以氣相層析質譜方法及氘同位素藥物為內部標準品應用於定量尿液中安非他命之探討   全文下載

GC/MS Analysis of Amphetamine in Urine with Amphetamine-d5(side chain) as an Internal Standard

本研究係探討以氘同位素安非他命為內部標準品於氣相層析質譜儀分析尿液中安非他命含量之適用性。本計畫所用之安非他命氘同位素藥物為amphetamine-d5其與原待測藥物amphetamine-d0經衍生化試劑trichloroacetic anhydride衍生化後探討全掃描質譜圖及指定離子碎片監測（SIM）圖譜之離子碎片強度是否相互干擾，並以迴歸方式分析其實際應用在定量上之可行性。研究結果顯示，由兩者之全掃描質譜圖來分析，兩者之離子碎片除m／z 91, 192外，其他主要離子之相互干擾甚低，其中amphetamine-d0之trichloroacetic衍生物（TCA-amphetamine-d0）適用之離子為118, 188, 190及91；而amphetamine-d5之trichloroacetic衍生物（TCA-amphetamine-d5）適用之離子則為123及196。由選定離子監測質譜圖（SIM）來分析，兩者之選定離子碎片相互干擾甚低，其中TCA-amphetamine-d0適用之離子為190, 188, 118及91，TCA-amphetamine-d5適用之離子為196, 194及123。由其適用之離子中，本研究選用TCA-amphetamine-d0之離子碎片190及TCA-amphetamine-d5之離子碎片194為定量用離子，以選定離子監測質譜中190與194之相對離子強度比值對標準品amphetamine-d0之序列濃度31.25, 62.5, 125, 250, 500, 1000, 2000 ng／mL 進行迴歸分析，於上述濃度範圍內，具有良好的線性關係（n = 7, SD ~ 0.03569, 相關係數為0.9987）。因此以trichloroacetic anhydride為衍生化試劑時，amphetarnine-d5應可供作氣相層析質譜法分析尿液中安非他命含量時的內部標準品。"

This study examined the accessibility of using amphetamine-d5 (side chain) as an internal standard in quantitative analysis of the trichloroacetyl derivative of urinary amphetamine by gas chromatography I mass spectrometry (GC/MS). In GC/MS analysis, fragments with minimal interference between the assay protocol and the internal standard were selected for quantitation. In the full-scan mode, no interference from TCA-amphetamine-d5, the trichloroacetyl derivative of amphetamine-d5, was observed for the fragments of trichloroacetylamphetamine (TCA-amphetamine) with m/z 190, 188, 118 and 91. On the other hand, no interference from TeAamphetamine was indicated in fragments of m/z 196 and 123 of TCA-amphetamine-d5. In the selective ion monitoring mode (SIM), the interference between m/z 190, 188, 118 and 91 of TCA-amphetamine and m/z 196, 194 and 123 of TCA-amphetamine-d5 was minimaL The fragments of m/z 190 from TCA-amphetamine and m/z 194 from TCA-amphetamine-d5 were thus selected for quantitation of urinary amphetamine by SIM. Calibration graph derived from the ratio of relative intensity of m/z 190 to m/z 194 versus the concentration of urinary amphetamine was linear over the concentration range of 31.25 - 2000 ng/mL (n=7, SD=0.0357, r=0.9987). Amphetamine-d5 was demonstrated to be a suitable internal standard in quantitative analysis of the trichloroacety1 derived urinary amphetamine by SIM analysis with GC/MS.