to five fold increase in the rate of reaction. Example. Storing b-lactam penicillins in a refrigerator reduces the hydrolysis rate by 90% of that at room temperature. K = A e Ea/RT or log K = log A (Ea/2.303R)/T. Plotting the rate of reaction (K) against 1/T K allows the calculation of rate at any temperature (Ea, activation energy; R, gas constant), and therefore a prediction of shelf-life (t90, time to 90% potency). This forms the basis of many accelerated stability tests!

Many pharmaceutical drug molecules are known to be
sensitive to light and degrade chemically under light exposure. During the development we have to evaluate this risk and how we can stabilise a potential photo-labile drug with packaging material, films and/or colorants. This evaluation of the drug molecule can be done by Isothermal Microcalorimetry (IMC) in real time. Photosensitivity of molecules is often studied in solutions, and the changes in concentrations have been examined by chromatography and spectrophotometry. Discoloration of powders has been examined by colorimetry.

Photostability studies
A systematic approach to photostability testing is recommended covering, as appropriate, studies such as: (i) Tests on the drug substance; (ii) Tests on the exposed drug product outside of the immediate pack; and if necessary, (iii) Tests on the drug product in the immediate pack; and if necessary, (iv) Tests on the drug product in the marketing pack.


Photostability As Per ICH

Two types of studies, exposure is cumulative from the light sources Forced degradation study to generate potential degradation products 2 X exposure to UV and fluorescent sources Confirmatory study to confirm product and package performance NLT 1.2 million Lux hours + 200 watts/hrs per sq. meter

Photostability studies
Light Sources: Option 1 Dual output light sources, such as D65/ID65 Simulates artificial day light fluorescent lamp Use Xenon or Metal Halide Option 2 Tandem exposure to single light source types Cool white fluorescent lamp + near UV fluorescent lamp (320-400 nm) Accumulate exposure under one, then the other

Solid state studies



Solid State Physical Stability & compatibility of

pharmaceutical Substances
Incompatibility is an undesirable interaction between two or more substances
drug drug, drug excipient, drug packing material, excipient packing material

Interaction can be chemical, physical or both Physical/chemical stability data of pure components obtained at the same time


Prerequisite of the stable and effective formulation Saves time - facilitate formulation process (exclude high-risk excipients) Lowers risk - support process and packaging development decisions Saves money - minimize the number of needless, formal (expensive) stability studies


Potential changes due to Incompatibility

Dissolution Crystallization Growth of crystals Crystallinity changes Phase transitions Degradation Degradation by nucleation via the gaseous phase {a contracting surface due to nucleation with coverage by the breakdown products} Degradation mediated by surface moisture Oxidation Photolysis

Applied procedures
Desktop working Traditional way Microcalorimetric way Application of Phys. analytical methods X-Ray Powder Diffraction, XRPD Differential Scanning Calorimetry, DSC Isothermal Microcalorimetry, IMC Inverse Gas Chromatography, IGC Chemical methods (HPLC, TLC,)

Desktop Working
PROCEDURE molecular formula of active substance and potential excipient are compared to each others potential interaction points are listed molecule modeler typically hunts compatible molecules with target molecule as a purpose getting strong interaction with the binding site of the target molecule pharmaceutical formulator decides on the basis of the list of excipients which have a minimum number of potential interaction points

Traditional Procedure
One option Binary Mix Compatibility Testing: samples or of drug and excipients are intimately mixed (1:1), but other mixtures used as well samples can be either powder blends or compressed discs one set of samples are moistened and sealed into ampoules to prevent moisture loss study design stored under room and accelerated conditions at least 1 week in accelerated conditions 40CRH75% or 60 C RH75% 1-3 month in room conditions (25CRH60%) analysed at various time points using HPLC, DSC and TGA (NIR, Raman, XRD,) and visually 113

Traditional Procedure
However, the binary mix approach takes time and resources is well known that the chemical compatibility of an API in a binary mixture may differ completely from a multi-component prototype formulation.
An alternative is to test prototype formulations. The amount of API in the blend can be modified according to the anticipated drug-excipient ratio in the final compression blend. Platform prototypes can be used for specific dosage forms, e.g. DC vs. wet granulation in tablets There is better representation of likely formulation chemical and physical stability

However, this is a more complex system to interpret


Example -Experiment Sheet


Drug excipient.
Can apply statistical models (e.g. 2n factorial design) to determine the chemical interactions in more complex systems such as prototype formulations, with a view towards establishing which excipients cause incompatibility within a given mixture.


Microcalorimetric Procedure
Most chemical or physical processes are due to changes in free energy and are usually accompanied by heat change.
ISOTHERMAL MICROCALORIMETRY (IMC) can monitor these changes ISOTHERMAL MICROCALORIMETRY (IMC) is exceptionally sensitive technique to see any reactions in the sample: - It has been estimated that a change that would take more than 200 years to run to completion could be easily detected with IMC - temperature difference sample versus reference < 10-6 C detectable - degradation rate 0.01%/annum detectable Non-specific method - for fully interpretation of the data it is necessary to investigate the origin of the thermal events by additional methods


about 1 gram of sample into the specific sample ampoule heat flow curves of pure substances are compared to those of the binary mixtures incompatibility is revealed if the features of the heat flow curve of the mixture doesnt resemble the sum curve of the components of the binary mixture studied.

Microcalorimetric Example


XRD Application/Methods
X-Ray Powder Diffraction powerful equipment to evaluate solid state properties
- polymorphism - solvation state (solvates, hydrates) - Crystallinity useful method to reveal any other phase transition or solid state impurities of the samples. non-ambient XRD can give valuable supporting data for stability prediction - humidity and temperature can be controlled

XRD Example


DSC Application Method

Differential Scanning Calorimetry- DSC
requires 5 mg of drug, in a 50% mixture with excipient mixtures should be examined under nitrogen to eliminate oxidative and pyrrolytic effects standard heating rate (2,5 or 10C min-1) over a temperature range on the DSC apparatus the melting range and any transitions of the drug will be known from earlier investigations into purity, polymorphism and solvates.


Excipient Compatibility

Excipients:API Interaction

Whereas excipients are usually biologically inactive, the same cannot be said from a chemical perspective. Excipients, and any impurities present, can stabilise and/or destabilise drug products.

Considerations for the formulation scientist: the chemical structure of the API the type of delivery system required the proposed manufacturing process


Excipients:API Interaction

Initial selection of excipients should be based on:

expert systems; predictive tools desired delivery characteristics of dosage form knowledge of potential mechanisms of degradation, e.g. Maillard reaction Prior development experience

The objective of drug/excipient compatibility considerations and practical studies is to delineate, as quickly as possible, real and possible interactions between potential formulation excipients and the API. This is an important risk reduction exercise early 126 in formulation development.

Excipient Compatibility Studies

One option.Binary Mix Compatibility Testing:

In the typical drug/excipient compatibility testing program, binary (1:1 or customised) powder mixes are prepared by triturating API with the individual excipients. These powder samples, usually with or without added water and occasionally compacted or prepared as slurries, are stored under accelerated conditions and analysed by stability-indicating methodology, e.g. HPLC. (The water slurry approach allows the pH of the drug-excipient blend 127 and the role of moisture to be investigated.)

Excipient Compatibility Studies

One option.Binary Mix Compatibility Testing:

Alternatively, binary samples can be screened using thermal

methods, such as DSC/ITC. No need for stability set-downs; hence cycle times and sample consumption are reduced. However, the data obtained are difficult to interpret and may be misleading; false positives and negatives are routinely encountered.

Also sensitive to sample preparation.


Excipient Compatibility Studies

However, the binary mix approach takes time and resources and it is well known that the chemical compatibility of an API in a binary mixture may differ completely from a multi-component prototype formulation. An alternative is to test prototype formulations. The amount of API in the blend can be modified according to the anticipated drug-excipient ratio in the final compression blend.
Platform prototypes can be used for specific dosage forms, e.g. DC vs. wet gran tablets

There is better representation of likely formulation chemical and physical stability However, this is a more complex system to interpret

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Drug excipient studies

Drug-excipient interactions can be studied using both approaches in a complementary fashion. The first tier approach is to conduct short-term (1-3m) stability studies using generic prototype formulations under stressed conditions, with binary systems as diagnostic back-up: Chemical stability measured by chromatographic methods Physical stability measured by microscopic, particle analysis, in vitro dissolution methods, etc. The idea is to diagnose any observed incompatibility from the prototype formulation work then hopefully identify the culprit excipients from the binary mix data. Hopefully, a prototype formulation can then be taken forward as a foundation for product 130 development.

Can apply statistical models (e.g. 2n factorial design) to determine the chemical interactions in more complex systems such as prototype formulations, with a view towards establishing which excipients cause incompatibility within a given mixture.

All starches are hygroscopic and rapidly absorb atmospheric moisture. Approximate equilibrium moisture content values at 50% relative humidity are: 11% FOR MAIZE (CORN) STARCH,

Between 30-80% relative humidity, corn starch is the least hygroscopic starch and potato starch is the most hygroscopic starch.

Oxidation and the Role of Excipients

Oxidation is broadly defined as a loss of electrons in a system, but it can be restated as an increase in oxygen or a decrease in hydrogen content. Oxidation always occurs in tandem with reduction; the so-called REDOX reaction couple. Oxidation reactions can be catalysed heavy metals, light, leading to free radical formation (initiation). Free radicals then react with oxygen to form peroxy radicals, which react with the oxidative substrate to yield further complex radicals (propagation), finally the reaction ceases (termination).


Oxidation and the Role of Excipients

Excipients play a key role in oxidation; either as a primary source of oxidants, trace amounts of metals, or other contaminants.
E.g. Peroxides are a very common impurity in many excipients, particularly polymeric excipients. They are used as initiators in polymerisation reactions, but are difficult to remove.


Hydrolysis and the Role of Excipients

One of the most common pathways of drug product degradation is hydrolysis. The source of the excipients can greatly influence hydrolytic reactions. For example, talc obtained from different sources impacts markedly on the overall stability of the aspirin tablet formulation. This is possibly attributable to the affect of different types and amounts of surface impurities, which are dissolved in the adsorbed moisture layer, where they subsequently react with the API. It could also influence the pH of the micro-environment.

Drug Excipient Incompatibilities

Ahlneck and Lundgren studied the compatibility of aspirin in the presence of 3 common diluents; lactose microcrystalline cellulose dicalcium phosphate Dicalcium phosphate despite having much lower moisture pick up levels than microcrystalline cellulose, had a greater de-stabilising effect. Attributed to the alkalinity of the dicalcium phosphate in the solid state (pH 7.4). The increase in the pH adversely affects the stability of the formulation, despite minimal solubility 136 in

Photolysis and the Role of Excipients

Sunlight (both in the UV and visible regions) may degrade drug products and excipients; and consequently photolabile APIs can raise many formulation (& phototoxicity) issues. The addition of light absorbing agents is a well known approach to stabilising photolabile products. Order of effectiveness: pigments > colorants > UV absorbers However, beware variable performance between grades/sources. e.g. Surface-treated titanium dioxide is inferior to the untreated excipient as an opacifier. 137

More examples.
Lactose can react with primary amines. Silicone dioxide acts as Lewis acid


Preformulation Study is Major tool for pharmaceutical product Development - it saves time as well as cost for development of Dosage form.


Pharmaceutical Preformulation and Formulation by Mark Gibson The theory and practice of Industrial pharmacy by Lachman Pharmaceutics by Aulton Handbook of Preformulation by S. Niazi


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