Hey...chems... Analytical chemistry complete notes for M.sc sem 1 to 4.

1. Body Fluids: Composition Of Body
Fluids And Detection Of Abnormal Levels
Of Glucose ,Creatinine

2. CONTENTS
COMPOSITION OF BODY FLUIDS
TYPES OF BODY FLUIDS.
DETECTION OF ABNORMAL LEVELS OF GLUCOSE.
DETECTION OF ABNORMAL LEVELS OF CREATININE.

3. Composition of body fluids
“Body fluids are the fluids such as blood, lymph, and saliva which are
produced in the body and then either circulated within the body or
secreted outside It.
Blood and lymph are the two most important body fluids in the human
body. Blood comprises plasma, white blood cells, red blood cells, and
platelets.
Lymph is a colorless fluid that circulates inside the lymphatic vessels

4. TYPES OF BODY FLUIDS
1.Blood
THE MAIN COMPONENTS OF BLOOD INCLUDE:
Plasma: Plasma is the liquid component of blood. It is a thick fluid .Plasma is
91% water and the rest of the total volume is made up of ions, proteins,
gases, nutrient molecules and waste products. . Albumin is the major protein
in plasma.
Red Blood Cells: 40% of the blood contains red blood cells. RBCs contain
protein hemoglobin that gives a red colour to the blood.

5. • White Blood Cells: The white blood cells are very few in number. They mainly
protect the body against infections. Many WBCs cross the walls of the vessels
and penetrate other tissues.
• Platelets: These are fewer in number than the red blood cells. The platelets
help in the clotting of blood at the site of a wound.
Lymph
Lymph is a colourless fluid present in the interstitial tissues. It circulates
throughout the lymphatic system.. The exchange of nutrients, hormones, and
gases occurs through this fluid. It consists of lymphocytes that play a major
function in the immune responses of the body.

6. INTRODUCTION
• Blood sugar, or glucose, is the main sugar found in your blood.
• It comes from the food you eat, and is your body's main source of
energy.
• Diabetes is a disease in which your blood sugar levels are too high.
• Over time, having too much glucose in your blood can cause serious
problems.

7. Normal values of blood glucose :
• Normal fasting level = 60-90mg/100ml of blood
• Glucose level half an hour (after meal) (post prandial) = 120-
150mg/100ml of blood
• In normal healthy individuals the peak glucose level (at any time of
the day) = 60-110 mg/100ml of blood is considered normal.

8. Clinical significance of blood sugar level
• Blood glucose level increases in diabetes mellitus, acute stress,
hyperthyroidism and chronic liver disease.
• Blood glucose level decreases in Addison’s disease, hypothyroidism and
cancer of the pancreas.
• The increase in the blood glucose level is called hyperglycaemia and
decrease in blood glucose level as hypoglycaemia.
• People suffering from diabetes mellitus need to get their blood glucose
tested frequently.

9. Methods used to measure blood glucose
level.
• Although a number of methods are used for glucose determination,
commonly used two methods are discussed here.
• These can be grouped into two categories- chemical and enzymatic.
Chemical method
• Folin-Wu method
• Ortho-Toluidine method
Enzymatic method
• GOD-POD method. (Glucose oxidase method)

10. Chemical method to estimate blood glucose;
 Folin-Wu method:
• It is based on the principle that glucose when heated with an alkaline copper solution, reduces
cupric ions to cuprous ions.
• The cuprous ions are then measured photometrically (colorimetry) by adding phosphomolybdic acid
which gets reduced to molybdenum blue.
• In this method, whole blood is used and the blood glucose value is determined by the intensity of
blue color.
 Ortho-Toluidine method:
• This is an ideal manual method used for its rapidity, sensitivity, accuracy, and relative simplicity.
• It is based on the principle that the aldose sugar i.e. glucose on condensation with ortho-toluidine in
glacial acetic acid gives a green color that can be measured spectrophotometrically.

11. Enzymatic method- GOD-POD method to estimate blood glucose
• Enzymatic methods provide maximum degree of glucose specificity, hence are
very good in estimating true blood glucose.
• For this method, only blood plasma or serum is used.
• The glucose remains stable for 24 hours if serum or plasma is prepared within 30
minutes after collection.
• The enzyme peroxidase catalyses the following reaction. The hydrogen peroxide
formed reacts with phenol to a red-violet dye as indicator.
• The intensity of the colour formed is measured colorimetrically (or
spectrophotometrically) which is directly proportional to the blood glucose level.
• This test is not influenced by the pressure of uric acid and ascorbic acid

12. WHAT IS CREATININE ..?
Creatinine is a nitrogenous waste product formed from the metabolism of
creatine in skeletal muscle. It has no useful function so excreted in urine
after it has been filtered by the glomerulus from the blood.
. With normal kidney function, then, the amount of creatinine in the
blood remains relatively constant and normal.
For this reason, and because creatinine is affected very little by liver
function, an elevated blood creatinine is a more sensitive indication of
impaired kidney function than the BUN.

13. • A BUN test may be done with a blood creatinine test. Blood urea nitrogen
(BUN) and creatinine tests can be used together to find the BUN-to-
creatinine ratio (BUN: creatinine).
• BUN-to-creatinine ratio 10:1–20:1
• High BUN-to-creatinine ratio occur with sudden (acute) kidney failure. A
blockage in the urinary tract (such as a kidney stone) can cause a high
BUN-to-creatinine ratio.
• A very high BUN-to-creatinine ratio may be caused by bleeding in the
digestive tract or respiratory tract.
• A low BUN-to-creatinine ratio may be caused by a diet low in protein, a
severe muscle injury and others.

14. Normal Value Of Creatinine
Serum creatinine is reported as milligrams of creatinine to a deciliter of
blood (mg/dL) or micromoles of creatinine to a liter of blood
(micromoles/L).
The typical range for serum creatinine is:
For adult men, 0.74 to 1.35 mg/dL (65.4 to 119.3 micromoles/L)
For adult women, 0.59 to 1.04 mg/dL (52.2 to 91.9 micromoles/L)

15. METHOD USED TO MEASURE CREATININE
• The principal of this assay is based on the reaction of creatinine with
sodium picrate as described by Jaffe.
• Creatinine reacts with alkaline picrate forming a red complex. The time
interval chosen for measurements avoids interferences from other serum.
• The intensity of the color formed is proportional to the creatinine
concentration in the sample.

16. REFERENCES
• https://byjus.com/biology/body-fluids-and-circulation/
• https://www.onlinebiologynotes.com/blood-urea-normal-value-clinical-
significance-and-methods-of-estimation/
• https://www.mayoclinic.org/tests-procedures/creatinine-test/about/pac-
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