CLINICAL SIGNIFICANCE
Decreased levels are observed In protein malnutrition, a b-lipoproteinemia, hyperthyroidism and cachectic states. Increased levels are commonly found in hyperlipoproteinemia, nephrotic syndrome, coronary artery disease, hypothyroidism, liver disorders and diabetes mellitus.
PRINCIPLE
Lipase Glycerol +
Triglycerides --- ------® Fatty acids
Glycerol-3-
Glycerol G K phosphate +
ATP ----------® ADP
Glycerol-3 GPO acetone
phosphate + O2--------® phosphate
+H2O2
H2O2 Coloured
ESPT Peroxidase
quinoneimine
Amino-4 --------® +H2O
antipyrine
COMPOSITION
Final concentration In the reactive medium
Pipes buffer >20mMol/L
4-Aminophenazone > 0.8 mMol/L
Adenosine-3-phosphate 1.32 mMol/L
Glycerol Kinase 0.04U/ML
Glycerol-3-phosphate oxidase > 4KU
Lipase 350 U/L
Peroxidase > 0.8 KU/L
Magnesium chloride 5 mMol/L
ESPT 2 mMol/L
Surfactant 1 g/L
Stabilizer______________________0.5 g/L
REAGENTS SUPPLIED
| Cat. No. LS 385 |
|
3x10mL |
| Reagent 1 |
10 mL |
3 Bottles |
| Standard (200 mg/dL) |
2 mL |
1
Bottle |
| Cat. No. LS 390 |
|
10x10mL |
| Reagent 1 |
10 mL |
10 Bottles |
| Standard (200 mg/dL) |
3 mL |
1 Bottle |
| Cat. No. LS 391 |
|
4x50mL |
| Reagent 1 |
50 mL |
4 Bottles |
| Standard (200mg/dl) |
3mL |
1 Bottle |
Preparation of Working Reagent
All reagent are ready to use.
STABILITY
The reagent is stable up to the expiry as
mentioned on the kit label.
A light yellow colour of blank does not affect
test performance.
SAMPLE
Sample can be serum or plasma which has no
sign of haemolysis. Triglycerides are affected by
food intake prior to the sample collection.
Therefore, keep the patient fasting for at least 8
hours prior to the sample collection. Common
anticoagulants have no interference on this assay. |
METHOD FOR AUTO ANALYSERS
Reagent...............................1000µL
Sample or Standard..................10µL
Mix well. Incubate at 37°C for 10 minutes. Read at 546 nM (540-560 nM) against Blank. The final colour is stable for at least 30 minutes away from bright light.
NOTE: Programme the analyser using system parameters. A specific programme data sheet may be provided for each analyser upon request.
MANUAL METHOD
1.
| Pipette into 3 Test Tubes |
BLANK |
STANDARD |
TEST |
| Distilled Water |
0.05 |
- |
- |
| Standard |
- |
0.05 |
- |
| Sample |
- |
- |
0.05 |
| Working Reagent |
1.00 |
1.00 |
1.00 |
2.Mix well. Incubate at 37 °C for 10 minutes.
3.Add 2mL distilled water, for a 3mL optical cuvette. Read at 546 nM (540-560 nM) or GREEN filter against Blank. The final colour is stable for at least 30 minutes, away from bright light.
SYSTEM PARAMETERS
| Reaction |
= |
End-point |
| Temperature |
= |
37°C |
| Wavelength |
= |
546nm |
| Standard Concentration |
= |
200 mgldL |
| Absorbance Range |
= |
0-2° A |
| Cuvette Path Length |
= |
1 cM |
| Reaction Time |
= |
10 Mins |
| Linearity |
= |
1000mg/dl |
| Max. limit of Blank Reagent |
= |
0.15 |
| Final Colour Stability |
= |
30 Mins |
| Reagent Volume |
= |
1000 µL |
| Sample Volume |
= |
10 µL |
RESULTS CALCULATION
ΔO.D.Test
Triglycerides in mGs/dL = ---------------- x 200
ΔO.D.STD
ΔO.D.Test
Triglycerides in mGs/dL = --------------- x 2.28
ΔO.D.STD
EXPECTED VALUES
40 -165 mGs/dL (0.4 to 1.86 mMol/L)
As with all diagnostic methods, the final diagnosis should not be made on the result of a single test as well as laboratory diagnosis must be confirmed with clinical manifestations.
LIMITATIONS
This reagent is highly specific to various compounds Invitro addition or contamination of glycerine, Hydroger peroxide, Hypochlorate. Formaldehyde or any such compound totally alters the results. Use of contaminatec pippet or water leads to elevated blank absorbance. This method Is linear up to 1000 mGs/dL For values higher than 1000 mGs/dL dilute the sample with 0.9% Sodium Chloride. Repeat the assay and multiply results by dilution factor i.e. by 2 for 1:1 dilution.
QUALITY CONTROL
To ensure adequate quality control, each kit should be tested against a standard control sera. It should be realised that the use of quality control material checks both instrument and reagent function together. Factors which might affect the performance of this test include proper instrument function, temperature control, cleanliness of glasswares and accuracy of pippetting.
It is appropriate to establish each laboratory's accuracy constant and interpret values accordingly. Similarly, laboratory findings should be established by clinical manifestations.
WARNING
This reagent system is for in vitro Diagnostic use only.
This reagent system is containing preservatives and components that have not established for safety if contacted on broken skin or eye or taken orally. In case of such incident wash off with plenty of water, or consult a physician.
BIBLIOGRAPHY
BUCOLO G., DAVID H.-Clin Chem.
1973,19,476
ESDER T.W., MICHRINA C.A.- J of
Biol.Chem. Philadelphia P.A.
|
