An LC-MS/MS Method for Determination of Imperatorin and Isoimperatorin in Rat Plasma and Application to a Pharmacokinetic Study


A highly sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for the de-termination of imperatorin and isoimperatorin in rat plasma using psoralen as internal standard. Analyses were performed on a Shimadzu HPLC system using a Shimadzu C18 column eluted with a mobile phase of methanol-water (25:75), containing 0.1% formic acid at a flow rate of 0.4 ml/min. The detection was performed by positive ion elec-trospray ionization in multiple reactions monitoring mode, monitoring the transitions m/z 203→147 and m/z 203→159 for imperatorin and isoimperatorin, respectively. The method was validated over the concentration range of 2.5 -1000.0 ng/mL for imperatorin and isoimperatorin. The limits of detection and quantification were 1.0 and 3.0 ng/mL, repectively for both analytes. The intra-day and inter-day accuracy and precision of the assay were less than 10.9%. This method has been applied successfully to a pharmacokinetic study involving the intragastric administration of extract of Angelicae Dahuricae Radix to rats.

Share and Cite:

J. Xing, H. Cao, Y. Chen, C. Shui, T. Shan and D. Hu, "An LC-MS/MS Method for Determination of Imperatorin and Isoimperatorin in Rat Plasma and Application to a Pharmacokinetic Study," Pharmacology & Pharmacy, Vol. 4 No. 5, 2013, pp. 417-424. doi: 10.4236/pp.2013.45059.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] China Pharmacopoeia Committee, “Section One,” In: China Pharmacopoeia Committee, Ed., Chinese Pharmacopoeia, China Medical Science Press, Beijing, 2010, pp. 97-98.
[2] J. Jin, C. S. Jin and D. L. Wu, “Historical Evolution of the Processing and Progress in Modern Research of Angelica dahurica,” Anhui Medical and Pharmaceutical Journal, Vol. 16, No. 1, 2012, pp. 1-4.
[3] Y. C. Wang, C. H. Guo and X. L. Chen, “Observation of the Curative Effect of Xuefu Zhuyu Decoction with Baizhi and Huangqi on Headache after Stroke,” Modern Journal of Integrated Traditional Chinese and Western Medicine, Vol. 18, No. 22, 2009, pp. 2628-2629.
[4] R. Hu, Z. L. Li and L. Yang, “Study on the Analgesic Effect of Imperatorin from Radix Angelicae Dahurica,” Journal of Sichuan of Traditional Chinese Medicine, Vol. 30, No. 4, 2012, pp. 58-61.
[5] X. L. Piao, H. Park and S. H. Baek, “Report the Isolation and the Antioxidative Activity against 2,2-Azobis (2-Aminodinopropane) Dihydrochloride (AAPH)-Induced Cellular Damage of 11 Furanocoumarins from Angelica dahuricae,” Journal of Ethnopharmacology, Vol. 93, No. 2-3, 2004, pp. 243-246.
[6] F. Wang and C. Wang, “Study on the Relationship between the Anti-Aging Function and Antioxidation of Ethanol Extract from Angelicoe dahuricae Radix,” China Pharmacy, Vol. 23, No. 7, 2012, pp. 599-602.
[7] W. H. Zhao, Y. X. Cao and J. Liu, “Research on Chronic Toxicology of Compound Radix Angelicae dahuricae Capsule,” Journal of Shanxi Medical University, Vol. 37, No. 2, 2006, pp.160-163.
[8] R. Hu, Z. L. Li and L. Yang, “Anti-Inflammatory Effect of Imperatorin from Radix Angelicae dahurica on Rats with Acute Pleurisy,” Journal of Emergency in Traditional Chinese Medicine, Vol. 21, No. 4, 2012, pp. 576-579.
[9] H. Zhang, G. F. Hai and C. Zhang, “Experimental Studies on Analgesia and Anti-Febrile Effects of the Different Extracts from Radix Angelicae dahuricae,” Journal of Xinxiang Medical College, Vol. 28, No. 4, 2011, pp. 431-434.
[10] M. Y. Wang, M. Jia and Y. Y. Ma, “Pharmacological Effect of the Total Coumarins in Radix Amgelicae dahuricae,” Lishizhen Medicine and Materia Medica Research, Vol. 16, No. 10, 2005, pp. 954-956.
[11] M. Y. Wang, M. Jia and Y. Y. Ma, “Pharmacological Effect of Four Linear Furocoumarins in Radix Angelicae dahuricae,” Natural Product Research and Development, Vol. 22, No. 3, 2010, pp. 485-489.
[12] D. C. Wang,K. Li and X. Y. Xu, “Experimental Study Oil the Antipyretic-analgesic and Anti-inflammatory of Total Conmarins from Radix Angellcae dahurica,” Chinese Journa1 of Information on TCM, Vol. 12 No. 11, 2005, pp. 36-38.
[13] K. Li, Q. W. Li and J. Liu, “Effect of AAD Anti-Tumor on Mice Bearing in U14 Cervical Cancer,” Journal of Yanshan University, Vol. 36, No. 1, 2012, pp. 89-94.
[14] X. H. Qin, Z. L. Li and C. F. Zhu, “Study on the Pharmacodynamics of Total Coumarin of Radix Angelicae dahuricae in Treating Migrain,” Joumal of Sichuan of Traditional Chinese Medicine, Vol. 26, No. 6, 2008, pp. 4-6.
[15] Y. B. Li, “Determination of Imperatorin in Rat Plasma by HPLC,” Journal of Pharmaceutical and Biomedical Analysis, Vol. 40, No. 5, 2006, pp. 1253-1256.
[16] K. Ishihara, M. Fukutakeb and T. Asanob, “Simultaneous Determination of Byak-Angelicin and Oxypeucedanin Hydrate in Rat Plasma by Column-Switching High-Performance Liquid Chromatography with Ultraviolet Detection,” Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 753, No. 2, 2001, pp. 309-314.
[17] Y. Xie, Y. Chen and M. Lin, “Developed and Validated for the Determination and Pharmacokinetic Study of Oxypeucedanin Hydrate and Byak-Angelicin after Oral Administration of Angelica dahurica Extracts in Mongrel Dog Plasma by HPLC,” Journal of Pharmaceutical and Biomedical Analysis, Vol. 44, No. 1, 2007, pp. 166-172.
[18] Z. Han, Y. L. Zheng and L. J. Luan, “Analysis of Ochratoxin A and Ochratoxin B in Traditional Chinese Medicines by Ultra-High-Performance Liquid Chromatography—Tandem Mass Spectrometry Using [13C20]-Ochratoxin A as an Internal Standard,” Journal of Chromatography A, Vol. 1217, No. 26, 2010, pp. 4365-4374.
[19] W. Yang, M. Ye and M. Liu, “Characterization of Coumarins in Radix Glehniae by Liquid Chromatography Coupled with Triple Quadrupole-Linear Ion Trap Mass Spectrometry,” Journal of Chromatography A, Vol. 1217, No. 26, 2010, pp. 4587-4600.
[20] S. C. Wang, Q. H. Chen and L. C. He, “Development and Validation of a Gas Chromatography—Mass Spectrometrymethod for the Determination of Isoimperatorin in Rat Plasma and Tissue: Application to the Pharmacokinetic and Tissue Distribution Study,” Journal of Chromatography B, Vol. 852, No. 1-2, 2007, pp. 473-478.
[21] L. Ling, Z. L. Li and Y. X. He, “GC-MS Analysis of Extraction Products from Angelica dahurica (Fisch. ex Hoffm.) Benth. et Hook,” Lishizhen Medicine and Materia Medica Research, Vol. 20, No. 2, 2009, pp. 306-307.
[22] L. Q. Zhu and H. G. Sheng, “GC-MS Analysis on Volatile Components of Radix Angelica dahuricate,” Guangzhou Chemical Industry, Vol. 40, No. 23, 2012, pp. 103-105.
[23] A. H. Zhao, X. W. Yang and X. B. Yang, “GC-MS Analysis of Essential Oil from Root of Angelica dahurica cv. Qibaizhi,” China Journal of Chinese Materia Medica, Vol. 36, No. 5, 2011, pp. 603-607.
[24] Y. C, Lee, C. Y. Huang and K. C. Wen, “Determination of Liquiritin, Glycyrrhizin, Hesperidin, Cinnamic Acid, Cinnamaldehyde, Honokiol and Magnolol in the Traditional Chinese Medicinal Preparation Wu-Ji-San by High-Performance Liquid Chromatography,” Journal of Chromatography A, Vol. 692, No. 1-2, 1995, pp. 137-145.
[25] X. G. Zheng, X. W. Zhang and X. N. Sheng, “Simultaneous Characterization and Quantitation of 11 Coumarins in Radix Angelicae dahuricae by HPLC-ESI-MS/MS,” Journal of Pharmaceutical and Biomedical Analysis, Vol. 51, No. 3, 2010, pp. 599-605.
[26] C. Y. Zhang, Z. G. Li and Q. X. Wang, “Study on HPLC Fingerprint of Processed Radix Angelicae dahuricae,” Journal of Chinese Medicinal Materials, Vol. 30, No. 11, 2007, pp. 1374-1378.
[27] T. T. Wang, X. H. Chen and Q. Q. Hu, “RP-HPLC Fingerprint for Quality Assessment of Radix Angelicae dahuricae,” Acta Pharmaceutica Sinica, Vol. 41, No. 8, 2006, pp. 747-751.
[28] J. Zhang, Y. S. Wang and X. R. He, “Studies on Identification of Radix Angelicae dahuricae by HPLC Fingerprint,” Chinese Journal of Experimental Traditional Medical Formulae, Vo1, 13, No. 7, 2007, pp. 1-4.
[29] G. H. Cui, L. Q. Huang and X. Li, “A New Molecular Identification Method: Anchored Primer Amplification Polymorphism DNA,” Acta Pharmaceutica Sinica, Vo1. 42, No. 3, 2007, pp. 329-335.
[30] J. Y. Deng, G. H. Gao and C. J. Zhao, “Determination of the Contents of Two Coumarins in Angelica dahurica from Different Regions by HPLC,” Journal of Shenyang Pharmaceutical University, Vol. 21, No. 5, 2004, pp. 354-357.
[31] Z. Q. Mao, X. L. Meng and Y. L. Li, “Determination of Xanthotoxol, Bergapten, Imperatorin and Isoimperatorin in Angelicae dahuricae Radix by HPLC,” China Pharmacist, Vol. 14, No. 2, 2011, pp. 205-207.
[32] Y. Y. Zhao, H. Sun and M. Wang, “Simultaneous Quantitative Determination of Five Coumarins in Angelicae dahuricae Introduced in Jiangsu by HPLC,” Chinese Wild Plant Resources, Vol. 32 No. 1, 2013, pp. 56-59.
[33] R. Deng, J. Zhang and W. Z. Luo, “Quantitative Determination of Six Coumarins in Angelica dahurica var. formosana by RRLC-UV,” China Journal of Chinese Materia Medica, Vol. 35, No. 23, 2010, pp. 3184-3187.
[34] Y. Y. Ma, S. H. Zhong and M. R. Jia, “Determination of Total Content of Coumarin in Angelica dahurica by UV Spectrophotometry,” West China Journal of Pharmaceutical Science, Vol. 20, No. 2, 2005, pp. 159-160.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.