For Urine as Sample

Pipette into a clean dry test tube labeled as Test (T):

Addition

Sequence

(T)

37°C

Amylase reagent (L1) 1.0 mL

Sample 0.01 mL

Mix well and read the initial absorbance A0 after 1

minute and repeat the absorbance reading after every 1, 2,

and 3 minutes. Calculate the mean absorbance change per

minute (∆A/min).

Calculations

α Amylase activity in U/L (Serum) = ∆A/min × 3954

α Amylase activity in U/L (Urine) = ∆A/min × 7830

Linearity

The procedure is linear up to 1000 U/L at 37°C. If the

absorbance change (∆A/min) exceeds 0.300, use only the

value of the first 2 minutes to calculate the result, or dilute

the sample 1 + 9 with normal saline (NaCL 0.9%) and

repeat the assay (Results × 10).

Note

Anticoagulants like oxalate and EDTA bind Calcium which

is needed for α Amylase activity and should not be used.

Heparin may be used. Saliva and sweat contain α Amylase.

Avoid contamination of reagent and sample during use.

For users to convert the values obtained by this method

to the EPS substrate methods, multiply the results obtained

by 2.45.

System Parameters

Reaction : Kinetic Interval : 60 sec.

Wavelength : 405 nm Sample vol : 0.02 mL (s)/

0.01 mL (u)

Zero setting : Distilled water Reagent vol : 1.00 mL

Incubation

temperature

: 37°C Standard :

Incubation

time

: — Factor : 3954/7830

Delay time : 60 sec Factor : Increasing

Read time : 180 sec Linearity : 1000 U/L

No. of read : 4 Units : U/L

Normal Values

Normal values urine

Mayoclonic method 10–80 Amylase U/h

Somogyi method 26–950 U/24 h

Beckman method 1–17 U/h

Normal values serum SI units

Somogyi method 50–180 U/dL 92–330 U/L

Beckman method 20–125 U/L

Over age 70 20–160 U/L

Panic level Three times the upper

limit of normal

Clinical Relevance

Increased levels are found in pronounced elevation

(5 or more times normal)

¾ Acute pancreatitis

¾ Pancreatic pseudocyst

¾ Morphine administration.

Moderate elevation (3 to 5 times normal)

¾ Pancreatic carcinoma affecting head of pancreas (late

manifestation)

¾ Mumps

¾ Salivary gland inflammation

¾ Perforated peptic ulcer (sometimes)

¾ Acute exacerbation of chronic pancreatitis

¾ Partial gastrectomy

¾ Obstruction of pancreatic duct

¾ Alcohol poisoning

¾ Acute cholecystitis

¾ Intestinal obstruction with strangulation

¾ Ruptured tubal ectopic

¾ Ruptured aortic aneurysm.

Decreased Levels are found in

¾ Acute pancreatitis subsidence

¾ Hepatitis

¾ Cirrhosis of liver

¾ Toxemia of pregnancy

¾ Severe burns

¾ Severe thyrotoxicosis.

LIPASE

Lipase Serum (Turbidimetric Method)

(Courstesy: Randox)

Intended Use

For the quantitative in vitro determination of lipase in

serum and plasma. This product is suitable for manual use.

Enzymology 521

Clinical Significance

A lipase test system is a device intended to measure the

activity of the enzyme lipase in serum and plasma. Lipase

measurements are used in the diagnosis and treatment of

diseases of the pancreas such as acute pancreatitis and

obstruction of the pancreatic duct.

Turbidimetric Method

Principle

 Lipase

Triolein + 2H2O Monoglyceride + 2 oleic acid

The decrease in turbidity is measured at 340 nm.

Sample

Serum or heparinized plasma.

Lipase is stable in the sample for 5 days at +2 to +8°C or

24 hours at +20° to +25°C.

Reagent Composition

Contents Concentrations in the test

1. Buffer

Tris buffer 26 mmol/L, pH 8.9

2. Substrate

Sodium deoxycholate 16.7 mmol/L

Calcium chloride 0.04 mmol/L

Triolein 0.3 mmol/L

Colipase 4 mg/L

3. Standard See assigned value on lot specific insert

Safety Precautions and Warnings

For in vitro diagnostic use only. Do not pipette by mouth.

Exercise the normal precautions required for handling

laboratory reagents.

Solution 1 contains sodium azide. Avoid ingestion or

contact with skin or mucous membranes. In case of skin

contact, flush affected area with copious amounts of water.

In case of contact with eyes or if ingested, seek immediate

medical attention.