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B A Middleton

Publications and source records attributed to B A Middleton.

5 recordsLinked to original sources

Development of an enzyme-linked immunosorbent assay for caffeine.

A competitive enzyme-linked immunosorbent assay suitable for the measurement of caffeine in plasma and serum has been developed. Sheep immunised with an immunogen prepared by coupling 7-(5-carboxypentyl)1,3-dimethylxanthine to egg albumin produced antibodies with little crossreactivity with the metabolites of caffeine. The enzyme label was prepared by coupling 7-(5-carboxypentyl)-1,3-dimethylxanthine to peroxidase using the mixed anhydride method. The assay, which has a sensitivity of 0.01 mumol/l, permits direct measurement of caffeine in plasma and serum samples. 50 plasma samples measured by ELISA and by an established radioimmunoassay showed a correlation of r = 0.97 (P less than 0.001).

Caffeine

The effect of storage temperature and physical state on the performance of antisera in radioimmunoassay after long-term storage.

The performance in radioimmunoassay of four antisera after storage at temperatures ranging from -40 degrees C to room temperature, in three physical states (frozen, liquid or freeze dried) was investigated over a 3-year period. No deterioration in antiserum performance in terms of precision and accuracy of quality control serum measurement or recovery of ligand was apparent under any of the storage conditions studied. Some lowering of titre became apparent in two of the antisera over the study period. Deterioration was most marked when antiserum was stored lyophilised at room temperature. Storage of antiserum frozen confers no advantage over storage at 4 degrees C provided precautions are taken to minimise bacterial contamination when storing antiserum in liquid form.

Humans