Improved enzymatic hydrolysis of hair

https://doi.org/10.1016/0379-0738(93)90271-BGet rights and content

Abstract

An enzymatic hair extraction method is proposed for drug analysis. Pronase digestion of various aliquots of hair from a cocaine abuser was preceded by a 2-h incubation with a dithiothreitol solution. The extraction solution was tested to identify possible interferences in the radioimmunoassay and was compared with other hydrolysis methods to assess the results of extraction.

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Cited by (25)

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    2013, Journal of Chromatography A
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    A second set of experiments were conducted to observe the effect of DTT during the enzymatic hydrolysis of hair. DTT is commonly used to enhance the enzymatic activity of Pronase E. Its action consists in breaking down certain disulphide bonds of the keratin matrix, allowing a faster action of the enzyme [50]. Fig. 1(b) shows results in triplicate obtained for MSPD-EH experiments and elution with 5 mL of a solution containing 2.0 mg mL−1 Pronase E and 12.5 mg mL−1 DTT in 1.4 M/1.4 M Tris/HCl, pH 7.3 (MSPD-EH with DTT), or with 5 mL of 2.0 mg mL−1 Pronase E in 1.4 M/1.4 M Tris/HCl, pH 7.3 (MSPD-EH without DTT).

  • Matrix solid-phase dispersion on column clean-up/pre-concentration as a novel approach for fast isolation of abuse drugs from human hair

    2010, Journal of Chromatography A
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    Reviews on this topic show numerous extraction treatments mainly based on methanol, aqueous acids or buffer solutions as extracting solvents, alkaline digestions with aqueous sodium hydroxide, or enzymatic digestions (hydrolysis) [1,11,12]. Methods based on acidic hydrolysis have been reported to offer high yields for cocaine, opiates and their metabolites [13], although enzymatic hydrolysis are also recommended, mainly because the moderate pH and temperature conditions inherent to these procedures [14–19]. In addition, the possibility of speeding up the enzymatic hydrolysis procedures by using ultrasound irradiation has offered important practical advantages for shortening the whole analytical procedure [20].

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