GC/MS analysis of LSD as Trimethylsilyl-Derivative

Dr. K. Besserer, Gerichtliche Medizin, Tbingen; Dipl.-Ing. P. Gerhards, SHIMADZU EUROPA GmbH, Duisburg

Many non-radioactive immunological tests for LSD in urine are available in the market place today. Because of this diversity of analytical techniques it becomes necessary to have control analysis available. GC/MS is a sensitive and specific method for the determination of LSD. Analysis of LSD in urine has several issues that must be addressed. Chromatographically the LSD-TMS derivative is difficult because of the very high retention index(3595). The deactivation of the sample path is an important concern. Column dimensions should be short and the liquid phase should be thin to minimize the possible formation of active sites. Injection port liner inertness must also be carefully monitored. The specimen samples are required to be free from the matrix. Chromatographic conditions Detector temperature: Gain: 300 C 2.0 kV

Reextraction is an option to ensure that samples are isolated from the matrix. Derivatization of the extremely dry residue is done in BSTFA/TMCS (Fluka 15238) 20 min. at 70 C. A mixture of BSTFA/TMSC was prepared at 30-50 % solution in a solvent such as acetic acid ethyl ether or dioxan. Sample analysis of the LSD derivatives using a GC-17A in combination with a QP-5000. A J&W column type DB-5MS with 0.25 mm I.D., 25 m length and 0.25 m film thickness was used for the chromatographic separation. Single-Ion-Monitoring (SIM) acquistion mode was used on the GC/MS. Using SIM the identification can be done with a short column, without endangering the quality of the results.

LSD/TMS

Libraries: 2 Scan range: Scan rate: Injection temperature: Injection volume: Column:

NIST 75.000 and PMW-Tox 55-499 AMU 0.45 Scans/sec 280 C 1 l DB-5MS, 25 m, 0.25 mm I.D., 0.25 m film (J&W) 200 C/3 min with 20 C/min to 300 C/10 min 100 kPa/3 min with 10 kPa/min to 150 kPa/10 min 1 min splitless 36 cm/sec

Temperature program: Pressure program: Split ratio: Linear velocity:

Fig. 1: Scan mode chromatogram and library search result of 20 ng LSD/TMS absolute measured in Full-Scan

Figure 1 demonstrates the scan mode chromatogram generated from a 20 ng LSD standard derivatized with BSTFA and the library search results. It should be noted that the derivatization with BSTFA requires a completely water free environment. If even a small amount of water is present the derivatization process will be negatively effected. To derivatize the sample add the BSTFA and place the vial on the hot injector for 45 minutes. The sample should be analyzed quickly because the peak area decreases with time. Figure 2 shows the extract of the urine spiked with LSD/TMS and LSD-d3/TMS. The samples were analyzed in SIM with the two significant masses for LSD and the internal standard LSD-d3. *Note* When analyzing LSD ensure that the samples are stored in brown glass vials or that the samples are protected from light. This precaution is necessary to prevent a loss of the compound due to reactive breakdown. Figure 3 shows the extract after SolidPhase-Extraction (SPE) of non spiked urine. This experiment was done as a control to demonstrate the influence of the matrix Fig. 3: Extract from a non spiked urine SPE shows some advantages when compared with the Liquid-LiquidExtraction: Purity of the extracts Concentration of the compounds

Conclusion GC/MS is a very good way to improve the results of the immuno-assay. Because of the very high specificity and sensitivity of the GC/MS in SIM, qualitative and quantitative determinations are achieved. The quality of the results is directly proportional to the sample preparation. To use BSTFA for derivatization requires special handling in an inert atmosphere. If the limitations of the BSTFA derivatization are considered good results can be achieved. Concentration during the extraction step extends the sensitivity of the analytical method.
SCA-280-029E