Takayama N, Tanaka S, Hayakawa K
Forensic Science Laboratory, Ishikawa Prefectural Police Headquarters, Kanazawa, Japan.
Biomed Chromatogr. 1997 Jan-Feb;11(1):25-8. doi: 10.1002/(SICI)1099-0801(199701)11:1<25::AID-BMC615>3.0.CO;2-Y.
Stimulants that are controlled by the Stimulant Drug Control Law of Japan are methamphetamine (MA) and amphetamine (AP). MA is used by most stimulant addicts, and AP is detected as its main metabolite. We have developed a high-performance liquid chromatography method with chemiluminescence detection (CL-HPLC), for determining trace levels of MA and its metabolites in a single human hair sample, in which bis(2,4,6-trichlorophenyl)oxalate and hydrogen peroxide are the postcolumn reagents. After washing a single hair sample with water and methanol, it was cut into pieces, extracted with a mixed solution of methanol and hydrochloric acid for 1 h under ultra-sonication and allowed to stand at room temperature overnight. Then the organic phase was evaporated to dryness. To the residues, 0.1 mL of carbonate buffer and 0.1 mL of dansyl chloride solution were added and the solution was heated at 45 degrees C for 1 h. An aliquot of the reaction mixture was then subjected to HPLC. MA and AP were chemiluminogenically detected as their dansyl derivatives from a sample of only a single hair. The detection limit was about 2 pg in an injected volume (20 microliters), and about 20 pg in a single hair sample. This detection limit was smaller than that by the gas chromatography/mass spectrometry (selective ion monitoring) method. Our method was useful as a screening test for stimulant users.
受日本《兴奋剂管制法》管制的兴奋剂是甲基苯丙胺（MA）和苯丙胺（AP）。大多数兴奋剂成瘾者使用MA，AP作为其主要代谢物被检测到。我们开发了一种采用化学发光检测的高效液相色谱法（CL-HPLC），用于测定单根人发样本中痕量水平的MA及其代谢物，其中双（2,4,6-三氯苯基）草酸酯和过氧化氢为柱后试剂。将单根头发样本用水和甲醇洗涤后，切成小段，用甲醇和盐酸的混合溶液在超声处理下提取1小时，并在室温下静置过夜。然后将有机相蒸发至干。向残留物中加入0.1 mL碳酸盐缓冲液和0.1 mL丹磺酰氯溶液，并将溶液在45℃加热1小时。然后取一份反应混合物进行HPLC分析。从仅一根头发的样本中，MA和AP作为其丹磺酰衍生物通过化学发光法检测。进样体积（20微升）时的检测限约为2皮克，单根头发样本中的检测限约为20皮克。该检测限低于气相色谱/质谱（选择性离子监测）法的检测限。我们的方法作为兴奋剂使用者的筛查试验很有用。
