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A potential application of this technology is the compliance monitoring of opioid addiction programmes, where methadone is used as a pharmacological treatment for drugs such as heroin. Here, we present the detection and quantification of methadone and 2\u2010ethylidene\u20101,5\u2010dimethyl\u20103,3\u2010diphenylpyrrolidine (EDDP) from water and clinical samples (saliva, urine, and plasma) from opioid dependent participants using pSi SALDI\u2010MS. A one\u2010step solvent phase extraction using chloroform was developed for the detection of methadone from clinical samples for analysis by pSi SALDI\u2010MS. Liquid chromatography\u2010mass spectrometry (LC\u2010MS) was used as a comparative technique for the quantification of methadone from clinical saliva and plasma samples. In all cases, we obtained a good correlation of pSi SALDI\u2010MS and LC\u2010MS results, suggesting that pSi SALDI\u2010MS may be an alternative procedure for high\u2010throughput screening and quantification for application in opioid compliance testing. 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