Chapter 1

The Clinical Biochemistry Laboratory the use and the

requirements of laboratory
Objective of the session
1.

Introduction :

The use of Biochemical tests :

•
!
 •

,
+ - . /
0 - 1" /
2 /

+ '

# $

The use of the laboratory :

% &

'
(! &

'
)

"
 !

" (

4 '
!

! )

Requirement of Blood Collection

" + 5
3 + 6.7
4

Blood collection :
• &
• + 8
• ( &
•

3
 • )
• '
+ 9
(
" :
3
;

Sampling errors

<
& +

• Blood sampling technique. *

&

• Prolonged stasis during venepuncture. '

'

• Insufficient specimen. % &

&

4
• Errors in timing.
"4

• Incorrect specimen container. #

#

>

%* (
!
5
• Inappropriate sampling site.
< < )
&
)
&
=7 0
?
&
• Incorrect specimen storage. (

=
 A (
(

, &
,

)
'

A
,
)*

Fig. 2 Circuit diagram of clinical biochemistry process.

Analysing the specimen
@
&
&
+
" (
&
@

1
B 9 *
% 9 ( &

6
 Chapter 2
The Interpretation Of Results
Objective of the session :
1. To teach the interpretation of results.

The laboratory report

)
?

CO'CE'TRATIO'
0 &
&
&

.. !
# ..
7
7 ..
7!
.. C ..
! 7 / ..
!

2
+ &
..

E+ ) E 0F) +
#

+) + ) !
G
& H
B
" 0

4. / +) 4. /
2. / .. +) . /
3 G

6 . / .. 6. /

D
Units of activity
0
5 5

( % 5

(
(
)
5
5

Reporting results
0 D= 7

.. / ..
.. .. I " / .. #

.
Significant figures
#

G
(
.. " / .. ( ."

0 5

6. 1 D= 6 .=
6 1= 6=

B 9 *
% 9 ( &
 Chapter 3
Quality Control
Objective of the session 9 &

Introduction :
Variation in results

Laboratory analytical performance

( 9
•
• +

/ (a) "
(b) (c).
• &
•

Precision and Accuracy

' (
(
3

"
 %
(
< < ) ?

Imprecise Precise but Precise and

inaccurate accurate

Fig. 3 Precision and accuracy

Sensitivity and specificity

+
(

&

Quality assurance
%

&

3
 <
&

) &
>

<

Reference ranges
(

3! B
D=7
=7
)
>

) < <

< < 4! (
false positive. (
< < false negative.

4
Biological factors affecting the interpretation of results

9
• Sex of the patient. ,

• Age of the patient.

• Effect of diet.

• Time when sample was taken.

• Stress and anxiety.

• Posture of the patient. ,
• Effects of exercise. + 5

• Medical history. ) / ?

• Pregnancy.
• Menstrual cycle. $

• Drug history. *

=
Others factors
G
(
@

&

Technical contents 9 &

Methodology of its teaching 9 *

Evaluation of the session 9 ( &

1
 Chapter 'o. 4
Photometery
Objective9

Introduction of the topic:

' &

'

B 9

" B # 9
#

Colorimetry:
B &

: ( : 9

6
" * @ *! ( (! % %!

@' ) (: *%F+) J :@K 7L: ../

Lambert Beer's Law:
:

)%
M
)
G
)% L )
) L )
M L (
L
L
L " 6 2!

)%/)@

/@ /% N%OL: )/ ! L :
..7 %L" : 7
: <

2
 ! @ :
! 5

G ( ! !

Parts of photo colorimeter

(! Light Sources : 4". 61. ! 08
$
! Monochromator/Filters :

Table :
# ' !
K , =2. 12.
J =". =2.
' K 4D. =".
, 46. 4D.
J 43. 46.
J 8 4.. 43.
)
K

! Cuvettes :

D
 K
& 5/
08
Glvanometer :
)

Prepration of solutions for measurement:

Blank : 5 (!
..7 7 !
" Standard :
!
3 (

Determination of absorpition maxima :

# /@ *

' @* J
P ; K

Verification of Beer Lambert's Law :

: <
@*!
@*
! J
P ) !

".
Reagents: - = 7
Procedure :
6 + +" +3 +4 +=
F 8 *
! !
. =
" + 4
3 +" " 3
4 +3 3 "
= +4 4
1 += = .
6 = ! .

F ; @*
' @*
#
; 7
Calculations :
)

%
L
%

%
L P

9
,
L P
,

"
+@0, % @# :)K$ #): %, 08% % '$@ @ %:: K(:8(F@B% %,
Fig.4 (diagrammatic) photoelectric colorimeter

9 '
B 9 *
% 9 ( &

""
 Chapter 5
Automation
Objective : (

&

Introduction
*
G

B
# G <

&
5
automation. )

"3
 & ?
& #

Continuous flow analysers

)
) )
&

B ?

discrete analysers

(! Batch analysers ! <Stat'

(A) Batch analysers

! "!

"4
 (1) Semi automated (batch) discrete analysers
)

,
! 5

Advantage of semiautoanalysers
!

"! + .=
.
3! % 5
3
4! 5

Fully automated batch analysers

!
"! & &

"=
3!

4!
=! ( &
!

1!

(B) Stat Analysers , !

) 2 ". !

$ ! #
F & >
!0 "! + 3! 4! ( =! +K'
1! +K@
• 5

•
+K' +K@

• & &

• 5
•

"1
•

< < ! 5

!
"! ) < <

3! ) 5

4! )
=! ) %
!

1!

Technical contents 9
Methodology of its teaching 9 * :

Evaluation of the session 9 (

"6
 Chapter 'o. 6
Estimation of Blood Glucose
Diabetes Mellitus: )

!;
! + +
;
1. Primary or Idiopathic or Essential Diabetes
!C * )* )
* B )**B!
: # &
@ =
*
' !
!B )) F
* F)**B!
B &
@
M

" +
)
!' *
'
$
B '

"2
 !)
$
$ -
$

!'
"! : ; & ' !
3! ' &
4! )
;
=! ) (
;
1! ) ;

6! ) &
; ;

Objective :

Introduction :
%

) &
G

"D
 . =7 (

= ".7!

a) Fasting blood Sugar (FBS) 9

"
b) Post Prandial Blood Sugar (P P B S) : ( "
..
! "
c) Random Sample :

Collection of Blood Sample :

F #!
930 4 &
F # , <
5 < <
Methods of Estimation :
% 5 9B
! Glucose Oxidase Method :
K
$" @"

3.
 K $"@"
Reaction 9 K K K ( I$"@"
@ @"

'
$"@" I I $"@
@"
B !

$
!$ B 9) I( ' 1 'I(*'

K-1-'
K - 1 - ' I F(* 1 I F(*$ I $ I

F(*$
34.

II. Reduction Methods

! Alkaline Copper Reduction Methods.
• ( ;M <
• # ;G
"! Ortho toludine method 9 @ &

Estimation of Blood Glucose by method of asatoor and King (modified)

Principle : ,
II I

@$ @$ "@ "@

' 9
I II

3
B
+@4 F "+@
,

+ ( M D=4
9
'
, + 4
%
1 .
Reagents :
! ) 9
F "+@4 )@$"@ - 3. +@4 =$" . 1

! + .7
! (
* "= F $ @3 ". F 3 @3 2 '
@ =.. ( "
:
! ' 9 3= =
@ < ".. .7 F @$ 3.
4. 3=.
"= 2=7! B =..

! + + 9 7 5

3"
 ! G 9* ..
) . 7 + 4

Procedure
' . ( 32
(
"... .< 0

+ + +" +3

+ .
+ K ." .4 .1
) +@4 .2 .1 .4 .
( . . . . .

B '
. 3 2
B @* 1 .
' @*

Calculation :
@* ( +
/ !L P ..
@* + 8
Interpretation : 1. ..
/ .. 4. /

33
Hyperglycagmia : 9
* -# 8 4. /
".. /
) .. /
8
K
" $
3 +

4 )

= * 9 5 ( $
*

Hypoglycaemia : G 1. /
B

" $ -
3 ) -
4 $ ( < !
= $
1 +
6 +
2
D -

34
( 9
, + -
" ( -*
3 -*
, &

Technical contents 9
Methodology of its teaching 9 *
Evaluation of the session 9 (

3=
 Chapter 'o. 7
Glucose Tolerance Test
Object :

Introduction :
K
F
13 .. 7 "
*B K

Oral G T T 9 ' 9( " 1

6= "=. 3..
3. "3
"
G$@
&
M ( (

Precautions :
! 3..
3
2) # . 1
@
3)

31
4) * !

Factors affecting GTT :

! + /)
"! %
3! '
4! )
"

=! '
1! %
6! * @
5

2! :

) 9
9
! F , 9# (
2.7 )
"
! * 9# 62 /: 4. :! "
".. / /:!
K

36
 ! ) K 9 " 4. /
".. / *B
! , K 9 *

! : /( 9# *
3.
2. / 2. !
"
! # 9

I ". 7

Extended GTT :
) 4=
*

Cortisone Stressed GTT :

*
Intrevenous GTT :
) K )
".7 .= /M
0 3.
D.
Methodology of its teaching 9 '
Evaluation of the session 9 ( &

32
 Chapter 'o. 8
Glycated hemoglobin
Object : K

Introduction : $ $
5

QK R $ (
$ ( $ ( $ (
$ (

Technical Contents 9
0 K
K

Principle and procedure:

Using kit of Glycated Hemoglobin an its measurement by semi
autoanalyser (
=

$ (.! (

4 = 4.= 4". !

3D
 F
1 .7 2 37
2. Using Ghb analyser.
)

4 =

% $ (
+ - D. =. = .7 6 .7
+ - =. 2. 6 .7 2 .7
+ - 2. 31. D .7 4 .7

Method of teaching : 0 K

Evaluation of session:

4.
 Chapter 'o. 9
Estimation Of Blood Urea
Kidney function tests : +

( &
(
$
B

(
@

!
"! +

Object :
Introduction :
0 )
0 4= =.7
2. D.7

K#,

4
Estimation : 8 9

! Enzymatic using urease :

a) 'eseler's Method : 0
5 ( F <
' !
4=. 5 ==
$6. 2.
*
F$3

! Berthelot Reaction :
) F$3

c) In the urease/glutamate dehydrogenose method

". .
34. @
F(*$I F(*I %
F(*I
! Kinetic Method: K:*$
0
@"
F(*$ K:*$ F(*I

4"
 F(*$

F L = 4= /
! Colorimetric Method : * B B
Principal : 0
..
=". 5

3.. 7 #

Reagents
! Reagent A : * = ".
..
2=7! B "=.
M 4
"! Reagent B : ( ".. $"+@4 2.. ":

3! Acid Reagent : ( .= ( :
4
4! Reagent C : * ". /: #
4
=! Reagent D : 5 = /:
1! Colour Reagent : B 16 16 *
... * $"@ 4
6! Stock urea standard : .. / ..
2! Working urea standard : * .. *$"@
/ ..

43
Procedure : . / .
9

B T S1 S2 S3 S4 S5
+ ! .
9 ..!
+ ! ." .4 .1 .2 .
* G ! " . 2 1 4 " .
, ! ". ". ". ". ". ". ".
( ! ". ". ". ". ". ". ".

B M 3.
=".

Calculation :
@* ( +
7! L ..
@* + 8

Interpretation :
F = =. 7
@F! 6 "= 7 )

44
 +
= ". / ..
Increase

Pre Renal : G
K#, +
, 9
( -
+
'
0
* M
+
) 9
#

Renal Disease : >

(
,
B

Post renal : *

%
+

4=
 0

Methodology of its teaching 9

Evaluation of the session 9 (

41
 Chapter 'o. 10
Estimation Of Serum Creatinine
Object 9

Introduction:

"7

@
)
"4 )
&

Estimation of serum creatinine by Jaffe's Alkaline Picrate Method

Principle :

=". C < 9
: 9 @ 2.7

+ '$

46
Reagents:
! ' - . .4B D 1 /:!
"! + - . 6=F
3! + - .7
4! "/3 F $" +@4
=! - .. 7
1! G -3 7

Procedure : ) " "

' " " "/3 F
+ .
0
B 9

B S1 S2 S3 S4 T
# 3
+ 3 7 .= . ". 3.
* G 3 "= ". .
F @$ . . . . .
' . . . . . .

B ( = =".

Calculation :
@* ( +
+ 7! L ..
@* + 8 +

42
Interpretation :
F 9 .6 4 / ..
# 9 .4 " / ..

) 5

!
)

C <
) M

$ )

Technical Contents: M
Methodology of its teaching 9
Evaluation of the session 9 (

4D
 Chapter 'o. 11
Estimation Total Protein
Object 9

Introduction:
+ B

Biuret Method
Principle :

Reagents
! , -3 =..
D =
"4
..

=.
 ! -

! + -
M?
1
/ .. ( 1 / ..

Procedure : 3 <0 < <+ < < < B =

G . <0 < . S
<>+ < .
< < B 3.
, <0 < <+ < < < =4.

Calculations
(
+ / .. ! L 1
(
#ote.
)
+ ! .
=
3. < < ( <+ >
<0

=
Interpretation
12 / ..
(
(

(
(

(
'

( ?
)

Technical contents : 0
Method of teaching 9 *
0

Evaluation 9 K

="
 Chapter 'o. 12
Estimation of Serum Albumin and Globulin
Object:

Bromocresol Green Method

Principle :
(
K K
$

Reagents
! + 2 2..
$ ? 4. . F

! K 4 D .

! K - "=. K 6=.
$ ? 4" . F
4 ? - 3= 3.7!
! + - &
4 / ..
+ 5 =.
.. ! '
5 !
4 / ..

=3
Procedure : : 3 <0 < <+ < < < B 4
K G . ."
<0 < . ." <+ < . ."
< < B =
, <0 < <+ < < < 13.

Calculations :
(
+ / .. ! L 4
(

Interpretation :
36=3 / .. +
2 31 / .. (9K "9
"9 " =9
*

( -

(9K

=4
 +

Technical contents : 0
Method of teaching 9 *
0
Evaluation 9 K

==
 Chapter 'o. 13
Estimation Of Aminotransferases (Transaminases)
Object 9
( ( (+ ! +K@
( ( (: ! +K'

Introduction :
5
αM
* 5 (+

B (+ (: 9

Colorimatric Methods :
, ;# D=6! 5 D6.! ; D64 !

Modified Reitman & Frankel Method :

Principle :

"4 5 *F'$!
5
=".

=1
Reagents :
-# 5 .. /:
" /: " ? .. /: : (+
".. /: *: (:
(+ =6 : 3" :

(: 62 *:
( ? $ 64 F @$ :

" "4 * 5 *F'$! ".. !/: /:

$ :
3 + 4.. 1 !/:
4 ' -" /: "" ..
!

Procedure : Mark test tubes and proceed as follows :

Blank Test S1 S2 S3 S4
.= .= .D .2 .6 .1
+
* G .= .= ." ." ." ."
+ ." ."
' + . ." .3 .4

) 36 (+ 1. (: - 3.
( .= *$'$ B
". F @$ . =

=6
 , =". =
M 0 !
(+ (:
+ "4 "=
+" 1 =6
+3 4 D6
+4 D. =.
M L. 423 ) 0
(+ )0! (: )0!
+ = 34
+" "D 3 "6 4
+3 =4 6 41 1
+4 D " 6" .

Interpretation :
(+ L 6 " )0
(: L 1 ". )0
(+ (:

% (+ ; (: 9
: 9
(+ (:
". "=
' 6 "
3 = (+

=2
 (+ (: /(+ * , !
T U
" % (+
(:
3 : (+
(:
4 '
(: 5 (+ (:

)) B % (+ 32
31 "4
(:
(:
(+
*
'
(
?
K
$
@ (+ (:

Technical contents : 0 +K@ +K'

=D
Method of teaching 9 * +K@
+K' 0

Evaluation 9 K +K@ +K'

1.
 Chapter 'O. 14
Estimation of Serum Alkaline Phosphatase
(King & King Method)
Object : To estimate serum alkaline phosphatase.
Introduction:
( )

!
5

Principle :
$ 4

=".

$ '
+
Reagents :
+ -* - .. /:!9 *
" 2 " =4 !
& > > 4

" + .. /:!9
* 1 31 3 31

1
3 B & , ,
" $ .
4 + /:! - *
.. /: $ : 24 !
M 4
= G / .. ! *
- ..
1 + - =.. /: ". /:!
6 + - =.. /: 4" /:!
2 4( - 1 /: +
D ' # - "4 /: +
Procedure : B 9
Reagent Blank Test S1 S2 S3 S4
(ml) (ml) (ml) (ml) (ml) (ml) (ml)
+ ". ". 1 .1 .
" G + .= . = ".
3 * G .
4 + .
) 36 =
= F @$ .2 .2 .2 .2 .2 .2
1 F $ @" " " " " " "
6 4( . . . . . .
2 ' . . . . . .
#
B =".

1"
Calculation :
( ( M( 0 !>
M (
..

@* ..
+ ( ' L ( +
@* + .
M( 0 / :!

Interpretations :
3 3 M(

) 9
' 9
-
'
' - 5

Pathological :
(! * 9 (

, - .. M( :
8 *
@ -

13
 ' < @ !
8
. "=
@ +
+
+

$ L
! : * 9
!
5

Technical contents : 0 ( '

Method of teaching 9 *
( ' 0

Evaluation 9 K ( '

14
 Chapter 'o. 15
Estimation of Serum Bilirubin
% B % D36 B !
Jaundice
C !

/ F
." /

) ?
"!
3!
• Haemolysis.
?
• Failure of the conjugating mechanism within the hepatocyte.
• Obstruction in the biliary system.
? ?
? 0 ?

Biochemical tests
)

1=
 5 (
G ?

:# ! (+ (:

Object 9
Introduction:
B
?

:
V ( 0*'
?
?
+
?

Principle: G 5 5 % < , !
5

? ? 5
5 5 ? &

11
* , ! ? &
) ,
? I ? !

Reagents :
* 5 9B .
( .3
+ (9 = $ :

+ 9.= / ..

" * 5 9 = $ :/
3 B
4 9 * .
= ! . F &
!
'
!
5
!

16
Procedure : : 9
Reagent Standard Total Bilirubin Conj. Bilirubin
+ + ! ! ?! ?!
+ ." ." ." ."
+ + ." ."
. / !
* G 2 2 2 2 43 43
* 5 , .= .= .=
* 5 .= .= .=
B "= "= "= "=

B 3.
, . =4. *

Calculation :

! ! ..
+ L (
/ :! + + 8 +

! ! ..
? + L (
/ :! + + 8 +

+ ?
?

12
Interpretations :
F / : $
3 / : ?
$ 9
I. Unconjugated Hyperbilirubinaemia (Retention Jaundice): *
?
(A) Haemolytic (Pre hepatic Jaundice) due to:
!) 9

,
K1
( )
@ '
!)
! F $ 9
! K <
!
9
HF ?? +
K < !
' ? '

? 5
)) Conjugated Hyperbilirubinaemia (Regurgitation Jaundice) :
* ?

1D
Technical contents : 0
Method of teaching 9 * +
0

Evaluation 9 K

6.
 Chapter 'o 16
Estimation of serum triglycerides
Lipid profile9 #
! 9'
F 4.. ... /
"! ' 9) 5
5 *
=.. F 1. "6. /
3! 9
4! 9
=! $*: 9
1! :*: 9 - $*: I 8:*:
6! 8:*: 9 K/=

Object: To estimate serum triglycerides.

Introduction :
%

Method:
(
'ormal range
. D. /

6
Principle:

#
(

4 .
!

Reagents:
! ( 9 9 !9 )
; ..
"! ) 9 (,
3! ( M@$ 9 ) =.
1.. 4..
4! B 9 ) 66
; 1=. D4.
1.
=! ( 9) 6=
".. 2..
1! 9 .. / )
!

Stability of the reagents

, " 3 4 ; = , 1
"2

6"
Procedure :

" ' .=
3 ( 4.
4 ( .
= ( . .. /

1 B

6 M = "= I= !

2
3... ,'B . F
9
+
# ! ".
# + ! ".
) ".
( M@$ .1 .1 .1
B 1. 6= =
B = = =
B =
( = = =
B 6. =
4".
!

63
Calculations :
@*
+ / L ..
@* +
F 9
+ + 1! "2
" (
..
Procedure limitations:
3.. / # 3.. /

Enzymatic determination of serum triglycerides:

%

Chemical principle of test

:
I $"@ K I
:

K
K I( ' K -3- I (*'
K
K -3 I @"
* I $"@"

'
$"@" I 4 5 I
=". !

64
Technical contents : 0
Method of teaching 9 *
0

Evaluation 9 K

6=
 Chapter 'o 17
Estimation of Serum Cholesterol (Total)
Object: To estimate serum cholesterol.

Introduction:

$
?
Sackett's Method:
Principle : '

Reagents
! ( -% D=7!
39
!
! ( (
".9 ?
! + "..
..

61
 ! G -
"= =
.4
Procedure : ' "
( ."
M 5
=.. = '
B =
: <0 <
: <+ < < <
= =
B =
( ? 12.
+ ..7 5 !
< < <0 < <+ <
Calculations

( .4
+ / .. ! L ..
( ."
(
L "..
(
Interpretation
+ =. "4. / ..
3.. / ..
( !
?
(

66
$
( !
!

, 5
+
5

* 5 (
!

Principle :

;
4 3
'
4 5 / !
= . =.= =3. !

Technical contents : 0
Method of teaching 9 *
0

Evaluation 9 K

62
 Chapter 'o 18
Estimation of Serum HDL cholesterol

Object : To estimation serum HDL cholesterol

Introduction :
) :*:
8:*: ; $*:
$*:

F
B 9 3. 1. /
G 9 4. 6. /

B 9
G !

Precipitating reagents:
! ' ' ( !9 )
" "= 2. )F
4" .
"! B 9 ". 34
) =.

6D
Procedure :
' 9

!+ .=
"! ' ( . .=
3! B " . ."
B 3... , ' B ". +

=..

Calculations :
@*
+ $*: / L 4
@* +

Additional Information:
$
$*:

Technical contents : 0 $*:

Method of teaching 9 * $*:
0

Evaluation 9 K $*:

2.
 Chapter 'o. 19
Estimation of Enzymes in diagnosis of Myocardial infraction

Object 9 :*$ M MB
Introduction:
5
B) 9

• M!
• ( (+ !
• : :*$!

"4
(+ :*$

Creatine kinase
M (
5
M BB! M 5 MB
5
M! B)

Aspartate and alanine aminotransterases

(+ B)
5
(: ! <

2
 5 < ! (:
(: (+
5
( "=7
B)
(+ :*$

Lacate dehydrogenase
: 5 B) :*$ )
(
:*$ ) 5 :*$=
5 :*$ :*$

:*$

Technical contents : 0 :*$ M MB

Method of teaching 9 *
Evaluation 9

2"
 Chapter no. 20
Estimation of Serum Calcium
Object 9

Introduction :
-
5
B 5
(
W

Principle:
+

Reagents:
! ( 47 4
..
"! ( "7 " + K . 22.! ..

3! + F "2 ...

23
 4! + . F 3 =2
...

=! ' .. F . . F
.. 5
.. F .. F
& ."
1! + . . F . 16
=
... .. F L .. F

Procedure:
B " " 47
B
B =.. = '

G ( 3
"7 ( " F
M
G
.. F
F ..
F + # "
F .. F

24
 ."
+ / .. !L ! ..
"
L ! .
Interpretation :
+ D - / ..
4.
=. (

(
*

)
3.

Technical contents : 0
Method of teaching 9 *
0

Evaluation 9 K

2=
 Chapter 'o. 21
Estimation of Serum Phosphorus (Inorganic)

Object 9

Introduction :
'

Method of Fiske and Subbarow*

Principle:
+ 5 '

" 4
!

Reagents:
! .7 .
..
"! B - "=
".. 3.. . F
...
3! ( - =D = "

21
 "=. 5 " 4
4..
4! + - . 3=
. .F
... =
.4
=! G -=
.. .7 =
. ."

Procedure :
B D .7 (
# =
<0 < ' =
<+ < = .7
< < 31 B
: =
, <0 < <+ < < < 12.

Calculation :
+ / .. !

( . ."
L ..
( .=

(
L 4
(

26
Interpretation :
"=-4= / ..
4-1 / ..
+ *

Technical contents : 0
Method of teaching 9 *
0

Evaluation 9 K

22
 Chapter 'o. 22
Estimation Of Serum Uric Acid
Object9
Introduction:
0 (
'
Caraway's Method of Estimation:
' 9 ' 5

6..

Reagents :
! + .7
"! "/3 F +
3! 9( =. .7 =. "/3 F $"+@4
2.. *
+
4! ' 9+ * =.
4.. ( 4. 2=7
" =.. *

=! F 3 @3 .7
1! + .. 7
6! G 7

2D
Procedure :
) .1 =4

B T S S2 S3
+ . ". 3.
7!
" + ! 3.
3 * G ! 3. ". .
4 F 3 @3 ! .1 .1 .1 .1 .1
= ' ! .1 .1 .1 .1 .1

B 3. ,
6..

Precautions :
! + ,

! +

! :

! +

D.
Calculations:

@* +
+ 0 L ..
@* + 8 +

Interpretation:
( 9 4=2" / :
# 9 3 1= / :
9 ".== / :
)

Hyperuricaemia :
+ %
)
? :
,

* B
+
* 5

D
Hypouricaemia :
: !
G < *
, # <
+
* 5

Other method of estimation :

Enzymatic Method:
! 5 5
5
@* "D3
"! (
$"@"

0 ( I $"@ I @ " 0 ( I $"@"

$"@
'
@! +

!
Technical contents : 0
Method of teaching 9 *
0
Evaluation 9 K

D"
 Chapter 'o. 23
Blood Gas Analyser
Object:

Introduction :

$
5 '@"
$ (
'@" $
$ ' @3

& "=
$ ' @ '@" $
@
$ @" + $ @" @"
$
)
$ ' @" '@"
)

+ & =. B
/ !
) .4

D3
 9 ' $
$ 1/2 $ 6 3X' @"
@" =7 .7 '@"
(
"7 ".7 !
( '@" '@"
@"

(
5 9
! ( 9 $ ' @"
$ $ &
! + 9) $ @3
& 36 ' @" 4. $ @"
$
! 9) $ 6 4!
' @" 4. $ 36 !
/: )

! @" 9 " @" $ @3

@"

D4
Reference Values :
-"3 "3 /:
( "4 3" /:
+ "" 4 "= 2 /:
( ' @3L34 4= $ @ ." 3= /:
$L6 31 6 4=

*
/ G

Precautions :
(
$ )
4= =. 4

"

3 #
4 )
36
$ L $ 36 I . . 4 36 !

D=
Type of possible acid base disturbances :
! B 9 ' $ @3

$ @3
9
* M
B M
: (
)
) F$4 F$3
$I 5 $ @3
)
))
$ @3 )
Respiratory acidosis : ) ' @"
@" @"
)

Metabolic alkalosis : ' $ @3

$" @3
F $ @3

Respiratory alkalosis : ' @"

D1
 %
$
#

+
I
5 3
I
5 " $

Combined respiratory and Metabolic acidosis causes are :

+
,

' @3
5 $ @3

Combined respiratory and metabolic alkalosis :

+

)
) ' @3
$ @3
$

D6
 Chapter 'o 24
Examination Of Cerbrospinal Fluid (CSF)
Object: +#

Introduction:
+#

..3 ..2 &

+#
Physical Examination
! F +#
"! ( F +#

Chemical Examination –
% +#
+#
B # G +
Test Std. Blank
(ml) (ml) (ml)
CSF ."
Std. Glucose ".
Sion. (10 mg%)
Distilled water 2 ".
Alkaline well Cuso4 ". ". ".
Reagent
Mix and keep the Folin Wu tubes in boiling
water bath for 8 minutes. Remove the tubes
and cool.
Phosphe molybdic acid ". ". ".

D2
 * "= B
B
Calculations :
@ + . "!
P P ..
+ % . "!

P .. L 7
+

Estimation of Protein in CSF

Test (ml) Std. (ml) Blank

(ml)
CSF Sample .
Std. (20 mg%) .
D.W. .
Sulphosalisylic 3. 3. 3.
acid Reagent

B , . ,

Calculations :

P ". L +# ' 7
+

Estimation of chlorides in CSF

, +#
* =7 =
( F 3 . "D7! % : J ,
, ... L 7

DD
 +#

Clinical Interpretation :

Constituents 'ormal range Clinical condition in which

variations may be found.
Sugar 1. 1= / .. , F+

Protein ". 4. / .. ,
conditions, # <
+
F
!
Chlorides 6.. 6=. / .. , :
% F !
". "6 %&/:

Colour
)
! ) ? -J
! * -J
$
! J +# P
! ) -
! , +# ?

! @ - !

..
Appearance –
) +#

.
 Chapter 'o. 25
Serum Amylase

Object:
Method : Caraway

Introduction:
( 5 )
?
+
5

Principle:
+

."
Procedure
' = =.
" ' < < 34 =

3 0 .
< < *

4
= ( 6= =

1 + =
< <
6 * =.

2 , D !
11.

Calculation
( - 2..
(
L ..

) 4.. .. .
= . ."

.3
(
. 3.

! + 6= <4<
3.
! + = "
<=<
.

! @ .
.. . ... ..
4/= ... L 2..

Precautions and sources of error.

1. SALIVA.
+ %
(
F
)
G

2. SUBSTRATE
0

< <

.4
 (
< <
3. E'ZYMES
5
$

4. PRECIPITATE

=.
< :

5. URI'E
(
"4

6. OTHER BODY FLUIDS

'
( ...
. DD .

Clinical interpretation
( ?
(

.=
)

"4

0 < <

+
)

(
& ( & < <

B
0

.1
 Chapter 'o. 26
Serum Electrolytes
Object :
Introduction :
+
@
B
G
9
F IM!- I $ @3 ! L 1 I "
?

&

Laboratory determination
Sodium and potassium -
&
# #
!

Principle of flame photometry :

(

.6
 &
!

'

Type of Flame Photometers :

* 9$

" ) + ) 9

@
(

Atomic absorption spectroscopy9 )

5
" Ion selective electrodes: &
+

.2
 &

F 9
+ 9 3= 4= B /:
' 93=-4= B /:
9 D= - .= B /:

.D
References
! ' $ 8 = D24

"! ' K DD4

3! 5 " DD4

4! B )*' G 1

=! ' , K 3 "..4

1! $ Y @ 4

6! : G , + "

2! ( K " DDD

.
 /
9

+) 0 @ 0
(: +K' ! 0 " 0/: 0 4. 0 /:
(+ +K@ ! 0 = 0/: 0 3= 0 /:
( 3= 46 /: 3 = = . / ..
( ". D. 0/: 3 3 M (
' 3=. 0/: / ..
( ' . . 3 0/: 3= M (
/ ..
( 6. 34. 0/: 1. ".. +
0 / ..
! 3 ". /: ." 3 / ..
F 2 16 /: . = 4 . / ..
" "= " 1 /: D / ..
F 2= 3 4= 6 4 3 2 / ..
/:
3 62 /: 4. "1. / ..
"3 3 /: "3 3 &/:
' @" 41 1 M' 3= 41 $
== . /: .6 4 / ..
% F 34 41 /: 34 41 &/:
M 31 =" /: 3 1 = " &/:
F 4. =D /: 4 . = D &/:
D1 .6 /: D1 .6 &/:
K T14 /: 1. .. / ..
) .2 = /: "= 4= / ..
'
' ! 1. 2. /: 1 2 / ..
0 ( ".1 41. /: "6 / ..
3= 32" /:
0 "= 2 /: = =. / ..
.= = /: . D. /
+ :*$ 6. "4. )0
+ 'M = 3. )0 B
= . )0 G
"