Ohta H, Seto Y, Tsunoda N
National Research Institute of Police Science, Chiyoda-ku, Tokyo, Japan.
J Chromatogr B Biomed Sci Appl. 1997 Apr 11;691(2):351-6. doi: 10.1016/s0378-4347(96)00471-9.
Determination of four toxic Aconitum alkaloids, aconitine, mesaconitine, hypaconitine and jesaconitine, in blood and urine samples has been established using high-performance liquid chromatography (HPLC) combined with ultraviolet absorbance detection, solid-phase extraction and mass spectrometry (MS). These alkaloids were hydrolyzed rapidly in alkaline solution (half lives (t1/2)<one day), were stable in solutions of acetonitrile, tetrahydrofuran and diluted hydrochloric acid (t1/2>five months) and were unstable in solutions of methanol and ethanol (t1/2<one month). These alkaloids were separated on an octadecylsilica column with isocratic elution using a solvent mixture of tetrahydrofuran and 0.2% trifluoroacetic acid (14:86, v/v), which was found to be the optimal solvent of the elution systems examined. Calibration curves with UV detection were linear on injection of amounts ranging from 2.5 to 500 ng, and the limit of detection was 1 ng (S/N=3). These four alkaloids in aqueous solution were recovered almost totally by solid-phase extraction using the styrene polymer resin, Sep-Pak Plus PS-1, and were eluted using a mixture of acetonitrile and hydrochloric acid. These Aconitum alkaloids were confirmed by HPLC coupled with fast atom bombardment MS, giving their protonated molecular ions as base peaks. These alkaloids were detected by HPLC with UV detection from blood samples spiked with more than 50 ng ml(-1) of alkaloids, but were not detectable from urine samples spiked with 5 microg ml(-1) of alkaloids because of severe sample interference.
已建立采用高效液相色谱（HPLC）结合紫外吸收检测、固相萃取和质谱（MS）法测定血液和尿液样本中四种有毒乌头生物碱，即乌头碱、中乌头碱、次乌头碱和杰斯乌头碱的方法。这些生物碱在碱性溶液中迅速水解（半衰期（t1/2）<1天），在乙腈、四氢呋喃和稀盐酸溶液中稳定（t1/2>5个月），在甲醇和乙醇溶液中不稳定（t1/2<1个月）。这些生物碱在十八烷基硅胶柱上采用等度洗脱，使用四氢呋喃和0.2%三氟乙酸（14:86，v/v）的混合溶剂，发现该混合溶剂是所考察洗脱系统中的最佳溶剂。紫外检测的校准曲线在进样量为2.5至500 ng范围内呈线性，检测限为1 ng（S/N = 3）。水溶液中的这四种生物碱几乎可通过使用苯乙烯聚合物树脂Sep-Pak Plus PS-1进行固相萃取完全回收，并用乙腈和盐酸的混合物洗脱。这些乌头生物碱通过HPLC与快原子轰击MS联用得到确认，其质子化分子离子作为基峰。通过HPLC紫外检测从添加了超过50 ng ml-1生物碱的血液样本中检测到了这些生物碱，但由于严重的样品干扰，从添加了5 μg ml-1生物碱的尿液样本中未检测到。
