CARBOHYDRATES-Part II

BILLY AFRICA RMT, MPH

 IMPAIRED PLASMA GLUCOSE/
IMPAIRED GLUCOSE METABOLISM

 Impaired glucose metab. refers to a condition

wherein the glucose homeostasis is disturbed, &
the serum glucose levels are not high enough to
be classified as diabetes.



 Syndrome X

 Cluster of condition that occurs

together
 1. Central obesity
 2. High BP
 3. ____________
 4. High triglyceride
 5. Low HDL Cholesterol
 TEST USED TO DIAGNOSE
DIABETES
 Fasting Blood Sugar (FBS)
 The test should be performed after an 8 hour
fast

 Criteria of fasting plasma glucose:

 1. Nondiabetic = ______ mg/dl
 2. Impaired plasma glucose = 100-125 mg/dl
 3. DM= _____ mg/dl
 Two-Hour Post Prandial Blood Sugar
(2HPPBS)
 The test is performed two hours after meal
 Non-DM-
 DM or Hyperglycemia-
Oral Glucose Tolerance Test
(OGTT)
 Requirements for OGTT:

1. patient should be ambulatory CHO depletion and

inactivity or bed rest impair glucose tolerance
2. Fasting of __________hours
3. Unrestricted diet of 150 g carbohydrates/day for 3 days
prior to testing to stabilize the synthesis of inducible
glycolytic enzymes.
4. The patient should not smoke and drink alcohol prior to
testing.
5. Glucose load
- 75 grams (WHO standard glucose load)
-100 grams
-1.75 g of glucose/kg body weight
 CATEGORIES OF ORAL GLUCOSE TOLERANCE TEST
 1. Normal/ non-diabetic = 2-hr PG < 140 mg/Dl
 2. Impaired GTT = 2-hr PG -____________ mg/dl
 3. DM = 2-hr PG > 200 mg/dl
 Intravenous Glucose
Tolerance Test (IVGTT)
 0.5 g of glucose/kg body weight (given within 3 minutes)
administered intravenously

INDICATIONS:
a． Those who are unable to tolerate a large carbohydrate load
b． Those with altered gastric physiology
c． Those who has undergone previous operation or surgery in the
intestine
d ． Those with chronic mal-absorption syndrome
 Monitoring Test for Diabetes
Mellitus

 Reliable method in the monitoring of long term
glucose control
 Determined once in 3 months- reflect the ave glucose
level over the previous 2-3 months
 Older red blood cells, IDA__________
 Not suitable for patients with shortened RBC lifespan
disorders_________
 Studies have shown that there is a linear relatioship between
ave. blood glucose and HbA1c
 eAG can be calculated from HbA1c reported value using
equation:
 eAG= 28.7 x HbA1c -46.7

 4%-6% HbA1c = normal glycosylation
 18%-20% HbA1c = prolonged hyperglycemia
 <7% HbA1c = cutoff value set by American Diabetes Association

 Dietary status on the day of the test has no effect on HbA1c

 Specimen: EDTA whole blood
 Methods: electrophoresis, immunoassay, HPLC, affinity
chromatography

 Once in ___ weeks lifespan of ALBUMIN is ___ days
 May be useful for monitoring diabetic individuals with
chronic hemolytic anemia and hemoglobin variant (Hb
S or Hb C) decreases RBC lifespan
 It should not be measured in cases of low plasma
albumin (< 30 g/L)  low fructosamine
 Reference value: ________ umol/L

 Methods: affinity chromatography, HPLC and

Photometric
 1,5-Anhydroglucitol
 determine glycemic control earlier than
HbA1c and Fructosamine
 completely reabsorbed by kidney tubules
 Normal: ______ug/mL
 Hyperglycemia: < 10 ug/mL
 CRITERIA USED TO DIAGNOSE
DIABETES
 HbA1c _____%using Nat’ l Glycohemoglobin
Standardization Program (NGSP) certified method
 A two-hour postload glucose of _____ mg/dL or greater
than in an OGTT.
 FBS level that is greater than or equal to _____ mg/dL
(7.0 mmol/L) on at least 2 occasions
 Two-hour postprandial glucose greater than ____
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)
 Exception to these criteria is the diagnosis of
gestational diabetes, which is a condition that develops
in approximately ___% of all pregnancies.
 Criteria for Testing & Diagnosis of GDM

 International Association of Diabetes &

Pregnancy Study Groups:
 recommends:
 2 Approaches for Diagnosis of GDM
 1. One-Step Approach
 2Hr OGTT using 75g glucose load
at 24-28 weeks’ of gestation
 2 Two Step Approach
 An initial measurement of plasma
glucose at 1Hr post load (50g
glucose load) is performed.
 A plasma glucose value > 140
mg/dL indicate the need to
perform a 3 hour OGTT using a
100 g glucose load.
 CHOICE OF SAMPLE

 Fasting glucose in WB is ____% lower than in serum or plasma

 Venous blood glucose is ____mg/dl lower than capillary blood due
to tissue metabolism; capillary blood glucose is same with arterial
blood glucose
 serum or plasma must be separated from the cells within one
hour to prevent losses of glucose (preferably within 30 minutes)
 WBC & RBC metabolize glucose resulting to decrease value in
clotted, uncentrifuged blood (leukocytosis can lead to excessive
glycolysis)
 @ RT (20-25 *C), glycolysis decreases glucose by 5%-7% hour
(5-10 mg/dl) in normal uncentrifuged coagulated blood
 Ref. Temp(4*C), glucose is metabolized at the rate of about 1-2
mg/dl/hr
 Plasma glucose level increase with age
 CHOICE OF SAMPLE

 NaF: 2mg/ml of blood or iodoacetate –

inhibit glycolysis and prevent most glucose
consumption by RBC (good for 24 hours)

 Stability: Serum>Plasma
 Hypoglycemia

 Result of an imbalance in the rate of

glucose appearance & disappearance from
the circulation.
 Causes: Treatment & Biological factors
 The warning S/Sx of hypoglycemia are all related to
CNS
 65-70 mg/dL:
 50-55%:
 < 70 mg/dL:
Methods of Glucose
Determination
 QUANTITATION OF BLOOD
GLUCOSE
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

a ． FOLIN WU METHOD
cuprous ions + phosphomolybdate  Phosphomolybdic acid or
Phosphomolybdenum blue

b ． NELSON SOMOGYI METHOD

cuprous ions + arsenomolybdate   arsenomolybdic acid or
arsenomolybdic blue
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

c. Neocuproine Method (2,9 dimethyl 1,10 phenantroline hydrochloride)

cuprous ions + neocuproine  cuprous-neocuproine complex
(yellow or yellow orange)
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
I. CHEMICAL METHOD
A. OXIDATION REDUCTION METHODS

2. Hagedorn Jensen)
principle: reduction of a yellow ferricyanide to a
colorless ferrocyanide by glucose (inverse colorimetry)

Ferricyaninde---Ferrocyanide
Yellow--- Colorless
I. CHEMICAL METHOD
B. CONDENSATION METHOD
Ortho-Toluidine (Dubowski) Method
-heating in a concentrated acetic acid solution
-reasonably specific (no known interference)

Glucose + aromatic amines  glacial HAC & Heat glycosylamine +

schiff base(green chromophore)
II. ENZYMATIC METHODS
-acts on glucose but not on other sugars and not on other reducing
substances.
1. GLUCOSE OXIDASE METHOD
Measures the B-D-Glucose
It also measures CSF glucose
a． Colorimetric glucose oxidase method (saifer gernstenfield
method)

Glucose + O2 glucose oxidase gluconic H2O2

H2O2 + chromogenic substance  peroxidase oxidized chromogenic
substance + H2O
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
- 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 + O2 glucos oxidase gluconic acid + H202

H2O2 + C2H5OH  catalase CH3CHO +2H20
H2O2 + 2H + 2I molybdate I2 + 2H2O
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.
 Effective in reducing the rate of development of diabetic
complications.
 Cellular strip impregnated with glucose oxidase, peroxidase, &
chromogen.
 Inborn Errors of Carbohydrate
Metabolism

 Galactosemia – congenital deficiency of one of three

enzymes involved in galactose metabolism
 1. galactose-1-phosphate uridyl transferase
 2. galactokinase
 3. uridine diphosphate galactose-4-epimerase (GALE)
LABORATORY FEATURES:
elevated blood and urine galactose
 Essential Fructosuria – fructokinase deficiency
 : the presence of fructose in urine

 Hereditary Fructose Intolerance – defect in Fructose

1,6-biphosphate aldolase B activity in the liver, kidney,
and intestine.
 Charac. by the inability to convert fructose-1-phosphate &
fructose-1,6-biphosphate intodihydroxyacetone
phosphate,glyceraldehyde-3-phosphate, & glyceraldehydes.
 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
 Glycogen storage disease
 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, VII