Use of liquid chromatography-atmospheric pressure chemical ionization-ion trap mass spectrometry for identification of oleanolic acid and ursolic acid in Anoectochilus roxburghii (wall.) Lindl.
Oleanolic acid (OA) and ursolic acid (UA) are the two important bioactive compounds in Anoectochilus roxburghii (wall) Lindl (A. roxburghii), which has been used as a traditional Chinese medicine. So far, there has been no report to indicate that A. roxburghii contains these two bioactive compounds. It is necessary to develop an effective method to extract and analyze OA and UA in A. roxburghii. In this paper, a quantitative method, consisting of supercritical fluid extraction (SFE) followed by liquid chromatography-atmospheric pressure chemical ionization-ion trap mass spectrometry (LC-APCI-IT-MS) analysis, was developed for identification of OA and UA in A. roxburghii. The extraction was carried out by using CO(2) as the supercritical fluid and ethanol as the modifier before LC separation. The mobile phase used for LC separation consisted of acetic acid (1%, v/v), water (15%, v/v) and methanol (84%, v/v), and the elution was performed at a flow rate of 0.8 ml/min. The mass spectrometer was operated in APCI(+) mode with selected ion monitoring (SIM) to quantify OA and UA at m/z 439.4. Under optimum conditions, the linear responses of OA and UA were obtained in the concentration range of 0.5-80 (r = 0.9992) and 0.5-50 microg/ml (r = 0.9989) with the detection limits of 0.125 and 0.085 microg/ml, respectively. The proposed method has been used for the identification and quantitation of OA and UA in a real A. roxburghii sample.
Huang
Liying
L
Ministry of Education Key Laboratory of Analysis and Detection Technology for Food Safety (Fuzhou University), and Department of Chemistry, Fuzhou University, Fuzhou, Fujian 350002, China.
Chen
Tianwen
T
Ye
Zhao
Z
Chen
Guonan
G
D002245:Carbon Dioxide / Q000737:chemistry; D002853:Chromatography, Liquid; D025924:Chromatography, Supercritical Fluid; D013058:Mass Spectrometry; D009828:Oleanolic Acid / Q000032:analysis*; D010944:Plants / Q000737:chemistry*; D015203:Reproducibility of Results; D012997:Solvents; D021241:Spectrometry, Mass, Electrospray Ionization; D014315:Triterpenes / Q000032:analysis*; D000097245:Ursolic Acid
D002245:Carbon Dioxide; D009828:Oleanolic Acid; D012997:Solvents; D014315:Triterpenes; D000097245:Ursolic Acid