COMMONWEALTH ASSOCATION FOR EDUCATION,
ADMINISTRATION AND MANAGEMENT
VOLUME 2 ISSUE 4
ISSN NO 2322-0147
APRIL 2014
ASSESSMENT OF SERUM TOTAL
PROTEINSTHROUGH BIURET METHOD IN
BILIOUS & PHLEGMATIC TEMPERAMENT
INDEXED WITH PARIS, DAIS.NET, DRJI, WORLDCAT, EBSCO-USA, J-GATE
(EDITOR-IN-CHIEF)
DR MUJIBUL HASAN SIDDIQUI
ASSISTANT PROFESSOR, DEPARTMENT OF EDUCATION,
ALIGARH MUSLIM UNIVERSITY,
ALIGARH-202002,
UTTAR PRADESH,
INDIA
www.ocwjournalonline.com
website: www.ocwjournalonline.com
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ASSESSMENT OF SERUM TOTAL PROTEINSTHROUGH BIURET
METHOD IN BILIOUS & PHLEGMATIC TEMPERAMENT
By
S M Ahmer
PG Scholar
Department of Kulliyat
Faculty of Unani Medicine
A.M.U Aligarh
E- mail: drsmahmer@yahoo.com
Dr. Ferasat ali
Associate Professor
Department of Kulliyat
Faculty of Unani Medicine
A.M.U Aligarh
ABSTRACT
The objectives of this study is to assess the value of total serum protein in bilious and
phlegmatic temperament individuals through Biuret Method, and to analyze whether serum
total protein levels in Bilious and Phlegmatic temperament (Mizaj) are similar or dissimilar
within normal healthy state.The mean and standard deviation of total serum protein levels
of bilious and phlegmatic subjects were calculated. It was revealed after this study that
the mean total serum protein of bilious subjects was 7.51 (g/dl) with standard deviation
of 0.83 while the mean total serum protein of phlegmatic subjects was 7.05 (g/dl) with
standard deviation of 0.63. In this way it can be said that the average total serum protein
of the healthy bilious subject is higher than that of a phlegmatic subject. The T-test was
applied to test the significance of the results and it was found that the difference of total
serum protein level of bilious and phlegmatic subjects was highly significant (p<0.001).
Keywords: Temperament (Mizaj), humor, bilious, phlegmatic, and serum total protein.
INTRODUCTION:
Research on the human serum proteins has been directed towards problems of
physiological and biochemical interest as well as towards those of medical and
genetical interest. From the physiological and biochemical point of view, the serum
proteins play important roles as biological buffer systems, in the maintenance of the
osmotic pressure of the blood, control of blood pH, transport of lipids, dietary nitrogen,
and other inorganic elements, and in immunological and other functions. Protein is a
proximate principle (Awamil-e-Qaribah) of the body because protein is an essential
substance of humors (Akhlat). Proteins are essential parts of organisms and participate in
virtually every process within cells. Proteins are the essential substances of which the protoplasm
of the cells and the intracellular substances are built. Proteins form the fundamental basis of the
structure and function of life. [1]
Temperament is an empirical expression describing the humoral composition that
regulates the physiological and pathological changes in the human body. Thus, there are
no two individuals are same in their temperament. So each individual due his specific
Temperament is said to possess certain innate strength and deficiencies, for which he
reacts differently to given set of environmental conditions. The temperament is formed
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by interaction of opposite qualities present in the elements (Arkan). These qualities are
primary four in number, namely hotness, coldness, moistness, and dryness. Thus, Unani
physicians broadly classified the people into four types of Temperament either on the
basis of humors. These four types of Mizaj (Temperament) are:
1).HOT & MOIST: DAMVI MIZAJ
(SANGUINEOUS)
2). COLD & MOIST: BALGHAMI MIZAJ (PHLEGMATIC)
3). HOT & DRY: SAFRAVI MIZAJ (BILIOUS)
4). COLD & DRY: SAUDAVI MIZAJ (MELANCHOLIC) [2 ]
The Hippocrates (460-370BC) gave the fundamental principles of Greco-Arab System of
Medicine with a belief that the body of the individual is composed of four basic elements, which
together, are termed as ‘Anasir-e-Arba’ or Arkan’(Elements) comprising earth, water, air and fire
as different states of matter and the building blocks of everything in the universe. These four
basic elements possess four different qualities, i.e. hot, cold, dry and wet. The admixture of these
four basic elements results in the formation of four biological fluids or Humors viz. Blood
(Dam), Phlegm (Balgham), Bile (Safra) and Black bile (Sauda); a right proportion, according to
quality and quantity constitutes health and upright proportion and irregular distribution,
according to their quantity and quality constitutes disease. Thus the basic principle of GreecoArabic Medicine is that the four humors represented in the human blood.[3]
The Literal Meaning of Temperament (Mizaj):
The literal meaning of mizajaccording to Nafis is “Intermixture” as he says “The word
mizaj originated from Arabic word imtizaj meaning intermixture ”.[4]
Mizaj is derived from Arabic word “mzj” which means mixing of humors. At other
places it is described that mizaj refers to the intermixture of four humors within human
body, it is the basis of body. (Lisan-ul-Arab)
The literal meaning of the word “temperament” as given in few authentic dictionaries
are as follows: [5]
Temperament – combination or predominance of humours. (Chamber dictionary)
Temperament is constitutionally based individual differences in emotion, attention,
activity level and self-regulation. (New Concise Medical Dictionary)
Temperament – the combination of intellectual, emotional, ethical a nd physical
characteristics of an individual. (Taber’s encyclopedia medical dictionary)
Temperament – the physical organization peculiar to the individual, which influences
one’s metabolic process, manner of thought and action and the general view of life.
(Stedman).
Temperament is defined as that part of the personality, which is genetical ly based.
Along with character and those aspects that are acquired through learning, the two
together are said to constitute personality. [6]
Definition of Mizaj:Galen(130-200 AD)says that: “Temperament is a quality produced by action
and reaction of opposite qualities of body fluids (Akhlat). When these components interact by
virtue of their respective powers (qualities) a condition is achieved which is found in equal
proportions in all the components of that intermixture; this is called temperament”.[7]
IbnSina(980-1037 AD) in his famous book “Al-Qanoon-fil-Tib”says:“The temperament is a
quality resulting from the interaction of opposite qualities present in elements
consisting of minute particles so that most of the particles of each of the elements may
touch most of the others. Thus when these particles act and react on one another with
their properties, there emerges from their total properties, a uniform quality which is
present in all of them. This is the temperament (Mizaj)” [8]
Similarly the normal temperament of an individual is defined as “a condition in which he
survives comfortably with all systems of healthy life”, which varies widely as per composition
and as well as other surrounding factors and circumstances in which he resides. [9]
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Therefore, in this universe all animates or inanimate, however alike in origin and
structure does not resemble one another perfectly. The Temperament fluctuates in particular
normal range of maximum and minimum limits. Whenever it crosses its normal limit, it leads to
the abnormality (Su-e-Mizaj) within the body.[10]
Characteristic of Safravi Mizaj (Bilious Temperament) Individuals
Humor: Yellow Bile Basic Qualities: Hot and Dry (Haar&Yabis).
The Bilious temperament is the hottest, most active and catabolic of all.
Physique: Compact, lean, moderately muscular and less fatty, Good muscle tone. Prominent
veins and tendons. Broad chest common. An active, sportive type. Weight gain usually in
chest, arms, belly, upper body.
Hair: Often thick and curly texture, black in color and rapid profuse growth.Balding common in
men.
Complexion: yellowish, reddish color if heat predominates; sallow or bright yellow if bile
predominates. Rough and dry, quite warm.
Appetite: Sharp and quick. Soon overcome by ravenous hunger. Fond of meat, fried foods,
salty or spicy foods, alcohol, intense or stimulating taste sensations.
Digestion: Sharp and quick. Tendency towards gastritis, hyperacidity, acid reflux. When
balanced and healthy, can have a "cast iron stomach", able to digest anything.
Metabolism: Strong, fast, active; catabolic dominant. Strong Innate Heat of metabolism. Liver
and bile metabolism can be problematic. Digestive secretions strong, bowel transit time short.
Adrenals, sympathetic nervous system dominant. Strong inflammatory reactions.
Predispositions: Fevers, infections, inflammation. Hives, rashes. Fatty liver, bilious
conditions. Hyperacidity, acid reflux, inflammatory and ulcerative conditions of middle GI
tract. Headaches, migraines, irritability. Eyestrain, red sore eyes. High cholesterol,
cardiovascular disorders. Bleeding disorders from excess heat. Hypertension, stress disorders.
Urine: Tends to be scanty, yellowish dark, and thin. Can be hot or burning.
Stool: Tends towards diarrhea, loose stools. Can have a yellowish color, foul odor.
Sweat: Profuse, especially in summer, or with vigorous physical activity. Strong body odor.
Sensitive to hot weather, suffers greatly in summer.
Sleep: Often fitful, restless, disturbed, especially with stress, indigestion. Often tends to wake
up early, or in the middle of the night.
Dreams: Often of a military or violent nature. Dreams of fire, yellow things common. Fight or
flight, confrontation.
Mind: Bold, daring, original, imaginative, visionary. Ideation faculty well-developed. Brilliant
intellect, sharp penetrating insight. The idea man who prefers to leave the details to others.
Personality: Prone to anger, impatience, irritability; short temper. Bold, courageous,
audacious; confrontive, contentious. Dramatic, bombastic manner; high powered personality.
The rugged individualist and pioneer; thrives on challenge. The fearless leader. Seeks
exhilaration, intense experiences.
Characteristic of Balghami Mizaj (Phlegmatic Temperament) Individuals:
Humor: Phlegm (Balgham) Basic Qualities: Cold and Wet (Barid&Ratab)
The Phlegmatic temperament is the coldest, most passive, energy conserving and anabolic.
Physique: Heavy frame, fatty and flabby, with flesh ample and well-developed. Often pudgy,
plump or overweight; obesity common. Joints dimpled, not prominent. Veins not prominent,
but can be bluish and visible. Lax muscle tone common. Feet and ankles often puffy,
swollen. Weight gain especially in lower body. Flat chest.
Hair: Thin and straight, slow scanty growth and distribution, brownish,light colored, blondish
hair common.
Complexion: Pale, pallid complexion; very fair. Soft, delicate, cool moist skin. Cool, clammy
perspiration common, especially in hands and feet.
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Appetite: Slow but steady. Craves sweets, dairy products, starchy glutinous foods.
Digestion: Slow but steady to sluggish. Gastric or digestive atony common. Sleepiness,
drowsiness after meals common.
Metabolism: Cold, wet and slow. Conserves energy, favors anabolic metabolism. Congestion,
poor circulation, especially in veins and lymphatics. Adrenals and thyroid tend towards
hypofunction; basal metabolic rate low. Metabolic Water drowning out metabolic Fire.
Predispositions: Phlegm congestion. Water retention, edema. Lymphatic congestion,
obstruction. Poor venous circulation. Gastric atony, slow digestion. Hypothyroid, myxedema.
Adrenal hypofunction. Weight gain, obesity. Frequent colds and flu. Chronic respiratory
conditions, congestion. Swollen legs, ankles, feet. Poor tone of skin, muscles.
Urine: Tends to be clear/pale and thick. Tends to be scanty in volume, with excess fluid
accumulation in the body.
Stool: Well-formed, but tends to be slightly loose, soft. Bowels tend to be sluggish.
Sweat: Cool, clammy sweat common, especially on hands and feet. Sweating can be easy and
profuse. Sensitive to cold weather; suffers greatly in winter.
Sleep: Very deep and sound. Tends towards excessive sleep, somnolence. Snoring common;
can be loud or excessive.
Dreams: Generally very languid, placid. Water and aquatic themes common.
Mind: Tends to be dull, foggy, slow. Slow to learn, but once learned, excellent and long
retention. Patient, devoted, faithful. Faculty of Empathy well-developed. Sentimental,
subjective thinking. A calm, good-natured, benevolent mental outlook.
Personality: Good natured, benevolent, kind. Nurturing, compassionate, sympathetic. Great
faith, patience, devotion; tends to be religious, spiritual. Sensitive, sentimental, emotional,
empathetic. Passive, slow, sluggish; averse to exertion or exercise. Calm, relaxed, takes life
easy. Excessive sluggishness, torpor can lead to depression.[11, 12, 13]
PURPOSE OF THE STUDY:
The present study has been attempted with two different categories Bilious and
Phlegmatic temperament individuals because these two categories show different types of signs
and symptoms as their physiological, physical, and psychological features and are opposite to
each other on the basis of their Hot and Cold temperament respectively. Basically these two
main states of hotness and coldness play vital role in determination of temperament.
The study was started with following aims and objectives.
1)
To assess the value of total serum protein in Bilious and phlegmatic temperament through
Biuret Method.
2) To analyze whether total serum protein levels in Bilious and Phlegmatic temperament are
similar or dissimilar within normal healthy state.
HYPOTHESES OF THE STUDY: Total serum protein would be relatively higher in Bilious
than Phlegmatic temperaments because as per Unani literature, The Bilious individuals are
energetic and hyperactive than phlegmatic individuals, they have excellent metabolism activities,
hence they have physiologically higher level of total protein because liver plays an important
role in metabolism and the metabolic rate of Safravi individuals are rapid than Balghami,
therefore the level of total protein should be higher in Safravi than Balghami individuals. In
comparison to Safravi, Balghami individuals are flaccid and obese body built with soft and
flabby muscles and they possess more fat in their body. Fat always denotes coldness. But
Safravi individuals are lean and thin they possess very little fat shows that the temperament is
hot and dry.[8]
METHOD OF THE STUDY:Selection of volunteers, Sample Size, and Inclusion criteria:
This study was carried out in PG Lab, Department of Kulliyat, Ajmal Khan Tibbiya
College, Aligarh Muslim University, Aligarh, during the period from 2010 -2013.
Volunteers were selected randomly from students of the Ajmal Khan Tibbiya College and
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some other faculties of Aligarh Muslim University and Aligarh city, preference was
given to students residing in hostels because their environment and nutritional status
remain almost static and constant. One hundred ten (110) male volunteers in the age
group of 20–35 years, having Bilious (Safravi) and Phlegmatic (Balghami) temperament
were selected for the study. The reason for selecting this age group is that the
volunteers in this age group are mostly healthy and disease free.
Exclusion Criteria: Female volunteers and volunteers below 18years and above 35
years were excluded,the persons of Melancholic and Sanguine temperament were
excluded from the study,the persons of the athletic activities, heavy exercise and
gymnastics were excluded from the study, thevolunteers having a history of
Radiotherapy, Chemotherapy, major accident and surgery or taking steroids for long
periods were excluded from this study andthe volunteers, suffering from Malnutrition,
Diabetes Mellitus, Poliomyelitis, Tuberculosis, Hyperthyroidism , Hypothyroidism or
any other chronic diseases, were excluded from the study.
For the selection of healthy volunteers, detailed clinical history, physical, general and
local examinations were done.
Determination of Temperament: The assessment of temperament (Mizaj) of the
volunteers was made on the basis of Ajnas-e-Ashra (ten determinants), mentioned in
classical Unani literature.
Categorization of individual: After determination of the temperament, selected
volunteers were divided into two groups according to their temperament.
Group A: Bilious temperament (Safravi Mizaj)
Group B: Phlegmatic temperament (BalghamiMizaj)
After determination of Bilious and Phlegmatic temperament, the volunteers were
randomly called for venipuncture. All volunteers were advised to remain at fast
overnight before their blood samples were drawn. The blood specimens were allowed to
clot at room temperature and then centrifuged within an hour of venipuncture. The sera
obtained were assayed within two hours of venipuncture.
ESTIMATION OF TOTAL PROTEIN: BIURETE METHOD was adopted in order to
estimate the total serum protein. The most widely used method of measuring serum protein is
the Biuret Reaction. ThePrinciple of this reaction is that Peptide bonds of protein form a
blue-violate colored complex with cupric ions an alkaline medium. The intensity of the color
is proportional to the number of peptide bonds and the color is read at 546 nm (530-570 nm).
Total protein
+
Biuret reagent
----->purple colored Cu-peptide bond complex
(alkalineCu++)
Reagents: (They were supplied by Siemens ltd)
Normal Values: Total Serum Protein: 6.6-8.3 g/dl.[14]
OBSERVATIONS & RESULTS:
Table–01: Showing Distribution of Volunteers according to the Temperament
Temperament
Frequency
Percentage
Bilious
65
59.10
Phlegmatic
45
40.90
Total
110
100
Graph-01: Showing Distribution of Volunteers according to the Temperament
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Frequency
Choleric, 65
Choleric
Phlegmatic
Total
Total, 110
Phlegmatic, 45
Out of 110 volunteers, 65 were Bilious temperament and 45 Phlegmatic temperaments
and their percentage of 59.10% & 40.90% respectively as displayed in Table -01and
Graph. 01.
Table-02:Indicating Total Serum Protein of volunteers expressed as Mean ± SD*
Temperament
Number of Volunteers
Mean ± SD*
Bilious
65
7.51 ± 0.83
Phlegmatic
45
7.05 ± 0.63
Total
110
Graph-02: Indicating Mean Difference of Total Serum Protein in different
Temperaments
Choleric
80
Phlegmatic
65
60
45
40
20
7.51
7.05
0
Number of Volunteers
Mean
The mean and standard deviation of total serum protein levels of bilious (Safravi)
and phlegmatic (Balghami) subjects were calculated. It was revealed after this study that
the mean total serum protein of bilious (Safravi) subjects was 7.51 (g/dl) with standard
deviation of 0.83 while the mean total serum protein of phlegmatic (Balghami) subjects
was 7.05 (g/dl) with standard deviation of 0.63. In this way it can be said that the average
total serum protein of the healthy bilious subject is higher than that of a phlegmatic
subject. (Table-02, Graph-02)
Table-03:Showing Characteristics of Volunteers Expressed As Mean ± SD
Temperaments
Safravi
Balghami
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No. of Subjects
65
45
Age (Mean ± SD)
Heigh(Mean ± SD)
Weight (Mean ± SD)
25.2±2.5
169.69±4.4
60.12±5.8
27.8±3.0
168.68±3.08
67.75±5.6
TP (Mean ± SD)
7.51±0.83
7.05±0.63
ISSN 2322-0147
SD: Standard deviation.
The mean and standard deviation of age, height, weight, and total protein of bilious and
phlegmatic temperaments subjects were calculated. It was revealed after this study that the
mean of age, height, weight and total protein of bilious subjects was 25.2 (years), 169.69 (cm),
60.12 (kg), and 7.51 (g/dl) with standard deviation of 2.5, 4.4, 5.8, and 0.83 respectively, while
the mean of age, height, weight and total protein of phlegmatic subjects was 27.8 (years), 168.68
(cm), 67.75 (kg) and 7.05 (g/dl) with standard deviation 3.0, 3.08, 5.6 and 0.63 respectively, as
shown in table 03.
Graph-03: Showing Total Serum Protein of Volunteer
Safravi
10
Balghami
TOTAL PROTEIN
8
6
4
2
0
1 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46 49 52 55 58 61 64
NO OF VOLUNTEERS
Total serum protein was higher in bilious volunteers as compared to phlegmatic volunteers and
the difference was statistically significant as shown in the graph-03.
Table-03: Showing the Results in Volunteers according to their Temperament
Parameters
Mizaj
Number of
Volunteers
Mean ± S.D
Bilious
65
7.51±0.83
Significance
P<0.001
Total Serum
Protein
Phlegmatic
45
7.05±0.63
Significant at the level of p<. 05
Present study after statistical analysis showed that the difference of total serum protein
level of bilious and phlegmatic temperament subjects was highly significant as shown in Table
03.
In this study of 110 volunteers, we have observed that a relation can be established in
between Total Serum Protein and temperament. The level of total serum protein was higher in
the bilious temperament volunteers as compared to volunteers of phlegmatic temperament.
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Result: The mean and standard deviation of total serum protein levels of bilious and
phlegmatic subjects were calculated. It was revealed after this study that the mean total
serum protein of bilious subjects was 7.51 (g/dl) with standard deviation of 0.83 while the
mean total serum protein of phlegmatic subjects was 7.05 (g/dl) with standard deviation of
0.63. In this way it can be said that the average total serum protein of the healthy bilious
subject is higher than that of a phlegmatic subject.
The T-test was applied to test the significance of the results and it was found that the difference
of total serum protein level of bilious (Safravi) and Phlegmatic (Balghami) subjects was highly
significant (p<0.001).
CONCLUSION: From the findings of this study following conclusion can be drawn.
The individuals having bilious (Safravi) temperament have higher levels of total protein
than the phlegmatic (Balghami) individuals. Thus a relationship can be established between the
temperament and blood protein which validates the Unani claims that The bilious (Safravi)
individuals are energetic and hyperactive than phlegmatic individuals, they have excellent
metabolism activities, hence they have physiologically higher level of total protein because liver
plays an important role in metabolism and the metabolic rate of Safravi individuals are rapid than
Balghami, therefore the level of total protein should be higher in Safravi than Balghami
individuals. In comparison to Safravi, Balghami individuals are flaccid and obese, they possess
more fat in their body as compared to protein. Fat always denotes cold temperament. But Safravi
individuals are lean and thin they possess very little fat shows that the temperament is hot and
dry.
In our study, it is clearly evident that temperament of an individual is influenced by his
blood composition and its constituents. Therefore, the blood compositions play an important role
in creation of temperament of an individual. The results show that there does exist a relationship
between blood protein and the temperament of an individual.
The need of the hour is to understand the concept & importance of temperament (Mizaj) and its
application in Unani as well as other health care.
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