Intended Use

For IN VITRO diagnostic use in the quantitative determination of β-Hydroxybutyrate in serum using manual or automated applications.

 

Method Principle

Several procedures have been reported in the literature for measuring β-HBA in serum. The Catachem procedure is based on the enzymatic procedure described by David D. Koch and Donald H. Feldbruegge. In this procedure β-Hydroxybutyrate Dehydrogenase (HBDH: EC 1.1.1.30) catalyzes the reaction where β-HBA is converted to acetoacetate with concomitant reduction of NAD to NADH. The increase in absorbance is monitored at 340nm. The delta absorbance produced is directly proportional to the concentration of        β-HBA in the serum sample. The reaction scheme below illustrates the reactions that take place in this β-HBA procedure.

Reagent Storage And Stability

Store the β-HBA Sample Diluent (R1) and the β-HBDH Enzyme Reagent (R2) at 2-8°C. When stored as directed these reagents are stable until expiration date stated on the label.

 

Working Reagent Preparation

Prepare the β-HBA Sample Diluent Working Reagent by diluting the contents of β-HBA Sample Diluent with the volume of distilled or deionized water indicated on the reagent container label. Mix by inversion for about 2-3 minutes until all materials are in solution. Upon reconstitution the Working Reagent is stable for 21 days at 2-8°C.

 

The β-HBDH Enzyme Working Reagent (R2) is ready for use. No preparation is required. Upon opening the Working Reagent (R2) is stable for 30 days at 2-8°C.

 

Method Performance Characteristics

Sensitivity: Using a path length of 1 cm, an D-absorbance of 0.003± 20% per mg/ml should be obtained.

Linearity: In this procedure there is not significant nonlinearity over the range of 0-75mg/dL.

Accuracy

Correlation studies were carried out between this automated β-Hydroxybutyrate method (Y) and a reference automated β-Hydroxybutyrate procedure based on the                           β-Hydroxybutyrate Dehydrogenase reaction (X). Canine serum samples were assayed and the results compared by the least square regression. The following statistics were observed.

TO CONTACT US:

 

CATACHEM, INC.

955 Connecticut Avenue

Suite No. 4106

Bridgeport, CT  06607

 

Phone:     203-335-9277

Fax:         203-335-9977

 

Email:

catachem@catacheminc.com

 

URL:

www.catacheminc.com