Rapid Identification and Simultaneous Quantification of Aristolochic Acids by HPLC-DAD and Confirmations by MS in Aristolochia chilensis Using a Limited Biomass.

ABSTRACT: Six aristolochic acids were identified in the Chilean species Aristolochia chilensis 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?µL of the extract that contains the aristolochic acids present in the Aristolochia chilensis 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^-1. The column InertSustain C18 250?×?4.6?mm and 3?µm was used, the injection volume was 20?µL, and the time of run was reduced to 15?min. Calibration curves were constructed with r²?being 0.9997. The quantification limit for AAI was 0.138?±?0.010?µg/mL, and for AAII, it was 0.558?±?0.042?µg/mL.

SUBMITTER: Araya M 

PROVIDER: S-EPMC6011054 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature