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K M Ahern

Publications and source records attributed to K M Ahern.

2 recordsLinked to original sources

An enzymatic method for erythrocyte acetylcholinesterase.

The acetylcholinesterase (EC 3.1.1.7) in 50 microL of a 61-fold dilution of erythrocytes in water hydrolyzes acetylcholine during a timed 20-min reaction at 37 degrees C. The resulting choline is measured by use of choline oxidase coupled to peroxidase, with phenol and aminoantipyrene to give a pink product that absorbs maximally at 500 nm. For calibration, a choline iodide standard is included in each batch of up to 19 samples. Accuracy was assessed by using specific inhibitors and measuring choline in the presence of excess erythrocyte solution. The standard curve for the assay is linear to threefold the normal enzyme activity. Between-batch precision was 0.40 kU/L at a mean of 11.5 kU/L (CV 3.5%), and comparison with an acetylthiocholine procedure (x) gave a good correlation: y = 1.02x - 0.27 kU/L (r = 0.991). Long-term precision (10 months), assessed from three sets of assays of samples from 17 individuals, was 0.71 kU/L at a mean of 11.7 kU/L (CV 6.1%).

Acetylcholine↗

A simple method for determining thiopental in plasma.

A simple rapid HPLC method for the measurement of thiopental is described. Plasma was deproteinised with 50% (v/v) acetonitrile in methanol containing flufenamic acid as an internal standard. Chromatography was carried out on a C-18 mu Bondapak reverse phase column using 220 mL of buffer (1.79 g of ammonium dihydrogen phosphate and 1.75 g of sodium sulfate adjusted to pH 7.9 with ammonium hydroxide) in 300 mL of methanol. Detection at 280 nm was near the absorption maxima of both thiopental and flufenamic acid. The method is linear up to 75 micrograms/mL and is therefore suitable for the measurement of thiopental in plasma from patients sedated by thiopental infusion. The sensitivity can be increased such that single dose studies can be investigated.

Chromatography, High Pressure Liquid↗