This document contains a lecture by Dr. Mohammed Bahgat Mohammed Sofyan on bilirubin metabolism. It discusses where bilirubin is produced in the body, how it is transported and conjugated in the liver, and excreted in bile, stool and urine. It also covers causes of increased bilirubin like hemolytic anemia and what happens in conditions like Gilbert's syndrome and Crigler-Najjar syndrome where bilirubin metabolism is impaired. The document contains the lecturer's permission for sharing and explaining his slides to others with his name included.
billirubin production billirubin transport and metabolism, different laboratory methods of billirubin estimation ,normal and abnormal levels of billirubin, different classification and types of jaundice and liver diseses, liver functioning, enterohepatic circulation, billirubin production and degradation, benefits and diseases of abnormal level of billirubin
It is characterized by a yellow appearance of the (1) Skin (2) Mucous membranes and (3) Sclera caused by bilirubin deposition. It is the most specific clinical manifestation of Hepatic dysfunction.
Jaundice is usually present clinically when the plasma bilirubin concentration reaches 2 to 3 mg/dl.
When bilirubin clearance from the Liver to the Intestinal tract is impaired (as in acute hepatitis and bile duct obstruction) it may be accompanied by alcoholic (Gray coloured) stools.Solubility increases in water , soluble conjugated bilirubin leads to Tea coloured urine.
billirubin production billirubin transport and metabolism, different laboratory methods of billirubin estimation ,normal and abnormal levels of billirubin, different classification and types of jaundice and liver diseses, liver functioning, enterohepatic circulation, billirubin production and degradation, benefits and diseases of abnormal level of billirubin
It is characterized by a yellow appearance of the (1) Skin (2) Mucous membranes and (3) Sclera caused by bilirubin deposition. It is the most specific clinical manifestation of Hepatic dysfunction.
Jaundice is usually present clinically when the plasma bilirubin concentration reaches 2 to 3 mg/dl.
When bilirubin clearance from the Liver to the Intestinal tract is impaired (as in acute hepatitis and bile duct obstruction) it may be accompanied by alcoholic (Gray coloured) stools.Solubility increases in water , soluble conjugated bilirubin leads to Tea coloured urine.
simple diagrammatic presentation of heme catabolism. highlighted the steps with explanation. Definition , causes, clinical features and biochemical investigation of various types of jaundice is explained in detail. congenital jaundice is included.
Catabolism of Heme | Jaundice | Hyperbilirubinemiakiransharma204
This PPT contain topics on Catabolism of heme; hyperbilirubinemia and jaundice
Books referred: https://www.amazon.in/Biochemistry-2019-Satyanarayana-Satyanarayana-Author/dp/B07WGHCTKZ/ref=sr_1_1?dchild=1&qid=1592209115&refinements=p_27%3AU+Satyanarayana&s=books&sr=1-1
LIVER FUNCTION TESTS BY DR. PREMJEET KAUR, ASSISTANT PROFESSOR BIOCHEMISTRY Premjeet Kaur
BY THE END OF THIS PRESENTATION YOU WILL BE ABLE TO ANSWER WHAT, WHY, WHICH ABOUT LIVER FUNCTION TESTS , WHAT IS JAUNDICE , METABOLISM OF HEME , FORMATION OF BILE PIGMENTS FROM HEME , TRASFER OF LILIRUBIN FROM BLOOD TO BILE , DETERMINATION OF SERUM BILIRUBIN, RETENTION JAUNDICE , REGURGITATION JAUNDICE ,DETERMINATION OF AMMONIA IN BLOOD ,ANTIPYRINE TEST, SERUM ENZYMES IN LIVER DISEASE, ASSESING EXTENT OF LIVER DAMAGE , DIAGNOSIS OF SUBCLINICAL JAUNDICE , BCG TEST , PLASMA PROTEINS , DETOXIFICATION FUNCTION OF LIVER
simple diagrammatic presentation of heme catabolism. highlighted the steps with explanation. Definition , causes, clinical features and biochemical investigation of various types of jaundice is explained in detail. congenital jaundice is included.
Catabolism of Heme | Jaundice | Hyperbilirubinemiakiransharma204
This PPT contain topics on Catabolism of heme; hyperbilirubinemia and jaundice
Books referred: https://www.amazon.in/Biochemistry-2019-Satyanarayana-Satyanarayana-Author/dp/B07WGHCTKZ/ref=sr_1_1?dchild=1&qid=1592209115&refinements=p_27%3AU+Satyanarayana&s=books&sr=1-1
LIVER FUNCTION TESTS BY DR. PREMJEET KAUR, ASSISTANT PROFESSOR BIOCHEMISTRY Premjeet Kaur
BY THE END OF THIS PRESENTATION YOU WILL BE ABLE TO ANSWER WHAT, WHY, WHICH ABOUT LIVER FUNCTION TESTS , WHAT IS JAUNDICE , METABOLISM OF HEME , FORMATION OF BILE PIGMENTS FROM HEME , TRASFER OF LILIRUBIN FROM BLOOD TO BILE , DETERMINATION OF SERUM BILIRUBIN, RETENTION JAUNDICE , REGURGITATION JAUNDICE ,DETERMINATION OF AMMONIA IN BLOOD ,ANTIPYRINE TEST, SERUM ENZYMES IN LIVER DISEASE, ASSESING EXTENT OF LIVER DAMAGE , DIAGNOSIS OF SUBCLINICAL JAUNDICE , BCG TEST , PLASMA PROTEINS , DETOXIFICATION FUNCTION OF LIVER
Management of child with neonatal jaundiceNEHA MALIK
Newborn jaundice is a yellowing of a baby's skin and eyes. Newborn jaundice is very common and can occur when babies have a high level of bilirubin, a yellow pigment produced during normal breakdown of red blood cells.
follow me on my YouTube channel :- medic o mania
What is the Difference Between Conjugated and Unconjugated Bilirubin?Sumit Sharma
Bilirubin is a biochemical parameter of your liver function test. Your doctor uses this tool to diagnose liver disease.
In the human body, bilirubin is a waste product of your hemoglobin produced during the red blood cells breakdown in the spleen.
Although bilirubin does not have a specific function, it is generally a component of bile juice. It is also called a bile pigment, which is yellow in color.
There are two forms of bilirubin in our blood.
One is unconjugated bilirubin, which forms during the breakdown of red blood cells, and the other is conjugated, which starts during metabolism in the liver.
The unconjugated bilirubin is a catabolic product of red blood cells. This non-conjugated compound is also known as indirect bilirubin.
Some possible reasons or diseases associated with unconjugated hyperbilirubinemia –
1. Hemolytic anemia
2. Neonatal Jaundice
3. Genetic factors
4. Drug induced unconjugated hyperbilirubinemia
On the other hands, Conjugated bilirubin is generally formed after the glucuronidation of the unconjugated bilirubin.
The conjugation process occurs in your liver and converts your unconjugated bilirubin into conjugated bilirubin. This type of glucuronidation compound is also known as direct bilirubin.
In simple words, direct bilirubin means conjugated bilirubin.
The elevated conjugated bilirubin indicates Conjugated hyperbilirubinemia.
This condition is most commonly seen in cholestatic liver disease.
Cholestatic is a medical condition where the flow of bile juice gets reduced or stopped. You may have this cholestatic liver disease in two forms – Intrahepatic and extrahepatic obstruction.
In this article, we will discuss bilirubin, metabolism of bilirubin, normal range of bilirubin, and its clinical relevance. Further, we will also know the difference between conjugated and unconjugated bilirubin.
Medical Education Curriculum :
1- General Need Assessment
2- Target Need Assessment
3- Goals and Objectives
4- Content and Educational Strategy
5- Implementation
6- Assessment and Evaluation
7- Maintainance, Enhancement, and Dissemination
June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
How to Make a Field invisible in Odoo 17Celine George
It is possible to hide or invisible some fields in odoo. Commonly using “invisible” attribute in the field definition to invisible the fields. This slide will show how to make a field invisible in odoo 17.
Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
Unit 8 - Information and Communication Technology (Paper I).pdfThiyagu K
This slides describes the basic concepts of ICT, basics of Email, Emerging Technology and Digital Initiatives in Education. This presentations aligns with the UGC Paper I syllabus.
Acetabularia Information For Class 9 .docxvaibhavrinwa19
Acetabularia acetabulum is a single-celled green alga that in its vegetative state is morphologically differentiated into a basal rhizoid and an axially elongated stalk, which bears whorls of branching hairs. The single diploid nucleus resides in the rhizoid.
Model Attribute Check Company Auto PropertyCeline George
In Odoo, the multi-company feature allows you to manage multiple companies within a single Odoo database instance. Each company can have its own configurations while still sharing common resources such as products, customers, and suppliers.
Biological screening of herbal drugs: Introduction and Need for
Phyto-Pharmacological Screening, New Strategies for evaluating
Natural Products, In vitro evaluation techniques for Antioxidants, Antimicrobial and Anticancer drugs. In vivo evaluation techniques
for Anti-inflammatory, Antiulcer, Anticancer, Wound healing, Antidiabetic, Hepatoprotective, Cardio protective, Diuretics and
Antifertility, Toxicity studies as per OECD guidelines
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
For more information, visit-www.vavaclasses.com
Palestine last event orientationfvgnh .pptxRaedMohamed3
An EFL lesson about the current events in Palestine. It is intended to be for intermediate students who wish to increase their listening skills through a short lesson in power point.
Instructions for Submissions thorugh G- Classroom.pptxJheel Barad
This presentation provides a briefing on how to upload submissions and documents in Google Classroom. It was prepared as part of an orientation for new Sainik School in-service teacher trainees. As a training officer, my goal is to ensure that you are comfortable and proficient with this essential tool for managing assignments and fostering student engagement.
Macroeconomics- Movie Location
This will be used as part of your Personal Professional Portfolio once graded.
Objective:
Prepare a presentation or a paper using research, basic comparative analysis, data organization and application of economic information. You will make an informed assessment of an economic climate outside of the United States to accomplish an entertainment industry objective.
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
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
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
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
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
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
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
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
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
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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
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