A Sensitive HPLC-MS/MS Analysis of Dencichine in Rat Plasma and Its Application to Pharmacokinetics


In order to quantitate dencichine in biological samples, a selective and sensitive method for the determination of dencichine in rat plasma based on high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed and validated. (l)-2-amino-3-(carboxymethylthio)propionic acid was used as the internal standard (I.S.). After a protein precipitation extraction with acetonitrile, dencichine and the I.S. were chromatographed on an Xterra MS-C18 column. The mobile phase was consisted of 20mmol/L ammonium acetate solution-acetonitrile (35:65, V/V) at a flow rate of 0.2 mL/min. The detection was performed on a triple quadrupole mass via electrospray ionization (ESI) source in the positive mode. The intra- and inter-day precision (relative standard deviation, R.S.D.) values of dencichine were below 6.7%. The extraction recoveries were up 85%. The lower limit of quantification was 20 ng/ml, which was sensitive enough to detect the analyte. The HPLC-MS/MS method was successfully applied to the pharmacokinetic study after an intravenous administration of dencichine in Sprague-Dawley (SD) rat.

Share and Cite:

C. Qian, Y. Yuan, X. He, J. Liu, Q. Shao, H. Wu and H. Qiao, "A Sensitive HPLC-MS/MS Analysis of Dencichine in Rat Plasma and Its Application to Pharmacokinetics," American Journal of Analytical Chemistry, Vol. 3 No. 8, 2012, pp. 596-603. doi: 10.4236/ajac.2012.38078.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] S. L. Rao, P. R. Adiga and P. S. Sarma, “The Isolation and Characterization of Beta-N-Oxalyl-L-α,β-Diaminopr-opionic Acid: A Neurotoxin from the Seeds of Lathyrus Sativus,” Bio-chemistry, Vol. 3, No. 3, 1964, pp. 432-436. doi:10.1021/bi00891a022
[2] M. Y. Quereshi, D. J. Pilbeam, C. S. Evans and E. A. Bell, “Neurolathyrogen, α-Amino-β-Oxalylaminopropionic Acid in Legume Seeds,” Phytochemistry, Vol. 16, No. 4, 1977, pp. 477-479. doi:10.1016/S0031-9422(00)94332-2
[3] Y. H. Kuo, F. Ikegami and F. Lambein, “Neuroactive and Other Free Amino Acids in Seed and Young Plants of Panax Ginseng,” Phytochemistry, Vol. 62, No. 7, 2003, pp. 1087-1091. doi:10.1016/S0031-9422(02)00658-1
[4] Y. Zhang, X. Y. Chen, X. Y. Li and D. F. Zhong, “Development of a Liquid Chromatographic-Tandem Mass Spectrometric Method with Precolumn Derivatization for the Determination of Dencichine in Rat Plasma,” Analytica Chimica Acta, Vol. 566, No. 2, 2006, pp. 200-206. doi:10.1016/j.aca.2006.03.011
[5] E. Rubenstein, “Biologic Effects of and Clinical Disorders Caused by Nonprotein Amino Acids,” Medicine, Vol. 79, 2000, pp. 80-89.
[6] H. L. Koh, A. J. Lau and E. C. Y. Chan, “Hydrophilic Interaction Liquid Chromatography with Tandem Mass Spectrometry for the Determination of Underivatized Dencichine (β-N-oxalyl-L-α,β-diaminopropionic acid) in Panax Medicinal Plant Species,” Rapid Communications in Mass Spectrometry, Vol. 19, No. 10, 2005, pp. 1237-1244. doi:10.1002/rcm.1928
[7] G. Q. Zhao and X. X. Wang, “He-mostatic Constituent of Sanchi (Panax Notoginseng)-Dencichine,” Chinese Traditional and Herbal Drugs, 1986, pp. 17-34.
[8] S. L. Rao, “A Sensitive and Specific Colorimetric Method for the Determination of α,β-Diaminopropionic Acid and the Lathyrus Sativus Neuro-toxin,” Analytical Biochemistry, Vol. 86, No. 2, 1987, pp. 386-396. doi:10.1016/0003-2697(78)90762-5
[9] A. Geda, C. J. Briggs and S. Venkataram, “Determination of the Neurolathyrogen Beta-N-Oxalyl-L-α,β-Diaminopropionic Acid Using High-Performance Liquid- Chromatography with Fluorometric Detection,” Journal of Chromatography A, Vol. 635, No. 2, 1993, pp. 338-341. doi:10.1016/0021-9673(93)80378-L
[10] J. K. Khan, N. Kebede, Y. H. Kuo, F. Lambein and A. Debruyn, “Analysis of the Neurotoxin Beta-Odap and Its Alpha-Isomer by Precolumn Derivatization with Pheny- lisothiocyanate,” Analytical Bio-chemistry, Vol. 208, No. 2, 1993, pp. 237-240. doi:10.1006/abio.1993.1038
[11] F. Wang, X. Chen, Q. Chen, X. C. Qin and Z. X. Li, “Determination of Neurotoxin 3-N-Oxalyl-2,3-diaminopro- pionic Acid and Non-Protein Amino Acids in Lathyrus Sativus by Precolumn Derivatization with 1-Fluoro-2,4- dinitrobenzene,” Journal of Chromatography A, Vol. 883, No. 1-2, 2000, pp. 113-118. doi:10.1016/S0021-9673(00)00264-8
[12] X. Chen, F. Wang, Q. Chen, X. C. Qin and Z. X. Li, “Analysis of Neurotoxin 3-N-Oxalyl-L-2,3-diaminopro- pionic Acid and Its Al-pha-Isomer in Lathyrus Sativus by High-Performance Liquid Chromatography with 6-Ami-no-quinolyl-N-hydroxysuccinimidyl Carbamate (AQC) Derivatization,” Journal of Agricultural and Food Chemistry, Vol. 48, No. 8, 2000, pp. 3383-3386. doi:10.1021/jf000033y
[13] Z. Y. Yang, C. J. Jiao, Y. P. Wang, F. M. Li, Y. M. Liang and Z. X. Li, “A Method for the SimμLtaneous Determination of Beta-ODAP, Alpha-ODAP, Homoarginine and Polyamines in Lathyrus Sativus by Liquid Chromatography Using a New Extraction Procedure,” Analytica Chimica Acta, Vol. 534, No. 2, 2005, pp. 199-205. doi:10.1016/j.aca.2004.11.051
[14] G. X. Xie, Y. P. Qiu, M. F. Qiu, X. F. Gao, Y. M. Liu and W. Jia, “Analysis of Dencichine in Panax Notoginseng by Gas Chromatography-Mass Spectrometry with Ethyl Chloroformate Derivatization,” Journal of Pharmaceutical and Biomedical Analysis, Vol. 43, No. 3, 2007, pp. 920-925. doi:10.1016/j.jpba.2006.09.009
[15] G. D. Zhang, J. F. Cui and H. Y. Liu, Chinese Journal of Pharmaceutical Analysis, Vol. 10, 1990, pp. 209-212.

Copyright © 2022 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.