Identification and Quantification of Heterologous Compounds Parthenin and Organic Acids in Parthenium Hysterophorus L. Using HPLC-PDA-MS-MS

Abstract

Parthenium hysterophorus L., is an obnoxious weed known for its environmental health hazards and medicinal uses. These characteristics are due to presence of sesquiterpene lactones and organic acids; therefore a rapid and sensitive analytical procedure using HPLC-PDA-MS-MS was developed and optimized for separation, identification, and quantification of parthenin and six organic acids. Separation and characterization of compounds was achieved on a RP-C18 column with 1% acetic acid in water (A) and acetonitrile (B) as a mobile phase at a flow rate of 0.6 mL min⁻¹ and by matching their UV and mass spectra with reference compounds. Six organic acids (ferulic acid, 0.1 mg g⁻¹ to coumaric acid, 13.6 mg g⁻¹) and parthenin (27.4 mg g⁻¹) were characterized within 26 minutes of chromatographic separation in plant extract. The calibration curves are linear with correlation coefficients from 0.985 to 0.998, limit of detection and quantification ranged between 1.0 µg mL⁻¹ (anisic acid) to 2.2 µg mL⁻¹ (parthenin) and 2.5 µg mL⁻¹ (coumaric acid) to 5.2 µ g mL⁻¹ (parthenin) and recovery ranged between 90.9% to 97.3%. To the best of our knowledge this is the first report for the simultaneous separation of parthenin and organic acids. The method is applicable for screening of commercial crops, medicinal plants, and their products which might be mixed with P. hysterophorus during harvesting period.

Keywords:

• HPLC-PDA-MS-MS
• Organic acids
• Parthenin
• Parthenium hysterophorus
• Validation

Acknowledgments

The study was supported by Task Force grant NWP-006 from Council of Scientific and Industrial Research (CSIR), New Delhi, India awarded to Dr. Chandra Shekhar Nautiyal.

Notes

^a The retention times (t_R) in minutes are the mean values of 10 replicates ± S.D.

^b Correlation coefficient.