The document discusses acid-base disorders and summarizes: 1) Acid-base disorders occur when the body's normal balance of acids and bases deviates from the normal pH range, causing acidosis or alkalosis. Disorders can be classified as respiratory or metabolic based on their etiology. 2) Biochemical findings include concepts of acids and bases donating and accepting protons. The strongest acid and base present in water are H3O+ and OH-. 3) Regulation of acid-base balance involves chemical buffer systems, respiratory mechanisms, and renal mechanisms working together to precisely control pH levels.

1. Sikkim Manipal University
Assignment BLOO43
Roll No-621132459(Vipin Pant)4rh semester
1) What are acid-base disorders? Explain its biochemical findings. Add a
note on regulation of acid-base balance
Ans- Acid–base imbalance is an abnormality of the human body's normal balance of
acids and bases that causes the plasma pH to deviate out of the normal range (7.35 to
7.45). In the fetus, the normal range differs based on which umbilical vessel is
sampled (umbilical vein pH is normally 7.25 to 7.45; umbilical artery pH is normally
7.18 to 7.38).[1] It can exist in varying levels of severity, some life-threatening.
Classification
A Davenport diagram illustrates acid–base imbalance graphically.
An excess of acid is called acidosis and an excess in bases is called alkalosis. The
process that causes the imbalance is classified based on theetiology of the disturbance
(respiratory or metabolic) and the direction of change in pH (acidosis or alkalosis).
This yields the following four basic processes:
process pH carbon dioxide compensation
metabolic acidosis down down respiratory
respiratory acidosis down up renal
metabolic alkalosis up up respiratory
respiratory alkalosis up down renal
]Mixed disorders
The presence of only one of the above derangements is called a simple acid–base
disorder. In a mixed disorder more than one is occurring at the same time.[2] Mixed
disorders may feature an acidosis and alkosis at the same time that partially counteract
each other, or there can be two different conditions affecting the pH in the same
direction. The phrase "mixed acidosis", for example, refers to metabolic acidosis in
conjunction withrespiratory acidosis. Any combination is possible, except concurrent
respiratory acidosis and respiratory alkalosis, since a person cannot breathe too fast
and too slow at the same time.

2. Explain its biochemical findings-:
Biochemists usually discuss acids and bases in terms of their ability to donate and
accept protons; that is, they use the Brønsted definition of acids and bases. A few
concepts from general chemistry are important to help organize your thoughts about
biochemical acids and bases:
1. A compound has two components — a conjugate acid and a conjugate base.
Thus, you can think of HCl as being composed of the proton-donating acidic
part (H+) and the proton-accepting basic part (Cl−). Likewise, acetic acid is
composed of H+ and the conjugate base (H3CCOO−).
2. The stronger the acid, the weaker its conjugate base. Thus, HCl is a stronger acid
than acetic acid, and acetate ion is a stronger base than chloride ion. That is,
acetate is a better proton acceptor than is chloride ion.
3. The strongest acid that can exist in appreciable concentration in a solution is the
conjugate acid of the solvent. The strongest base that can exist in a solution is
the conjugate base of the solvent. In water, the strongest base that exists is OH−.
If a stronger base, such as NaOCH3, is added to water, the methoxide ion rapidly
removes protons from the solvent:
4.
leaving the base OH- as the strongest base in solution. (Don't try these reactions
at home; they are highly exergonic!) The strongest acid that can exist in water in
appreciable amounts is H3O+, the conjugate acid of H2O:
5. Weak acids and bases — those less strong than H+ or OH− — exist in equilibrium
with water:
Regulation of acid-base balance. –
1. Chemical Buffer system:
– Responds within seconds
– Does not eliminate or add H+ from body
– Operates by binding or to tied up H+ till balance is reestablished.
a. In ECF:
– Mainly HCO-3/CO2 Buffer system
– Plasma Proteins
– HPO–4/H2PO-4 Buffer system
b. In ICF:
– Proteins Mainly e.g.: Hb in RBCs
– HPO–4/H2PO-4 Buffer system
Routes of excretion of acids; lungs & kidneys
2. Respiratory Mechanisms:
– Responds within minutes
– Takes 6-12 hours to be fully effective
– Operates by excreting CO2 or (adding H2CO3/HCO-3)

3. 3. Renal Mechanisms:
• Responds slowly (effectively in 3-5 days)
• Eliminates excess Acids or Base from body
• The most powerful mechanism
e.g. i. HCO-3/CO2 Buffer system
ii. NH3/NH+4 Buffer system
iii. HPO–4/H2PO-4 Buffer system
Chemical Buffer System
• Consists of a ‗pair of substances‘ present in a mixture of a solution that ‗minimizes
pH changes‘ when an ‗acid or base‘ is ‗added or removed‘ from the solution.
• Consists of;
1. Carbonic Acid – Bicarbonate Buffer System
2. Phosphate Buffer system
3. Protein Buffer system
Chemical Buffer System of ECF
1. Bicarbonate Buffer System: H2CO3/NaHCO3
consists of H2CO3 (weak Acid) + NaHCO3 (Bicarbonate salt)
– CO2 + H2O ↔H2CO3 ↔ H+ + HCO-3
– NaHCO3 ↔ Na+ + HCO-3 → H2CO3 → CO2 + H2O
Bicarbonate buffer system is quantitatively the most powerful ECF buffer system
Its two components HCO-3 & CO2 are precisely regulated by kidneys & lungs.
2. Phosphate Buffer System:
– Not of major importance in ECF
– Only 8% of the conc. of HCO-3 Buffer system
– Comprised of HPO–4/H2PO-4
– Plays major role in ICF & in Renal tubules
3. Proteins: (ICF proteins, Hb, Plasma proteins)
– Excellent buffers as proteins contain both Acidic & Basic groups.
– More important in ICF H2CO3 ← H2O + CO2
HCO-3 + H+ + HbO2 ↔ H.Hb + O2
– In RBCs, Hb is important
– 60-70% of total chemical buffering of body fluids inside the
cells & in ICF is by proteins.
– Hb buffers H+ ions generated by H2CO3
– Proteins are the most abundant buffers in cells & in blood
– Histidine and Cysteine are the two A. Acids that contribute
most of the buffering capacity of proteins
Respiratory Mechanisms in Regulation of Acid-Base
• Second line of defense against acid base disturbances
• Operates through regulation of ECF CO2 concentration by lungs
• Effectiveness between 50-75% [feedback gain is 1-3 i.e. fall in pH
from 7.4 to 7.0 is returned by Resp System to 7.2 to 7.3 within 3-12
minutes]
2) Discuss the advantages of automation in clinical biochemistry
laboratory. Make a list of few (at least five) automated instruments
available for biochemical analysis. Discuss the principle of each .

4. Ans- Automation in clinical biochemistry laboratory- Clinical pathology or
laboratory medicine has a great influence on clinical decisions and 60–70% of the
most important decisions on admission, discharge, and medication are based on
laboratory results.1 As we learn more about clinical laboratory results and incorporate
them in outcome optimization schemes, the laboratory will play a more pivotal role in
management of patients and the eventual outcomes.2 It has been stated that the
development of information technology and automation in laboratory medicine has
allowed laboratory professionals to keep in pace with the growth in workload.3 In a
paper on ―robotics into the millennium,‖ the various types of automation have been
outlined4 while other authors have classified laboratory automation into total
laboratory automation, modular laboratory automation, and workcell/workstation
automation.5,6
This article evaluates the relationship of scientific staff, automation, and expert
systems in clinical chemistry with particular reference to the core laboratory and
ascertains staff requirements. The changes in work practices due to the introduction of
automation and computers in other industries are discussed and similarities with
clinical chemistry elucidated as it has been noted that the original total laboratory
automation was based on the manufacturing/factory model of production.7 Others
have also written on automation in various industries over the last century and how
the patterns of its implementation and effects can be applied to pathology.8 The goal
of a successful automation must be to change the way in which work is done in the
laboratory and this involves changing not only the tools and processes, but also the
job structure and ultimately the way people think about their work.7 The progress in
automation and convergence of technologies are two key factors, which particularly
affect how we think about the future of clinical chemistry.9 The role of the scientific
staff, use of automation and expert systems shall be discussed for a core laboratory
focusing on the Monash Medical Centre, Melbourne, Victoria, Australia where one of
the authors is based. It is our belief that to consolidate changes that are advocated,7 it
is important to look at skill requirements and training of the operatives in clinical
chemistry.
Four, flow type automatic biochemical analyzer
Flow type automatic biochemical analyzer can be divided into air staging system and
non segmented system. The former is a most typical flow analyzer.
(a) air staging system
The analyzer is through proportional pump extrusion elastic sample tube, air pipe and
the reagent tube (commonly known as the "tube"), the sample sequentially inhalation
and transported along the sample tube, on the other hand, by the air pipe into the
bubble will be determined by the same principle of inhalation and reagent
continuously flowing in pipeline is divided into segments of the reagent uniform,
sample and reagent flow in the process of continuous flow forward encounter, mixing,
absorption through (when necessary), thermal insulation, reaction and measured. The
analysis process is the flow process of continuous flow in the pipeline.
(two) non segmented system

5. Non segmented system is the reaction liquid, depending on the reagent blank or buffer
to interval of each sample, the continuous flow in the pipeline liquid can not be
segmented. Non segmented system can be divided into system and space system for
flow.
1, flow injection system
The system composition and air segmentation system is similar, but some structure
and working principle of different, air staging system is the use of the bubble to
prevent cross contamination of the reaction liquid in the pipeline in the flow process
of segmentation, and flow injection system is through the sample are injected to
prevent cross contamination to continuous flow reagent flow in a pipe.
2, clearance system
The system structure, composition and working principle and flow injection system is
similar, but its characteristic is each sample must be in the analysis process after the
end of the previous samples (including pipeline cleaning) to start, but not
continuously in turn into the sample, a time gap between each sample, it is person is
not continuous flow analyzer.
Principle-:
air staging system
With increasing use of biomass in combustion processes, the reduction of the
related NOx
emissions which originate mainly from the fuel nitrogen becomes more and
more important.
Efficient primary measures for NOx reduction are staged combustion
techniques. Air staging
has been investigated earlier and has found its way into practice. Since fuel
staging has not
been applied with nonpulverized biomass yet, the aim of the present work was
to investigate the
potential of fuel staging for NOx reduction in fixed bed systems. For this
purpose, a prototype
understoker furnace of 75 kW thermal input with two fuel beds in series was
developed.
Experiments were performed with wood chips (low nitrogen content) and UF-
chipboards (high
nitrogen content) to investigate the influences of the main process parameters,
i.e., stoichiometric
ratio, temperatures, residence time, and fuel properties on the conversion of
fuel nitrogen to
N-species. The most important parameters were found to be the temperature
and the stoichiometric ratio in the reburn zone. The potential of fuel staging was
measured and compared with
air staging and unstaged combustion. The experiments show that low NOx
emissions are already

6. achievable with fuel staging at lower temperatures than with air staging, i.e.,
900-1000 °C, and
at a stoichiometric ratio of 0.85 in the reduction zone. The NOx reduction
achieved under optimum
conditions for UF-chipboard as main fuel was 78% which is higher than with
air staging, where
72% NOx reduction was measured. For wood chips both measures attained
about 66%. The
nitrogen conversion during air and fuel staging has also been simulated using
a furnace model
based on ideal flow patterns as perfectly stirred reactors and plug flow
reactors. A detailed reaction
mechanism including the nitrogen chemistry (GRI-Mech 2.11) was
implemented. The trends found
with this model are in good agreement with the experiments and they indicate
that even higher
NOx reduction may be reached with improved process design. The
investigations show that fuel
staging is a promising technology for NOx reduction also for fixed bed
biomass furnac
3) What is quality control? What are its components? Add a note on
implementation of external quality control in various biochemical tests
Ans- quality control-:
The automated analyzers in clinical laboratories Nowadays, the overwhelming
majority of laboratory results in clinical laboratories is being generated by automated
analyzers. Modern automated analyzers are highly sophisticated instruments which
can produce a tremendous number of laboratory results in a very short time. This is
achieved thanks to the integration of technologies from three different scientific
fields: analytical chemistry, computer science and robotics. The combination of these
technologies substitutes a huge number of glassware equipment and tedious, repetitive
laboratory work. As a matter of fact, the laboratory routine work has diminished
significantly. Today laboratory personnel‘s duties have been shifted from manual
work to the maintenance of the equipment, internal and external quality control,
instrument calibration and data management of the generated results.
Components-:
The purpose of a control is to aid the operator in deciding whether an analytical
system is producing reliable results for a given assay, and ultimately whether to
release the results. This unit presents information on the techniques for determining
when results are in control or out of control.
External quality assessment

7. This is the evaluation by an outside agency of the performance by a number of
laboratories on specially supplied samples. Analysis of performance is retrospective.
The objective is to achieve between lab and between method compatibility, but this
doesn‘t guarantee accuracy unless the specimens have been assayed by a reference lab
alongside a reference preparation of known value. Schemes are usually organized on a
national or regional basis. Hence, EQA is mainly concerned with analytical part of the
test.
4) List various methods used for estimation of serum calcium. Explain
the principle and procedure of any one method
Ans- estimation of serum calcium-:
Calcium is the most abundant mineral1
and fifth
most common element in the body.2 Almost all blood calcium is present in
plasma and reference range
is 2.10 to 2.65 mmol/L. It is present as free or ionized (50%), protein bound
usually with albumin
(40%) and complexes with small anions (10%). Calcium is needed for bone
mineralisation, blood coagulation and influences the permeability and excitation
of plasma membranes. It is usually monitored
for hypoparathyroidism, hyperparathyroidism, vitamin D deficiency,
malnutrition, cancers, enhanced
renal retention, osteoporosis, etc.
There are many different methods for estimation of serum calcium like
spectrophotometeric, ion
selective electrode (ISE) and atomic absorption methods. The
spectrophotometric techniques use metallochromic indicators which change
color when
they bind to calcium. Arsenazo III and o-Cresolphthalein Complexone (CPC)
methods are the two spectrophotometric techniques frequently used.
Aim of our study was to compare serum calcium
estimation by CPC method using direct colorimetric
and volume / volume colorimeteric (v/v) methods.
The principle of CPC method is that calcium reacts
with CPC in an alkaline medium to form a red coloured complex. This complex
is measured at a wavelength at 570 nm. Sample is diluted with aci to release
protein bound and complexed calcium. Diethylamine, 2-amino-2-methyl-1-
propranolol or 2-
ethylaminoethanol is added to buffer the solution
and provide an alkaline medium.3 Effect of magnesium can be minimised either
by adding 8-hydroxyquinolone, buffering the solution to pH of around
12 or by measuring absorbance at 580 nm.
MATERIALS AND METHODS
The study was performed in a tertiary care laboratory in Rawalpindi from
March to June ‘2011. It was

8. a prospective comparative study. Seventy quality
control samples of Randox laboratories were used of
these thirty five were normal controls while thirty
five were abnormal controls. Controls were tested
simultaneously on both the kits provided by SS diagnostics using a fully
automated chemistry analyser
(Selectra E). Data was recorded using specially designed proformas and results
were analysed using
SPSS version 17.
RESULTS
Our results showed that in normal control samples
with a target of 2.33 mmol/L where the range of
calcium was 2.10 to 2.56 mmol/L. The v/v method
kit gave the mean result of 2.34 mmol/L ± 0.04
with a CV of 1.70%. With the direct calorimetric
Principle of GOD-POD-
Intended Use
The reagents are used for the quantitative determination of
Glucose in serum or plasma. For in-vitro diagnostic use only.
Introduction
Glucose is the reducing monosaccharide that serves as the
principal source of cellular energy in the body. It enters into the
cell under the influence of insulin and undergoes a series of
chemical reactions to produce energy. Lack of insulin or
resistance to its action at the cellular level causes diabetes.
Therefore, in diabetes mellitus the blood glucose level are
very high. However, high blood glucose level is also observed
in the pancreatitis, pituitary or thyroid dysfunction, renal
failure and liver disease whereas low glucose level is
associated with starvation, hyperinsulinaemia, neopalasms
or insulin induced hypoglycemia.
Method
GOD-POD method, End Point.
Principle
Glucose is oxidized by glucose oxidase(GOD) to produce
gluconate and hydrogen peroxide. The hydrogen peroxide is
then oxidatively coupled with 4 amino- antipyrene(4-AAP)
and phenol in the presence of peroxidase(POD) to yield a red
quinoeimine dye that is measured at 505nm. The absorbance
at 505 nm is proportional to concentration of glucose in the
sample.
Glucose +2H2O + O2
Gluconate + H2O2
2H O + 4-AAP + Phenol Quinoeimine Dye 2 2
Absorbance of the colored solution is directly proportional to
the glucose concentration, when measured at 505nm