Abstract
In recent years, photodiode array detectors (DADs) have been much improved with respect to wavelength accuracy and resolution, sensitivity, linearity and software operation. UV spectra of drugs measured with up-to-date DADs from different manufacturers are in excellent agreement, have the same quality as measured by a conventional UV spectrometer and are highly reproducible. The calculation of similarity parameters by the DAD software includes the entire range of the spectra compared and allows recognition of very small differences. It was shown in a systematic study of more than 2500 toxicologically relevant substances that UV spectra have a very high specificity with respect to substance structure. Therefore, HPLC-DAD in combination with a comprehensive database of UV spectra and retention parameters is one of the most efficient techniques used in systematic toxicological analysis (STA). Furthermore, the method is advantageous for the identification of metabolites, since in many cases they have the same or very similar UV spectra compared with their respective parent substances and their retention times on reversed-phase columns are shifted in a manner typical of the particular biotransformation reaction. Beside these general aspects, practical applications of HPLC-DAD for STA are reviewed with respect to sample preparation and chromatographic conditions. The efficiency of the method is demonstrated for an example of a routine procedure using liquid-liquid extraction with CH2Cl2 and protein precipitation for sample preparation, a system of three isocratic mobile phases with different acetonitrile/phosphate buffer ratios and RP8 columns for chromatography and a database of 2682 UV spectra and relative retention times for substance identification.
References
1 Bogusz M, Wu M. Standardized HPLC/DAD system, based on retention indices and spectral library, applicable for systematic toxicological screening. J Anal Toxicol 1991; 15:188–97.10.1093/jat/15.4.188Search in Google Scholar
2 Bogusz M, Erkens M. Reversed-phase high-performance liquid chromatographic database of retention indices and UV spectra of toxicologically relevant substances and its interlaboratory use. J Chromatogr A 1994; 674:97–126.10.1016/0021-9673(94)85220-0Search in Google Scholar
3 Elliott SP, Hale KA. Applications of an HPLC-DAD drug-screening system based on retention indices and UV spectra. J Anal Toxicol 1998; 22:279–89.10.1093/jat/22.4.279Search in Google Scholar
4 Gaillard Y, Pépin G. Use of high-performance liquid chromatography with photodiode-array UV detection for the creation of a 600-compound library. Application to forensic toxicology. J Chromatogr A 1997; 763:49–63.Search in Google Scholar
5 Koves EM. Use of high-performance liquid chromatography-diode array detection in forensic toxicology. J Chromatogr A 1995; 692:103–19.10.1016/0021-9673(94)00840-6Search in Google Scholar
6 Lai C-K, Lee T, Au K-M, Chan AYW. Uniform solid-phase extraction procedure for toxicological drug screening in serum and urine by HPLC with photodiode-array detection. Clin Chem 1997; 43:312–25.10.1093/clinchem/43.2.312Search in Google Scholar
7 Lambert WE, Meyer E, De Leenheer AP. Systematic toxicological analysis of basic drugs by gradient elution of an alumina-based HPLC packing material under alkaline conditions. J Anal Toxicol, 1995; 19:73–8.10.1093/jat/19.2.73Search in Google Scholar PubMed
8 Logan BK, Stafford DT, Tebbett IR, Moore CM. Rapid screening for 100 basic drugs and metabolites in urine using cation exchange solid-phase extraction and high-performance liquid chromatography with diode array detection. J Anal Toxicol 1990; 14:154–9.10.1093/jat/14.3.154Search in Google Scholar PubMed
9 Maier RD, Bogusz M. Identification power of a standardized HPLC-DAD system for systematic toxicological analysis. J Anal Toxicol 1995; 19:79–83.10.1093/jat/19.2.79Search in Google Scholar PubMed
10 Politi L, Groppi A, Polettini A, Montagna M. A rapid screening procedure for drugs and poisons in gastric contents by direct injection-HPLC analysis. Forensic Sci Int 2004; 141:115–20.10.1016/j.forsciint.2004.01.010Search in Google Scholar PubMed
11 Tracqui A, Kintz P, Mangin P. Systematic toxicological analysis using HPLC/DAD. J Forensic Sci 1995; 40:254–62.10.1520/JFS15353JSearch in Google Scholar
12 Turcant A, Premel-Cabic A, Cailleux A, Allain P. Toxicological screening of drugs by microbore high-performance liquid chromatography with photodiode-array detection and ultraviolet spectral library searches. Clin Chem 1991; 37:1210–5.10.1093/clinchem/37.7.1210Search in Google Scholar
13 Pragst F, Herzler M, Herre S, Erxleben B-T, Rothe M. UV spectra of toxic compounds. Database of photodiode array UV spectra of illegal and therapeutic drugs, pesticides, ecotoxic substances and other poisons. Book and CD. Heppenheim: Dieter Helm, 2001:1034 pp.Search in Google Scholar
14 Herzler M, Herre S, Pragst F. Selectivity of substance identification by HPLC-DAD in toxicological analysis using a UV spectra library of 2682 compounds. J Anal Toxicol 2003; 27:233–42.10.1093/jat/27.4.233Search in Google Scholar
15 Maurer HH. Liquid chromatography-mass spectrometry in forensic and clinical toxicology. J Chromatogr B Biomed Sci Appl 1998; 713:3–25.10.1016/S0378-4347(97)00514-8Search in Google Scholar
16 Venisse N, Marquet P, Duchoslav E, Dupuy JL, Lachatre G. A general unknown screening procedure for drugs and toxic compounds in serum using liquid chromatography-electrospray-single quadrupole mass spectrometry. Anal Toxicol 2003; 27:7–14.10.1093/jat/27.1.7Search in Google Scholar
17 Van Bocxlaer JF, Clauwaert KM, Lambert WE, Deforce DL, Van den Eeckhout EG, De Leenheer AP. Liquid chromatography-mass spectrometry in forensic toxicology. Mass Spectrom Rev 2000; 19:165–214.10.1002/1098-2787(200007)19:4<165::AID-MAS1>3.0.CO;2-YSearch in Google Scholar
18 George SA. Historical and modern developments. In: Huber L, George SA, editors. Diode array detection in HPLC. New York: Marcel Dekker, 1993:1–40.Search in Google Scholar
19 Milano MJ, Lam S, Grushka ME. Rapid scanning diode array as multi-wavelength detector in liquid chromatography. J Chromatogr 1976; 125:315–26.10.1016/S0021-9673(00)93828-7Search in Google Scholar
20 Dessy RE, Nunn WG, Titus CA. Linear photodiode array spectrometers as detectors in automated liquid chromatography. J Chromatogr Sci 1976; 14:195–200.10.1093/chromsci/14.4.195Search in Google Scholar
21 Sievert H-J P, Drouen ACJH. Spectral matching and peak purity. In: Huber L, George SA, editors. Diode array detection in HPLC. New York: Marcel Dekker, 1993:51–126.Search in Google Scholar
22 De Zeeuw RA, Hartstra J, Franke JP. Potential and pitfalls of chromatographic techniques and detection modes in substance identification for systematic toxicological analysis. J Chromatogr A 1994; 674:3–13.10.1016/0021-9673(94)85214-6Search in Google Scholar
23 Moffat AC, Smalldon KW, Brown C. Optimum use of paper, thin-layer and gas-liquid chromatography for the identification of basic drugs I. Determination of effectiveness for a series of chromatographic systems. J Chromatogr 1974; 90:1–7.10.1016/S0021-9673(01)94768-5Search in Google Scholar
24 Schepers PG, Franke JP, de Zeeuw RA. System evaluation and substance identification in systematic toxicological analysis by the mean list length approach. J Anal Toxicol 1983; 7:272–8.10.1093/jat/7.6.272Search in Google Scholar
25 Demme U, Ahrens B, Klein A, Werner R. Discriminating power nearly one – ion-trap GC-MS as an ideal tool for STA. In: Rasanen I, editor. Proceedings of the 38th meeting of TIAFT. Helsinki: Tallina Raamatutrükikoja, 2001:19–26.Search in Google Scholar
26 Pragst F, Maurer HH, Hallbach J, Staerk U, Külpmann WR, Degel F, Gibitz HJ. Suchverfahren (General unknown). In: Külpmann WR, editor. Klinisch-toxikologische Analytik – Verfahren, Befunde, Interpretation. Weinheim: Wiley-VCH, 2002:49–124.Search in Google Scholar
27 Herre S, Pragst F. Shift of the high performance liquid chromatographic retention times of metabolites in relation to the original drug at an RP-8 column with acidic mobile phase. J Chromatogr B Biomed Sci Appl 1997; 692:111–26.10.1016/S0378-4347(96)00472-0Search in Google Scholar
28 Herre S, Pragst F, Gillner E. Todesfälle nach Verapamil-Überdosis. Rechtsmedizin 1994; 4:103–6.Search in Google Scholar
29 Franke JP, de Zeeuw RA. Solid-phase extraction procedures in systematic toxicological analysis. J Chromatogr B Biomed Sci Appl 1998; 713:51–9.10.1016/S0378-4347(97)00536-7Search in Google Scholar
30 Polettini A. Systematic toxicological analysis of drugs and poisons in biosamples by hyphenated chromatographic and spectroscopic techniques. J Chromatogr B Biomed Sci Appl 1999; 733:47–63.10.1016/S0378-4347(99)00264-9Search in Google Scholar
31 Drummer OH. Chromatographic screening techniques in systematic toxicological analysis. J Chromatogr B Biomed Sci Appl 1999; 733:27–45.10.1016/S0378-4347(99)00265-0Search in Google Scholar
32 Decaestecker TN, Coopman EM, Van Peteghem CH, Van Bocxlaer JF. Suitability testing of commercial solid-phase extraction sorbents for sample clean-up in systematic toxicological analysis using liquid chromatography-(tandem) mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci 2003; 789:19–25.10.1016/S1570-0232(03)00208-3Search in Google Scholar
33 Stimpfl T, Jurenitsch J, Vycudilik W. General unknown screening in postmortem tissue and blood samples: a semi-automatic solid-phase extraction using polystyrene resins followed by liquid-liquid extraction. J Anal Toxicol 2001; 25:125–9.10.1093/jat/25.2.125Search in Google Scholar
34 Degel F. Comparison of new solid-phase extraction methods for chromatographic identification of drugs in clinical toxicological analysis. Clin Biochem 1996; 29:529–40.10.1016/S0009-9120(96)00096-3Search in Google Scholar
35 De Zeeuw RA, Wijsbeek J, Franke JP. SPEC disc solid-phase extraction for rapid broad-spectrum drug screening in urine. J Anal Toxicol 2000; 24:97–101.10.1093/jat/24.2.97Search in Google Scholar
36 Pragst F, Herzler M, Herre S. Use of high performance liquid chromatography with photodiode array detection (HPLC-DAD) in clinical toxicology. Klin Biochem Metab 2000; 8:13–6.Search in Google Scholar
37 Herzler M, Witteweg M, Pragst F. Nachweis und quantitative Bestimmung einiger Lösemittel und andere flüchtiger Substanzen durch HPLC mit Photodiodenarray-Detektor. Toxichem Krimtech 2002; 69:13–22.Search in Google Scholar
38 Demme U, Becker J, Bussemas H, Daldrup T, Erdmann F, Erkens M, et al. Zur Extrahierbarkeit toxikologisch relevanter Verbindungen mit 1-Chlorbutan. In: Pragst F, Aderjan R., editors. Proceedings of the GTFCh Symposium 1999. Heppenheim: Dieter Helm, 1999:213–8.Search in Google Scholar
39 Von Meyer L, Schmoldt A, Külpmann WR. Hypnotika/Sedativa: Benzodiazepine. In: Külpmann WR, editor. Klinisch-toxikologische Analytik – Verfahren, Befunde, Interpretation. Weinheim: Wiley-VCH, 2002:287–97.Search in Google Scholar
40 Titier K, Bouchet S, Pehourcq F, Moore N, Molimard M. High-performance liquid chromatographic method with diode array detection to identify and quantify atypical antipsychotics and haloperidol in plasma after overdose. J Chromatogr B Anal Technol Biomed Life Sci 2003; 788:179–85.10.1016/S1570-0232(02)01003-6Search in Google Scholar
41 Binder SR, Regalia M, Biaggi-McEachern M, Mazhar M. Automated liquid chromatographic analysis of drugs in urine by on-line sample cleanup and isocratic multi-column separation. J Chromatogr 1989; 473:325–41.10.1016/S0021-9673(00)91317-7Search in Google Scholar
42 Chiap P, Rbeida O, Christiaens B, Hubert P, Lubda D, Boos KS, Crommen J. Use of a novel cation-exchange restricted-access material for automated sample cleanup prior to the determination of basic drugs in plasma by liquid chromatography. J Chromatogr A 2002; 975:145–55.10.1016/S0021-9673(02)01334-1Search in Google Scholar
43 Rbeida O, Christiaens B, Hubert P, Lubda D, Boos KS, Crommen J, et al. Evaluation of a novel anion-exchange restricted-access sorbent for on-line sample clean-up prior to the determination of acidic compounds in plasma by liquid chromatography. J Chromatogr A 2004; 1030:95–102.10.1016/j.chroma.2003.11.031Search in Google Scholar
44 Vielhauer S, Rudolphi A, Boos KS, Seidel D. Evaluation and routine application of the novel restricted-access precolumn packing material Alkyl-Diol Silica: coupled-column high-performance liquid chromatographic analysis of the photoreactive drug 8-methoxypsoralen in plasma. J Chromatogr B Biomed Appl 1995; 666:315–22.10.1016/0378-4347(94)00578-SSearch in Google Scholar
45 Yamamoto E, Murata K, Ishihama Y, Asakawa N. Methyl-cellulose-immobilized reversed-phase precolumn for direct analysis of drugs in plasma by HPLC. Anal Sci 2001; 17:1155–9.10.2116/analsci.17.1155Search in Google Scholar
46 Walles M, Borlak J, Levsen K. Application of restricted access material (RAM) with precolumn-switching and matrix solid-phase dispersion (MSPD) to the study of the metabolism and pharmacokinetics of verapamil. Anal Bioanal Chem 2002; 374:1179–86.10.1007/s00216-002-1614-2Search in Google Scholar
47 Yu Z, Westerlund D, Boos KS. Evaluation of liquid chromatographic behavior of restricted-access media precolumns in the course of direct injection of large volumes of plasma samples in column-switching systems. J Chromatogr B Biomed Sci Appl 1997; 704:53–62.10.1016/S0378-4347(97)00471-4Search in Google Scholar
48 Saint-Marcoux F, Lachatre G, Marquet P. Evaluation of an improved general unknown screening procedure using liquid chromatography-electrospray-mass spectrometry by comparison with gas chromatography and high-performance liquid-chromatography-diode array detection. J Am Soc Mass Spectrom 2003; 14:14–22.10.1016/S1044-0305(02)00801-2Search in Google Scholar
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