Araya Michael, García Samantha, González-Teuber Marcia
Centro de Investigación y Desarrollo Tecnológico de Algas (CIDTA), Facultad de Ciencias del Mar, Universidad Católica del Norte, Coquimbo, Chile.
Departamento de Química, Facultad de Ciencias, Universidad de La Serena, La Serena, Chile.
J Anal Methods Chem. 2018 Jun 6;2018:5036542. doi: 10.1155/2018/5036542. eCollection 2018.
Six aristolochic acids were identified in the Chilean species using high-performance liquid chromatography coupled to a diode array detector (HPLC-DAD) and subsequent confirmation with mass spectrometry (MS). The fractions of each signal were collected and injected directly into an Orbitrap mass detector model Q Exactive Focus (Thermo Scientific). The acids extraction was done with 0.10-0.50 g of lyophilized and pulverized sample and concentrated in Soxhlet extraction equipment. The liquid-liquid separations and a subsequent solid phase extraction (SPE) C18 were performed using 100 L of the extract that contains the aristolochic acids present in the plant. The HPLC conditions used a single mobile phase acetonitrile : water (1 : 1) acidified with 0.1% acetic acid and an isocratic elution to 1 mL·min. The column InertSustain C18 250 × 4.6 mm and 3 m was used, the injection volume was 20 L, and the time of run was reduced to 15 min. Calibration curves were constructed with being 0.9997. The quantification limit for AAI was 0.138 ± 0.010 g/mL, and for AAII, it was 0.558 ± 0.042 g/mL.
使用高效液相色谱仪与二极管阵列检测器（HPLC-DAD）联用，并随后通过质谱（MS）进行确认，在智利的物种中鉴定出了六种马兜铃酸。收集每个信号的馏分，并直接注入Q Exactive Focus型轨道阱质谱检测器（赛默飞世尔科技公司）。酸的提取是用0.10 - 0.50克冻干并粉碎的样品进行的，并在索氏提取设备中浓缩。使用含有植物中存在的马兜铃酸的100微升提取物进行液-液分离以及随后的C18固相萃取（SPE）。HPLC条件使用单一流动相乙腈∶水（1∶1），用0.1%乙酸酸化，并采用等度洗脱至1毫升·分钟。使用InertSustain C18 250×4.6毫米、3微米的色谱柱，进样体积为20微升，运行时间缩短至15分钟。构建的校准曲线相关系数为0.9997。AAI的定量限为0.138±0.010微克/毫升，AAII的定量限为0.558±0.042微克/毫升。
