title: Fast gradient elution reversed-phase liquid chromatography with diode-array detection as a high-throughput screening method for drugs of abuse. II. Data analysis.
abstract: In Part I of this work, we developed a method for the detection of drugs of abuse in biological samples based on fast gradient elution liquid-chromatography coupled with diode array spectroscopic detection (LC-DAD). In this part of the work, we apply the chemometric method of target factor analysis (TFA) to the chromatograms. This algorithm identifies the target compounds present in chromatograms based on a spectral library, resolves nearly co-eluting components, and differentiates between drugs with similar spectra. The ability to resolve highly overlapped peaks using the spectral data afforded by the DAD is what distinguishes the present method from conventional library searching methods. Our library has a mean list length (MLL) of 1.255 and a discriminating power of 0.997 when both retention index and spectral factors are considered. The algorithm compares a library of 47 different compounds of toxicological relevance to unknown samples and identifies which compounds are present based on spectral and retention index matching. The application of a corrected retention index for identification rather than raw retention times compensates for long-term and column-to-column retention time shifts and allows for the use of a single library of spectral and retention data. Training data sets were used to establish the search and identification parameters of the method. A validation data set of 70 chromatograms was used to calculate the sensitivity (correct identification of positives) and specificity (correct identification of negatives) of the method, which were found to be 92% and 94%, respectively.
authors.affiliation: Virginia Commonwealth University, Department of Chemistry, 1001 W. Main Street, Box 842006, Richmond, VA 23294-2006, USA.
authors.forename: Sarah E G
authors.initials: SE
authors.lastname: Porter
authors.forename: Dwight R
authors.initials: DR
authors.lastname: Stoll
authors.forename: Changyub
authors.initials: C
authors.lastname: Paek
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Rutan
authors.forename: Peter W
authors.initials: PW
authors.lastname: Carr
mesh_terms: D000465:Algorithms; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004563:Electrochemistry; D053593:Forensic Toxicology / Q000379:methods; D006801:Humans; D013287:Illicit Drugs / Q000097:blood*; D008962:Models, Theoretical; D015813:Substance Abuse Detection / Q000379:methods*
chemical_list: D013287:Illicit Drugs