BY: MARIA CRISTIN A PAMINTUAN, RMT,

MPH
 C ARBOHYDRATES

• These are hydrates of aldehyde or ketone

derivatives based on the location of the C O
functional group.
• Monosaccharides, oligosaccharides
disaccharides,
polysaccharides – carbohydrates ,

• Glycol aldehyde – is the simplest carbohydrate (CHO)

• Glucose is the only carbohydrate to be directly used
for energy or stored as glycogen.
Reducing Aldoses Ketoses
Sugar

R H CH2OH

C=O C=O C=O

(CHOH)n (CHOH)n (CHOH)n

CH2OH CH2OH CH2OH

Aldoses Ketoses

CHO
CH2OH

(CHOH)4 C=O

CH2OH (CHOH)3

Glucose
CH2OH

Fruct
ose
 IMPORTANCE OF CARBOHYDRATES
• Provides a significant fraction of energy in
the diet of most organism
• Provides the storage form of energy in the
body
• Serves as cell membrane components
• Ribose and deoxyribose sugars form part of
the structural framework of RNA and DNA

• EMPIRICAL FORMULA- Cx(H2O)y

 Glucose, maltose, fructose, lactose and galactose –
reducing substances/sugars.
 The presence of a double bond and a negative charge in
the enol anion makes glucose an active reducing substance.
 Sucrose - most common non reducing sugar.
 Nonreducing sugar do not contain an active ketone
or
aldehyde group.
 The brain is completely dependent on blood glucose for
energy production – 2/3 of glucose utilization in resting adults
occurs in the central nervous system (CNS)
CLASSIFICATION
AND
NOMENCLATURE
• M O N O SACCHARIDES- simple
sugars
GE N ER IC N A M E S E X AMP L E S

3 CARBONS-triose GLYCERALDEHYDE

4 CARBONS-tetrose ERYTHROSE

5 CARBONS- pentose RIBOSE, DEOXYRIBOSE

6 CARBONS- hexose FRUCTOSE, GLUCOSE

7 CARBONS- heptose SEDOHEPTULOSE

9 CARBONS- nonose NEURAMIDIC ACID

GLYCOSIDE
S
• Monosaccharides can be linked by glycosidic linkage to form larger
stucture

. Disaccharides – 2 monosaccharides joined by glycosidic

linkage

Ex 1. Sucrose- glucose and fructose

2. Maltose- 2 glucose

3.Lactose- glucose and galactose

. Oligosaccharides- 3-12 monosaccharides

.Polysaccharides

Homopolysaccharides- Ex. Glycogen

HETEROPO LYSACC HARIDES- Ex.

Glycosaminoglycans

CELLULO SE

STARCH
DIETARY SOURCES

• MONOSACCHARIDES-
GLUCOSE- fruits, sweet corn, corn syrup and
honey
sucrose in
FRUCTOSE- found together wit free glucose honey
and and fruit

• DISACCHARIDES
SUCROSE- Ordinary table sugar abundant in molasses and maple
syrup LAC TO SE- principal sugar found in milk
MALTOSE- In beer
Regulation of Blood Glucose Levels
1. The liver plays a major role in the regulation
of blood glucose level.

2. The liver accomplish this major task by :

A. The release of absorbed glucose for the
cells’ immediate energy needs: GLYCOLYSIS
glucose lactate CO2 + H2O + Energy
B.Conversion of non-carbohydrate substances
such as amino acids, fatty acids and glycerol
to glucose: GLUCONEOGENESIS

C.Conversion of excess glucose into glycogen for

storage: GLYCOGENESIS

D. Breaking down of stored glycogen to glucose:

GLYCOGENOLYSIS
HORMONES SECRETED BY THE ENDOCRINE
PANCREAS THAT REGULATE GLUCOSE
HOMEOSTASIS
Pancreas
-Functions as both an endocrine and exocrine organ in the control of
carbohydrates metabolism.
- As an endocrine gland, it produces and secretes the hormones insulin,
glucagon and somatostatin from different cells residing in the islets of
langerhans in the pancreas.

Insulin
• The primary hormone responsible for the entry of glucose into the cell.
• It is synthesized by the β-cells of the islets of langerhans in the pancreas.
• It is normally released when glucose levels are high.
• It is the only hormone that decreases glucose levels – hypoglycaemic agent.
• Stored from sources such as liver, fat and muscle.
• Has a reciprocal relationship with glucagon.
• It promotes glycogenesis, lypogenesis, and glycolysis, decrease
glycogenolysis
Glucagon
• Is the primary hormone responsible for
increasing glucose – hyperglycemic agent.
• It is synthesized by the α-cells of the islets of
langerhans in the pancreas.
• It is released during stress and fasting states.
• It enhances catabolic functions during fasting
period; promotes glycogenolysis.
 Other hormones that tend to increase glucose
concentrations
1.) Cortisol and corticosteroids (glucocorticoids)
 Secreted by the cells of the zona fasciculata and zona reticularisof the
adrenal cortex.
 Decreases intestinal entry of glucose into the cell.
 It promotes gluconeogenesis and lypolysis.

2.) Catecholamines
 It is released from the chromaffin cells of the adrenal medulla.
 Inhibits insulin secretion and promotes glycogenolysis and lypolysis.

3.) Growth hormone (somatotrophic)

 Secreted by the anterior pituitary gland.
 Decreases entry of glucose into the cell.
 It promotes glycogenolysis and glycolysis.
4.)Thyroid hormones
 It promotes glycogenolysis, gluconeogenesis and
intestinal adsorption of glucose.

5.)Adrenocorticotropic hormone (ACTH)

 It stimulates release of cortisol from the adrenal
cortex.
 It promotes glycogenolysis and gluconeogenesis.

6.)Somatostatin
 It is produced by the delta cells of the islets of
langerhans in the pancreas.
 It primarily inhibits the action of insulin and
glucagon.
 Clinical Conditions of Carbohydrates
Metabolism:
1. Hyperglycemia
 Is an increase in blood glucose levels.
 Causes: stress, severe infection, dehydration or pregnancy,
pancreatectomy, hermochromatosis, insulin deficiency or abnormal
insulin receptor.
 FBS level = > 126 mg/dL.
 All adults older than 45 years should have a measurement of FBS every
3 years unless the individual is diabetic.
Laboratory findings in hyperglycemia:
1) Increase glucose in plasma and urine
2) Increase urine specific gravity
3) ketones in serum and urine
4) Decrease blood and urine pH (acidosis).
5) Electrolyte imbalance. (Decrease sodium and Bicarbonate;
increase
Potassium)
2. Hypoglycemia
 It results from an imbalance between glucose utilization and production.
 Involves decreased glucose levels and can have many causes.
 The warning signs and symptoms of hypoglycemia are related to central
nervous system.
 50mg/dL (2.8-3.0 mmol/L) – observable symptoms of hypoglycemia occur
< 50mg/dL - diagnostic hypoglycemia value.
 A diagnosis of hypoglycemia should not be made unless a patient meets
the criteria of Whipple’s triad – low blood glucose concentration with
typical symptoms alleviated by glucose administration.
 Diagnostic test-5 hour glucose tolerance test, (hypoglycemic dip is often
not seen until after 3 hours)
Symptoms of hypoglycemia:
1) Neurogenic – tremors, palpitations, anxiety, diaphoresis
2) Neuroglycopenic – dizziness, tingling, blurred vision, confusion,
behavioural changes.
• Classification:
1. Drug administration – insulin, alcohol, salicylates,
sulphonamides, pentamidine
2. Critical illnesses – hepatic failure, sepsis, renal
failure,
cardiac failure, malnutrition
3. Hormonal deficiency – epinephrine, glucagons, cortisol,
growth hormone
4. Endogenous hyperinsulinism – pancreatic beta cell
disorders
5. Autoimmune hypoglycemia – insulin antibodies
6. Non-beta cell tumors – leukemia, hepatoma,
pheochromocytoma, lymphoma
7. Hypoglycemia of infancy and childhood –
galactosemia,
GSD, Reye’s syndrome.
8. Alimentary (reactive) hypoglycemia – post
Diabetes Mellitus (DM)
- Is a group of metabolic disorders characterized
by hyperglycemia resulting from defects in
insulin secretion, insulin receptors or both.
- Fasting plasma glucose concentrations >
126mg/dL (7mmol/L) on more than one
testing are diagnostic or diabetes.
- Glucosuria occurs when the plasma glucose
level exceeds 18.3mg/dL (9.99 mmol/L) with
normal renal function.
DM:
- Ketosis develops in DM from excessive synthesis of
acetyl-CoA, as the body attempts to obtain
required energy from stored fat in the absence of
an adequate supply of carbohydrate metabolites
– the presence of ketone bodies is a frequent
finding in individuals with severe, uncontrolled
diabetes.
- In severe DM, the ratio of β-hydroxybutyrate to
acetoacetate is 6:1.
- The entire process of ketosis can be reserved by
insulin administration.
 Criteria for the Diagnosis of DM
1. Obesity
2. Family history of diabetes in a first degree
relative
3. Membership in a high risk minority
population (African, American, Hispanic
Americans, Native American, Asian
American)
4. History of GDM or delivering baby >9
lbs
5. Hypertension (>140/90)
6. Low HDL(<35mg/dL)
7. Elevated triglycerides (>250 mg/dL)
8.History of impaired glucose/ impaired glucose
tolerance.
 Classification of Diabetes:
A.Type 1 Diabetes
Formerly known as: Insulin Dependent Diabetes Mellitus (IIDM)
Juvenile Onset Diabetes Mellitus
Brittle Diabetes
Ketosis-prone Diabetes
 Is a result of cellular-mediated autoimmune destruction of the β-cells of
the pancreas.
 Diabetic individuals have insulinopenia (absolute insulin deficiency) due to
loss of pancreatic β-cells, and depend on insulin to sustain life and prevent
ketosis.
 80-90% reduction in the volume of the β-cells is required to induce
symptomatic typw 1 diabetes.
 There is genetic association between type 1 diabetes and HLA DR3
and
DR4 – the major locus is the major histocompatibility complex on
chromosome number 6.
 Individuals at greater risks of developing this type of diabetes have high
titers of multiple autoantibodies (IAA).
• Microalbuminuria of 50-200mg/24 hours (diabetic
nephropathy) – small changes in the blood filtering
parts of the kidneys allow very small amount of
albumin to leak through usually as a result of having
diabetes.
• Signs and Symptoms: polydipsia,
polyphagia,
polyuria, rapid weight loss, hyperventilation, mental
confusion and possible loss of consciousness.
• Complications: nephropathy, neurophaty and
retinophaty – microvascular disorders.

• Idiopathic type 1 DM – is a form of type 1 diabetes

that has no known etiology; it is strongly inherited; it
does not have β-cell autoantibodies and have episodic
requirements for insulin replacement.
B. Type 2 Diabetes
• Formerly known as:
– Non-insulin Dependent Diabetes Mellitus
- Adult type/maturity Onset Diabetes Mellitus
- Stable Diabetes
- Ketosis-Resistant Diabetes
- Receptor-Deficient Diabetes Mellitus
 Is characterized by hyperglycemia due to an
individual’s resistance to insulin; there is relative
insulin deficiency.
 It is associated with strong genetic predisposition
and not related to an autoimmune disease.
- It has been described as a geneticist’s nightmare.
- The individuals are at risk of developing
macrovascular and microvascular complications.
- It has milder symptoms as compared to type 1,
however, untreated type 2 DM will result to
nonketotic hyperosmolar coma – overproduction
of glucose (>500mg/dL) severe dehydration,
electrolyte imbalance and increased BUN and
creatinine.

• Risk factors: obesity, family history, advanced age,

hypertension, lack of exercise GDM, impaired
glucose metabolism.
 TYPE 1 diabetes TYPE 2 Diabetes

Pathogenesis β-cells destruction Insulin resitance

Incidence rate 10-15% 90-95%

Onset Any; childhood/teens Any; over 40 years old

Genetics, obesity,
Risk Factors Genetic, autoimmune
lifestyle
Decreased or
C-peptide levels undetectable Detectable

Pre-diabetes Autoantibodies (+) Autoantibodies (-)

Symptoms develop
Symptoms gradually (some patients
Symptomatology develop Asymptomatic)
abruptly

Common; poorly
Ketosis controlled Rare

Medication Insulin absolute Oral Agents

C.) Other specific types of Diabetes
1. Pancreatic disorders

2. Endocrine Disorders – Cushing’s syndrome,

Klinefelter’s syndrome, Rabson-Mandengall
syndrome, Leprechaunism
D.) Gestational Diabetes Mellitus (GDM)
 A disorder characterized by impaired ability to
metabolize carbohydrate usually caused by a deficiency
of insulin, metabolic or hormonal changes, occurring in
pregnancy and disappearing after delivery but, in some
cases, returning years later.
 Glucose intolerance with onset or first recognition
during pregnancy (diabetic women who become
pregnant are not included in this category).
 Screening should be performed between 24 and 28
weeks of gestation (1-hr glucose Challenge Test – 50g
glucose load).
 A plasma glucose concentration of 140mg/dL or
greater requires a full diagnostic glucose
tolerance test (3-hour GTT with 100g glucose).

• OGTT Results:
FBS - ≥95 mg/dL
1-hour - ≥180
2- mg/dL
hour - ≥155
3-hour mg/dL
 GDM is diagnosed
- if 2≥140mg/dL
plasma values or more of
the above glucose levels are exceeded.
 GDM is diagnosed if 2 plasma values or more
of the above glucose levels are exceeded.
 Infants born to diabetic mothers are at
an
increased risk for distress
respiratory syndrome, and
hypocalcemia
hyperbilirubinemia.
 After giving birth, women with GDM
should be evaluated 6 to 12 weeks
postpartum.
 GDM converts to DM within 10 years in 30%-
40% of cases.
E.) Impaired Fasting Glucose
- It is characterized by fasting blood
glucose concentrations between normal and
diabetic values.
F.) Impaired glucose Tolerance
- It is by blood
characterized
fasting glucose concentrations less
than for the diagnosis of diabetes, but
required those
the
OGTT is between normal and diabetic values.
 Inborn error of Carbohydrates
Metabolism:
1. Galactosemia
 It is a congenital deficiency of one of three enzymes involved in
galactose metabolism.
 It is cause by failure to thrive syndrome in infant.
 3 enzymes: galactose-1`-phosphate uridy 1 transferase (most common
deficiency), galactokinase (GALK) and uridine diphosphate galactose-4-
epimirase (GALE).
 Galactose-1-phosphate urydil transferase converts galactose-1-
phosphate to glucose.

Laboratory features: elevated blood and urine galactose.

Clinical features: jauncide, hepatomegally, easy bruisability,
galactosuria,
E. Coli, sepsis, cataract,hypotonia and sensory neural deafness.
Diagnostic test: Erythrocyte, Galactose-1-PO₄ uridyl transferase
activity.
2. Essential Fructosuria
• An autosomal recessive disorder characterized by
fructokinase deficiency.
• Fructokinase catalyzes the conversion of fructose to
fructose-1-phosphate.
• Diagnostic indicator: the presence of fructose in urine
(fructosuria)

3. Hereditary fructose intolerance

• A defect of fructose 1-6-biphosphate aldolase B activity in
the liver, kidney and intestine.
• Inability to convert fructose-1-phosphate and fructose-1,6-
biphosphate into dihydroxyacetone phosphate,
glyceraldehydes-3-phosphate and gluceraldehydes.
• Clinical features: irritability, lethargy, seizures and
hepatomegaly.
4. Fructose-1,6-biphosphate deficiency
• A defect in fructose-1,6-biphosphate results in
failure of hepatic glucose generation by
gluconeogenic precursors such as lactate and
glycerol.
• Clinical features: hypoglycemia, lactic acidosis,
convulsions and coma.
5. Glycogen Storage Disease

• Deficiency of a specific enzyme involve in the

metabolism of glycogen.
• It is an inherited autosomal recessive trait.
• Several blood specimens are collected for
2 hours at 15 minutes interval for testing.
• Von Gierke disease – most GSD,
common associated with
lyperlipidemia.
• Liver Damage – types I, III, IV, IV, VI, IX, 0
• Muscular Defect – types V. VII
• Other GSDs: deficiencies of LDH,
PK, phosphoglycerate kinase and mutase
• Intravenous galactose tolerance test – test
 ENZYME
TYPES of GSD SYNONYMS DEFFICIENCY CLINICAL FEATURES
(Tissues Affected)

Hepatomegaly,
Glucose-6- retarded growth
Ia Von Gierke phosphatase Seizures

Glucose-6- Sames as Ib;

Ib phosphatase recurrent bacterial
translocase Infections

Cardiomegaly,
II Pompe 1,4-Glucosidase infantile death

Hepatomegaly,
De Brancher muscle weakness,
IIIa Cori; Forbes (liver and muscle) retarded growth
cardiomyopathy

Same as IIIa except

IIIb De brancher (liver) muscle weakness
 Muscle Myoglubinoria,
V Mc ardle
Phosphorylase muscle cramps

Liver Hepatomegaly,
VI Hers
Phosphorylase hypoglycemia

Phosphofructokin Pain and stiffness

VII Tarui
ase on exertion

Urinary excretion
VIII Adenyl kinase of cathecolamines

Hepatomegaly,
Phosphorylase hypoglycemia,de
IXa kinase (liver) l ay
In motor
development
 Hepatomegaly,
Phosphorylase retarded growth,
IXb (liver and muscle) muscle hypotonia

Cyclic AMP-
X dependent kinase Hepatomegaly
Glucose Hepatomegaly,
XI Fanconi Bickel Transporter-2 rickets
No
Hepatomegaly;
hypoglycemia
Glycogen Symptoms in
0 Thomson synthase morning; mild
growth
delay
 Glucose methodologies
 Fasting glucose in whole blood is 15% lower than
in serum or plasma.
 Venous blood glucose is 7mg/dL lower
capillary
than due to tissue metabolism;
blood
capillary glucose is same with plasma
blood
glucose.
 Peritoneal glucose same with plasma
 Plasma glucose levels
is increase with age – fasting,
fluid
2 mg/dL/decade; postprandial, 4mg/dL/decade;
glucose.
glucose challenge, 8-13mg/dL/decade.
Specimen Consideration:
 Serum or plasma must be separated from the cells
within one hour to prevent losses of glucose
(preferably within 30 minutes).

 At room temperature (20-25°C), glycolysis decreases

glucose by 5-7%hour (5-10mg/dL) in normal
centrifuged coagulated blood.

 At refrigerated temperature (4°C), glucose is

metabolized at the rate of about 1-2 mg/dL/hr.

 WBC and RBC metabolize glucose resulting to decrease

value in clotted, uncentrifuged blood (leukocytosis can
lead to excessive glycolysis).
 I. Chemical Methods
A. Oxidation Reduction Methods
1. Alkaline Copper Reduction Methods
Principle: Reduction of ions
cupric ions forming cuprous oxide
cuprous top
in alkaline solution by glucose.
hot
Alkaline Copper Tartrate Glucose Cuprous Ions
Heat
A. Follin Wu Method
Cuprous Ions + Phosphomolybdate

Phophomolybdic Acid or
Phosphomolybdenum Blue
B. Nelson Somogyl Method

Cuprous Ions + Arsenomolybdate

Arsenomolybdic Acid or
Arsenomolybdenum Blue
C. Neocuproine Method (2, 9 Dimethyl 1,10
phenantroline hydrochloride)

Cuprous Ions + Neocuproine

Cuprous Neocuproine Complex

(Yellow or yellow orange)
D. Benedict Method (Modification of Folin-Wu)
-Used for the detection and quantitation
of reducing substances in body fluids like
blood and urine.
- Use citrate or tartrate as stabilizing agent.
2.) Alkaline ferric Reduction Method
(Hagedorn Jensen)

- Reduction of a yellow ferrycyanide to a

colorless ferrocyanide by glucose (Inverse
colorimetry).
B. Condensation Method
 Ortho toluidine (Dubowsli Method)

Glucose + Aromatic
Amines

Glacial HAC + heat

glycosylamine + schiff’s base

II. Enzymatic Methods
• Acts on glucose but not on other sugars and not
on other reducing substances.

1.) Glucose Oxidase Method

- Measures the β-D glucose
- It also measures CSF glucose
Disadvantages: This reaction is inhibited by high
concentrations of uric acid, vitamin C, bilirubin, glutathione,
creatinine, L-cysteine, L-dopa, dopamine, methyldopa, and
citric acid
a. Colorimetric glucose Oxidase Method (Saifer
Gernstenfield Method)
glucose oxidase
Glucose + O₂ Gluconic Acid + H₂O₂

H₂O₂ + Chromogenic
Substances
peroxidase

Oxidized Chromogenic Substances + H₂O

b. Polarographic Glucose Oxidase
• The enzymatic conversion of glucose is
quantitated by the consumption of oxygen on
an oxygen-sensing electrode.
• Measure rate of oxygen consumption which is
proportional to glucose concentration.
• Glucose oxidase in the reagent catalyzes the
oxidation of glucose by oxygen under first
order conditions, forming hydrogen peroxide.
• The hydrogen peroxide is prevented from re-
forming oxygen by adding molybdate, iodide,
catalase and ethanol.
Glucose oxidase
Glucose + O₂ Gluconic Acid +
H₂O₂
H₂O₂ + C₂H₅OH catalase
CH₃CHO
H₂O₂ + 2H + 2 I I₂ + 2H₂O
+

2H₂O
2.) Hexokinase Method
- Most specific glucose method,
method.
reference
- Plasma collected using heparin, EDTA,
fluoride, oxalate or citrate may be used for
this test.
– Other Samples: urine, CSF and serous fluids
– Disadvantage: Hemolyzed samples can pose
problem because contents RBC’s may interfere
with the stoichiometric relationship between
glucose and NAD(P)H accumulation.
a.) hexokinase
Glucose + Glucose-6-phosphate + ADP
ATP
b.) Glucose-6-phosphate + NADP

glucose-6-phosphate dehydrogenase (G-6-PD)

6-Phosphogluconolactone +
NADPH
3)Glucose Dehydrogenase Method
- The amount of NADH generated is
proportional to the glucose concentration.
- It provides results in close agreement with
hexokinase procedures.

• Mutarotase is also added to shorten the time

necessary to reach equilibrium.
Glucose is reduced to form a chromophore.
α- D – glucose (mutarotase) β-D-glucose

β-D-glucose+ NAD (Glucose dehydrogenase)

D-gluconolactone + NADH

Chromogen (MTT) + NADH (diaphorase)

reduced chromogen (MTT colored blue)
+ NAD
4)Dextrostics (cellular strip)
-Important is establishing correct
insulin amount for next dose.

-Effective in reducing the rate

of development of diabetic
complications
 Samples for Glucose measurement:
1. RBS - Random Blood Sugar
-requested during insulin shock,
hyperglycemic ketonic coma
2. FBS - Fasting Blood Sugar
-NPO (Non-per Orem) at least 8 hours
before the test
3. 2-HR PPBS - 2 Hour Postprandial Blood
Sugar
4. GTT - glucose tolerance test
- multiple blood sugar test
-to determine how well the body metabolizes
glucose over a given time required.
 Kinds of Glucose Tolerance Tests:
I. Oral Glucose Tolerance Test
a.Janney Isaacson Method (single Dose
Method) – most common.
b.Exton Rose Method (Divided Oral Dose or
Double rose method)
• Added plasma Glucose after OGTT:
30 mins = 30-60mg/dL above fasting
1 hour = 20-50mg/dL above fasting
2 hour = 5-15mg/dL above fasting
3 hour = fasting level or below
b. Intravenous Glucose Tolerance Test (IVGTT)
-Itis used for diabetes patients
with gastrointestinal disorders.
-0.5 g of glucose/kg body weight
(given w/in 3minutes) administered
intravenously.
 Fasting sample is also required.
 The first blood collection is after 5 minutes of IV
glucose.
• Indications (IVGTT):
a. Those who are unable to tolerate a large
carbohydrate load.

b. Those with altered gastric physiology.

c. Those who had undergone previous operation

or surgery in the intestine.

d. Those with chronic melabsorption syndrome.

• Requirements for OGTT:
a.)Patient should be ambulatory
-CHO depletion and inactivity or bed rest impair
glucose tolerance.
b.) Fasting of 8-14 hours.
c.)Unrestricted diet of 150g carbohydrate/day for 3
days prior to testing.
-to stabilize the synthesis of inducible glycolytic
enzymes.
d.) The patient should not smoke and drink
alcohol prior to testing.
e.) Glucose load
- 75gms (WHO standard glucose load)
- 100gmsn
-1.75g of glucos/kg body weight (children).
Procedure for OGTT
• The patient should avoid exercise, eating,
drinking (except water) and smoking during the
test.
• For non pregnant women and adults, only the
fasting and the 2 hour sample may be measured
(or according to the physician’s request).
1. Collect the fasting blood sample (urine may
also be collected)
2. Instruct the patient to drink the glucose load.
3. Collect blood sample after 30 minutes, 1
hour, 2 hour, 3 hour, respectively.
• Criteria of Fasting Plasma Glucose (FPG)

Non-Diabetic (Prediabetes) = <100mg/dL

Impaired Plasma Glucose (PG)= 100-125/mg/dL
Diabetes Mellitus = ≥126 mg/dL
• Categories of Oral Glucose Tolerance Test
Normal/Non-Diabetic = 2-hr plasma glucose
(PG)<140mg/dL
Impaired GTT = 2-hr PG 140-199 mg/dL
Diabetes Mellitus = 2-hr PG ≥ 200 mg/dL

• Diagnostic Criteria for diabetes Mellitus

RBS = ≥200mg/dL (with symptoms of DM)
FBS = ≥126 mg/dL (7.0mmol/L)
2-hr Post glucose Load = ≥200 mg/dL (11.1mmol/L)
5. Glycosylated haemoglobin (HbA)
• Also called glycated hemoglobin
• It is the largest subfraction of normal
hemoglobin A in both diabetic and non-
diabetic individuals.
• It is a glucose molecule attached to one or
both N- terminus valines of the beta
polypeptide chains ofnormal
hemoglobin. adult
• A reliable method in the monitoring of
long- term glucose control.
• It reflects the average blood glucose level over
the previous 2-3 months.
• For every 1% change in the HbA₁c value, 35 mg/dL is
added to plasma glucose.
• Older red blood cells and iron deficiency anemia – high
HbA₁c levels.
• Not suitable for patients with shortened RBC
lifespan disorders – low HbA₁c value.
• Dietary status on the day of the test has no effect on
HbA₁c.
• 3%-6% HbA₁c = normal glycosilation; 18%-20% HbA₁c is
prolonged hyperglycemia.
• 7% HbA₁c = cut off value set by American
Diabetes Association (ADA).
• Specimen: EDTA whole blood.
• Methods: electrophoresis, immunoassay, HPLC, Affinity
Chromatography.
6. Fructosamine
• Also called glycosylated or glycated albumin
plasma protein ketoamine.
• It is a reflection of short term glucose control (2-3
weeks).
• May be useful for monitoring diabetic individuals
with chronic haemolytic anemias and hemoglobin
variants (Hb S Hb C) – decreased RBC lifespan.
• It should not be measured in cases of low plasma
albumin (<30 g/L) – low fructosamine.
• Reference Values: 205-285μmol/L
• Methods: Affinity chromatography, HPLC and
photometric
Notes to Remember
1. CSF Glucose
• It is about to 40-60% of the blood plasma glucose level.
• Any changes in the blood sugar are reflected in the CSF
approximately one hour later because og the lag in CSF
glucose equilibrium time.
• For comparison, a blood glucose specimen should be collected
at least 60 minutes before the lumbar puncture.
• A markedly decreases CSF glucose (<40mg/dL)
&increased WBC count (neutrophils) is seen in bacterial
meningitis.
• Increased levels: diabetes
• Decreased levels: bacterial meningitis, tuberculosis, fungal and
amebic meningitis, subarachnoid hemmorhage; systemiv
hypoglycemia
• Reference values: 40-70 mg/dL (child)
• 60-80 mg/dL (adult)
• Normal CSF to Glucose ratio: < 0.5
2. C-Peptide Test
• C-peptide is formed during the conversion of pro-
insulin to insulin
• The amount of circulating C-peptide provides
reliable indicators for pancreatic and insulin
secretions (β-cell function).
• It can be used to monitor individual responses to
pancreatic surgery.
• This test mainly evaluates hypoglycemia and
continuous assessment of β-cell function
• Specimen: fasting blood (serum)
• Decreased: type 1 DM
• Reference values: 0.90-4.3 ng/mL (CF:0.333)
• Normal ratio of C-peptide: insulin= 5:1 to 15:1
Test for Ketone Bodies
• Normal ratio of β-hydroxybutyrate and
acetoacetic acid 1:1 (0.5-1.0 mmol/L)
• The ratio of β-hydroxybutyrate to acetoacetate is
greatly increased in diabetic ketoacidosis (DKA)
due to the altered redox state and elevated levels
of NADH in the hepatic mitochondria.
• An increased in serum acetone is indicative of a
defect in the metabolism of carbohydrate.
• The entire process of ketosis can be reversed
through restoration of an adequate level of
carbohydrate metabolism.
1. Gerhardt’s ferric chloride test – reacts
only with acetoacetate
2. Nitroprusside test – 10x more sensitive
to acetoacetate than to acetone
3. Acetest tablets – detects acetoacetate
(lesser degree)
4. Ketosis – detects acetoacetate better
than acetone.
5. KetoSite Assay – detects β-
hydroxybutyrate; not widely used