LIVER FUCTION TEST (LFT)

LIVER FUCTION TEST (LFT)

Liver function test is done to examine the healthy status of the liver, the test is mostly requested for those suspects of liver disease, or those infected with Hepatitis virus, mostly Hepatitis B and C.

PARAMETER FOR THE ANALYSIS

1.      Total Bilirubin           (TB)

2.      Conjugated Blirubin  (CB)

3.      Total protein               (TP)

4.      Albumin                     (AL)

5.      Globulin                     (GL)

ESTIMATION OF BILIRUBIN USING VAN DEN BERGH DIAZO REAGENT

REAGENT

1.      SULPHANILIC ACID REAGENT

COMPOSITION

Sulphanilic acid                                   5.0g

Hydrochloric acid concentration         15ml

Distilled water                                     1litre

PREPARATION

Dissolved 5.0g of Sulphanilic acid in 600ml warm distilled water, when dissolved, add 15ml hydrochloric acid concentration and make up the solution to 1litre with distilled water.

2.      SODIUM NITRATE SOLUTION

COMPOSITION

Sodium nitrate                      5.0g

Distilled water                      100ml 

PREPARATION

Dissolved 5.0g of Sodium nitrate in 80ml warm distilled water, and make up the solution to 100ml with distilled water.

When store at 4ºC the reagent is active for 2 weeks.

3.      DIAZO REAGENT SOLUTION

COMPOSITION

Reagent 1                            10.0ml

Reagent 2                            0.25ml

PREPARATION

Mix 10ml of reagent 1 with 0.25ml of reagent 2. The reagent is stable for 3 hour so it must be prepared for every use.

4.      CAFFEINE REAGENT SOLUTION

COMPOSITION

Sodium benzoate                                 75g

Caffeine                                              50g

Sodium acetate anhydrous                 75g

EDTA                                                  1g   

Distilled water                                     1litre

PREPARATION

Dissolved 75g of sodium benzoate, 50g caffeine, 57g sodium acetate anhydrous, and 1g EDTA in 600ml of distilled water, and make up the solution to 100ml with distilled water

5.      ALKALINE TARTARATE

COMPOSITION

Sodium hydroxide                                100g

Sodium potassium tartarate                  350g

Distilled water                                      1litre

PREPARATION

Dissolved 100g of sodium hydroxide, and 350g of sodium potassium tartarate, in 750ml of distilled water, and make up the solution to 1litre with distilled water

6.      ASCOBIC ACID SOLUTION

COMPOSITION

Ascorbic acid                                  0.02g

Distilled water                                5ml

PREPARATION

Dissolved 0.20g of ascorbic acid, in 5ml of distilled water

Make freshly for use every day

7.      STANDARD BILIRUBIN SOLUTION (200µmol/l)

COMPOSITION

Hepatits antigen free non icteric human serum

Pure bilirubin                                                            11.7mg

Dimethylsulphoxide                                                 4ml

PREPARATION

Dissolved 11.7mg of pure bilirubins, and in 4ml of Dimethylsulphoxide by warming and 35ºC, shake the solution at interval until total dissolved.

Make up the volume to 100ml with serum.

Distribute 1ml amounts in small vials or containers and store in deep freeze until required.

NB: There are commercially available of this reagent in the store selling laboratory reagent.

METHOD

Set up four tubes as shown bellow for total bilirubin (TB);

REAGENT	TOTAL BILIRUBIN	BLANK	STANDARD	STANDARD BLANK
Serum	0.2ml
Standard 200µmol/l			0.2ml
Distilled water	0.8ml	0.8ml	0.8ml	0.8ml
Diazo reagent	0.5ml		0.5ml
caffeine	2.0ml		2.0ml

   Leave at room temperature for 10 minute and continue as shown bellow