Radioimmunoassay of nicotine

doi:10.1016/0006-291X(75)90360-5

Biochemical and Biophysical Research Communications

Volume 64, Issue 2, 19 May 1975, Pages 574-580

https://doi.org/10.1016/0006-291X(75)90360-5 Get rights and content

Abstract

A sensitive and specific radioimmunoassay of nicotine was developed using antisera raised against 6-(p-aminobenzamido) nicotine coupled to bovine serum albumin. Inhibition studies with various nicotine analogues revealed that the antisera are highly specific for both the N-methylpyrrolidine ring and the pyridine ring of nicotine, and especially for the structural changes of the former. The use of these antisera in an assay of nicotine in biological fluids is desirable, since the pyrrolidine ring of nicotine is first metabolized in vivo and antibodies must, therefore, discriminate nicotine from other nicotine metabolites.

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Citation Excerpt :
This work includes a series of linkers of different lengths and flexibility that were coupled to the 5- or 6-position of the nicotine molecule and conjugated to keyhole limpet hemocyanin (KLH, Fig. 1). The pyridine ring was chosen for linker attachment in order to increase the antigenicity of the pyrrolidine ring, in which nicotine is first metabolised in the body, and thereby minimising cross-reactivity of generated antibodies with the metabolites of nicotine [22]. Since in tobacco, (S)-nicotine is the natural occurring enantiomer, some immunogens comprising the (S)-enantiomer of nicotine were also prepared and tested.
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A new simple method for the conjugation of nicotine and cotinine to protein has been developed. The pyridine alkaloids were coupled with ethyleneimine to form their aminoethylpyridinium derivatives, which were conjugated to bovine serum albumin (BSA) in the presence of 1-cyclohexyl-3-(2-morpholinoethyl)-carbodiimide metho-p-toluenesulfonate. About forty molecules of S-nicotine and thirty-five of S-cotinine per molecule of starting BSA were respectively recognized to be present in the conjugates by determining the liberated aminoethylpyridinium compounds after hydrolysis.
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Effects of cigarette smoking on dopamine β-hydroxylase activity (DBH), cyclic AMP, cyclic GMP and nicotine in plasma and urine during water diuresis were examined in 19 healthy men. After smoking, plasma DBH decreased and plasma cyclic AMP increased significantly, followed by the increases in urinary cyclic AMP and nicotine excretion. No significant change in both plasma and urinary cGMP was observed.
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Radioimmunoassay of nicotine

Abstract

Biochem. Biophys. Res. Comm

Anal. Chem

Z. Anal. Chem