Carbohydrates

- one of the biomolecule of our body

Biomolecule - these are the substances
that are very important in diff body process
Biomolecule - these Biomolecules
are the substances that - Carbohydrates
are very important in diff - Proteins
bodies processes
- Lipids
- Nucleic Acid

Among the four biomolecules, the main source of

the energy is the carbohydrates

Next is the Lipids

 CARBOHYDRATES
 Primary source for brain erythrocytes and retinal
cells in human
 Major food source and energy supply of the body
 Stored primarily as liver and muscles glycogen
 Building blocks for many processes of metabolism
 “Central ingredient for life”
 Carbohydrates
Storage form of
carbohydrate is
glycogen
Glycogen is stored in
the muscle and liver
 CARBOHYDRATES

 GENERAL DESCRIPTION: compounds

containing C, H and O
 GENERAL FORMULA: Cx(H20)y
 Contain C=O and –OH functional groups
 Derivatives: phosphates, sulfates and
amines
CHO (carbohydrates)
 Carbohydrates
Classification
 CLASSIFICATION is based on 4 different
properties
 Size of the base carbon chain
 Location of the CO function group
 Number of sugar units
 Stereochemistry of the compound
 CLASSIFICATIONS OF
CARBOHYDRATES:
 Based on the number of carbons in the
molecule (Generic classifications)
 3 carbons compound (Trioses) e.g.
glyceraldehyde
 4 carbons compound (Tetroses)
 5 carbons compound (Pentoses)* (Note: *most
important) because DNA and RNA is composed of pentose
 6 carbons compound (Hexoses)*
 CLASSIFICATIONS OF
CARBOHYDRATES:
 Based on the location of CO functional group
 Hydrates of aldehyde or ketone
 Aldose – functional group: aldehyde
 Ketose – functional group: ketone

 anomeric carbon – carbon in the functional

group
 MODELS TO REPRESENT
CARBOHYDRATES
 FISCHER PROJECTION
 Has the aldehyde or ketone at the top of the
drawing
 Carbons are numbered starting at the
aldehyde or ketone end
 Straight chain (linked in hemiacetal form)
 MODELS TO REPRESENT
CARBOHYDRATES
 HAWORTH PROJECTION
 Cyclic form
 More representative of the actual structure
 Formed when the functional group (ketone or
aldehyde) reacts with an alcohol group on the
same sugar to form a ring (hemiacetal ring)
 CLASSIFICATIONS OF
CARBOHYDRATES:
 Based on the stereoisomers of the compound
 Various spatial arrangements of carbohydrates
of carbohydrate molecule
 Have the same order and types of bonds, but
different spatial arrangements and different
properties
 For each asymmetric carbon there are 2
possible isomers
 Mirror images
 CLASSIFICATIONS OF
CARBOHYDRATES:
 Based on the number of sugar units in the
chain
 Relies on the formation of glycoside bonds that
bridges of oxygen atoms
 Based on Sugar Units

 Monosaccharide
 Simple sugars that cannot be hydrolyzed to a
simpler form
 Small molecules; they are extremely vital to the
proper functioning of living systems
 Most important are the pentoses and hexoses
Example of
monosaccharides

- Glucose - Most
important
- Fructose
- Galactose
 Monosaccharides

 GLUCOSE /blood sugar/dextrose

 Central, pivotal point of carbohydrate
metabolism Simpliest
the body
sugar that can provide energy to

 Use to assess total CHO use by the body when

we measure blood glucose level
To know if the patient has a:
diabetes
hyperglycemic
hypoglycemic
 Monosaccharides
- fruits sugar
 FRUCTOSE - abundant in fruits
 Formed from the glucose and from the
breakdown of the sucrose
 Intermediate in the utilization of the
monosaccharides
 Monosaccharides

 GALACTOSE
 Must be converted to glucose before it can be
used by the body
 Less significant from a metabolic point of view
 GALACTOSEMIA – have difficulty in carrying
out this tranformation
Galactose - came form
lactose
 Disaccharides

 Importance: Nutrients they provide

 Biochemical markers for certain disorders of
CHO metabolism
 Formed on the interaction of group between
2 monosaccharides with the production of
molecule of water
 Ex. of disaccahrides
 example of disaccharies
 1. Sucrose
 2. Lactose
 3. Maltose
 SUCROSE

 Glucose + Fructose
 ‘Common table sugar’
 Best known of the disaccharides
 Obtained from beets, sugar cane
 Provides a major portion of CHO intake for
many individual
 LACTOSE

 Glucose + Galactose
 ‘Milk sugar’
 Found in dairy products such as milk and
cheese
 MALTOSE

 Glucose + Glucose
 Good sources are cereals, wheat and malt
products
 Polysaccharides

 Formed by the linkage of many

monosaccharides units

Hydrolysis
 Polysaccharides -------->10
monosaccharides
 Polysaccharides
 STARCH (glucose molecules)
 Primary CHO in the diet and is found in most plants
 GLYCOGEN
 Storage form of CHO
 Formed from glucose by the liver
 CELLULOSE
 Another polysaccharide in plants
 Not digested by humans, it does not provide bulk for
proper intestinal functioning
 Oligosaccharides

 Chaining of 2-10 sugar units

 Glucose Metabolism

 Glycolysis - Break down of glucose to produce energy

 Gluconeogenesis -other production of glucose from
sources aside from
carbohydrate
- it is the production of glycogen out of
 Glycogenesis the glucose molecule
 Glycogenolysis - it is the production of glucose form the
gylcogen
 Hormones related with
glucose
 Insulin is the primary hormone responsible for the
entry of glucose into the cell. It is synthesized by
the b-cells of islet of Larngerhans in the pancreas.
When these cells detect an increase in body
glucose , they release insulin,
 Increases glycogenesis and glycolysis
 Increases lipogenesis
 Decreases glycogenolysis
 Hormones related with
glucose
 Glucagon is the primary hormone responsible for
increasing glucose levels. It is synthesized by the
alpha cells of islet of Langerhans. When these
cells detect a decrease in body glucose, they
release glycogen.
 Increases glycogenolysis
 Glycogen-glucose
 Increses gluconeogenesis
 Fatty acids-Acetyl CoA- ketone
 Proteins-amino acids
 Other hormones that tend to
increase glucose concentration:
1. Cortisol and corticosteroid
2. Catecholamines
3. Growth hormones
4. Thyroid hormones
5. Adrenocorticod hormone
6. Somatostatin
 Hypoglycemia

 Glucose level below the normal range

 Whipple’s triad – low blood glucose
concentration with typical symptoms
alleviated by glucose administration
 Symptoms of
Hypoglycemia
 Neurogenic – tremors, palpitations, anxiety,
diaphoresis
 Neuroglycopenic – dizzines, tingling, blurred
vision, confusion, behavioral changes
 Classification

 Drug Administration – insulin, alcohol,

salicylates, sulfonamide, pentamidine
 Critical Illnesses – Hepatic failure, sepsis,
renal failure, cardiac failure, malnutrition
 Hormonal deficiency – epinephrine,
glucagon, cortisol, growth hormone
 Endogenous Hyperinsulinism – Pancreatic
beta-cell disorder
 Clasification

 Autoimmune hypoglycemia – insulin ab

 Non-beta cell tumors – leukemia, hepatoma,
pheochromocytoma, lymphoma
 Hypoglycemia of infancy and childhood –
galactosemia, GSD, Reye’s syndrome
 Alimentary (reactive) – post-gastric surgery
 Idiopathic (Functional) – postprandial
hypoglycemia
 Diabetes Mellitus

 Diabetes mellitus is a group of diseases in

which blood glucose levels are elevated due
to deficiency in insulin secretion or due to
abnormal insulin action
 Glucosuria occurs when the plasma glucose
level exceeds 180 mg/dL
 Diabetes
- Obese
- Family history of diabetes
- Membership in a high risk
minority population
Ex. African-American
Hispanic America
Native American
Asian American
- History of GDM (Gestational
diabetes melittus)
- Hypertension
- Low HDL cholesterol (High
density lipoprotein)/ low level of
good cholesterol
- Elevated triglycerides
- History of impared fasting
glucose / impared glucose
tolerance test
 Ketosis

 Excessive synthesis of Acetyl CoA –

obtaining energy from stored fat
 Frequent finding in patients with severe,
uncontrolled DM
 Ratio of β-hydroxybutyric acid to
acetoacetate – 6:1
 Can be reversed by insulin administration
 COMPLICATIONS OF DM
 End-stage renal disease
 Non-traumatic amputation
 New blindness in adults aging 20-74 y/o
 Diabetic neuropathy in 60-70 % of diabetic
patients
 Atherosclerotic disease
 2-4x predisposition to heart and cardiovascular
disease
 Type I Diabetes
- insulin-dependent diabetes mellitus
 IDDM
 Juvenile Onset DM
- thin
 Brittle diabetes
 Ketosis-Prone diabetes

Type 1 / I means bata palang

may problem na sa insulin
 Type I

 Diabetic individuals have insulinopenia

(absolute insulin deficiency)
 80-90% reduction in the volume of the β-cell
is required to induce symptomatic type 1 DM
 Genetic association between type 1 and
HLA DR3 and DR4 – MHC on chromosome
6
 TYPE 1
 Beta-cell destruction
 Absolute insulin defiency
 Autoantibodies
 Islet cell autoantibodies
 Insulin autoantibodies
 Glutamic acid decarboxylase autoantibodies
 Tyrosine phosphatase IA-2 and IA2B
autoantibodies
 Signs and Symptoms

 Polyuria, polydipsia, polyphagia, rapid

weight loss, hyperventilation, mental
confusion, possible loss of consciousness
 Idiopathic type

 No known etiology, strongly inherited, does

not have β-cell autoantibodies and requires
episodic insulin replacement
 Microalbumin

 Early indicator of kidney disorder/glomerular

degeneration
 Microalbuminuria of 50-200mg/24 hrs
(diabetic nephropathy)
 Albumin excretion of >30 mg/g crea-<300
mg/g crea
 Type 2

 NIDDM
 Adult type/Maturity onset
 Stable DM
 Ketosis-resistant DM
 Receptor-deficient DM
 Type 2

 “Geneticist’s nightmare”
 Has milder symptoms but may lead to
hyperosmolar coma – overproduction of
glucose (>500 mg/dL) severe dehydration,
electrolyte imbalance, increased BUN, Crea
 TYPE 2

 Insulin resistance with insulin secretory

defect
 Relative insulin deficiency
 Type 1 Type 2

Pathogenesis β-cell destruction Insulin resistance

Incidence Rate 10-15% 90-95%

Onset Any; childhood/teens Any; over 40 y/o

more on life style and heredity

Risk Factors Genetic, autoimmune

undetected
C-peptide levels Detectable

Pre-diabetes Autoantibodies (+) Autoantibodies (-)

Symptoms develop
Symptomatology Symptoms develop abruptly
gradually/asymptomatic
Common, poorly Very rare to have ketosis but
Ketosis they can develop ketosis
controlled

Medication Insulin Oral agents

 OTHER TYPES OF DM

 Associated with secondary conditions

 Genetic defects of beta-cell function
 Pancreatic diseases
 Drug or chemical induced
 Insulin receptor abnormalities
 Other genetic syndromes
 Other types
 Pancreatic disorders
 Endocrine diorders – Cushing’s syndrome,
pheochromocytoma, acromegaly, hyperthyroidism
 Drugs/chemical inducers of β-cell dysfunction –
dilantin pentamidine
 Impaired insulin action – thiazides, glucocorticoids
 Genetic syndromes – Down syndrome, Klinefelter
syndrome, leprechaunism
 GESTATIONAL DM
- develop among pregnant women

 Glucose intolerance during pregnancy

 Due to metabolic and hormonal changes
 Diabetes
Group of metabolic
disorders
 TEST USED TO DIAGNOSE
DIABETES
 Fasting Blood Sugar (FBS)
 The test should be performed after an 8 hour
fast
 TEST USED TO DIAGNOSE
DIABETES
Criteriaof fasting plasma glucose:
1. nondiabetic = < 100 mg/dL
2. impared plasma glucose = 100-
125 mg/dL
3. DM (for hyperglycemia)= > 126
mg/dL
 TEST USED TO DIAGNOSE
DIABETES
 Two-hour post prandial test (PPBS)
 The test is performed two hours after meal
Purpose:

To challenge pancreas to release insulin

and also the action of insulin to store or
remove the glucose form the blood

Results for the two-hour post prandial:

for diabetic it is more than 140mg/dL

for non diabetic less than 140mg/dL
for hypoglycemic less than 50mg/dL
 Impaired plasma
glucose/glucose metabolism
this
insulin resistant state is also referred to as
"SYNDROME X"

referres to a condition where in glucose

homeostasis is disturpted and the serum glocuse
levels are not high enough to be classified as
diabetes.
 TEST USED TO DIAGNOSE
DIABETES
 Oral Glucose Tolerance Test (OGTT)
 Patient to be tested should ingest at least 150 g of
CHO 3 days prior to testing
 Patient must fast the night before the testing is
performed
 The patient is challenged orally with 75 g glucose
during the test
 Patient should not eat food, drink tea, coffee or
alcohol, vigorously exercise, or smoke cigarettes
during the test
 Venous blood samples collected in gray-top tubes
containing fluoride anticoagulant
 Test used to diagnose
diabetes
Oral Glucose Tolerance Test (OGTT)
The patient is challenge orally with a glucose load during the
test
Patient should not eat food, drink tea, coffee or alcohol,
vigorously exercise, or smoke cigarretes during the test
Venous blood samples collected in gray-top tubes
containing flouride anticoagulant
 Requirements for OGTT:
1. Patient should be ambulatory
2. Fasting i needed between 8-14 hours
3. Unrestricted diet of 150g carbohydrates/day
for 3 days prior to testing ---> is required to
stabilize the synthesis of inducible glycolytic
enzymes.
4. The patient should not smoke and drink
alcohol prior to testing
 Glucose Load for OGTT
Glucose loads
- 75 grams (is the WHO
standard glucose load) for
adults
-100 grams is used for DX
OF GDM
- 1.75g of glucose/kg body
weight is used for dx type 1
diabetes among children
 Categories of oral glucose tolerance test
 1. normal/non-diabetic = 2-hr PG <140mg/dL
 2. impaired GTT = 2-hr PG 140-190mg/dL
 3. DM = 2 - hr PG > 200mg/dL

Conversion factor
0.0555
 Miscellaneous
Intravenous Glucose Tolerance Test
• Here the glucose load is administered through the Vein, directly to
the circulation.
• The glucose load is 0.5 g of glucose/kg body weight (given within
3 minutes)
 Indicators:  d. those with chronic
• a. those who are mal-absorption
unable to tolerate a syndrome
large carbohydrate
load
• b. those with
altered gastric
physioloy
• c. those who has
undergone previous
operation or surgery
Medic Orange/Glucose load- used to
challenge patient for the oral glucose
tolerance test
 TEST USED TO DIAGNOSE
DIABETES
 A 50 g oral glucose load is recommended as basis of initial
diagnosis. If the 1 hour postload glucose level is 140 mg/dL
(7.8 mmol/L), a complete 100 g three-hour oral glucose
tolerance test should be performed
 Gestational diabetes is diagnosed if the woman is at or
exceeds two of the following four plasma glucose levels
during the complete OGTT
 fasting – 105 mg/dL
 one hour – 190 mg/dL
 two hours – 165 mg/dL
 three hours – 145 mg/dL
 CRITERIA USED TO
DIAGNOSE DIABETES
 FBS level that is greater than or equal to 126 mg/dL (7.0
mmol/L) on at least 2 occasions
 Two-hour postprandial glucose greater than 140 mg/dL
(7.8 mmol/L).
 Symptoms of hyperglycemia which include: polyuria,
polyphagia, polydipsia, unexplained weight loss plus a
casual or RBS level of greater than or equal to 200 mg/dL
(11.1 mmol/L)
 A two-hour postload glucose of 200 mg/dL or greater than
in an OGTT.
 Exception to these criteria is the diagnosis of gestational
diabetes, which is a condition that develops in
approximately 4% of all pregnancies.
 Symtoms of hyperglycemia which include:
the 3 Ps or polyuria, polyphagia, polydipsia, also
unexplained weight loss, and RBS level of greater
than or equal to 200 mg/dL (11.1 mmol/L)
 Glycosylated Hemoglobin (HbA1c)
 For glucose monitoring
 Older red cells, IDA > Iron deficency anemia
 Not suitable for patients with shortened RBC lifespan disorders > Low HbA1c
level
 Determined once in 3 months and it reflect the ave glucose level over the
previous 2-3 months
 Uses column chromatography
 Fructosamine
 Once in 3 weeks
 Glycosylated albumin
Glycosylated Hemoglobin
- every 1% change in the HbA1c value, 30-35 mg/dL is added to plasma
glucose
- 3-6 % HbA1c = normal glycosylation
-18-20% HbA1c = prolonged hyperglycemia
- 7 % HbA1c = cuoff value set by American Diabetes Association

- Specimen: EDTA whole blood and involves the lysis rbc

- Methods: electrophoresis, immunoassay, HPLC, and affinity
chromatography
Fructosamine
- Is measured once in 3 weeks > the basis is the
lifespan of albumin which is 19 days
- may be useful for monitoring diabetic individuals with
chronic hemolytic anemia and hemoglobin variant (Hb S
or Hb C)
- It should not be measured in cases of low plasma
albumin (<30g/L)

- Reference value: 205 - 285 umol/L

- Methods of determination is affinity chromatography,
HPLC and bi
 CHOICE OF SAMPLE
 Precautions in sample collection to prevent
glucose utilization by leukocytes (WBC)
 RBC have all the enzymes necessary to
metabolize glucose
 The sample should be kept cool; loss on
standing in a warm room maybe as high as
10 mg/dL (0.6 mmol/L)/ hr
 Choice of sample
 Fasting glucose in WB is 15% lower than in serum
or plasma
 Venous blood glucose is 7mg/dL lower than
capillary blood due to tissue metabolism; capillary
blood glucose is same with arterial blood glucose
 Serum or plasma must be separated form the cells
within 30 minutes
 WBC & RBC metabolize glucose resulting to
decrease value in clotted, uncentrifuged blood
 CHOICE OF SAMPLE

 Serum should be separated from the clot

within 0.5 to 1 hour
 NaF: 2mg/ml of blood or iodoacetate – inhibit
glycolysis and prevent most glucose
consumption bt RBC (good for 24 hours)
 Stability: Serum>Plasma
 Choice of sample
 Leukocytosis can lead to excessive glycolysis
 At Roon temperature (20-25C), glycolysis decreases glucose by 5-7% per hour (5-10mg/dL) in
normal uncentrifuged coagulated blood
 At Refrigerator temp (4C), glucose is metabolized at the rate of about 1-2 mg/dL/hr
 Plasma glucose level increase with age

 NaF: 2mg/ml of blood or iodoacetate - inhibit glycolysis and prevent most glucoe consumption by
RBC (good for 24 hours)

 Stabilty: Serum > Plasma, Because interferance may be absent to serum as compare to plasma
Methods of Glucose
Determination
 Methods of Glucose
Determination
1. Chemical Method
Oxidation Reduction Method
- Alk. Copper Reduction Method
- Alk. Ferric Reduction Method

Condensation method
- Ortho-Toluidine Method or Dubowski Method

2. Enzymatic Method

Glucose Oxidase Method
Hexokinase Method
Glucose Dehydrogenase Method
Cellular Strip (Dextrostics)
 I. CHEMICAL METHOD
 A. OXIDATION REDUCTION METHODS
 1. ALKALINE COPPER REDUCTION METHODS
 PRINCIPLE: reduction of cupric ion to cuprous ions forming
 cuprous oxide in hot alkaline solution by glucose

 Alkaline copper tartrate > glucose& heat > cuprous ions

 d. Benedict's method (modification of Folin Wu)
 -used for the detection and quantitation of reducing substance in body fluids
 like blood and urine
 -use citrate or tartrate as stabilizing agent
 CHEMICAL METHOD
 A. OXIDATION REDUCTION METHODS

 2. ALKALINE FERRIC REDUCTION METHOD (Hagedorn Jensen)

 principle: reduction of a yellow ferricyanide to a colorless
 ferrocyanide by glucose (it is an inverse colorimetry)because instead
 of producing a color, the reaction produced a colorless product.

 Ferricyaninde-›Ferrocyanide
 Yellow-> Colorless
 I. CHEMICAL METHOD
 B. CONDENSATION METHOD
 Ortho-Toluidine (Dubowski) Method
 -heating in a concentrated acetic acid solution
 Glucose + aromatic amines > glacial HAC & Heat > glycosylamine +
 schiff base (green chromophore)
 I. ENZYMATIC METHODS
 -acts on glucose but not on other sugars and other reducing
 substances.
 1.GLUCOSE OXIDASE METHOD
 Measures the B-D-Glucose.
 It also measures SF glucose
 a.Colorimetric glucose oxidase method (safer gernstenfield
 method)
 Glucose + 02 glucose oxidase > gluconic acid + H202
 H202 + chromogenic substance
 > peroxidase -> oxidized chromogenic
 substance + H20
 QUANTITATION OF BLOOD
GLUCOSE
 Folin Wu Method – uses PMA
 NELSON-SOMOGYI METHOD
 Accurate but labor intensive and difficult to automate
 Cu2+ ->Cu+
 Cu+ reduces AMA to molybdenum blue
 QUANTITATION OF BLOOD
GLUCOSE
 Dubowski Method – o-toluidine method
 Most sensitive method
 Uses acetic acid
 O-toluidine is carcinogenic and poisonous
 630 nm

Glucose + O-toluidine -> Glycosylamine

(green)
 QUANTITATION OF BLOOD
GLUCOSE
 Enzymatic Method - very specific to a substrate
 Glucose oxidase method – first enzymatic reaction used
Glucose + O2→gluconic acid + H2O2
H2O2 measurement:
a. Trinder’s method - H2O2 + Organic dye → colored
rxn.
b. Peroxidase Method
H2O2 →H2O + O2
O2 is measured by:
a. Clark electrode
b. Ortho-dianisidine method – initially colorless but
when exposed to O2 becomes orange-brown
 QUANTITATION OF BLOOD
GLUCOSE
 Disadvantage of Glucose Oxidase Reaction
 Glucose oxidase can only measure beta glucose
 2 types of glucose
 Alpha-glucose = 35% cannot be oxidized
 Beta glucose = 65% the only type which can be
oxidized
 II. ENZYMATIC METHODS
 -acts on glucose but not on other sugars and not on other reducing
 substances.
 1.GLUCOSE OXIDASE METHOD
 b. Polarographic glucose oxidase
 The enzymatic conversion of glucose is quantitated by the
 consumption of oxygen on an oxygen-sensing electrode
 It measures rate of oxygen consumption which is proportional to
 glucose concentration.
 Glucose oxidase in the reagent catalyzes the oxidation of glucose by
 oxygen forming hydrogen peroxide.
 The hydrogen peroxide is prevented from re-forming oxygen by adding
 molybdate, iodide, catalase, and ethanol
 II. ENZYMATIC METHODS
 -acts on glucose but not on other sugars and not on other reducing
 substances.
 1.GLUCOSE OXIDASE METHOD
 b. Polarographic glucose oxidase
 Glucose + 02 > glucos oxidase > gluconic acid + H202
 H202 + C2H50H > catalase > CH3CHO +2H20
 H202 + 2H + 21 molybdate > 12 + 2H20
 Enzymatic methods
 HEXOKINASE METHOD
 Fewer substances interfere than other methods
 Generally accepted method for measuring glucose
 Disadvantage: hemolyzed samples can pose problem
because contents from RBCs may interfere with the
stoichiometric relationship between glucose and
NAD(P)H accumulation
 2. HEXOKINASE METHOD
 -most specific glucose method, reference method
 -plasma collected using heparin, EDTA, flouride, oxalate or citrate may be
 used for this test.
 -other samples: urine, CSF, & serous fluid
 a. Glucose + ATP > hexokinase -> glucose-6-phosphate +ADP
 b. G6P + NADP >G6PD-> 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 + NAD > glucose dehydrogenase > D-gluconolactone + NADH

 4. DEXTROSTICS (CELLULAR STRIP)
 Important in establishing correct insulin amount for next dose
 Cellular strip impregnated with glucose oxidase, peroxidase, &
 chromogen
 QUANTITATION OF BLOOD
GLUCOSE
 Hexokinase Method
Glucose + ATP → G6PO4 + ADP
Measurement of G6PO4:
G6PO4 → Phosphogluconic acid + H
NAD →NADH (colored product)
 Measures both alpha and beta glucose
because of MUTAROTASE
 QUANTITATION OF BLOOD
GLUCOSE
 Autoanalyzer method
 Uses alkaline ferricyanide reagent
 FERRICYANIDE METHOD
 Gave falsely high results because several
compounds in serum alse were oxidized by
ferricyanide
 400 nm
 Enzymatic methods

 Spectrophotometric assay
 Disadvantage: This reaction is inhibited by
high concentrations of uric acid, vitamin C,
blirubin, glutathione, creatinine, L-cysteine, L-
dopa, dopamine, methyldopa, and citric acid
 Enzymatic methods

 GLUCOSE DEHYDROGENASE METHOD

 Spectrophotometric assay
 Glucose is reduced to form a chromopore
 Inborn Errors of
Carbohydrate Metabolism
 Galactosemia – congenital deficiency of one of
three enzymes involved in galactose metabolism
 Essential Fructosuria – fructokinase deficiency
 Hereditary Fructose Intolerance – defect in
Fructose 1,6-biphosphate aldolase B activity
 Fructose 1,6-biphosphate deficiency – a defect
in Fructose 1,6-biphosphate results in failure of
hepatic glucose generation
 Glycogen Storage diseases – deficiency of
enzymes involved in the metabolism of glycogen
 Galactosemia
 1. galactose-1-phosphate uridyl transferase
 - 2. galactokinase
 - 3. uridine diphosphate galactose-4-epimerase
(GALE)

 LABORATORY FEATURES:
 elevated blood and urine galactose
Essential Fructosuria -is charac by fructokinase
deficiency
-diagnostic indicator: the presence of fructose in urine

- Hereditary Fructose Intolerance - defect in Fructose 1,6-

biphosphate aldolase B activity in the liver, intestine, and
kidney.

Fructose 1,6-biphosphate deficiency - is a defect in

Fructose 1,6-biphosphate which will results in failure of
hepatic glucose generation by gluconeogenic precursors
such as lactate and glycerol
Glycogen Storage
Diseases
GLYCOGEN STORAGE
DISEASES
 Glycogen storage disease
 1 Deficiency of a specific enzyme involved in the
metablolism
 Von Gierke Disease- most common GSD,
associatedwith
 hyperlipidemia
 - LIVER DAMAGE: types I, III, IV, VI, IX, O
 - MUSCULAR DEFECT: types V, VIl
 Types of GSD Synonyms Enzyme Deficiency
Ia Von Gierke Glucose-6-phosphatase
Ib Glucose-6-phosphatase translocase
II Pompe 1,4-Glucosidase
IIIa Cori-Forbes De brancher (liver and muscles)
IIIb De brancher (liver)
IV Andersen Brancher
V Mc Ardle Muscle phosphorylase
VI Hers Liver phosphorylase
VII Tarui Phosphofructokinase
VIII Adenyl kinase
IXa Phosphorylase Kinase
IX b Phosphorylase (liver and muscle)
X Cyclic-AMP-dependent kinase
XI Fanconi-Bickel Glucose transporter 2
0 Glycogen Synthase
 Coverage of Midterm Exam
 Anticoagulants, Reagents, & chemicals
 Quality Assurance and Quality Control,
Computation
 Carbohydrates

 Goodluck and Godbless…