Introduction:

In the early stages of any project it is important to adequately characterize both the drug substance and excepientsselection of an appp. This reduces the risk of undesirable findings during clinical and manufacturing stages. Any alteration in the drug substance would require fresh investigations including bioequivalence studies. This is the key issue a pharmaceutical scientist has to keep in mind always when dealing with new chemical entities( new drug substance) and the corresponding products. Alternatively, although there is lot of information available for generic drugs, new polymorphs are always investigated to improve the properties. In these two issues the study of physical properties of drug substance becomes essential. To develop a solution, a suspension, a syrup, an emulsion the information of physico chemical properties such as solubility, pka and partition coefficient would be required. To develop a tablet, a capsule or any other solid dosage form solid state properties the information such as specific surface area, flowability, particle size, bilk density etc would be required. Thus the study of these properties is essential to develop a decent formulation for a novel chemical entity, right from the beginning to the end of drug development. The following reasons for the evaluation of the physical properties of early developmental candidates could be furnished: Reducing the time and cost of introducing a molecule into the market. Selection of an appropriate form of the drug substance, such as salt form, prodrugs etc. Selection of application type ( eg: oral, dermal, injectable). Selection of the form of delivery ( eg: quick acting or slow release). Increasing the ease of product development. Reducing undesirable findings during clinical phases. Release of best dug into the market.

Physical properties:
Specific surface area, hygroscopicity, bulk density, flow properties, crystallization are the physical properties to be investigated for new drug substances, whether flexible or stubborn.

Specific surface area:

Surface area properties of a drug particle affect the dissolution and chemical reactivity of a drug substance. These properties include size, shape and surface morphology of a drug substance. The smaller the particles, the better are the bilk flow and formulation homogeneity. The simplest way to measure the particle size is to use a microscope. However it is tedius to measure the average particle size with such techniques. The best way is to use photomicrographs and hemocytometer slides. Particles with a larger specific area are good absorbents for the absorption of gases and of solutes from solution. The other factor that is also important is the

particle shape. Generally a sphere has minimum surface area per unit volume. The more asymmetric a particle, is the greater surface area per unit volume. For a collection of particles that are not spherical, which is the case with drug powders, the diameter that is related to the surface area or volume through a correction factor is to be considered. Since these surface properties affect their homogeneity, content uniformity and dissolution properties of a tablet form, which ultimately affect the bioavailability, these properties have to be thoroughly evaluated during toxicological stages before clinical trails are preceded so that perfect correlation is obtained between the bioavailability data with a formulation. When the studies are transferred from toxicology studies to clinical studies. Accordingly, sophisticated methods are currently used. These include adsorption methods and air permeability methods. Quantasorb, an instrument used to obtain specific surface area measurements. A mixture of helium and nitrogen is passed through the sample; helium is inert and is not absorbed on the powder surface while nitrogen is absorbed on the powder. A thermal conductivity instruments attached to the instrument measures the conductivity associated with the absorption, which in turn indicates the size of the particles. In air permeability technique, the resistance to the flow of a fluid, such as air through a plug of compacted powder is used to determine the surface area of the powder. The greater the surface area of the powder the greater is the resistance offered to the flow of the air.

Hygroscopicity:
The amount of water absorbed on the surface of drug particle influences the solid state stability as well as the flow properties and compactibiliy of a drug substance. Most drugs are partially hygroscopic. Sometimes drugs exists as different crystal structures with different properties. Hygroscopicity is one such character, provided the opportunity, the first property to be determined for a new drug characterization is to measure its hygroscopicity. Alternatively, these properties could be studied later, after preformulation is accomplished. Hygroscopicity depends on the synthetic techniques and the recrystallisation methods. Judicious selection of a suitable crystal form for further development is the essential step in the development of solid dosage forms. The stability of a solid drug depends on the hygroscopicity of a particular solid state of a drug, which in turn depends on the type of the crystal or physical form of the drug thai in turn depends on the synthetic techniques or the recrystallization method for that particular drug the hygroscopicity of a substance is determined by exposing the compound to different humidity conditions for a specific time intervals and then assaying for water content using karl fisher reagent etc. other methods that could be used to measure the hygroscopicity is the gas chromatography. Dynamic water sorption ( DWS) that requires very little amount of compound for handling is also used in the hygroscopicity measurements at above 25c.

Hygroscopicity most of the times affects the compatibility of new drug substances. A rosy picture would be when hygroscopicity of an NCE would be very less or void. Compatibility as a property is affected by compressibility, adhesive/cohesive interactions and mechanical properties of the components. Water content also influences the compactibility, suggesting that hygroscopicity is one of the key issues in the development of tablet dosage forms. The mechanism of water absorption in most of the cases is either hydrate formation or site specific adsorption. The greater the compactibility, the better are the tablet properties. Many attempts were tried to increase the compactibility of tablet substance. In this regard the reduction of hygroscopicity of drug substance is very crucial. This can be achieved by obtaining drug crystals by using altered synthesis or recrystallization techniques.

Bulk density and porosity:

Bulk density is an essential pharmaceutical property to be thoroughly investigated for a new chemical entity . this is because of its importans in capsule filling and tablet compression. Apparatus high bulk density will notallow a capsule to be filled in the specific volume and in addition during tablet compression, the tablets would not be compressed either because of the rebounded effect or because of the bulk volume occupied by the tablet powder in the die. Bulk density along with flow properties of a drug substance occupied major investigation problems, which have to be sorted out as early as possible in new drug chemical entity investigations experimentally, the true density is determined by suspending drug particles in solvents of various densities and in which the compound is insoluble. In these measurements, wetting and pore penetration are enhanced by the addition of a small quantity of surfactant to the solvent mixtures. After vigorous shaking, the sample are centrifuged briefly and then lift to stand undisturbed until flocculation or settling has reached equilibrium the sample that retains suspended corresponds to the true density of the material. One way of avoiding this density problem for a new chemical entity is to use wet granulation and then punch the tablet or fill the granules in a capsule. If a drug has very high bulk density, it may not be used in a direct compression process. The drug has to be modified so as to obtain bulk drug with good compressibility properties. In modern solid dosage form technology, the current practice is to prepare dosage forms with reduced excepient content. Technology hat reduces the size of the dosage form, improve the compressibility of the solid drug, its flowability and enhances the aesthetics as desirable.

Crystallization:
Crystallization is a common phenomenon in pharmaceutical processing right from the manufacturing of active pharmaceutical ingredient to the storage of final formulation approved. Crystallization process can be termed as a meta stable thermo dynamic state. This occur because any substance or events tend to stabilize to reach the lowest possible thermodynamic state. This state of any substance is termed as a metastable state. This metastable state is either intentionally

or unintentionally created either by supersaturation, in the crystallization of desired solid state modifications and in the control of solid phase conventions during isolation, manufacturing, storage and dissolution. Examples of metastable state include solid solutions, freeze concentrated solutions, solutions of weak acids/bases exposed to a PH changes, solutions prepared by dissolving a solid state modification with a higher solubility, residual solutions during filteration, granulation and drying. The factors that can appear in the affect crystallization include molecular or ionic transport, viscosity, supersaturation, solubility, solid liquid interfacial tension and temperature. Nuclear kinetics is experimentally, determined from measurement of nucleation rates, induction time and metastability zone width as a function of initial supersaturation. Currently, molecular simulations from the data obtained from the solution and crystal structure of drug substance is used in establishing the crystal structure of new chemical entity. Molecular association process in super saturated systems is obtained by laser raman spectroscopy and laser light scattering is used in the identification of pre-nucleation clusters and growth units well defined experimental conditions. Raman fluorescence spectroscopic technique used are capable of providing information about the solution structures are the species present in the solutions.

Physico chemical properties :

Several physic-chemical properties of new leads have to be investigated very early on these could include Pha , solubility analysis, partition co-efficient, dissolution rate , solid state stability , solution state stability.

Pka
Pka determination of anew chemical entity are important because this controls solubility and consequently the oral absorption of a molecule in a given solution, formulation or body fluids. In ph range from 1-10 , the solubility and consequently oral absorption could be altered by orders of magnitude with changing ph. Pka is the ph at which 50% of the substance is ionized. Buffer, temperature, ionic strength and cosolvents effect the pka values. Incorpolation of cosolvents in pka measurements instrument methods is important because of the likely poor solubility and possible precipitation of these compounds in aqueous media. Potentiometric and spectrophotometric methods are the p[opular methods used in the determination of pka of new chemical entity. Currently, glpka instrument is in the market for the determination of pka of new chemical entities. The instrument measures the potentiometric pka of a compound. The advantage offered by the current glpka instrument is that, the assays are fully automated; temparatures and ionic strength are monitored during the runs and four line cosolvent options available. The advantage is that using organic solvents help in determination ionization constants of poorly soluble compounds. As per indications of manufacturers, the functions of the instruments include: Pkas is measured from 2 to 12.

Log p measurements from -2 to +8. Overlapping and multiple pkas routinely measured. Easily handles protogenic counter ions. Sparingly soluble compounds titrated in either possible supported cosolvents. Typical sample concentrations of 0.25 to 0.5 m M( 1 2 mg of 400 MW compounds in 10ml). Fast ( typical titration = 25mins). Accurate and precise.

In spectrophotometric method of determination, at a given PH, if the ion concentrations are determined using beers law one can calculate the approximate pka of a drug. For example, if the drug is a free acid [HA] in equilibrium with its base[A], then Pka = PH + log [HA] / [A] When [HA] = [A], as determined by their respective absorbence in the spectrophotometric determinations, pka = PH.

Solubility analysis:
Solubility analysis is of new chemical entity is essential for further processing of a compound. The factors that would effect the solubility of a new chemical entity are PH, temperature, ionic strength and buffer concentrations. For equilibrium solubility determinations, different methods are employed. To determine the aqueous solubility, the drug is solubilised in which it is highly soluble and this solution is slowly added to the distilled water and agitated. At the end of agitation, the suspension is filtered to obtain a filterate that is then assayed using techniques like spectrophotometry and HPLC. In this retard temperature also plays a role some times. Usually, the solubility of drugs is more in high temperature conditions. The principle can be used to saturate the aqueous suspension containing a drug. The compound that is not soluble is precipitated out. This is filtered and submitted for analysis to determine the solubility of a drug substance. The simplest technique that is routinely used to add excess of drug to water and this is then agitated overnight to obtain maximum solubility of the drug in the media and then filtered and assayed to obtain the desired aqueous solubility. The other aspect of solubility is dissolution. To determine the solubility of a poorly soluble compound in water, generally 24hrs equilibrium time is given. During the time the drug slowly dissolves in water. It is a similar phenomenon with the dissolution of the drug in gastric fluid or dissolution media from a solid powder or a capsule or from a tablet dosage form. The drug is slowly dissolved and the drug dispersed by agitation to form a uniform solution. It is then analysed to obtain the concentration of the drug in the dissolution medium. Drugs with limited solubility( < 1%) in the fluids of gastro intestinal tract often exhibit poor or eratic absorption unless dosage forms are specifically tailored for the drug.

Partition coefficient:
Octanol water partition coefficient is the ratio of concentration of a chemical in octanol and in water at equilibrium and at a specified temperature. Octanol is an organic solvent that is used as a surrogate for natural organic matter. The octanol water partition coefficient has been correlated to water solubility; therefore the water solubility of a substance can be used to estimate octanol water partition coefficient. As mentioned previously, the octanol water partition coefficient ( Kow ) is defined as the ratio of chemicals concentration in the octanol phase to its concentration in the aqueous phase of a two phase octanol water system.

K ow

Concentration in octanol phase / Concentration in aqueous phase.(I 1)

Values of K ow are thus unit less. The parameter is measured using low solute concentrations, values of Kow are usually measured at room temperature ( 20 - 25c). The effect of temperature on K ow is not great. Usually on the order of 0.001 0.01 log Kow / c and may be either + or ve. The octanol / water partition coefficient is not the same as the ratio of the chemicals solubility in octanol to its solubility in water, because organic and aqueous phases of the binary octanol / water system are not pure octanol and pure water. Kow is often found to be a function of solute concentration. The chemical in question is added to a mixture of octanol and water whose volume ratio is adjusted according to the expected value of Kow. Very pure octanol and water must be used, the concentration of the solute in the system should be less then 0.01 mole / litre. The system is shaken gently until equilibrium was achieved (15mins 1hr). centrigugation is generally required to separate the two phases, especially if an emulsion is formed. An appropriate analytical technique is then used to deterimine the solute concentration to each phase. A rapid laboratory estimate of Kow may be obtained by measuring the retension time in HPLC , the logarithm og retension time and the logarithm of Kow have been found to be linearly correlated. Conversely chemicals with high Kow (>104) are very hydrophobic.

Dissolution rate :
Dissolution rate is the predictable measure of time required for a given dug or active ingredient in an oral solid dosage form to go into solution under the specified set of conditions. Since absorption and physiological availability of any nutritional supplement is largely dependent upon having in a dissolved state, a suitable dissolution rate is crucial. Calculating intrinsic dissolution rate makes comparison of the individual drug substances and the effect of different conditions on drug dissolution. The intrinsic dissolution rate is generally defined as the dissolution rate of a pure drug substance under the conditions of constant surface area. The true

intrinsic dissolution rate may be better described as the rate of mass transfer from the solid surface to the liquid phase. Intrinsic dissolution is generally determined by measuring the dissolution of a non disintegrating disc made by compressing pure powder drug substance under high pressure using a specially constructed punch and die system. The test material is compressed with a bench top punch tablet press for 1 minute at the minimum compression pressure necessary to form a non disintegrating compacted tablet. Changes in the crystal form may occur during compression , conformation of the solid form should be verified by powder x ray diffraction or anot her similar technique. Compression plays an important role in the test, if it is too low, a non disintegrating tablet may not be obtained and if its too high it may change the crystal form. Compression pressure should be high enough to produce a translucent pellet with no powder or flakes on the surface. It is important to study the effct of compression pressure on intrinsic dissolution rates as it has been observed for several drug substances that the intrinsic dissolution rate varies with compression pressure. Dissolution rate determines the availability of the drug for absorption when slower than the absorption , dissolution becomes the rate limiting step. Overall selection of an appropriate formulation can control absorption. Dissolution rate is effected by whether the drug is in salt, crystal or hydrated form. The sodium salt of weak acids ( ex: barbiturates, salicylates ) dissolve faster than their corresponding free acids regardless of the PH of the medium.

Solid state stability:

This involves stability of the drug substance as a solid and stability of a drug substance in a solid dosage form. Drug instability in pharmaceutical formulation may be detected in some instances by a change in the physical appearance, colour, odour, taste or texture of the formulation where as the chemical stability of the drug substance is determined by chemical analysis. The second study is termed reaction kinetics. A kinetic study on a drug substance is examined by subjecting an NCE in several physical and chemical and stressed condition. The samples are withdrawn at periodic times and assayed for the drug content using a HPLC or other techniques. Then the active chemicals and degradants are mathematically dissected to obtain chemical kinetics of the drug substance. These reaction kinetics could be zero order, first order, second order and some times inverse reaction kinetics. Inverse kinetics are determined when there is a transition of one impurity to other or one degradant to the drug, which may help in long run in the formulation movement predictions and during storage. As a standard stability protocol, the utilization of exaggerated conditions such and high temperature and high light intensity and high humidity are investigated for the stability determination. Accelerated temperature studies, for example, may be conducted for 6 months at 40c and 75% RH. If a significant change occurs in the drug or drug product under these conditions, lesser temperature and humidity may be used such as 30c and 60% RH. Product container, closures, and other packaging conditions features are also to be considered in stability testing during this stage.

Solution state stability:

Solution state stability of a drug is valid for stability testing of liquid formulations and for HPLC method development. NCE is generally mixed in aqueous media at different PH conditions. The samples are withdrawn at regular time intervals and are submitted for analysis. Once the data is obtained, the active amount present is mathematically fitted to obtain the reaction kinetics in the solution state. Different PH conditions, different humidity conditions, different temperature conditions, different packaging conditions can be used in the solution state stability determination. The reaction kinetics is the same and is zero order, first order, second order, multi order and inverse kinetics. In solid state characterization apart from the stability impurity, polymorphs, racemates etc are determined as a first step in the physical characterization of a new chemical entity. The following discussions reveal the requirement for physical characterization as per the regulatory agencies.

Enantiomers and racemates:

Enantiomer is one of two stereoisomers that are mirror images of each other that are non-superposable (not identical), much as one's left and right hands are the same except for opposite orientation. Organic compounds that contain an asymmetric (chiral) Carbon usually have two nonsuperimposable structures. These two structures are mirror images of each other and are, thus, commonly called enantiomorphs (enantio = opposite ; morph = form) Hence, optical isomerism (which occurs due to these same mirror-image properties) is now commonly referred to as Enantiomerism Enantiomers have, when present in a symmetric environment, identical chemical and physical properties except for their ability to rotate plane-polarized light (+/) by equal amounts but in opposite directions (although the polarized light can be considered an asymmetric medium). A mixture of equal parts of an optically active isomer and its enantiomer is termed racemic and has zero net rotation of plane-polarized lightbecause the positive rotation of each (+) form is exactly counteracted by the negative rotation of a () one. Enantiomers of each other often show different chemical reactions with other substances that are also enantiomers. Since many molecules in the body of living beings are enantiomers themselves, there is often a marked difference in the effects of two enantiomers on living beings. In drugs, for example, often only one of a drug's enantiomers is responsible for the desired physiologic effects, while the other enantiomer is less active, inactive, or sometimes even responsible for adverse effects (unwanted side-effects).

The following table lists pharmaceuticals that have been available in both racemic and singleenantiomer form. Racemic mixture Single-enantiomer

Amphetamine (Benzedrine) dextroamphetamine (Dexedrine)

Bupivacaine (Marcain)

levobupivacaine (Chirocaine)

Cetirizine (Zyrtec / Reactine)

levocetirizine (Xyzal)

Impurities:
Impurities in new drug substances are addressed from two perspectives: Chemistry aspects include classification and identification of impurities, report generation, listing of impurities in specifications, and a brief discussion of analytical procedures. Safety aspects include specific guidance for qualifying those impurities that were not present, or were present at substantially lower levels, in batches of a new drug substance used in safety and clinical studies.

The studies conducted to characterize the structure of actual impurities present in a new drug substance at a level greater than 1% the identification threshold many batch manufactured by the proposed commercial process should be identified. In addition, any degradation product observed in stability studies at recommended storage conditions at a level greater than 1% the identification threshold should be identified. Identification of impurities present at apparent level of not more than 1% the identification threshold is generally not considered necessary.

Polymorphs:
Any pharmaceutical solids can exist in different physical forms. Polymorphism is often characterized as the ability of a drug substance to exist as two or more crystalline phases that have different arrangements and or conformations of the molecules in the crystal lattice. Amorphous solids consist of disordered arrangements of molecules and do not possess a distinguishable crystal lattice. Solvates are crystalline solid adducts containing either stoichiometric or nonstoichiometric amounts of a solvent incorporated within the crystal

structure. If the incorporated solvent is water, the solvates are also commonly known as hydrates.polymorphism refers to the occurrence of different crystalline forms of the same drug substance. Polymorphs and solvates of a pharmaceutical solid can have differen chemical and physical properties such as melting point, chemical reactivity, apparent solubility, dissolution rate, optical and electrical properties, vapour pressure, and density. The properties can have a direct impact on the processability of drug substances and the quality / performance of drug products, such as stability, dissolution, and bioavailability. A metastable pharmaceutical solid form can change crystalline structure or solvate / desolvate in response to changes in environmental conditions, processing, or over time. Based on the above physical properties the different instruments used for analyzing a drug molecule are: