Sample Material

Serum, plasma. Triglycerides is reported to be stable in the

sample for 5 days when stored at 2–8°C.

Procedure

Wavelength/filter : 505 nm (Hg 546 nm)/green

Temperature : 37°C/RT

Light path : 1 cm

Pipette into clean dry test tubes labeled as blank (B),

standard (S), and test (T):

Addition

Sequence

B

(mL)

S

(mL)

T

(mL)

Working reagent 1.0 1.0 1.0

Distilled water 0.01

Triglycerides standard (S) - 0.01

Sample - - 0.01

Mix well and incubate at 37°C for 5 minutes. or at RT

(25°C) for 15 minutes. Measure the absorbance of the

standard (Abs. S), and test sample (Abs. T) against the

blank, within 60 minutes.

Calculations

 Abs T

Triglycerides in mg/dL = _ _______ × 200 Abs S

Linearity

This procedure is linear upto 1000 mg/dL. If values exceed

this limit, dilute the serum with normal saline (NaCI 0.9%)

and repeat the assay.

Note

Fasting samples of 12 to 14 hours are preferred. Fatty meals

and alcohol may cause elevated results. Patient should not

drink alcohol for 24 hours before the test.

System Parameters

Reaction : End point Interval : —

Wavelength : 505 nm Sample

volume

: 0.01 mL

Zero setting : Reagent blank Reagent

volume

: 1.00 mL

Incubation

temperature

: 37°C/RT Standard : 200 mg/dL

Incubated

time

: 5 mm/15 min Factor :

Delay time : — React slope : Increasing

Read time : — Linearity : 1000 mg/dL

No. of read : — Units mg/dL

Normal Values

SI units

Adult females

Age 20–29 10–100 mg/dL 0.11–1.13 mmol/L

Age 30–39 10–110 mg/dL 0.11–1.24 mmol/L

Age 40–49 10–122 mg/dL 0.11–1.38 mmol/L

Age 50–59 10–134 mg/dL 0.11–1.51 mmol/L

Age > 59 10–147 mg/dL 0.11–1.66 mmol/L

Adult males

Age 20–29 10–157 mg/dL 0.11–1.77 mmol/L

Age 30–39 10–182 mg/dL 0.11–2.05 mmol/L

Age 40–49 10–193 mg/dL 0.11–2.18 mmol/L

Age 50–59 10–197 mg/dL 0.11–2.22 mmol/L

Contd...

490 Concise Book of Medical Laboratory Technology: Methods and Interpretations Age > 59 10–199 mg/dL 0.11–2.24 mmol/L

Children

Female,

age 1–19 10–121 mg/dL 0.11–1.36 mmol/L

Male,

age 1–19 10–103 mg/dL 0.11–1.16 mmol/L

Note: Plasma values are lower by about 3%.

Classification of Triglyceride Levels

Borderline 200–400 mg/dL 2.26–4.5 mmol/L high

High 400–1000 mg/dL 4.5–11.3 mmol/L

Very high >1000 mg/dL > 11.3 mmol/L

Clinical Implications

1. Increased triglyceride levels are believed to be a factor

in increased risk for atherosclerosis

 A. Increased levels occur in

 1. Types I, IIb, III, IV, and V hyperlipoproteinemias

 2. Liver disease

 3. Nephrotic syndrome

 4. Hypothyroidism

 5. Poorly controlled diabetes

 6. Pancreatitis

 7. Glycogen storage disease

 8. Myocardial infarction (increases may last one

year)

 9. Metabolic disorders related to endocrinopathies.

 B. Many of the clinical conditions that cause an

increase in cholesterol levels also cause increase

in triglycerides

 1. Nephrotic syndrome

 2. Pancreatic dysfunction

 3. Toxemia

 4. Hypothyroidism.

2. Decreased levels occur in malnutrition and congenital

alpha-beta lipoproteinemia.

BLOOD GLUCOSE

Glucose (GOD/POD Method)

(Courtesy: Tulip Group of Companies)

For the determination of glucose in serum, plasma, and

CSF (for in vitro diagnostic use only).

Summary

Glucose is the major carbohydrate present in blood.

Its oxidation in the cells is the source of energy for the

body. Increased levels of glucose are found in diabetes

mellitus, hyperparathryroidism, pancreatitis, renal failure.

Decreased levels are found in insulinoma, hypothyroidism,

hypopituitarism and extensive liver disease.

Principle

Glucose is oxidised to gluconic acid and hydrogen peroxide

in the presence of glucoseoxidase. Hydrogen peroxide

further reacts with phenol and 4-aminoantipyrine by the

catalytic action of peroxidase to form a red colored quinoneimine dye complex. Intensity of the color formed is directly

proportional to the amount of glucose present in the sample.

 Glucose oxidase

Glucose + O2 + H2O Gluconate + H2O2

 Peroxidase

H2O2 + 4 Aminoantipyrine Red + Phenol

quinoneimine

dye + H2O

Normal Reference Values

Serum/plasma (fasting) : 70–110 mg/dL

(2 hours PP) : upto 140 mg/dL

CSF : 50–80 mg/dL

It is recommended that each laboratory establish its

own normal range representing its patient population.

Contents 2 × 150 mL 1000 mL

L1: Glucose Reagent 2 × 150 mL 1000 mL

S: Glucose Standard (100 mg/dL) 5 mL 5 mL

Storage/Stability

Contents are stable at 2–8°C till the expiry mentioned on

the labels. Upon storage the glucose reagent may develop

a slight pink color. This does not affect the performance of

the test.

Reagent Preparation

Reagents are ready to use.

Sample Material

Serum, plasma, CSF. Glucose is reported to be stable in the

sample for 7 days when stored at 2–8°C.

Procedure

Wavelength/filter : 505 nm (Hg 546 nm)/green

Temperature : 37°C/RT

Light path : 1 cm

Pipette into clean dry test tubes labeled as blank (B),

standard (S), and test (T):

Contd...

Clinical Chemistry 491

Addition B S T

Sequence (mL) (mL) (mL)

Glucose reagent (L1) 1.0 1.0 1.0

Distilled water 0.01

Glucose standard(s) - 0.01

Sample - - 0.01

Mix well and incubate at 37°C for 10 minutes or at RT

(25°C) for 30 minutes. Measure the absorbance of the

standard (Abs S), and test sample (Abs T) against the

blank, within 60 minutes.

Calculations

 Abs T

Total glucose in mg/dL = ________ × 100 Abs S

Linearity

This procedure is linear upto 500 mg/dL. If values exceed

this limit, dilute the serum with normal saline (NaCL 0.9%)

and repeat the assay. Calculate the value using the proper

dilution factor.