ACTIVITY No.

5 Blood Glucose Test

Human placenta -Increase glucose ( glycolysis)

Carbohydrates
Placental - Insulin antagonist
 Primary source of energy for brain, Lactogen
erythrocyte, and retinal cells in human Somatostatin Delta cell -Increase plasma glucose by
 Major food source and energy supply of the inhibiting insulin ,glucagon
body and are stored primarily as liver and muscle glycogen
,growth hormone & other
 Provide structural integrity to the cell membrane
endocrine hormone
Source of CHO Epinephrine Adrenal -Increase glucose ( glycolysis)
Sources of CHO
medulla -stimulates glycogenolysi
Carbohydrates are found in a wide array of both healthy T3 and T4 Thyroid -Increase glucose
and unhealthy foods—fruit, vegetables, bread, beans, glands (gluconeogenesis )
milk, popcorn, potatoes, cookies, spaghetti, soft drinks, -stimulates glycogenolysis
corn, and cherry pie. They also come in a variety of -Promote intestinal absorption of
forms. The most common and abundant forms are glucose
sugars, fibers, and starches
CLINICAL CONDITIONS OF CARBOHYDRATE
Forms of CHO METABOLISM
2 Forms of Carbohydrates:
a. sugars (such as fructose, glucose & lactose)
b. starches 1. HYPERGLYCEMIA
-An abnormally high level of glucose concentration in the blood
Role of Pancreas  FBS level: ≥126 mg/dL (>7.0 mmol/L), occurs when
After a meal, the blood sugar (glucose) level rises as the body does not produce enough insulin.
carbohydrate is digested. This signals the beta cells of
the pancreas to release insulin into the bloodstream. Causes: stress, severe infection, dehydration or pregnancy,
Insulin helps glucose enter the body's cells to be used pancreatectomy, hemochromatosis, insulin deficiency or
for energy. abnormal insulin receptor

SIGN and SYMPTOMS

ORGANS RESPONSIBLE IN CONTROLLING ü Frequent urination
GLUCOSE CONCENTRATION: ü Increased thirst
Liver ü Blurred vision
ü Fatigue
Pancreas
ü Headache
Endocrine glands ü Tiredness
Hormone Source Action
Insulin Beta -Decrease serum glucose 1. HYPORGLYCEMIA
cells ( glycolysis)
-Stimulates glucose uptake by -Characterized by low or decreased plasma glucose level
cells of 2.5 mmol/L
- Increase Glycogenesis,
lipogenesis -It results from imbalance between glucose utilization
-Decrease glycogenolysi and production
Glucagon Alpha -Increase serum glucose(
cells glycogenolysis) The warning signs and symptoms are related to CNS:
-Increase gluconeogenesis o Nervousness
,lipogenesis o Sweating
ACTH Anterior -Increase glucose ( glycolysis) o Intense hunger
pituitary -Insulin antagonist o Dizzy
-Gluconeogenesis o Trembling
Growth Anterior -Increase glucose ( glycolysis) o Weakness
Hormone pituitary -Insulin antagonist o Palpitations
-Acromegaly=hyperglycemia o Often have trouble speaking
Cortisol Adrenal -Increase glucose ( o Pale
cortex glycogenolysis) o Trouble concentrating
-Stimulates gluconeogenesis
(glucose from non
CHO sources)
-lipolysis
 ACTIVITY No. 5 Blood Glucose Test

HYPOGLYCEMIA- WHIPPLE’S TRIAD Type 2 diabetes relative insulin deficiency to

predominantly an insulin secretory defect with insulin
resistance) Adult- onset, Maturity- onset DM, Stable
Hypoglycemia attack precipitated by fasting Diabetes,
Ketosis- resistant Diabetes, Receptor Deficient DM
Plasma glucose level of <45 mg/dL
 accounts for 80–90%
 individuals do not need insulin treatment to
Symptoms relieved promptly by survive.
administration of glucose  Age of onset 35 insidious or gradual
 Acute complication Hyperosmolar coma
 Responsive to oral hypoglycemic drugs
DIABETES MELLITUS
Gestational Diabetes
a group of metabolic disease characterized by
Disorder characterized by impaired ability to
hyperglycemia resulting from defects in insulin
metabolize CHO usually caused by insulin
secretion or due to abnormal insulin action or both. deficiency, metabolic or hormonal changes
The chronic hyperglycemia of diabetes is associated
with long-term damage, dysfunction, and failure of occurs during pregnancy and disappears after
various organs, especially the eyes, kidneys, nerves, delivery, in some cases, after 5-10 years it
heart, and blood vessels. develops into Type II DM

Screening should be performed between 24 to 28

COMPLICATIONS OF DM weeks of gestation

The goal of screening is to reduce maternal and

The chronic complications of diabetes are broadly fetal complications such as preeclampsia,
divided into microvascular and macrovascular, with cesarean delivery, congenital malformations,
the former having much higher prevalence than the jaundice, and infant death
latter.
a. Microvascular complications include nephropathy, LABORATORY TESTS
neuropathy and retinopathy
b. Macrovascular complications consist of Diagnosis of hyperglycemia and hypoglycemia
cardiovascular disease, stroke, and peripheral artery includes:
disease (PAD). 1. Fasting blood glucose
2. 3-Hours Oral glucose tolerance test (OGTT)
3. Two-hours post-prandial glucose test
THREE CLASSIC MANIFESTATION OF DM 4. Hemoglobin A1c test.
1. Polyuria
excessive urination
2. Polydipsia
excessive thirst
3. Polyphagia
excessive hunger

CLASSIFICATION OF DIABETES MELLITUS

Type 1 diabetes (β-cell destruction) Immune-mediated
diabetes. Juvenile onset, Brittle diabetes, Ketosis-
prone DM
 form of diabetes which accounts for 10-20%
prevalence,
 results from a cellular-mediated autoimmune
destruction of the β-cells of the pancreas
usually leading to absolute insulin deficiency.
 Age of onset usually Childhood/Puberty
 Acute complication Ketoacidosis
 Unresponsive to oral hypoglycemic drugs
 ACTIVITY No. 5 Blood Glucose Test

FASTING BLOOD GLUCOSE (FBG) 3-Hours Oral Glucose Tolerance Test Normal
 The patient has not eaten or taken in reference range
calories for 8 -10 hours prior to
testing FBS < 95 mg/dL (5.3 mmol/L)
 A fasting blood sample is drawn from One-hours < 180 mg/dL (10 mmol/L)
the patient. Two hours < 155 mg/dL (8.6 mmol/L)
 Allowed to clot and centrifuged. Three hours < 140 mg/dL (7.8 mmol/L)
 The serum or plasma should be
separated from clotted blood 2-hours postprandial glucose
 Glucose analysis is done on the
 blood sample taken from the patient 2-hours
serum or plasma free from hemolysis.
after eating for diagnosis of diabetes.
 The fasting glucose level indicates  Postprandial tests are most reliable if the
the glucose level at the specific time patient is tested following a standard glucose
the specimen is collected. dose (50 or 75gms) – ADA
Fasting Blood Glucose (FBG) Normal Reference How to perform 2-HOURS POSTPRANDIAL
Range GLUCOSE
Normal : <100 mg/dL (<5.5 mmol/L)
Prediabetic/impaired Plasma Glucose : 100-125 1. Patient must fast for 8-10 hrs. then eat a meal with
mg/dL (5.5-6.9 mmol/L) at least 75 gms of carbohydrates
Diabetic : ≥126 mg/dL (>7.0 mmol/L) diabetes
2. Two hours after the meal blood sample will be
drawn for glucose analysis.
3-HOURS ORAL GLUCOSE TOLERANCE TEST
(OGTT) Modification method for 2-HPPBS
= gold standard for the diagnosis of type 2 diabetes. 1. Patient must fast for 8-10 hrs
= used during pregnancy for diagnosing gestational 2. A standard glucose dose (50 or 75 gms) will be
diabetes. consumed rather than a
= the person fasts overnight (at least 8 hours, but random meal.
not more than 16 hours). the next morning, the 3. Two hours after blood sample will be drawn for
fasting plasma glucose is tested. glucose analysis.
= the person receives a dose of oral glucose 2-HOUR POSTPRANDIAL GLUCOSE Reference
= Blood samples are taken up to four times at Normal range
different time points after consumption of the sugar Normal/Non-diabetic 2- hr plasma glucose : < 140
to measure the blood glucose. mg/dL
How to perform 3-HOURS ORAL GLUCOSE Impaired GTT 2- hrs PPG : 140-199 mg/dL
TOLERANCE TEST (OGTT) Diabetes mellitus 2- hrs PPG : > 200 mg/dL
1. The patient should consume a normal diet or one
that contains approximately 150 grams of
carbohydrates per day.
2. The patient should arrive fasting for a minimum of
eight hours before the test If the patient does not
arrive fasting, the test should be rescheduled.
3. The fasting sample should be taken and the time
point should be noted.
4. The patient should then consume the correct
amount of glucose (established by weight, up to 75
grams) over a maximum of a 5-minute period.
5. Additional blood samples should be taken at 1
hour, 2 hours and 3 hours
6. The blood samples should be processed for
analysis.
 ACTIVITY No. 5 Blood Glucose Test

4. HEMOGLOBIN A1C

 glycated hemoglobin
 is hemoglobin with glucose attached.
 evaluates the average amount of glucose in the
blood over the last 2 to 3 months by measuring the
percentage of glycated hemoglobin in the blood.
 reliable method for diagnosing diabetes.

HEMOGLOBIN A1C Normal reference range

HbA1c : > 6.5 %

Blood Sample Collection requirements and considerations

 Standard clinical specimen: fasting serum
 Fasting glucose in whole blood is 15% lower than
in serum/plasma
 Serum is appropriate for glucose analysis when it is F
separated from the cells within 30-60 minutes
 Venous blood glucose is 7 mg/dL lower than
capillary blood due to tissue metabolism; capillary
blood glucose is same with arterial blood glucose.
 Glucose is metabolized at room temperature at a
rate of 7 mg/dL/hr
 At 4 0C glucose decreases by approximately 2
mg/dL/hr
 2 mg of NaF per mL of whole blood prevents
glycolysis for up to 48 hrs.

I. GLUCOSE ENZYMATIC TESTS: FBS, OGTT, 2-HPPBS

Glucose analysis methods use the enzymes glucose

oxidase, hexokinase, or glucose dehydrogenase. These
enzyme tests are simple, quick, and specific for glucose
and have been adapted for use in many types of glucose
analyzers, both large and small. In general, tests that use
hexokinase or glucose dehydrogenase are more specific
and have fewer interferences than those using glucose
oxidase.
 ACTIVITY No. 5 Blood Glucose Test

II. CHEMICAL METHODS- OXIDATION REDUCTION

METHOD

1. ALKALINE COPPER REDUCTION

= oldest method
Principle: Reduction of cupric ions to cuprous ions in the
presence of hot alkaline solution by glucose.
𝑔𝑙𝑢𝑐𝑜𝑠𝑒
Alkaline copper tartrate cuprous ions
ℎ𝑒𝑎𝑡

Folin-Wu :
- uses tungstic acid to remove protein from serum
- measure sugar (FBS)
phosphomolybdic
cuprous ions + phosphomolybdate acid /phosphomolbdenum
blue

Nelson-Somogyi:
- uses Zn (OH)2
- measure true glucose (FBG)
GLUCOSE OXIDASE ENZYMATIC METHOD arsenomolybdic
cuprous ions + arsenomlybdate
acid/arsenomolybden
Conversion factor: mg/dL x 0.0555= mmol/L. um blue

Measuring range (Linearity): Neocuproine method :

The assay is linear between 10mg/dl and 600 mg/dl.
If the results obtained were greater than 600mg/dl, dilute cuprous neocuproine
cuprous ions + neocuproine
the sample with NaCl and multiply the result by 2. complex ( yellow to yellow
orange)

Benedict’s method
used for detection and quantitation of reducing
substance in body fluids

2. ALKALINE FERRIC REDUCTION METHOD

= Hagedorn Jensen Method

= reduction of yellow ferricyanide to a colorless
ferrocyanide by glucose

II CHEMICAL METHODS – CONDENSATION METHOD

O–toluidine Method
= chemical color reaction; most sensitive, but carcinogenic
―Dubowski Reaction‖
𝐻𝐴𝑐
Glucose + O-toluidine 𝑏𝑜𝑖𝑙 Glycosylamine