以氘相層析質譜分析方法及氘同位素藥物為內部標準品應用於定量尿液中6-乙醯嗎啡之探討   全文下載

Evaluation of Isotopic Analogs as Internal Standard for Quantitative Determination of Urinary 6-Acetylmorphine by Gas Chromatography/Mass Spectrometry

本研究係探討以6-乙醯嗎啡（6AM）之氘同位素藥物6-AM-d3為內部標準品及不同衍生化試劑於氣相層析質譜儀分析尿液中6AM含量之適用性。本研究所採用之衍生化試劑為trifluoroaceticanhydride （TFAA）, pentafluoropropionic anhydride （PFPA）及hexamethyldisilazane （HMDS），分別進行衍生化後，探討全掃描質譜圖（Full Scan）及選定離子監測（SIM）圖譜之離子碎片強度是否相互干擾，並以迴歸分析其實際應用在定量上之可行性。研究結果顯示，由全掃描質譜圖來分析， 6-TMS-AM之質譜圖對於6-TMS-AM-d3於m／z 287處有些許干擾；但以TFAA及PFPA為衍生化劑時，並無明顯干擾。由選定離子監測質譜圖分析，6-AM-d3與6-AM經TFAA及PFPA衍生化後之選定離子碎片相互干擾甚低；另， 6-TMS-AM之質譜圖對於6-TMS-AM-d3 於m／z 343 處有明顯干擾。本研究再分別以TFAA' HMDS 及PFPA 為衍生化試劑，選擇適用之離子碎片為定量用離子，進行迴歸分析，於50, 100, 200, 300, 500, 1000 ng／ml濃度範園內，具有良好的線性關係。因此以TFAA，HMDS及PFPA為衍生化試劑時，6-AM-d 3應可供作氣相層析質譜法分析尿液中6- 乙醯嗎啡定量時的內部標準品。但以HMDS衍生化定量時，由於有明顯干擾，應慎選適用之離子。"

This study examined the accessibility of using 6-acetylmorphine-d3 (6-AM-d3) as the internal standard and trifluoroacetic anhydride, pentafluoropropionic anhydride, and hexamethyl-disilazane as the derivatizing agents in the quantitative analysis of 6-acetylmorphine (6-AM) by full scan mode and selective ion monitoring (SIM) mode gas chromatography I mass spectrometry (GC/MS). In this study, fragments with minimal interference between the analyte and the internal standard were evaluated and selected for quantitative determination. Calibration curves and precision analyses were further performed using these selected ions. In the full scan mode, there was a certain interference between the mass spectra of trimethylsilyl derivative of 6-AM and those of 6-AM-d3 at some fragment ions. This requires that there be carefull selection of fragment ions for quantitative determination due to the potential crosscontribution and subsequent inaccurate results. On the other hand, 6-AM-d3 might be suitable as the internal standard in quantitative analysis of the trifluoroacetyl or pentafluoropropionyl derivative by GC/MS since the interference between the mass spectra of these derivatives of 6- AM and those of 6-AM-d3, was minimal. In the SIM mode, results were similar with those of the full scan mode. The suitable fragments were thus selected for the calibration curve and precision analysis by SIM GC/MS. Excellent linearity was obtained over the concentration range of 50-1000 ng/ml of 6-AM for their TFA, PFP, and TMS derivatives. Good precision was also obtained from within-run and between-run CVs for 6-TFA-AM, 6-PFP-AM and 6-TMSAM at a concentration of 200 ng/ml. 6-AM-d3 was demonstrated to be a suitable standard in quantitative analysis of TFA-, PFP-, or TMS- derived urinary 6-AM in SIM mode GC/MS analysis. 本研究係探討以6-乙醯嗎啡（6AM）之氘同位素藥物6-AM-d3為內部標準品及不同衍生化試劑於氣相層析質譜儀分析尿液中6AM含量之適用性。本研究所採用之衍生化試劑為trifluoroaceticanhydride （TFAA）, pentafluoropropionic anhydride （PFPA）及hexamethyldisilazane （HMDS），分別進行衍生化後，探討全掃描質譜圖（Full Scan）及選定離子監測（SIM）圖譜之離子碎片強度是否相互干擾，並以迴歸分析其實際應用在定量上之可行性。研究結果顯示，由全掃描質譜圖來分析， 6-TMS-AM之質譜圖對於6-TMS-AM-d3於m／z 287處有些許干擾；但以TFAA及PFPA為衍生化劑時，並無明顯干擾。由選定離子監測質譜圖分析，6-AM-d3與6-AM經TFAA及PFPA衍生化後之選定離子碎片相互干擾甚低；另， 6-TMS-AM之質譜圖對於6-TMS-AM-d3 於m／z 343 處有明顯干擾。本研究再分別以TFAA' HMDS 及PFPA 為衍生化試劑，選擇適用之離子碎片為定量用離子，進行迴歸分析，於50, 100, 200, 300, 500, 1000 ng／ml濃度範園內，具有良好的線性關係。因此以TFAA，HMDS及PFPA為衍生化試劑時，6-AM-d 3應可供作氣相層析質譜法分析尿液中6- 乙醯嗎啡定量時的內部標準品。但以HMDS衍生化定量時，由於有明顯干擾，應慎選適用之離子。