1.

Introduction:

1.1. Helminthes:

Helminthes may also be called as worms. They typically have a long cylindrical tube-like body and no
limb. Usually worms include annelids(earthworms and marine polychaete or bristle
worms), nematodes(roundworms), platyhelminthes (flatworms), marine nemerteanworms(bootlace
worms), marine Chaetognatha (arrow worms),  Cestodes (tapeworms), priapulid worms,
and insect larvae such as grubs and maggots.They vary in size from microscopic to over 1 metre
(3.3 ft) in length for marine polychaete worms (bristle worms),6.7 metres (22 ft) for the African giant
earthworm and 58 metres (190 ft) for the marine nemertean worm (bootlace worm).worms like
hookworms, tapeworms, pinworms, flukes, roundworms, ascariasis, trichinella, threadworm,
whipworms and other worms live as parasite.
 Figure 1.1:Different Helminthes

1.1.1. Types of helminthes or worm:

There are many different phyla of worms. The following three are the best-known. Some worms live
on land in burrows, feeding on plant matter; others live in the sea or fresh water, filtering food from
water.

Phylum: Platyhelminthes

(Flatworms)

Features: about 20,000 species flat, unsegmented bodies, with a mouth but no anus, many live in
water

Phylum: Annelida

(Segmented worms)

Features: about 15,000 species segmented bodies, mostly burrowing, gut with mouth and anus, live on
land and in water

Phylum: Nematoda

(Roundworms)

Features: about 25,000 species unsegmented bodies, gut with mouth and anus

FLATWORMS
There are about 20,000 species of flatworm. They have a solid, flat body that does not contain blood.
Most flatworms are parasitic, but some are free-living.

MARINE FLATWORM

Marine flatworms absorb oxygen through the surface of their very thin, flattened body. They creep
along, rippling their body to help them move. Eyespots enable them to find their way around. Most
are predators, eating tiny animals with the mouth situated on the underside of their body.

TAPEWORM

Tapeworms are parasites that live in other animals, including humans. They have hooks and suckers
on their head to attach themselves to the animal’s gut wall. They have no digestive system but absorb
food through the surface of their body. They are hermaphrodites – they produce both eggs and sperm.

ROUNDWORM

Roundworms, or nematodes, are found almost anywhere and exist in huge numbers. As many of the
roundworms are transparent, few people are aware of them.

The roundworm has a long, round body that tapers towards the tail. The outer layer, or cuticle, is
smooth. Muscles run along its body, but not around it. To move along, the worm contracts these
muscles, thrashing backwards and forwards in a single plane, making C or S shapes.

SEGMENTED WORMS

This group divides into earthworms, bristleworms, and leeches. All have segmented bodies. The
worms’ bodies are fluid-filled, but the leeches are solid.

EARTHWORM

Earthworms are formed from many segments. Only the gut runs through the whole body from head to
tail. Worms have a circulatory system with blood vessels but no heart. The thickened area towards the
front of their body secretes mucus, which binds mating worms together and forms a cocoon for eggs.

LEECH

Leeches are parasites that live on the outside of other animals. They have specialized cutting jaws to
bite through skin so that they can suck the animal’s blood. Substances in their saliva prevent the blood
from clotting and make the bite painless so that the animal is unaware it has been bitten. Leeches
move by shifting one sucker forwards and then bringing the other one up behind it.

1.1.2. Helminthes in human body:

Different types of worm can cause problems in humans

1.1.2.1. Threadworms: Sometimes called pinworms, are the only common worm infects human.
They usually occur in children and large number of children may be infected at some time. Signs of
an infestation include an itchy bottom, disturbed sleep, irritability, tiredness and a lack of interest in
eating.

Threadworms are small parasites that live in the intestines of humans. They are particularly common
in children under the age of 10. The worms are white, about 8mm long, with a blunt head and a
pointed tail. They can live for up to 6 weeks.

The female worm lays many tiny eggs around the anus. This usually happens at night while the person
is asleep since the female worms only come out at night. While laying the eggs, the worm also
produces a chemical that creates an itchiness and causes the person to scratch the area.

Eggs then stick under the fingernails and on fingertips and can be transferred to the mouth. They may
then be swallowed and cause a re-infestation.

The swallowed eggs hatch in the intestine. After a couple of weeks, the worms reach adult size and
begin to reproduce.
 Figure 1.1.2.1: Threadworm

Threadworms can’t be catched from animals with worms. The only way animals may be responsible
for spreading human threadworms is by transporting them on their fur after human contact.

Treatment for threadworms is designed to get rid of the parasites and prevent re-infestation. To
successfully treat threadworms, you can see your doctor or pharmacist who will be able to advice on
medication for you or your child. It is commonly advised to treat the entire family at the same time to
successfully get rid of the infection.

When taking medicine, strict hygiene practices should be followed to prevent re-infestation.

1.1.2.2. Roundworm: Roundworms can live in human and can cause many problems. They are
usually found in soil and stool. They can enter the body through the mouth or direct contact with the
skin.

They can live in the human intestine for a very long time. There are several types of roundworms and
they can all be quite harmful.

Roundworms are able to pose a significant risk to humans. Contact with contaminated soil or dog
feces can result in human ingestion and infection. Roundworm eggs may accumulate in significant
numbers in the soil where pets deposit feces. Once infected, the worms can cause eye, lung, heart and
neurologic signs in people

Figure 1.1.2.2: Roundworm

Medications orally are recommended for treating roundworm infections in adults and children over
the age of one. It works by stopping the roundworms using glucose (sugar). Without glucose, the cells
of the roundworms lose their energy supply and quickly die.

1.1.2.3. Tapeworm: Tapeworms are flat, segmented worms that live in the intestines of some animals
Eating undercooked meat from infected animals is the main cause of  tapeworm infection in people.
Although tapeworms in humans usually cause few symptoms and are easily treated, they can
sometimes cause serious, life-threatening problems.
 Figure 1.1.2.3: Tapeworm

Tapeworms are usually treated with a medicine taken by mouth. The medications paralyze

the tapeworms, which let go of the intestine, dissolve, and pass from your body with bowel
movements.

1.1.2.4. Hookworm: Hookworms are parasites. This means they live off other living
things. Hookworms affect lungs, skin, and small intestine. Humans
contract hookworms through hookworm larvae found in dirt contaminated by feces.

As their name suggests, hookworms have hooks on their mouth that help them attach better to the
intestines of their host. Humans affected can also develop a skin condition called cutaneous larva
migrans.

Figure 1.1.2.4: Hookworm

Anthelminthic medications orally are the drugs of choice for treatment of hookworm infections.
Infections are generally treated for 1-3 days. The recommended medications are effective and appear
to have few side effects. Iron supplements may also be prescribed if the infected person has anemia.

1.1.2.5. Whipworm: Whipworm (Trichuris trichiura) is an intestinal parasite of humans. The

larvae and adult worms live in the intestine of humans and can cause intestinal disease. The name is
derived from the worm's distinctive whip-like shape.

Trichuriasis is infection with the parasite whipworm, is a very common intestinal helminthic

infection worldwide. About one quarter of the world's population is thought to carry the parasite.
Principally a problem in tropical Asia and, to a lesser degree, in Africa and South America, a lack of a
tissue migration phase and a relative lack of symptoms characterize whipworm infection. Trichuris is
also notable for its small size. Only individuals with heavy parasite burden become symptomatic.
Vitamin A deficiency has been seen in patients with trichuriasis.

Figure 1.1.2.5: Whipworm

Poor hygiene is associated with whipworm transmission and children are especially vulnerable
because of their high exposure risk. This is especially true in developing countries, where sanitary

conditions are poor.

The most common and effective treatment for a whipworm infection is an antiparasitic medication.

This type of medication gets rid of any whipworms and whipworm eggs in the body. The medication
usually needs to be taken for one to three days orally.

1.2. Anthelminthic Drugs:

1. Benzimidazoles (Albendazole, Mebendazole, Thiabendazole)

2. Piperazine (Diethylcarbamazine citrate, Piperazine citrate)

3. Levamisole, pyrantel and morantel

 4. Paraherquamide

5. Ivermectin (macrocylic lactones and milbemycins)

6. Emodepside (cyclodepsipeptides, PF1022A)

7. Nitazoxan

1.2.1:Albendazole: Albendazole is a broad-spectrum antihelminthic agent of the benzimidazole type.

It is useful for giardiasis, trichuriasis, filariasis, neurocysticercosis, hydatid disease, pinworm disease,
and ascariasis It has good activity in multiple dosesagainst cutaneous larva migrans and
strongyloidiasis, but single-dose therapy with ivermectin is more effective for these two helminthic
infections. Successful treatment with albendazole of visceral larva migrans has been reported,
although ivermectin is the drug of choice. Efficacy against the microfilarial stage ofWuchereria
bancrofti, Brugia malayi, and Loa loa is well documented, although diethylcarbamazine(DEC)
remains the parasite adulticidal drug of choice for the treatment of patients with these filarial
infections.DEC plus albendazole in combination has superior activity against adult W.
bancrofti compared with either drug alone.Albendazole may be of benefit in patients with
gnathostomiasis, baylisascariasis, trichinellosis, toxociariasis, capillariasis, and angiostrongyliasis.

Prolonged high-dose regimens of albendazole constitute the most effective medical treatment of larval
cestode disease caused by Echinococcusgranulosus, E. multilocularis, and E. vogelii.Dosage is not
defined for children younger than 6 years of age. Treatment is tailored to response and normally is
required for a minimum of several weeks or months. Albendazole should be started several days
before surgery to minimize the effect of intraoperative spillage of cyst contents. Albendazole also is
used in the treatment of patients withcysticercosis, often with concomitant steroids. Albendazole
binds irreversibly to the nematodal isoform of β-tubulin, blocking microtubule assembly, disrupting
tegumental integrity, inhibiting motility, and impeding glucose uptake by the worm. Detectable levels
are achieved in serum,cerebrospinal fluid, cyst fluid, and bile, with aserum half-life of 8 to 15 hours
depending on the dose. Rapid, extensive hepatic biotransformation to the active
metabolite, albendazole sulfoxide, occurs, but the major route of excretion is not clear.
Side effects of low-dose albendazole therapy are minimal, consisting of diarrhea, abdominal pain,
migration of Ascaris through the mouth or nose, and rare hypersensitivity. With high-dose or
prolonged therapy, elevated levels of hepatic transaminases, dizziness, neutropenia, and alopecia are
most common.Serum hepatic enzyme levels and the blood count should be monitored every 2 weeks
during high-dose therapy.

Figure 1.2.1:Albendazole

Lack of data for children younger than 2 years of age has precluded specific US labeling for or
establishment of dosage for this age group, although the label leaves the physician discretion in
electing to treat children younger than 2 years of age. The pediatric dosage is identical to the adult
dosage. Embryotoxic potential usually precludes the use of albendazole during pregnancy.

Albendazole is manufactured as a suspension and as tablets that can be crushed with food, chewed, or
taken whole. Albendazole is poorly absorbed and should be ingested with food, preferably a fatty
meal, to maximize absorption.

` 1.2.2: Mebendazole: Mebendazole is a broad-spectrum antihelmintic drug indicated for

the treatment of  nematode infestations, including roundworm, hookworm, whipworm, threadworm,
pinworm, and the intestinal form of trichinosis prior to its spread into the tissues beyond the digestive
tract. Other drugs are used to treat worm infections outside the digestive tract, as mebendazole is
poorly absorbed into the bloodstream. Mebendazole is used alone in those with mild to moderate
infestations. It kills parasites relatively slowly, and in those with very heavy infestations, it can cause
some parasites to migrate out of the digestive system, leading to appendicitis, bile duct problems, or
intestinal perforation. To avoid this, heavily infested patients may be treated with piperazine, either
before or instead of mebendazole.
Mebendazole and other benzimidazole antithelmetics are active against both larval and adult stages of
nematodes, and in the cases of roundworm and whipworm, kill the eggs, as well. Paralysis and death
of the parasites occurs slowly, and elimination in the feces may require several days.

Mebendazole works by selectively inhibiting the synthesis of microtubules via binding

to colchicine binding site of β-tubulin, thereby blocking polymeration of tubulin dimers in intestinal
cells of parasites. Disruption of cytoplasmic microtubules leads to blocking the uptake of glucose and
other nutrients, resulting in the gradual immobilization and eventual death of the helminths.

Figure1.2.2: Mebendazole

Poor absorption in digestive tract makes mebendazole an efficient drug for treating intestinal parasitic
infections with limited adverse effects. However mebendazole has impact on mammalian cells
mostly by inhibiting polymeration of tubulin dimers, thereby disrupting essential microtubule
structures such as mitotic spindle. Disassembly of mitotic spindle then leads to apoptosis mediated via
dephosphorylation of  Bcl-2 which allows pro-apoptotic protein  Bax to dimerize and initiate
programmed cell death.

1.2.3. Piperazine: Piperazine belongs to the family of medicines called anthelmintics.

Anthelmintics are used in the treatment of worm infections.
Piperazine is used to treat:

 common roundworms (ascariasis) and

 pinworms (enterobiasis; oxyuriasis).

Piperazine works by paralyzing the worms. They are then passed in the stool.

1.2.4. Ivermectin: Ivermectin is a broad-spectrum anti-parasite medication. It is a

medication used to treat many types of parasite infestations. This includes head lice, scabies, river
blindness (onchocerciasis), strongyloidiasis, trichuriasis, and lymphatic filariasis. It can be taken by
mouth or applied to the skin for external infestations.

Ivermectin causes an influx of Cl- ions through the cell membrane of invertebrates by activation of
specific ivermectin-sensitive ion channels. The resultant hyperpolarization leads to muscle paralysis.

Ivermectin stimulates excessive release of neurotransmitters in the peripheral nervous system of

parasites. It is thought to work by paralyzing the parasite or inactivating the parasite gut. A protective
barrier, called the blood-brain barrier, blocks ivermectin from reaching the human brain.

1.2.5. Pyrantel: Pyrantel is a pyrimidine-derivative anthelmintic agent for the oral

treatment of various parasitic worm infections including ascariasis, hookworm infections, enterobiasis
(pinworm infection), trichostrongyliasis, and trichinellosis.
 1.2.6. Nitazoxanide: Nitazoxanide is a broad-spectrum antiparasitic and broad-spectrum
antiviral drug that is used in medicine for the treatment of various helminthic, protozoal, and viral
infections.

The antiprotozoal activity of nitazoxanide is believed to be due to interference with the

pyruvate:ferredoxin oxidoreductase (PFOR) enzyme-dependent electron transfer reaction which is
essential to anaerobic energy metabolism.

1.2.7. Paraherquamide: Paraherquamide is a novel natural anthelmintic product with a

mode of action that is incompletely characterized. Nicotine and cholinergic-anthelmintic agonists of
different chemical classes were used to produce contraction in Ascaris muscle strips. Paraherquamide
and a semisynthetic derivative, 2-deoxy-paraherquamide, antagonized these responses.

1.2.8. Emodepside: Emodepside is the only drug in this class currently prescribed to dogs
or cats. It has been evaluated for oral (dogs) or topical (cat) administration and is considered safe and
effective for use in the management of several nematode infections in dogs or cat..Emodepside is
combined with praziquantel.

The emodepside mechanism of action is considered unique to the drugs with anthelmintic activity.

The drug binds to the latrophilin receptor presynaptically, which activates a complex signal
transmission cascade that eventually results in the release of inhibitory neuropeptides into the synaptic
gap. The resulting ion influx that occurs post synaptically results in the inhibition of pharyngeal pump
function that leads to paralysis and death of susceptible nematodes.

1.3. Combination of Albendazole and Mebendazole:

Albendazole is used to treat such conditions,

 Ascariasis
 Capillariasis
 Cutaneous Larva Migrans
 Cysticercus cellulosae
 Echinococcus
 Enterocolitis
 Filariasis, Elephantiasis
 Giardiasis
 Gnathostomiasis
 Hookworm Infection (Necator or Ancylostoma)
 Hydatid Disease
 Liver Fluke
  Loiasis
 Microsporidiosis
 Neurocysticercosis
 Pinworm Infection (Enterobius vermicularis)
 Strongyloidiasis
 Trichinosis
 Trichostrongylosis
 Visceral Larva Migrans, Toxicariasis
 Whipworm Infection

Mebendazole is used to treat following conditions,

 Angiostrongylosis

 Capillariasis

 Dracunculiasis

 Echinococcus

 Filariasis, Elephantiasis

 Hookworm Infection (Necator or Ancylostoma)

 Ascariasis Hydatid Disease

 Pinworm Infection (Enterobius vermicularis)

 Trichinosis

 Trichostrongylosis

 Visceral Larva Migrans, Toxicariasis

 Whipworm Infection

The combination of Albendazole and Mebendazole can treat all the condions above. So the
combination is highly efficient than a single drug and can provide a full coverage against soil
transmitted helminth and some other infections.
1.3.1. Different Methods: Different methods can be used to formulate the combination of
Albendazole and Mebendazole.

1.3.1.1. Non aqueous Granulation: Molecules that need wet granulation are those not suited for
dry granulation process – high dose, poor flow, low in bulk density, without binding properties.
Depending on molecule sensitivity, non-aqueous(organic) solvents are used for
the granulation process.

1.3.1.2. Aqueous Granulation: The process of adding a liquid solution to powders involves the
massing of a mix of dry primary powder particles using a granulating fluid. The fluid contains a
solvent that must be volatile, so that it can be removed by drying, and be non-toxic. Aqueous
granulation forms by binding the powders together with an adhesive, instead of by compaction.
bridges are developed between the particles and the tensile strength of bonds increases as the amount
of liquid added is increased.

Step 1: Weighing and mixing of formulation ingredients (excluding the lubricant).

Step 2: Preparing the damp mass.

Step 3: Wet screening/ Screening the dampened powder into pellets or granules.

Step 4: Drying of moist granules.

Step 5: Sizing the granulation by dry screening

Step 6: Lubrication of granules
Step 7: Compression of granules into tablets

1.3.1.3. Direct Compression: The term “direct compression” is defined as the process by which
tablets are compressed directly from powder mixture of API and suitable excipients. No pretreatment
of the powder blend by wet or dry granulation procedure is required. The processing of drug
with excipients can be achieved without any need of granulation and related unit operations. By
simply mixing in a blender, formulation ingredients can be processed and compressed
into tablets without any of the ingredients having to be changed. Direct compression method requires
fewer processing steps (unit operations) and less equipment. Therefore, the method is potentially less
expensive than other methods used in tablet manufacture. High-dose drugs may present problems with
direct compression if it is not easily compressible by itself. The choice of excipients used in the
manufacture of tablets by direct compression technology is highly restricted since most materials do
not have inherent binding properties

1.4. Tests For Quality:

1.4.1. Carr’s Index:  Carr's index is an indication of the compressibility of a powder.

Carr's Compressibility Index is named after the scientist Ralph J. Carr, Jr.
The Carr index is calculated by the formula, 

Tapped density−Bulk density

Car r ' s Index = ×100 % where  is the freely settled bulk density of
Tapped density
the powder, and is the tapped bulk density of the powder after "tapping down".  
The Carr’s index is frequently used in pharmaceutics as an indication of the flowability of a powder.
In a free-flowing powder, the bulk density and tapped density would be close in value, therefore, the
Carr’s index would be small. On the other hand, in a poor-flowing powder where there are greater
interparticle interactions, the difference between the bulk and tapped density observed would be
greater, therefore, the Carr index would be larger. A Carr’s index greater than 25 is considered to be
an indication of poor flowability, below 15 is considered to be an indication of good flowability.

1.4.2. Hausner Ratio: The Hausner ratio is a number that is correlated to the flowability of a powder
or granular material. It is named after the engineer Henry H. Hausner (1900–1995).

The Hausner ratio is calculated by the formula,

Tapped density
Hausner ratio=
Bulk density

where  is the freely settled bulk density of the powder, and is the tapped bulk density of the powder.
The Hausner ratio is not an absolute property of a material; its value can vary depending on the
methodology used to determine it.

The Hausner ratio is used in a wide variety of industries as an indication of the flowability of a
powder. A Hausner ratio greater than 1.25 is considered to be an indication of poor flowability. The
Hausner ratio (H) is related to the Carr’s index(C), another indication of flowability . Both the
Hausner ratio and the Carr index are sometimes criticized, despite their relationships to flowability
being established empirically, as not having a strong theoretical basis. Use of these measures persists,
however, because the equipment required to perform the analysis is relatively cheap and the technique
is easy to learn.

1.4.3. Angle Of Repose: The angle of repose, or critical angle of repose, of a granular material is the
steepest angle of descent or dip relative to the horizontal plane to which a material can be piled
without slumping. At this angle, the material on the slope face is on the verge of sliding. The angle of
repose can range from 0° to 90°.

The equation for calculating the angle of repose is: tan-1(2h/d). Using your scientific calculator,

multiply height by 2 and divide this value by the distance.

The size of the particles is a factor. Other factors being equal, fine grained material will form a
shallower pile, with a smaller angle of repose than coarser grains. Moisture affects the angle of
repose, as anyone who has ever built a sand castle can confirm.

Theoretically, the maximum angle at which grains in a heap of sediment can remain in place
without becoming unstable. The angle of repose of granular materials is relevant in various
applications of science and engineering, such as sedimentology, geomorphology, hydraulic, and
chemical engineering. It can be used in the descriptions of initial motion of sediment grains and
sediment transport process, and in the investigations of bank stability, riprap protection, and
reservoir sediment removal.The angle of repose is measured using different methods.

 Tilting box method. This method is appropriate for fine-grained, non-cohesive materials,

with individual particle size less than 10 mm.
 Fixed funnel method. The material is poured through a funnel to form a cone.
 Revolving cylinder method. The material is placed within a cylinder with at least one
transparent face.

1.4.4. Hardness Test: A hardness test is a method employed to measure the hardness of a material.
Hardness refers to a material's resistance to permanent indentation. There are numerous techniques to
measure hardness and each of these tests can identify varying hardness values for a single material
under testing.

Hardness is a characteristic of a material, not a fundamental physical property. It is defined as the

resistance to indentation, and it is determined by measuring the permanent depth of the indentation.

More simply put, when using a fixed force (load)* and a given indenter, the smaller the indentation,
the harder the material. Indentation hardness value is obtained by measuring the depth or the area of
the indentation using one of over 12 different test methods.

Hardness testing is used for two general characterizations

1.Material Characteristics
• Test to check material
• Test hardenability
• Test to confirm process 
• Can be used to predict Tensile strength

2. Functionality
• Test to confirm ability to function as designed.
• Wear Resistance
• Toughness
• Resistance to impact

Hardness Testing Considerations

The following sample characteristics should be consider prior to selecting the hardness testing method
to use:

• Material
• Sample Size
• Thickness
• Scale
• Shape of sample, round, cylindrical, flat, irregular
• Gage R & R

Material
The type of material and expected hardness will determine test method. Materials such as hardened
bearing steels have small grain size and can be measured using the Rockwell scale due to the use of
diamond indenters and high PSI loading. Materials such as cast irons and powder metals will need a
much larger indenter such as used with Brinell scales. Very small parts or small sections may need to
be measured on a microhardness tester using the Vickers or Knoop Scale.

When selecting a hardness  scale, a general guide is to select the scale that specifies the largest load
and the largest indenter possible without exceeding defined operation conditions and accounting for
conditions that may influence the test result.  

Sample Size 
The smaller the part, the lighter the load required to produce the required indentation. On small parts,
it is particularly important to be sure to meet minimum thickness requirements and properly space
indentations away from inside and outside edges. Larger parts need to be fixtured properly to ensure
secure placement during the test process without the chance for movement or slippage. Parts that
either overhang the anvil or are not easily supported on the anvil should be clamped into place or
properly supported. 

Cylindrical Samples
A correction to a test result is needed when testing on cylinder shapes with small diameters due to a
difference between axial and radial material flow. Roundness correction factors are added to your
testing result based on the diameter of convex cylinder surfaces. Additionally, it is important to
maintain a minimum spacing equal to 2~1/2 times the indentation's diameter from an edge or another
indentation. 

Sample Thickness 
Your sample should have a minimal thickness that is at least 10x (ten times) the indentation depth that
is expected to be attained. There are minimum, allowable thickness recommendations for regular and
superficial Rockwell methods 

Scales 
Sometimes it is necessary to test in one scale and report in another scale. Conversions have been
established that have some validity, but it is important to note that unless an actual correlation has
been completed by testing in different scales, established conversions may or may not provide reliable
information. Refer to ASTM scale conversion charts for non-austenitic metals in the high hardness
range and low hardness range. Also refer to ASTM standard E140 for more scale conversion
information. 

Gage R&R 
Gage Repeatability and Reproducibility Studies were developed to calculate the ability of operators
and their instruments to test accordingly within the tolerances of a given test piece. In hardness
testing, there are inherent variables that preclude using standard Gage R&R procedures and formulas
with actual test pieces. Material variation and the inability to retest the same area on depth measuring
testers are two significant factors that affect GR&R results. In order to minimize these effects, it is
best to do the study on highly consistent test blocks in order to minimize these built-in variations.

Newage Testing Instruments hardness testers operate are ideally suited for these studies.
Unfortunately, since these studies can only be effectively done on test blocks, their value does not
necessarily translate into actual testing operations. There are a host of factors that can be introduced
when testing under real conditions. Newage testers excel at testing in real-world conditions by
reducing the effects of vibration, operator influence, part deflection due to dirt, scale, a specimen
flexing under load.

Types of Hardness Test:

 Brinell Hardness Test.
 Rockwell Hardness Test.
 Rockwell Superficial Hardness Test.
 Vickers Hardness Test.
 Knoop Hardness Test.
 Shore Scleroscope Hardness Test.

Brinell Hardness Test:

In this test a hardened steel ball of 2.5, 5 or 10 mm in diameter is used as indenter.

The loading force is in the range of 300N to 30000N (300N for testing lead alloys, 5000N for testing
aluminum alloys, 10000N for copper alloys, 30000N for testing steels). The Brinell Hardness Number
(HB) is calculated by the formula:
HB = 2F/ (3.14D*(D-(D² - Di²)½))
Where

F- applied load, kg
D – indenter diameter, mm
Di – indentation diameter, mm.
In order to eliminate an influence of the specimen supporting base, the specimen should be seven
times (as minimum) thicker than indentation depth for hard alloys and fifteen times thicker than
indentation depth for soft alloys.

Rockwell Hardness Test:

In the Rockwell test the depth of the indenter penetration into the specimen surface is measured. The
indenter may be either a hardened steel ball with diameter 1/16”, 1/8” or a spherical diamond cone of
120º angle (Brale).
Loading procedure starts from applying a minor load of 10 kgf (3kgf in Rockwell Superficial Test)
and then the indicator, measuring the penetration depth, is set to zero. After that the major load (60,
100 or 150 kgf)is applied. The penetration depth is measured after removal of the major load.

Hardness is measured in different scales (A, B, C, D, E, F, G, H, K) and in numbers, having no units

(in contrast to Brinell and Vickers methods).

Aluminum alloys, copper alloys and soft steels are tested with 1/16” diameter steel ball at 100 kgf
load (Rockwell hardness scale B).
Harder alloys and hard cast iron are tested with the diamond cone at 150 kgf ( Rockwell hardness scale
C).
An example of Rockwell test result: 53 HRC. It means 53 units, measured in the scale C by the
method HR (Hardness Rockwell).

Rockwell Superficial Hardness Test

Rockwell Superficial Test is applied for thin strips, coatings, carburized surfaces.
Reduced loads (15 kgf, 30 kgf, and 30 kgf) as a major load and deduced preload (3kgf) are used in the
superficial test.

Depending on the indenter, two scales of Rockwell Superficial method may be used: T (1/16” steel
ball) or N (diamond cone).

62 R30T means 62 units, measured in the scale 30T (30 kgf, 1/16” steel ball indenter) by the
Rockwell Superficial test.
Vickers Hardness Test:

The principle of the Vickers Hardness method is similar to the Brinell method.

The Vickers indenter is a 136 degrees square-based diamond pyramid.

The impression, produced by the Vickers indenter is clearer, than the impression of Brinell indenter,
therefore this method is more accurate.

The load, varying from 1kgf to 120 kgf, is usually applied for 30 seconds. 
The Vickers number (HV) is calculated by the formula:
HV = 1.854*F/ D²
Where

F-applied load, kg
D – length of the impression diagonal, mm
The length of the impression diagonal is measured by means of a microscope, which is usually an
integral part.
Knoop Hardness Test:

A diamond pyramid indenter with angles 130º and 170º30’ is used in this method.
The Knoop Hardness Test is applied for testing soft material and thin coating, since the penetration
depth is very small (about 1/30 of the impression length).

The loading force in the Knoop method are usually in the range of 10 gf to 1000gf (micro-hardness
range).

The Knoop number (HK) is calculated by the formula:

HK = 14.229*F/L²
Where

F-applied load, kg
L – long diagonal of the impression, mm

Shore Scleroscope Hardness Test:

The Shore Scleroscope hardness is associated with the elasticity of the material.

The appliance consists of a diamond-tipped hammer, falling in a graduated glass tube from a definite
height. The tube is divided into 140 equal parts.
The height of the first rebound is the hardness index of the material.

The harder the material, the higher the rebound.

The Shore method is widely used for measuring hardness of large machine components like rolls,
gears, dies, etc.

The Shore scleroscope is not only small and mobile, it also leaves no impressions on the tested
surface. 

1.4.5. Dissolution Test: Dissolution testing is an in vitro method that characterizes how an API is
extracted out of a solid dosage form. It can indicate the efficiency of in vivo dissolution but does not
provide any information on drug substance absorption.

When administered through oral route, the drug must be absorbed from the GI tract, and for the
absorption the drug should be in solution form, thus assessment of dissolution characteristics. nature
of the solvent including its pH and temperature, and stirring and mixing environment.

In the pharmaceutical industry, drug dissolution testing is routinely used to provide critical in vitro
drug release information for both quality control purposes, i.e., to assess batch-to-batch consistency of
solid oral dosage forms such as tablets, and drug development, i.e., to predict in vivo drug release
profiles.

Factors affecting dissolution

A dissolution test measures the amount of drug that goes into solution over a period of time under
standardized conditions. Factors that affect the dissolution of a drug product include the intrinsic
properties of the API (e.g., solubility, wettability, particle size, surface area, morphology,
polymorphs), the formulation composition and characteristics (e.g., excipients, hardness,
manufacturing process), and the dissolution method used for its assessment (e.g., apparatus, medium,
test conditions, sampling, and sample analysis) .

Method development
In dissolution testing, the aim is to develop a discriminatory method that is sensitive to variables that
affect the dissolution rate, and consequently, the in-vivo performance of the drug product. The method
must be able to distinguish between drug products manufactured under target conditions and
formulations with meaningful variations for the most relevant critical manufacturing variables, such
as drug substance particle size, compression force, and tablet hardness, for example. The dissolution
method should also be sufficiently rugged and reproducible for daily operations as well as transferable
between laboratories .
Dissolution apparatus
The United States Pharmacopeia and European Pharmacopoeia describe four different dissolution
apparatuses that can be used to develop an appropriate dissolution method for oral solid-dosage forms
based on the drug product characteristics .

Apparatus 1 (basket) and Apparatus 2 (paddle) are most commonly used methods in dissolution
testing. Apparatus 1 consists of a vessel made of glass or other inert, transparent material and a
cylindrical basket attached to the lower part of a rotating stirrer. The set up for Apparatus 2 is
generally the same except that a paddle formed from a blade and a shaft is used as the stirring
element. Sinkers may be used for dosage forms that would otherwise float. These apparatuses are
simple, robust, well standardized, and flexible enough to allow dissolution testing for a wide variety
of oral solid-dosage forms. Baskets and paddles are recommended for dissolution method
development unless they have been shown to be unsatisfactory (1). One disadvantage with the basket
and paddle methods is that the volume for dissolution is fixed. In addition, simulation of
gastrointestinal transit condition is not easily possible.

Apparatus 3 (reciprocating cylinder) was develop in recognition of the need for a system that
sequentially alters the dissolution conditions to mimic the gastrointestinal tract, so that in vitro-in
vivo correlations can be established. Apparatus 3 consists of a set of cylindrical, flat-bottomed glass
vessels, a set of glass reciprocating cylinders with inert fittings and screens at the top and bottom of
the cylinders, and a motor and drive assembly to reciprocate the cylinders vertically inside the vessels;
and if desired, the reciprocating cylinders can be moved horizontally to different rows of vessels (8,
9). Unlike the baskets and paddles, agitation in Apparatus 3 comes from dipping within the vessel,
rather than through a stirred media approach. The advantages of the reciprocating cylinder include the
ability for pH profiling, which enables simulation of pH changes in the gastrointestinal tract; and the
system can be programmed to run dissolution in different media and at different speeds at various
times (i.e., programmable dip speeds at each interval and programmable interval time). Such
flexibility allows for better in vitro-in vivo modeling.  
Apparatus 4 (flow-through cell) consists of a reservoir containing the dissolution medium, a pump
that forces the medium upwards through the vertically positioned flow-cell, and a water bath to
maintain the temperature of the dissolution medium (8, 9). The flow-through cell apparatus can
operate in two different modes—an open system with continuous flow of solvent passing through the
cell from the reservoir, and a closed system where a fixed volume of media is recirculated. The open
system is suitable for poorly soluble drugs, which require a high volume of media for dissolution. The
closed system, on the other hand, is used when a low volume of medium is required.
1.4.6. Disintegration Test: This test is provided to determine whether tablets or capsules
disintegrate within the prescribed time when placed in a liquid medium under the experimental
conditions presented below.

For the purposes of this test disintegration does not imply complete dissolution of the unit or even of
its active constituent. Complete disintegration is defined as that state in which any residue of the unit,
except fragments of insoluble coating or capsule shell, remaining on the screen of the test apparatus
or adhering to the lower surface of the discs, if used, is a soft mass having no palpably firm core.

Use apparatus A for tablets and capsules that are not greater than 18 mm. For larger tablets and
capsules use apparatus B.

Test A. Tablets and capsules of normal size

This text is based on the internationally-harmonized texts developed by the Pharmacopoeial

Discussion Group (PDG). Some editorial modifications have been made in order to be in line
with the style used in The International Pharmacopoeia.

Apparatus. The apparatus (Figure 1) consists of a basket-rack assembly, a 1000 mL, low-form
beaker, 138–160 mm in height and having an inside diameter of 97–115 mm for the immersion fluid,
a thermostatic arrangement for heating the fluid between 35 °C and 39 °C and a device for raising and
lowering the basket in the immersion fluid at a constant frequency rate between 29 and 32 cycles per
minute, through a distance of not less than 53 mm and not more than 57 mm. The volume of the fluid
in the vessel is such that at the highest point of the upward stroke the wire mesh remains at least 15
mm below the surface of the fluid and descends to not less than 25 mm from the bottom of the vessel
on the downward stroke. At no time should the top of the basket-rack assembly become submerged.
The time required for the upward stroke is equal to the time required for the downward stroke and the
change in stroke direction is a smooth transition rather than an abrupt reversal of motion. The basket-
rack assembly moves vertically along its axis. There is no appreciable horizontal motion or movement
of the axis from the vertical.
Basket-rack assembly . The basket-rack assembly consists of six open-ended transparent tubes each
75.0–80.0 mm long and having an internal diameter of 20.70–23.00 mm and a wall 1.0–2.8 mm thick;
the tubes are held in a vertical position by two plates, each 88–92 mm in diameter and 5.00–8.50 mm
in thickness, with six holes, each 22–26 mm in diameter, equidistant from the centre of the plate and
equally spaced from one another. Attached to the lower surface of the lower plate is a woven stainless
steel wire mesh which has a plain square weave with 1.8–2.2 mm apertures and with a wire diameter
of 0.570–0.660 mm. The parts of the apparatus are assembled and rigidly held by means of three bolts
passing through the two plates. A suitable means is provided to suspend the basket-rack assembly
from the raising and lowering device using a point on its axis.

The design of the basket-rack assembly may be varied somewhat provided the specifications for the
glass tubes and the screen mesh size are maintained. The basket-rack assembly conforms to the
dimensions shown in Figure 1.

Discs. The use of discs is permitted only where specified or allowed. Each tube is provided with a
cylindrical disc 9.35–9.65 mm thick and 20.55–20.85 mm in diameter. The disc is made of a
suitable, transparent plastic material having a specific gravity of

1.18–1.20. Five parallel 1.9–2.1 mm holes extend between the ends of the cylinder. One of the holes
is centered on the cylindrical axis. The other holes are centered 5.8–6.2 mm from the axis on
imaginary lines perpendicular to the axis and parallel to each other. Four identical trapezoidal-shaped
planes are cut into the wall of the cylinder nearly perpendicular to the ends of the cylinder. The
trapezoidal shape is symmetrical; its parallel sides coincide with the ends of the cylinder and are
parallel to an imaginary line connecting the centres of two adjacent holes 6 mm from the cylindrical
axis. The parallel side of the trapezoid on the bottom of the cylinder has a length of 1.5–1.7 mm and
its bottom edges lie at a depth of 1.50–1.80 mm from the cylinder’s circumference. The parallel side
of the trapezoid on the top of the cylinder has a length of 9.2–9.6 mm and its centre lies at a depth of
2.5–2.7 mm from the cylinder’s circumference. All surfaces of the disc are smooth. If the use of discs
is specified add a disc to each tube and operate the apparatus as directed under procedure. The discs
conform to the dimensions found in Figure 1.
The use of automatic detection employing modified discs is permitted where the use of discs is
specified or allowed. Such discs must comply with the requirements of density and dimension given
in this chapter.

Procedure. Place one dosage unit in each of the six tubes of the basket and if specified add a disc.
Operate the apparatus using water as the immersion fluid unless another liquid is specified and
maintain its temperature at 35–39 °C. At the end of the specified time lift the basket from the fluid
and observe the dosage units: all of the dosage units have disintegrated completely. If one or two
dosage units fail to disintegrate repeat the test on 12 additional dosage units. The requirements of the
test are met if not less than 16 of the 18 dosage units tested are disintegrated.

Test B – Large tablets and large capsules

This test is reproduced from The European Pharmacopoeia with permission and with appropriate
editorial modifications.

Apparatus. The main part of the apparatus (Figure 2) is a rigid basket-rack assembly supporting 3
cylindrical transparent tubes 77.5 ± 2.5 mm long, 33.0 mm ± 0.5 mm in internal diameter and with a
wall thickness of 2.5 ± 0.5 mm. Each tube is provided with a cylindrical disc 31.4 ± 0.13 mm in
diameter and 15.3 ± 0.15 mm thick, made of transparent plastic with a relative density of1.18–1.20.
Each disc is pierced by 7 holes, each 3.15 ± 0.1 mm in diameter, 1 in the centre and the other 6 spaced
equally on a circle of radius 4.2 mm from the centre of the disc. The tubes are held vertically by 2
separate and superimposed rigid plastic plates 97 mm in diameter and 9 mm thick with 3 holes. The
holes are equidistant from the centre of the plate and equally spaced. Attached to the underside of the
lower plate is a piece of woven gauze made from stainless steel wire 0.63 ± 0.03 mm in diameter and
having mesh apertures of 2.0 ± 0.2 mm. The plates are held rigidly in position and 77.5 mm apart by
vertical metal rods at the periphery. A metal rod is also fixed to the centre of the upper plate to enable
the assembly to be attached to a mechanical device capable of raising and lowering it smoothly at a
constant frequency of between 29 and 32 cycles per minute, through a distance of 55 ± 2 mm.
The assembly is suspended in the specified liquid medium in a suitable vessel preferably a 1 litre
beaker. The volume of the liquid is such that when the assembly is in the highest position the wire
mesh is at least 15 mm below the surface of the liquid, and when the assembly is in the lowest
position the wire mesh is at least 25 mm above the bottom of the beaker and the upper open ends of
the tubes remain above the surface of the liquid. A suitable device maintains the temperature of the
liquid at 35–39 °C.

The design of the basket-rack assembly may be varied provided the specifications for the tubes and
wire mesh are maintained.

Method. Test 6 tablets or capsules either by using 2 basket-rack assemblies in parallel or by

repeating the procedure. In each of the 3 tubes place 1 tablet or capsule and, if prescribed, add a disc;
suspend the assembly in the beaker containing the specified liquid. Operate the apparatus using
water as the immersion fluid unless another liquid is specified for the prescribed period, withdraw
the assembly and examine the state of the tablets or capsules. To pass the test all 6 of the tablets or
capsuls must have disintegrated.

1.4.7. Friability Test:

Friability (the condition of being Friable) testing is a method, which is employed to determine
physical strength of compressed and uncoated tablets upon exposure to mechanical shock
andattrition. In simple words,friability test tells how much mechanical stress tablets are able to
withstand during their manufacturing, distribution and handling by the customer. Throughout
pharmaceutical industry, friability testing has become an accepted technology and the instrument
used in to perform this process is called Friabilator or Friability Tester. The mechanical strength
of tablet or granules can be determined by its hardness and through friability test. The strength
of a tablet plays a very important role in its marketing and dissolution.