byakangelicol and isoimperatorin

Rapid, simple and reliable HPLC/UV and LC-ESI-MS/MS methods for the simultaneous determination of five active coumarins of Angelicae dahuricae Radix, byakangelicol (1), oxypeucedanin (2), imperatorin (3), phellopterin (4) and isoimperatorin (5) were developed and validated. The separation condition for HPLC/UV was optimized using a Develosil RPAQUEOUS C(30) column using 70% acetonitrile in water as the mobile phase. This HPLC/UV method was successful for providing the baseline separation of the five coumarins with no interfering peaks detected in the 70% ethanol extract of Angelicae dahuricae Radix. The specific determination of the five coumarins was also accomplished by a triple quadrupole tandem mass spectrometer equipped with an electrospray ionization source (LC-ESI-MS/MS). Multiple reaction monitoring (MRM) in the positive mode was used to enhance the selectivity of detection. The LC-ESI-MS/MS methods were successfully applied for the determination of the five major coumarins in Angelicae dahuricae Radix. These HPLC/UV and LC-ESI-MS/MS methods were validated in terms of recovery, linearity, accuracy and precision (intra- and inter-day validation). Taken together, the shorter analysis time involved makes these HPLC/UV and LC-ESI-MS/MS methods valuable for the commercial quality control of Angelicae dahuricae Radix extracts and its pharmaceutical preparations.

Topics: Angelica; Chromatography, High Pressure Liquid; Coumarins; Furans; Furocoumarins; Linear Models; Plant Roots; Reproducibility of Results; Sensitivity and Specificity; Spectrophotometry, Ultraviolet; Tandem Mass Spectrometry; Temperature

Fractionation of the MeOH extract of Angelica dahurica Benth et Hook resulted in the isolation of six furocoumarins, imperatorin (1), isoimperatorin (2), (+/-)-byakangelicol (3), (+)-oxypeucedanin (4), (+)-byakangelicin (5), and (+)-aviprin (6). Among these, compounds 1 and 5 exhibited strong hepatoprotective activities, displaying EC(50) values of 36.6 +/- 0.98 and 47.9 +/- 4.6 microM, respectively. Compounds 3 and 4 showed moderate activities with EC(50) values of 112.7 +/- 5.35 and 286.7 +/- 6.36 microM, respectively. Silybin as a positive control showed the EC(50) value with 69.0 +/- 3.4 microM. Comparison of hepatoprotective activities for six furocoumarins 1 - 6 suggested that oxy-substitution at the C-9 position increased the hepatoprotective activity.

Topics: Angelica; Carcinoma, Hepatocellular; Coumarins; Furans; Furocoumarins; Humans; Medicine, East Asian Traditional; Molecular Structure; Plant Extracts; Plant Roots; Tacrine; Tumor Cells, Cultured