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An HPLC Method for the Determination of Adenosine Diphosphate: An Important Marker of Hexokinase Activity in Metabolic Diseases

An HPLC Method for the Determination of Adenosine Diphosphate: An Important Marker of Hexokinase Activity in Metabolic Diseases

Rodrigo Brito Santos, Railmara Pereira da Silva, Felipe Akihiro Melo Otsuka, Danielle de Jesus Trindade, Aline Costa Santos, Humberto Reis Matos

Biomed Chromatogr. 2019 Apr;33(4):e4473.

PMID: 30567013

Abstract:

Hexokinases play a critical role in the cellular uptake and utilization of glucose. As such, they are of fundamental importance to all cells. By catalyzing glucose to produce glucose-6-phosphate, hexokinases control the first irreversible step of glucose metabolism and initiate all major pathways of glucose consumption. Our objective was to develop and validate highly sensitive and selective high-performance liquid chromatography with photodiode array detector (HPLC-PDA) assays allowing the determination of adenosine diphosphate, which was used for the determination of hexokinase activity. Samples were analyzed by HPLC-PDA using a C18 analytical column (250 × 4.6 mm) for chromatographic separation. Optimal detection was achieved based on isocratic elution with a mobile phase consisting of a mixture of sodium phosphate monobasic buffer and methanol. This method met all of the requirements of specificity, sensitivity, linearity, precision, accuracy and stability generally accepted in bioanalytical chemistry and was successfully applied to a study of hexokinase activity in an alloxan-induced diabetic rat model. Determination of hexokinase activity will permit characterization of cellular metabolic state in many diseases, such as cancer and diabetes.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP7558807-B	Sodium phosphate monobasic		7558-80-7	Price

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