رايط الشرح علي اليوتيوب https://youtu.be/gBg1Kn05WfY

1. BY
dr: Mohammed Bahgat Mohammed Sofyan
Assistant lecturer of medical biochemistry,
Faculty of medicine, Al-Azhar university
(Assiut branch)
1

2. 2

3. ‫هللا‬ ‫رسول‬ ‫صدق‬
‫وسلم‬ ‫عليه‬ ‫هللا‬ ‫صلى‬
3

4. ‫صورة‬ ‫كل‬ ‫على‬ ‫اضغط‬
‫عرض‬ ‫وضع‬ ‫في‬
‫على‬ ‫وادعمنا‬ ‫الشاشة‬
‫اإلنترن‬ ‫على‬ ‫قنواتنا‬
‫ت‬

5. ‫على‬ ‫ملفي‬ ‫في‬ ‫هنا‬ ‫أرفعها‬ ‫التي‬ ‫شرائحي‬ ‫ومذاكرة‬ ‫بالتعلم‬ ‫وأسعد‬ ‫وأرحب‬ ‫أسمح‬
Slide share website
‫وال‬
‫مانع‬
‫أيضا‬
‫من‬
‫نسخ‬
‫شريحة‬
‫او‬
‫اثنين‬
‫عند‬
‫الضرورة‬
،
‫وال‬
‫مانع‬
‫من‬
‫شرح‬
‫البوربو‬
‫ينت‬
‫الخاصة‬
‫بي‬
‫للغير‬
‫بشرط‬
‫عدم‬
‫إزالة‬
‫اسمي‬
‫من‬
‫البوربوينت‬
،
‫فال‬
‫أسمح‬
‫أبدا‬
‫بإزالة‬
‫اسمي‬
‫من‬
‫على‬
‫الباوربوينت‬
‫ووضع‬
‫اسمك‬
‫بدال‬
‫منه‬
‫لتصبح‬
‫وكأنك‬
‫من‬
‫صممتها‬
‫فهذه‬
‫سرقة‬
‫ال‬
‫أسمح‬
‫بها‬
‫وتضييع‬
‫لحق‬
‫من‬
‫تعب‬
‫في‬
‫عملها‬
.
‫وفقكم‬
‫هللا‬
‫وإياي‬
‫للتعلم‬
‫ونفع‬
‫اآلخرين‬
I allow, welcome, and be happy to learn and study my slides that I
upload here in my profile on Slide share website
There is also no objection to copying one or two slides when
necessary, and there is no objection to explaining my PowerPoint to
others on the condition that my name is not removed from the
PowerPoint. I never allow my name to be removed from PowerPoint
and to replace it with yours, Make it look like you designed it. This is
theft that I do not allow and a waste of the right of those who are
tired in this work. May God bless you and me for learning and
benefiting others
5

6. Dr Thomas sowel
If you wanna be a skilled
doctor you must be
good at chemistry
whether you like it or
not.
6

7. Subclinical
jaundice
‫عدت‬ ‫لو‬ ‫الجسم‬ ‫علي‬ ‫وتظهر‬ ‫الدم‬ ‫في‬ ‫تكون‬ ‫ممكن‬ ‫الصفرا‬
2 mg/dl.
‫وتسمي‬ ‫الجسم‬ ‫علي‬ ‫تظهر‬ ‫ال‬ ‫كده‬ ‫من‬ ‫أقل‬
Subclinical jaundice.
Jaundice becomes clinically evident when
serum bilirubin level exceeds 2.0 mg/dl.
7

8. Bilirubin metabolism
‫مكانين‬ ‫في‬ ‫بتتكسر‬ ‫الحمراء‬ ‫الدم‬ ‫كرات‬ ‫بداية‬
:
‫بيمثل‬ ‫وده‬ ‫األول‬ ‫المكان‬
١٥
%
‫الناتج‬ ‫البلوربين‬ ‫من‬
Ineffective erythropoiesis.
‫ال‬ ‫في‬ ‫بيحصل‬ ‫وده‬
BM
،
‫حاالت‬ ‫زي‬ ‫بتتكسر‬ ‫الحمراء‬ ‫الدم‬ ‫كرات‬ ‫أمهات‬ ‫إن‬ ‫نتيجة‬
:
Megaloblastic anaemia, myelofibrosis and Myelodysplasia
Ineffective hematopoiesis may occur in these diseases leads to jaundice.
‫من‬ ‫وكمان‬
and from myoglobin, cytochromes, and peroxidases.
8

9. ‫وهو‬ ‫بالطبع‬ ‫األكبر‬ ‫وهو‬ ‫الثاني‬ ‫السبب‬
:
Destruction of RBCs by reticuloendothelial cells especially spleen
‫بيمثل‬ ‫وده‬
٨٥
%
‫الناتج‬ ‫البلوربين‬ ‫من‬
‫بيحصل‬ ‫اللي‬ ‫ايه‬
Hb ➡ degraded into haem and globin.
Globin degraded into A.A. that recycled again
Haem ➡ degraded into iron and protoporphyrin
Iron recycled again in the body
Protoporphyrin converted into ➡ Bileverdin ➡ indirect bilirubin
Bilirubin metabolism
9

10. 10

11. 11

12. 12

13. ‫ال‬ ‫اتكون‬ ‫هنا‬
indirect bilirubin and it is water insoluble
‫وله‬ ‫البول‬ ‫في‬ ‫ينزل‬ ‫ال‬ ‫ولذلك‬ ‫األلبيومين‬ ‫علي‬ ‫ويحمل‬ ‫الدم‬ ‫في‬ ‫السير‬ ‫يستطيع‬ ‫ال‬
‫أسماء‬
Indirect bilirubin = unconjugated bilirubin = hemobilurbin = Alfa
bilirubin
Bilirubin metabolism
13

14. 14

15. ‫له‬ ‫جدا‬ ‫مهمة‬ ‫خطوات‬ ‫ثالث‬ ‫ويحدث‬ ‫للكبد‬ ‫يذهب‬ ‫ذلك‬ ‫بعد‬
1 _uptake of unconjugated bilirubin by liver.
2_ Bilirubin is conjugated with glucuronic acid to form bilirubin
monoglucuronide and diglucuronide(conjugated bilirubin); this process is
mediated by the enzyme UDP glucuronyl transferase.
Conjugated bilirubin is water-soluble.
3_ secretion of Conjugated bilirubin into bile canaliculi ➡ hepatic duct ➡ bile
duct ➡ hepatopancreatic ampulla ➡ second part of the duodenum
Bilirubin metabolism
15

16. 16

17. ‫ال‬ ‫كده‬ ‫بعد‬
:
Conjugated bilirubin is deconjugated by glucuronidase then reduced by intestinal bacteria
into urobilinogen .
‫ال‬
urobilinogen
‫اما‬
:
1_ convert into stercobilinogen ➡ oxidized into stercobilin ➡ escapes into the stool and
gives its color.
2 _absorbed from Porta circulation to the liver and again to the bile in the form of
urobilinogen ( enterohepatic circulation).
3_ A small amount of urobilinogen escapes in circulation and excreted in the urine.
Bilirubin metabolism
17

18. 18

19. ‫الصفرا‬ ‫عن‬ ‫نتكلم‬ ‫بقه‬ ‫نتعالوا‬
:
‫الحمراء‬ ‫الدم‬ ‫كرات‬ ‫تكسير‬ ‫في‬ ‫زيادة‬ ‫عندي‬ ‫أنا‬ ‫لو‬
Hemolytic anaemia
‫مثل‬ ‫تحصي‬ ‫وال‬ ‫تعد‬ ‫ال‬ ‫األسباب‬ ،‫كان‬ ‫سبب‬ ‫أي‬
Hemoglobinopathy, microangiopathic hemolytic anemia, neonatal
jaundice, hemolytic disease of newborn, hemolytic reactions,
ineffective erythropoiesis,.... etc.
19

20. 20

21. ‫هيحصل؟‬ ‫اللي‬ ‫ايه‬ ‫غيرها‬ ‫أو‬ ‫األسباب‬ ‫هذه‬ ‫من‬ ‫سبب‬ ‫عندي‬ ‫انا‬ ‫لو‬
1 _Increased RBCs destruction ➡ ⬆ indirect bilirubin
2_ liver doubles its actions ➡ more uptake Conjugation and secretion of bilirubin ➡ ⬆
urobilinogen in urine and stercobilin in the stool and becomes dark
‫ال‬ ‫ألن‬ ‫طبيعي‬ ‫البول‬ ‫وطبعا‬
Indirect bilirubin water insoluble don't escape in urine
‫ال‬ ‫لزيادة‬ ‫داكن‬ ‫والبراز‬
stercobilinogen
‫ال‬ ‫مواصفات‬ ‫كل‬ ‫فيها‬ ‫وبيبقي‬ ‫صفراء‬ ‫فيه‬ ‫والجسم‬
Hemolytic anemia as Reticulocytosis, high LDH and low Haptoglobin.
NB ➡ bilirubin doesn’t exceed 5_6 mg/dl
‫في‬ ‫طبعا‬ ‫ده‬ ‫الكالم‬
hemolytic anemia, not others. 21

22. This is a transient increase of unconjugated bilirubin which is observed in almost all
newborns. It usually develops during the 2nd to 4th day after birth with the return to
normal bilirubin level by the 7th to10th day.
It is because of deficiency of glucuronyl transferase and increased RBCs in the
neonate leading to impaired conjugation during the first few days of life.
‫الكبد‬ ‫علشان‬ ‫جامد‬ ‫هنا‬ ‫بيعلي‬ ‫البلوربين‬ ‫طبعا‬
immature
‫يعمل‬ ‫بيعرف‬ ‫فمش‬
Conjugation of increased bilirubin
‫يخترق‬ ‫تجعله‬ ‫قد‬ ‫كبيرة‬ ‫بمعدالت‬ ‫زيادته‬ ‫إن‬ ‫المشكلة‬ ،‫كبيرة‬ ‫بمعدالت‬ ‫البلوربين‬ ‫فيزداد‬
Blood brain barrier and causes kernicterus
22

23. 1- Gilbert’s syndrome :
‫مثل‬ ‫جينية‬ ‫امراض‬ ‫فيه‬
Gilbert’s syndrome ➡ isolated defect uptake of bilirubin by the liver
‫يتعدي‬ ‫ال‬ ‫البلوربين‬ ‫الغالب‬ ‫في‬
3
‫لما‬ ‫انه‬ ‫سوي‬ ‫حاجة‬ ‫اي‬ ‫فيه‬ ‫يبقاش‬ ‫وما‬ ‫طبيعي‬ ‫بيعيش‬ ‫والشخص‬
‫عنده‬ ‫بيعلي‬ ‫بيصوم‬
unconjugated bilirubin
all liver function tests are normal
‫جدا‬ ‫مشهور‬ ‫المرض‬ ‫بالمناسبة‬
23

24. 2- Crigler-najjar syndrome
‫بعد‬ ‫الغالب‬ ‫في‬ ‫وبيموت‬ ‫جدا‬ ‫عالي‬ ‫البلوربين‬ ‫وبيبقي‬ ‫الوالدة‬ ‫بعد‬ ‫بيظهر‬ ‫ده‬
٣
‫سنين‬
24

25. 25

26. 26

27. ‫هيحصل؟‬ ‫اللي‬ ‫ايه‬ ‫المرارية‬ ‫القنوات‬ ‫في‬ ‫انسداد‬ ‫عندي‬ ‫بقه‬ ‫انا‬ ‫لو‬
‫ال‬
bile
‫وبالتالي‬ ‫أخرى‬ ‫مرة‬ ‫للدم‬ ‫وهيرجع‬ ‫األمعاء‬ ‫في‬ ‫هينزل‬ ‫مش‬
1- negative urobilinogen in urine and stercobilin in stool leads to clay
stool
2_ Conjugated bilirubin increased in blood ➡ excreted in urine
a_ ⬆ Conjugated bilirubin in the blood
b_ ⬆ bilirubin in urine
27

28. Causes of obstructive jaundice
a- drugs as estrogen, oral contraceptives, anabolic steroids, oral anti-diabetics
Phenothiazines and erythromycin.
b- acute hepatitis
C- pregnancy
D- primary biliary cirrhosis ➡it's autoimmune destruction of intrahepatic bile ducts.
It predominantly occurs in middle-aged females and is characterized by chronic
elevation of alkaline phosphatase and positive anti mitochondrial antibody in serum.
There is an association with other autoimmune disorders.
28

29. a_ gall stones
b_ cancer head pancreas
c_ carcinoma of the bile duct and
ampulla of Vater
d_ biliary strictures or atresia
e_ primary sclerosing cholangitis ➡
is an autoimmune disorder occurring in young to middle-aged men in whom there is
inflammation and destruction of both intrahepatic and extra-hepatic bile ducts.
Associated inflammatory bowel disease is often present.
Serum alkaline phosphatase is elevated, and many patients have circulating
perinuclear antineutrophil cytoplasmic antibodies (ANCA) .
29

30. 1_ Absent urobilinogen in urine and stercobilin in stool leads to clay stool.
2_ Conjugated bilirubin increased in blood ➡ excreted in urine
a_ ⬆ Conjugated bilirubin in the blood
b_ ⬆ bilirubin in urine ➡ tea colored urine
‫ال‬ ‫بيزداد‬ ‫كمان‬
Elevation of Serum cholesterol, serum transaminases, serum alkaline phosphatase, serum Gamma
GT, 5 'nucleotidase and serum bile salt
‫وهنا‬
‫البلوربين‬
‫بيعلي‬
‫ألرقام‬
‫عالية‬
‫قد‬
‫تصل‬
‫ل‬
50
‫والجسم‬
‫بيصفر‬
‫جامد‬
‫والعين‬
‫ممكن‬
‫تبقي‬
‫اصفر‬
‫مائل‬
‫للون‬
‫األخضر‬
‫والبول‬
‫بيغمق‬
‫ألن‬
‫ف‬
‫يه‬
‫بيلوربين‬
‫والبراز‬
‫بيبقي‬
‫فاتح‬
‫ألن‬
‫الدنيا‬
‫مسدودة‬
‫وطبعا‬،
‫امتصاص‬
‫الدهون‬
‫بيبقي‬
‫فيه‬
‫مشاكل‬
‫ويقل‬
‫امتصاص‬
‫الفيتامينات‬
‫التي‬
‫تذوب‬
‫في‬
‫الدهون‬
‫مثل‬
A,K,E,D
‫من‬ ‫هيعاني‬ ‫المريض‬ ‫وطبعا‬
Itching and bradycardia from high bile salt
‫علي‬ ‫محمال‬ ‫للدم‬ ‫يرجع‬ ‫العالي‬ ‫والفوسفوليبيد‬ ‫الكوليسترول‬
Lipoprotein X
‫نف‬ ‫نعرف‬ ‫األهم‬ ‫لكن‬ ‫تجيله‬ ‫ما‬ ‫قبل‬ ‫النتائج‬ ‫عارف‬ ‫بيبقي‬ ‫الطبيب‬ ‫إن‬ ‫حتي‬ ‫الحاالت‬ ‫هذه‬ ‫في‬ ‫جدا‬ ‫جدا‬ ‫سهل‬ ‫الصفرا‬ ‫تشخيص‬ ‫صراحة‬
‫بين‬ ‫رق‬
Extrahepatic and intrahepatic biliary obstruction
‫باألشعة‬ ‫بيبقي‬ ‫الغالب‬ ‫في‬ ‫وده‬
Us, CT scan and ERCP.
30

31. Normal eye Olive green eye in obstructive jaundice
31

32. ‫فيها‬ ‫بيبقي‬ ‫أمراض‬ ‫فيه‬
Isolated defect in secretion as
Dubin-johnson syndrome or Roter syndrome
‫ال‬ ‫بيزداد‬ ‫دي‬ ‫الحاالت‬ ‫في‬
Conjugated bilirubin
‫فيها‬ ‫وبيبقي‬
No evidence of obstruction nor LCF.
32

33. 33

34. ‫او‬ ‫السونار‬ ‫في‬ ‫عالمة‬ ‫أو‬ ‫األلبيومين‬ ‫في‬ ‫نقص‬ ‫او‬ ‫الكبد‬ ‫انزيمات‬ ‫في‬ ‫ارتفاع‬ ‫معاها‬ ‫بيبقي‬ ‫الكبد‬ ‫سببها‬ ‫اللي‬ ‫الصفرا‬ ‫حاالت‬ ‫في‬
‫الم‬ ،‫مرضي‬ ‫تاريخ‬
‫هم‬
‫الكبد‬ ‫بسبب‬ ‫انه‬ ‫المرض‬ ‫سبب‬ ‫واضح‬ ‫بيبقي‬
Hepatocellular disease:
Liver enzymes (aspartate aminotransferase and alanine aminotransferase) are markedly elevated,
and serum bilirubin is usually in the range of 4.0 to 8.0 mg/dl. Conjugated bilirubin is 20-50% of total
bilirubin.
In hepatocellular injury, both conjugated and unconjugated bilirubin are increased.
‫حالتين‬ ‫في‬ ‫بتزداد‬ ‫الصفرا‬ ‫الكبد‬ ‫في‬ ‫المشكلة‬ ‫لو‬ ‫أقول‬ ‫عاوز‬
:
1- Early in Acute hepatitis due to intrahepatic biliary obstruction due to liver cell swelling
2- Late in liver cell failure due to inability of liver to uptake, conjugate and secretion of bilirubin
‫و‬ ‫غيبوبة‬ ‫في‬ ‫وبيخش‬ ‫واأللبيومين‬ ‫استسقاء‬ ‫وعنده‬ ‫للكبد‬ ‫مرضي‬ ‫تاريخ‬ ‫عنده‬ ‫بيبقي‬ ‫الثاني‬ ‫جدا‬ ‫سهله‬ ‫بينهم‬ ‫التفرقة‬ ‫طبعا‬
‫متقد‬ ‫حالة‬ ‫وفي‬ ‫دم‬ ‫بيرجع‬
‫مة‬
‫الكبد‬ ‫أمراض‬ ‫من‬
.
34

35. Estimation of both types of bilirubin is helpful in the differential diagnosis of
jaundice.
In the post hepatic type of jaundice, direct bilirubin is the predominant form
(> 50% of the total).
In hepatocellular jaundice, direct bilirubin is usually between 20-50% of the
total.
Indirect bilirubin predominates in hemolysis, Gilbert’s syndrome, and Crigler
Najjar syndrome (direct bilirubin is < 15% of total).
35

36. 36

37. 37

38. 38

39. 39

40. 1- hemolytic disorders as ( G6PD, spherocytosis)
2- Hemolytic disease of the newborn (RH/ABO incompatibility)
3- congenital TORCH infection.
40

41. 1- physiological jaundice in a full-term baby
2- sepsis
3- polycythemia
4- hemolytic disorders
5- criggler Najjar syndrome
41

42. 1.physiological jaundice in preterm
2.neonatal sepsis
3.hemolytic anemia (G6PD difciency_spherocytosis)
4.cephalohematoma
5.TORCH
42

43. 1. breast milk
jaundice
2. congenital
biliary atresia
3. galactosemia
4. neonatal
hepatitis or
sepsis
5. congenital
hypothyroidism
6. criggler
Najjar
syndrome
43

44. 44

45. 1_ Total and direct bilirubin
2_ ABO, RH of baby and mother
3_ CBC, Blood film, reticulocytic count and hematocrit :
4_ coombs test
5_ Liver function tests
6_ blood sugar
7_ TORCH screen
8_ Blood culture
9_ G6PD Assay, LDH, Uric acid
45

46. Speicment
collection
Specimen ➡ fresh
Serum
Normal total bilirubin
➡ up to 1.2 mg/ dl
Normal direct bilirubin
➡ up to 0.3 mg/ dl
46

47. Specimen collection
NOT HEMOLYSED ➡ AS IT
⬇BILIRUBIN
PREFERRED IN THE
MORNING
NOT LIPAMIC
47

48. Bilirubin
measurement
avoid light it’s photosensitive
➡ direct sunlight may cause up
to 50% decrease in direct
bilirubin within 1 hour.
theophylline and propranolol
drugs may cause artificial low
bilirubin
NB➡ bilirubin and creatine
kinase are photosensitive
48

49. transcutaneous
bilirubin meter
49

50. Estimation of bilirubin (diazo reaction)
Depend on the diazo reaction
،‫أنابيب‬ ‫أربع‬ ‫نحضر‬
٢
‫لل‬ ‫منهم‬
Total bilirubin
‫لل‬ ‫منهم‬ ‫واحدة‬
sample
‫لل‬ ‫واألخري‬
blank
‫و‬
٢
‫لل‬
Direct bilirubin
‫لل‬ ‫منهم‬ ‫واحدة‬
sample
‫لل‬ ‫واألخري‬
blank
‫ملحوظة‬
:
We made the sample blank as bilirubin itself has color in serum, we should subtract this color
from change in color produced by reactions.
50

51. ‫حاجتين‬ ‫نعرف‬ ‫عاوزين‬
:
‫عاوزين‬
‫نعرف‬
‫حاجتين‬
:
‫األولي‬
:
‫إن‬
‫الفرق‬
‫بين‬
Direct and indirect bilirubin :
Direct bilirubin ( cholebilirubin ) is water-soluble, react directly with diazo reagent
but indirect bilirubin ( hemobilirubin ) binds to albumin, it can't react with diazo reagent
until putting an accelerator as methanol or caffeine, dimethyl sulfoxide,..... that breaks
the bond between indirect bilirubin and albumin ➡ allow it to react with diazo reagent➡
So it reacts with diazo reagent indirectly.
Estimation of bilirubin
51

52. ‫الثانية‬
:
‫إن‬
‫الفرق‬
‫بين‬
Sample and sample blank
‫إن‬
‫في‬
‫ال‬
In the Sample we allow the reaction of diazo with bilirubin to occurs by
adding Na nitrites
‫إنما‬
‫في‬
‫ال‬
Sample blank not allow the reaction to occurs
‫فقط‬
‫بشوف‬
‫لون‬
‫ال‬
Serum and reagent
Estimation of bilirubin
52

53. ‫أنابيب‬ ‫األربع‬ ‫أحضرنا‬ ‫ما‬ ‫بعد‬
‫ال‬ ‫نضع‬
serum
‫وهحط‬ ،‫أنابيب‬ ‫األربع‬ ‫في‬
Diazo reagent ( sulfinilic acid ) ( R1)
‫أنابيب‬ ‫األربع‬ ‫في‬
‫ال‬ ‫نضع‬
(R2 ) Na nitrite
‫ال‬ ‫أنبوبتي‬ ‫في‬ ‫فقط‬
sample
‫سواء‬
Total or direct
‫ال‬ ‫وليس‬
blank
Na nitrite is placed only on sample tubes, either total or direct, and not
blank
‫ال‬ ‫في‬ ‫يحدث‬ ‫التفاعل‬ ‫يقوم‬
sample
‫في‬ ‫يحدث‬ ‫وال‬
blank
Estimation of bilirubin
53

54. Diazo reagent ( sulfinilic acid + Na nitrite ) + bilirubin ➡ azobilirubin ( red color at acidic pH ).
‫ال‬ ‫وهحط‬
accelerator ( R3 )
‫ال‬ ‫أنبوبتي‬ ‫في‬ ‫فقط‬
Total bilirubin
‫كانت‬ ‫سواء‬
Sample or blank
The accelerator is placed only on total bilirubin tubes either sample or blank
Accelerator breaks the bond between indirect bilirubin and albumin allow indirect bilirubin to react with the
diazo reagent.
‫سواء‬ ‫البيلوربين‬ ‫كل‬ ‫هقيس‬ ‫كده‬ ‫يبقي‬
Direct and indirect ( total bilirubin)
‫فيه‬ ‫إن‬ ‫أخيره‬ ‫حاجة‬ ‫فيه‬
Alkaline tartrate ( R4 ) in the reagent, reacts with azo bilirubin and provides alkaline pH to give blue-green
color ( intense the color of diazo reagent)
‫في‬ ‫بنعملها‬ ‫ودي‬
Total bilirubin
Estimation of bilirubin
54

55. 1- put the serum specimen in the 4 tubes
2- put diazo reagent ( sulfinilic acid )( R1) in the 4 tubes
3- put Na nitrite ( R2 ) in sample tubes not blank to allow the
reaction to occurs
4- put accelerator ( R3) in the total bilirubin tubes only to allow
indirect bilirubin to react with diazo reagent, and put saline instead
of it in direct bilirubin tubes
55

56. Direct bilirubin :
Mix and incubate 5 minutes at room temperature or as mentioned in the pamphlet, then read
Absorbance of direct bilirubin sample against blank at wavelength 546 nm ( 530 _ 555 nm )
In total bilirubin :
Mix and incubate 10 minutes then
5 _ add alkaline tartrate ( R4 ), Alkaline tartrate reacts with azo bilirubin and provide alkaline pH
to give blue-green color ( intense the color of diazo reagent)
Mix and incubate 5 minutes at room temperature or as mentioned in the pamphlet , then read
Absorbance of total bilirubin sample against blank at wavelength 578 nm ( 560 _ 600 nm )
Then multiply this Absorbance in specific factor mentioned in the pamphlet
EX ➡ Total bilirubin ( mg/ dl ) = Absorbance X 10.8
Direct bilirubin ( mg/ dl ) = Absorbance X 14.4
Estimation of bilirubin
56

57. REFERENCES
• _ https://labtestsonline.org
• _ https://www.medscape.com
• _ https://www.wikipedia.org
• _ https ps://www.labcorp.com
• _ https://www.uptodate.com
• _ https://www.ncbi.nlm.nih.gov Home - PubMed – NCBI
• _TIETZ textbook of clinical chemistry and molecular diagnostics, sixth edition 2018.
• _Essential of clinical pathology book; 1st edition; Shirish M Kawthalkar; 2010.
• _Essential of biochemistry book ;1st edition; 2012.
• _ Harper's illustrated biochemistry 30th edition 2015.
• _ Lippincott's illustrated review of biochemistry sixth edition 2014.
• _ Lecture Notes Clinical Biochemistry, 9th Edition Walker, Simon, 2103.
• _Many audios and videos from Well-known, trusted professors who study from accredited
books.
• _ Clinical chemistry from principles to practice 2nd Edition dr Ola H. Demerdash, second edition
57

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