CHEMISTRY REAGENT KITS----->TRIGLYCERIDES
                                                  (GPO - PAP)End-Point
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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 37C 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.