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Sharanabasappa B. Patil et al /J. Pharm. Sci. & Res. Vol. 12(2), 2020, 271-278
Extraction of Amino acids from Human Hair “Waste” and
Used as a Natural Fertilizer
Sharanabasappa B. Patil1, Shreya K2 and Kruti S2
1
Department of Chemistry, Ramaiah Institute of Technology, Bangalore-560054, India
2
Department of Chemical Engineering, Ramaiah Institute of Technology, Bangalore-560054, India
Abstract
Hair is a protein filament that grows from follicles found in the dermis. Hair is one of the defining characteristics of
mammals. The human body, apart from areas of glabrous skin, is covered in follicles which produce thick terminal and
fine vellus hair. Most common interest in hair is focused on hair growth, hair types, and hair care, but hair is also an
important biomaterial primarily composed of protein, notably alpha-keratin. Attitudes towards different forms of hair, such
as hairstyles and hair removal, vary widely across different cultures and historical periods, but it is often used to indicate a
person's personal beliefs or social position, such as their age, sex, or religion. Human hair is a material considered useless in
most societies and therefore is found in the municipal waste streams in almost all cities and towns of the world.
Key Words: Human Hair, Amino acids and Fertilizer
INTRODUCTION: Hair growth begins inside the hair follicle. The only
1.1Discription: The word “hair” usually refers to two "living" portion of the hair is found in the follicle. The hair
distinct structures: that is visible is the hair shaft, which exhibits no
1. The part beneath the skin, called the hair follicle, or, biochemical activity and is considered "dead". The base of
when pulled from the skin, the bulb. This organ is located a hair's root (the "bulb") contains the cells that produce the
in the dermis and maintains stem cells, which not only re- hair shaft. Other structures of the hair follicle include the
grow the hair after it falls out, but also are recruited to re- oil producing sebaceous gland which lubricates the hair
grow skin after a wound. and the arrector pili muscles, which are responsible for
2. The shaft, which is the hard filamentous part that causing hairs to stand up. In humans with little body hair,
extends above the skin surface. A cross section of the hair the effect results in goose bumps.
shaft may be divided roughly into three zones.
The hair’s structure can be divided into three distinct
parts:
3. Medulla: innermost layer of the hair shaft
composed of an amorphous, soft, oily substance.
4. Cuticle: thin protective outer layer that contains
the nourishing portion essential to hair growth. It is
highly keratinized, composed of cells shaped like
scales that are layered one over the other, measuring
about 60 micrometers long and about 6 micrometers
wide.
5 Cortex: main component of the hair, containing
long keratin chains that add elasticity, suppleness and
resistance to the hair. The cells of the cortex are
joined together by an intercellular cement rich in Cross Section of Hair Strand
lipids and proteins. Each cell is composed of bundles
that lie in the direction of the hair length: these are
macro-fibrils which are made up of micro-fibrils,
which in turn contain proto-fibrils.
The shape of the follicle determines the shape of the
cortex, and the shape of the fiber is related to how straight
or curly the hair is. People with straight hair have round
hair fibers. Oval and other shaped fibers are generally
more wavy or curly. The cuticle is the outer covering. Its
complex structure slides as the hair swells and is covered
with a single molecular layer of lipid that makes the hair
repel water. The diameter of human hair varies from 0.017
to 0.18mm (0.00067 to 0.00709 in). There are two million
small, tubular glands and sweat glands that produce watery
fluids that cool the body by evaporation. The glands at the
opening of the hair produce a fatty secretion that lubricates Root of the hair
the hair.
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1.2 Natural Color are made up of C,O,H,N and S elements, (Carbon,
All natural hair colors are the result of two types of hair Oxygen, Hydrogen, Nitrogen and Sulfur).
pigments. Both of these pigments are melanin types,
produced inside the hair follicle and packed into granules 2.2 The percentage of COHNS elements in hair is as
found in the fibers. Blonde hair is the result of having follows:
little pigmentation in the hair strand. Gray hair occurs
when melanin production decreases or stops, Element Percentagein Normal hair
while poliosis is hair (and often the skin to which the hair Carbon 51%
is attached), typically in spots, that never possessed Oxygen 21%
melanin at all in the first place, or ceased for natural Nitrogen 17%
genetic reasons, generally, in the first years of life. Hydrogen 6%
Sulfur 5%
1.3 Texture These elements form bonds called side bonds which link
Hair exists in a variety of textures. Three main aspects of together the long chain of amino acids known as the
hair texture are the curl pattern, volume, and consistency. polypeptide chain. This chain forms a helix by creating
The derivations of hair texture are not fully understood. spiral movement that intertwines.
All mammalian hair is composed of keratin, so the make-
up of hair follicles is not the source of varying hair 2.3 The following are the amino acids and the
patterns. There are a range of theories pertaining to the percentage found in hair fiber:
curl patterns of hair. Scientists have come to believe that Percentage in Normal
the shape of the hair shaft has an effect on the curliness of Amino acid
hair
the individual's hair. A very round shaft allows for Cysteine 17.50%
fewer disulfide bonds to be present in the hair strand. This
Serine 11.70%
means the bonds present are directly in line with one
Glutamic Acid 11.10%
another, resulting in straight hair.
Threonine 6.90%
The flatter the hair shaft becomes, the curlier hair gets,
because the shape allows more cystines to become Glycine 6.50%
compacted together resulting in a bent shape that, with Leucine 6.10%
every additional disulfide bond, becomes curlier in Valine 5.90%
form. As the hair follicle shape determines curl pattern, the Arginine 5.60%
hair follicle size determines thickness. While the Aspartic Acid 5.00%
circumference of the hair follicle expands, so does the Alanine 4.80%
thickness of the hair follicle. An individual's hair volume, Proline 3.60%
as a result, can be thin, normal, or thick. The consistency Isoleucine 2.70%
of hair can almost always be grouped into three categories: Tyrosine 1.90%
fine, medium, and coarse. This trait is determined by the Phenylalanine 1.40%
hair follicle volume and the condition of the strand. Fine Histidine 0.80%
hair has the smallest circumference, coarse hair has the Methionine 0.50%
largest circumference, and medium hair is anywhere
between the other two. Coarse hair has a more open cuticle Millions of polypeptide chains reside in the cortex layer.
than thin or medium hair causing it to be the most porous. Side bonds such as hydrogen bonds, salt bonds and
disulfide bonds link together these polypeptide chains.
2.1Hair Composition Hair fibers are held in place by the side bonds which
Human hair is an appendage which grows from follicles, attribute to the elasticity and strength of hair.
tube like sacs in the scalp or skin containing the hair root. A hydrogen bond can easily be broken by water or heat,
The hair that we cut, relax, color and style is a non-living and is a physical side bond. Collectively, hydrogen bonds
fiber comprised of keratinized protein. Within the hair account for one-third of hair’s strength. Salt bonds are also
follicle cells are produced. These cells mature in an physical side bonds. Strong acidic or alkaline solutions
upward moving process through the follicle. This break salt bonds because they are affected by changes in
maturing process is known as keratinization. During pH. Like hydrogen bonds, salt bonds also account for
keratinization cells absorb keratin, a fibrous protein. As approximately one-third of hair’s strength. Disulfide bonds
the cells continue to move upward they lose their nucleus differ from hydrogen and salt bonds because they are not
and die off, producing the non-living keratinized cells physical side bonds. Disulfide bonds are chemical side
(appendage) that emerge from the scalp. bonds. Disulfide bonds link together two sulfur atoms
Hair is comprised of many contributing factors. Proteins, attached to cysteine amino acids within the polypeptide
raw elements, amino acids and bonds work together in chains. Chemical hair relaxers and permanent waves
forming hair fiber. The dominant contributor in the chemically alter the hair’s disulfide bond. Disulfide bonds
composition of hair is protein, accounting for 91 percent cannot be broken by water or heat1.
of hair fiber. Amino acids, the building blocks of protein,
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cosmetic brushes4. Almost all kinds of straight hair can be
used for brushes.
4.2 Agriculture
4.2.1.As Fertilizer. Human hair is one of the highest
nitrogen containing (∼16%) organic material in nature
because it is predominantly made up of (nitrogen-
containing) proteins. For comparison, cattle dung contains
only ∼0.2-0.3% nitrogen. In addition, human hair also
contains sulfur, carbon, and 20 other elements essential for
3. Uses of Human Hair plants5. In the atmosphere, hair decomposes very slowly,
The unique properties of human hair such as its unique but moisture and keratinolytic fungi present in soil, animal
chemical composition, slow degradation rate, high tensile manure, and sewage sludge can degrade hair within a few
strength, thermal insulation, elastic recovery, scaly months6. In traditional Chinese agriculture, human hair
surface, and unique interactions with water and oils, along was mixed with cattle dung to prepare compost that was
with its socio-cultural roles, have led to many diverse uses. applied to the fields in the winter season. In some
These uses also depend on the variety of hair available, communities in India, hair has been used directly as
varying in terms of five parameters: length, color, fertilizer for many fruit and vegetable crops and in making
straightness or curliness, hair damage, and contamination. organic manures. Recent experiments on horticulture
This variation depends on the culture, ethnicity, hair plants show that direct application of human hair to soil
styles, and the hair care practices in the region. For provides the necessary plant nutrients for over two to three
example, in areas with malnutrition or in areas where hair cropping seasons. A company named Smart Grow has
treatments such as permanent waving, dyeing, and popularized the fertilizer use of human hair in the USA by
chemical shampoos are common, hair is more damaged. selling it in the form of hair mats for potted plants. Small
Chemical contamination in hair is observed due to use of entrepreneurs in the USA are also promoting hair as
toxic dyes and chemicals in hair care, or due to the fertilizer by packaging it in various user-friendly forms
presence of toxic chemicals in the atmosphere or food such as in tea bags. By mixing human hair with cattle
chain of the area. For example, in many areas of the world dung and feeding worms on the mixture, it is also possible
where electronic waste recycling is carried out, hair is to make good quality vermin compost within a period of
found contaminated with brominated flame retardants2. about 2 months. Non composted hair, however, has
This section describes uses for different kinds of human advantages than composted hair because composting can
hair according to the field of application. lead to some loss of nitrogen. While the biological
decomposition pathways take a few months, human hair
4. Applications of Human Hair can also be decomposed within a few hours by chemically
4.1 Fashion, Theatre, and Cosmetics Industry hydrolyzing it at high temperatures in acid or base
4.1.1. Wigs, Hair Extensions, Eyelashes, Moustaches, solutions. The hydrolyzed solution, which mainly consists
Beards, and Other Beauty Accessories. This is one of the of amino acids with some fatty acids and nucleotides, can
most ancient and currently the largest of the human hair- be used as a liquid fertilizer after neutralization.
based industries, with a constantly increasing scale due to Experiments using this hydrolyzed as foliar spray show
global expansion of the fashion industry. The oldest enhancement of the chlorophyll content as well as biomass
known wigs are from1400 B.C. Egypt, some of which are in spinach and wheat plants. Application of the solution to
still intact today after3400 years3. In the eastern countries, soil also shows improvements in the color and size of
such products primarily catered to the theatre world, but in Amarant husdubius and hot pepper plants. Experiments on
the west they evolved significantly with the fashion the hot pepper plants also show increased diversity of soil-
industry. This application predominantly uses good intrinsic bacteria, which significantly reduces the spread of
quality, long hair of almost all colors. In addition, hair in a wilt disease in these plants caused by the bacterium
which all strands have surface scales in the same direction Ralstoniasolanacearum. Long term impacts of this use,
(similar to hair on the human body), known as Remy hair, however, need to be assessed. Any kind of hair without
is preferred because it tangles much less during working. toxic contamination can be used for fertilizers. Finely
Non-Remy hair is often used after chemically removing shredded hair, however, is better for faster decomposition.
the outer scale containing cuticle layer. Pure Remy hair 4.2.2.Pestcontrol. Human hair is also known to address
products are expensive, while those with non-Remy hair or problems arising from many animals as well as insect
human hair mixed with other fibers are cheaper. The users pests, although by different mechanisms. Among large
include hair care researchers, product manufacturers, hair animals, it has been used to repel rabbits in Mauritius,
stylists, and trainees in both labs and salons. These tests rodents, and wild boar in India, and deer in the USA.
use hair of different colors, range of curliness, and Typically, the hair is spread along the boundary of the
different levels of damage. fields/farms or near rat holes in the field. Rabbits, rodents,
4.1.2. For Making Cosmetic Brushes. Scales on hair can and wild boar finds their food by sniffing, and hair
hold cosmetic powder particles and apply it uniformly on supposedly causes them discomfort during sniffing by
skin or a surface. Therefore, human hair is used in making coming into their nostrils. In the case of deer, repulsionis
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supposedly caused by human smell emanating from the spills in the Philippines and the USA. In this method, oil
hair because hanging hair in nylon bags also works, and can be recovered by wringing out the hair, which then can
the technique does not seem to work well in the areas be reused up to 100 times—advantages not present in
where deer are unafraid of humans. Among insects, human other oil spill remediation methods. With this method, up
hair is used for deterring rhinoceros beetles in India. Small to 98% of the spilled oil can be recovered. The oily hair
balls of human hair are placed at the nodes of the affected can then be used to grow oyster mushrooms, which
plant such as coconut tree. Beetles get tangled in the hair decompose the oil. The hair then left can be composted.
becoming unable to move. By using human hair mats, Human hair can also separate emulsified oil in water,
farmers in Florida(USA) were able to save ∼$45,000 on which is very expensive to clean by other methods.
pesticides on about 1million plants in the year 2007 in 4.4.2. Removing Phenols, Aldehydes, Dyes, and Heavy
addition to the labor savings and benefits as fertilizer. Metal Pollutants from Water. Human hair absorbs several
chemicals from aqueous solutions. Experiments show that
4.3 Composite Materials human hair can absorb organic pollutants such as
4.3.1. Reinforcement of Construction Materials. Due to formaldehyde and phenol, and heavy metals such as
high tensile strength and high friction coefficient, human mercury (Hg),copper (Cu), cadmium (Cd), and silver (Ag)
hair has been used for reinforcing clay-based from aqueous solution. The capacity for metal absorption
constructions. In rural areas in Uttar Pradesh and Madhya can be enhanced by pre-treating the hair with an alkali.
Pradesh, India, Bangladesh, Syria, and in European Powdered human hair has good absorptive capacity for Ni
countries, human hair/clay mixture (along with other (II) and Cr(VI) ions at acidic pH, and partially burned
binders) is used in plastering house walls, lining ovens, human hair9 shows selective absorption of mercury (Hg2+)
making wheels, and so forth. The addition of hair and silver(Ag+) ions over cobalt (Co2+), copper (Cu2+),
significantly reduces cracking and prolongs the life of and iron (Fe3+).Thus, human hair can be a low-cost
these structures. Research shows that human hair absorbent for purification of polluted waters. The
reinforcement enhances the structural strength as well as recycling/disposal of contaminated hair, however, can
the thermal insulation capacity of the claystructures7-8. become a problem. Partial recovery of absorbed metals
Although such clay-based constructions are now from the hair has been explored, but further research is
decreasing in rural areas, they are gaining importance in needed to develop recovery/recycling methods for such
sustainable architecture. Human hair reinforcement also chemicals from the contaminated hair.
reduces cracks in cement mortar caused by plastic
shrinkage by as much as 92%and increases the 4.5 Pharmaceuticals and Biomedical Applications
compressive strength of fly ash/cement concrete by over 4.5.1. Pharmaceuticals. Human hair proteins typically
three times. Any kind of hair can be used in these contain 20 essential amino acids, which can be extracted
applications. by complete hydrolysis of hair. Some of the amino acids
4.3.2. Molded Furniture and Objects. A UK-based obtained in good yield from human hair are L-cysteine,
entrepreneur, Ronald Thompson, has developed a method Leucine,L-isoleucine, and L-valine. L-cysteine and its
for making composite materials which includes first chemical derivatives are used in many cosmetics and
weaving human hair into a web or mat and then adding a pharmaceutical formulations. For example, L-cysteine is
structural additive like resin or flexible polymer used for permanent wave lotions and wound healing
(preferably a recyclable or biodegradable material). The formulations, while one of its derivatives, N-acetyl L-
composite has good strength and can be used for making cysteine (NAC),is used to treat conditions such as chest
molded structures such as furniture and mannequins. A congestions and acetaminophen poisoning. Hair from
similar composite with unwoven hair has also been used certain demographic regions is better for extracting certain
for making biodegradable eye glasses. amino acids. For example, black hair from Asia has more
4.3.3. Composites for Superconducting Systems. cysteine than blonde hair. In addition, hair that is
Superconducting power equipmentoften use fiber-glass- chemically not altered by any styling treatment is better,
based composites for cryogenic insulations. Michael et al because some of these treatments can change the chemical
have that composite laminate of human hair (and several structure of the hair. For example, increasing trend of
other natural fibers) with epoxy resin has dielectric permanent waving in some parts in China has made
breakdown properties suitable for insulation in cryogenic cysteine extraction from this hair difficult.
systems. Compared to currently used glass-fiber 4.5.2.HydrolyzedHair Keratin. Mixture of amino acids and
composites, these composites can significantly lower the polypeptides obtained by the hydrolysis of keratin protein
production costs of cryogenic equipment. from human hair, known as hydrolyzed human hair keratin
protein (HHKP), is used in hair care products by
4.4 Pollution Control and Remediation manycompanies10-11. It is reported to repair hair damage
4.4.1. Oil-Water Separation and Oil Spill Remediation. caused by various hair styling treatments supposedly
Human hair surface has a high affinity for oils—much because its constituents are similar to the native hair
higher than its affinity for water. This property is very protein.
useful in oil-water separation. After the pioneering work 4.5.3. Ethno medicinal Uses. Several cultures have been
of Phillip A. McCrery from Alabama, USA, booms and using human hair for preparing traditional medicines.
mats of human hair have been used to clean up coastal oil Carbonized human hair has been used in Traditional
274Sharanabasappa B. Patil et al /J. Pharm. Sci. & Res. Vol. 12(2), 2020, 271-278
Chinese Medicine for treating hemorrhage, burns, wounds, microsensors for small biomolecules. Furtherexperiments
and scars. It is also used in veterinary medicine to stop are needed to test their efficacy in variousbiological
bleeding and to promote urination. In rural communities in systems.
Chhattisgarh, India, hair ash is applied to open wounds for
immediate pain reliefs well as long term recovery. In 4.6 Food Industry
addition, these communities use formulations made from Many amino acids obtained from human hair such as L-
powdered hair, hair ash, and hair decoction for the cysteine are also used in the food industry as leavening
treatment of mouth ulcers, ringworm, and blisters due to agent for pizza dough and doughnuts, for artificial meat
burns. Another ethno medicinal studying India has flavor, in nutritional supplements, and so forth. The use of
revealed therapies using human hair for treatments of human hair derived amino acids in the food industry,
anemia, asthma, urinary calculus, piles, rat bite poisoning, however, is a big source of concern in many countries
foot sprains, sexual problems, and childbirth pain. The (videinfra).
quality and purity of hair is of essence in these
preparations. 4.7 Scientific Instrumentation
4.5.4. Suturing Material in Surgery. Human hair has Human hair expands in length on absorbing moisture, and
sufficiently high strength for use as suture in most this expansion is reversible. This property was used by
surgeries. It is relatively easy to tie knots with and is Horace-Benedict de Saussure in1783 to make a hair
noninfectious (because of its slow decomposition rate and hygrometer for humidity measurements. Since then, the
high compatibility with the human body). Its use as suture instrument has undergone improvements and
was known in European the Middle Ages. Studies have modifications in design12-13. Although more sophisticated
now established the potential of human hair sutures in electronic instruments are now available for more precise
cataract and conjunctiva wound repair surgeries, general measurements, hair hygrometers are still cheap and simple
surgeries on humans and animals, and in microsurgery. It for reasonably good estimations of humidity and are still
can be easily sterilized by autoclaving. Long, undamaged used in many metrological stations in the world. This
hair of medium thickness (not too coarse or too thin) is application requires long hair (∼12 inches or longer).
best for suturing.
4.5.5. Keratin-Based Engineering Biomaterials.In 2002, 4.8 Textiles, Fiber stuffing, and Other Artifacts
Nakamura etal. developed the method to extract proteins 4.8.1. Stuffing Toys, Mattresses, and Other Household
rapidly and efficiently from human hair, opening Items. Due to its elastic and cushiony nature and good
possibilities of reengineering human hair proteins into new thermal insulation properties, human hair has been used to
materials. Based on this method, many materials with stuff household items such as hair-pin cushions and toys in
novel properties have been developed such as hair protein the USA14-15. And toys, furniture, mattresses, quilts,
based thin films, hairprotein gellan chitosan hybrid fibers, jackets, and so forth, in India. In pin cushions, natural oil
and protein scaffolds and hydro gels for tissue of the hair prevents the pins and needles from rusting. For
engineering. These materials have several potential toys and mattress stuffing, human hair is usually mixed
applications in bioengineering and medical science such as with cotton or other fibers.
for wound dressing and soft tissue regeneration with the 4.8.2. Fabrics. High thermal insulation, elasticity, and
advantage ofgood biocompatibility with the human body. good tensile strength also make human hair useful for
Investigations on effects of these materials in surgical making various kinds of fabrics. In Arunachal Pradesh,
applications are under progress. India, people have traditionally been making fabrics by
4.5.6. Human Hair Follicle Cell Cultures and Tissue mixing human hair with yak hair, nettle fiber, and cotton.
Regeneration. Biomedical studies show that certain cells In China, human hair, yak hair, and cotton are used to
from human hair follicles such as outer root sheath cells make interlining cloth for coats and jackets. In these
also are useful in wound treatments, antilogous grafting of applications, mostly cotton yarn is used as warp and the
chronic wounds, and treatment of alopecia. The follicles hair is used as weft.Human hair blended with animal fibers
are collected from fallen hair or hair plucked from is used for making blankets in Panipat, India. A company
volunteers.Therefore, this application is not likely to al. Kishore’s (Chennai, India) has also started making
utilize much of the hair waste. purely human hair fabric and clothing. Making felt from
4.5.7. Flexible Microelectrodes. Human hair by itself is human hair is more difficult than from animal hair, but it
not a good conductor of electricity, but Xu et al. from is made by many for use as doormats, thermal padding of
China have developed a human hair microelectrode by furniture, artwork, and so forth. A Serbian artist produced
coating its surface with an ultrathin layer of gold. 1200 sq. m. of felt from human hair in 2009.
Currently used carbon fiber microelectrodes have good 4.8.3. Oil Filters. Tightly woven human hair cloths were
electrical conductivity, chemical stability, and low cost but used in the 1920s as filters for heavy oils in refineries and
are brittle and have weaksignal strength and limited distilleries because these processes involved high pressure
biocompatibility. The humanhair microelectrode is that many natural fibers could not withstand16. Haircloths,
flexible and its signal strength canbe tailored by changing by contrast, were tough and almost untreatable. Later,
the hair length. The gold coating ischemically stable and synthetic fibers with higher tensile strength and smaller
the electrode is likely to be compatiblewith biological diameters became available. Filters of these fibers could
systems. In addition, these electrodes can alsobe useful as
275Sharanabasappa B. Patil et al /J. Pharm. Sci. & Res. Vol. 12(2), 2020, 271-278
filter even smaller particles and therefore replaced the 5. Materials Used
human hair filters. • Human hair
4.8.4. Ropes. Due to good tensile strength, human hair • NaOH
has-been used to make ropes in many cultures, for • HCl
example, to lift heavy beams and bells in the construction • Ethanol
of Japanese temples and for household purposes by Native • Acetone
Americans17.These ropes are still valued for horse riding. • Ninhydrin
• H2SO4
4.9 Artwork 6. Extraction of Amino acids
Two art traditions evolved in the world around human hair
Collecting the initial weighed hair samples and dissolved
as the key material. The first—hair embroidery flourished
in 0.05 N (20 ml) HCl in a beaker. Mix properly and
in China between the7th and 13th centuries, when women
manually stirred for 15 minutes. After 15 minutes 0.1N
made images of Buddha with their own hair. This art
NaOH solution added drop wise until the solid separates
revived again in the late 20thcenturies and now has
and residue is washed with 20 ml distilled water. After
surpassed its past in color and variety. Earlier, natural washing the residue is dissolved in 20 ml of ethanol and
shades of hair were used, but now dyed hair is also used.
heated for 10 min then filtered.
The tradition has even developed a rich school of portrait
embroidery. The second tradition hair work evolved in the
19thcentury Scandinavian countries and then spread to
England and America18-19. Hair work included intricate
jewelry, flowers, buttons, brooches, and so forth, made of
human hair with gold and resins. It also included the craft
of hair embroidery. Dissolved hair and finely chopped hair
mixed with oil were used as paint. The 1853 Crystal
Palace Exposition in New York included a full line of hair
work jewellery, buttons, flowers, and even a tea-set made
completely of hair. Socially, hair work pieces were mainly
associated with special affection such as between spouses
and in memories of a dead person (with hair work piece
made from the deceased person’s hair). Increasing
symbolism of hair work with death and mourning led to
fading of this tradition by the early 20th century; however,
recently, there is increasing interest in reviving this art and
a society has been established to connect and promote hair After it is dried, test the residue for the presence of amino
work artists in theworld20. acids using following methods.
7. Amino Acid Tests
4.10 Miscellaneous Uses 7.1 Ninhydrin Test
Human hair is placed with other fibers as nesting material Ninhydrin (triketohydrindene hydrate) is a chemical used
to increase breeding of birds in places where bird to detect ammoniaor primary and secondary amines.
populations are declining. Long fibers can entangle and Amino acids also react with ninhydrin at pH=4. The
cause injuries to the birds; therefore, short hair (3 inches or reduction product obtained from ninhydrin then reacts
less) is recommended for the use. A stringed musical with NH3 and excess ninhydrin to yield a blue colored
instrument named “git-git” is also made using human hair substance. This reaction provides an extremely sensitive
as strings21. test for amino acids. Apply this test to any of the amino
acids you choose.
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Procedure: - Note that this is a test for phenols, and the ninhydrin
- To 0.1 gram of amino acid dissolved in water, add test should also be positive if it is to be concluded that
200mg of ninhydrin dissolved in 10ml acetone. the substance is a phenolic amino acid.
- Notice color changes.
- Measure the absorbance of the solution using calorimeter 7.4 Hopkin’s Cole Test
at 550nm. The indole group of tryptophan reacts with glyoxylic acid
Observation: Direct reaction of ninhydrin solution with (glacial acetic acid, which has been exposed to light,
amino acid gave dark blue color. always contains glyoxylic acid CHOCOOH as an
Inference :Amino acids are detected. impurity) in the presence of concentrated H2SO4 to give a
purple color. Apply this test to glycine, tryptophan and
7.2 Xanthoproteic Test tyrosine.
Some amino acids contain aromatic groups that are
derivatives of benzene. These aromatic groups can
undergo reactions that are characteristics of benzene and
benzene derivatives. One such reaction is the nitration of a
benzene ring with nitric acid. The amino acids that have
activated benzene ring can readily undergo nitration. This
nitration reaction, in the presence of activated benzene
ring, forms yellow product. Apply this test to tyrosine, Procedure:
tryptophan, phenylalanine and glutamic acid. - To a few mL of glacial acetic acid containing glyoxylic
acid, add 1-2 drops of the amino acid solution.
- Pour 1-2 mL H2SO4 down the side of the sloping test
tube to form a layer underneath the acetic acid.
- The development of a purple color at the interface proves
a positive reaction.
Observation :Purple color wasn’t detected
Inference :Negative test
Procedure:
- To 2 mL amino acid solution in a boiling test tube, add 7.5 Lead-Sulfide Test
equal volume of concentrated HNO3. When cystine is boiled with 40% NaOH, some of sulfur in
- Heat over a flame for 2 min and observe the color. its structure is coverted to sodium sulfide (Na2S). The
- Now cool thoroughly under the tap and cautiously run in Na2S can be detected by using sodium plumbate solution
sufficient 40% NaOH to make the solution strongly which causes the precipitation of PbS from an alkaline
alkaline. solution. In order to apply this test, first the sodium
- Observe the color of the nitro derivative of aromatic plumbate solution should be prepared. Apply this test to
nucleus. cysteine and cystine.
Observation: Yellow color is seen.
Inference : Amino acid is detected.
7.3 Millon’s Test
Millon’s test is specific to phenol containing structures Procedure:
(tyrosine is the only common phenolic amino acid). - Sodium Plumbate Solution Preparation:
Millon’s reagent is concentrated HNO3, in which mercury - Add 5 mL dilute NaOH to 2 mL dilute lead acetate.
is dissolved. As a result of the reaction ared precipitate or - A white precipitate of lead hydroxide forms.
a red solution is considered as positive test. A yellow - Boil until the precipitate dissolves with the formation of
precipitate of HgO is NOT a positive reaction but usually sodium plumbate.
indicates that the solution is too alkaline. Apply this test to - Boil 2 mL amino acid solution with a few drops of 40%
tyrosine, phenylalanine, glycine and β-naphtol. NaOH for 2 min.
- Cool and add a few drops of the sodium plumbate
solution.
- A brown color or precipitate is a positive test for
sulfides.
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