HPLC determination of the major active flavonoids and GC-MS analysis of volatile components of Dysphania graveolens (Amaranthaceae).

Introduction: Dysphania graveolens is used mainly in Mexican traditional medicine against gastrointestinal ailments. Previous investigations revealed that its flavonoids are important active principles; however, there is not a reliable and accurate analytical method for determining these compounds in the crude drug or preparations of the plant. In addition, its volatile chemical composition remains unknown.

Objective: The main goals were to develop a validated HPLC method for quantifying the active flavonoids (pinostrobin (1), pinocembrin (2) and chrysin (3)) of D. graveolens and to establish its volatile composition.

Methodology: Separation was carried out on a Licrospher100 RP18 column with a linear gradient acetonitrile 0.1% formic acid and aqueous 0.1% formic acid. Accuracy was determined by spiking the crude drug with the standards, the recoveries were between 99% and 101%. A systematic description of the volatile components of D. graveolens was assessed via GC-MS using headspace solid-phase microextraction (HS-SPME) and hydrodistillation extraction methods.

Results: The developed HPLC method represented a powerful technique for the quality control of D. graveolens allowing the quantification of the three active flavonoids. For each compound a linear response was evaluated within the range of 0.5-2.0 mg/mL for pinostrobin (1), 0.25-1.25 mg/mL for pinocembrin (2) and 0.05-0.5 mg/mL for chrysin (3). According to SPME the major components in D. graveolens were p-cymene (84.85%) and eucalyptol (11.26%). On the other hand, the essential oil had eucalyptol (42.89%) and p-cymene (16.51%) and did not contain ascaridol. Thus the most relevant volatile components in the species were monoterpenoids.