Natural Product Sciences

The Korean Society of Pharmacognosy (한국생약학회)
권호 표지

HPLC-tandem Mass Spectrometric Analysis of the Marker Compounds in Forsythiae Fructus and Multivariate Analysis

  • Received : 2011.04.17
  • Accepted : 2011.06.12
  • Published : 2011.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

A high-performance liquid chromatography-electrospray ionization-tandem mass spectrometric method was developed to determine simultaneously eight marker constituents of Forsythiae fructus, and subsequently applied it to classify its two botanical origins. The marker compounds of Forsythia suspensa were phillyrin, pinoresinol, phillygenin, lariciresinol and forsythiaside; those of F.viridissima were arctiin, arctigenin and matairesinol. Separation of the eight analytes was achieved on a phenyl-hexyl column (150${\times}$2.0 mm i.d., 3 ${\mu}M$) using gradient elution with the mobile phase: (A) 10% acetonitrile in 0.5% acetic acid, (B) 40% aqueous acetonitrile. A few fragment ions specific to the types of lignans, among the product ions generated by collisonally induced dissociation (CID) of molecular ion clusters, such as [M-H]$^-$ or [M+OAc]$^-$ were used not only for fingerprinting analysis but for the quantification of each epimer by using multiple-reaction monitoring mode. It was shown good linearity ($r^2{\geq}$ 0.9998) over the wide range of all analytes; intra- and inter-day precisions (RSD, %) were within 9.14% and the accuracy ranged from 84.3 to 115.1%. The analytical results of 40 drug samples, combined with multivariate statistical analyses - principal component analysis (PCA) and hierarchical cluster analysis (HCA) - clearly demonstrated the classification of the test samples according to their botanical origins. This method would provide a practical strategy for assessing the authenticity or quality of the herbal drug.

Keywords

References

  1. Chen, C.C., Chen, H.Y., Shiao, M.S., Lin, Y.L., Kuo, Y.H., and Ou, J.C., Inhibition of Low Density Lipoprotein Oxidation by Tetrahydrofurofuran Lignans from Forsythia suspensa and Magnolia coco. Planta Med. 65, 709-711 (1999). https://doi.org/10.1055/s-1999-14093
  2. Chiba, M., Hisada, S., and Nishibe, S., Studies on the chinese crude drug "Forsythiae fructus." III. On the constituents of fruits of Forsythia viridissima and F. suspense. Shoyakugaku Zasshi. 32, 194-197 (1978).
  3. Chiba, M., Tsukamoto, H., Hisada, S., and Nishibe, S., Studies on the Chinese crude drug "Forsythiae fructus". IV. On the constituents of fruits of Forsythia koreana and forsythiae fructus from Korea on the market, Shoyakugaku Zasshi. 33, 150-154 (1979).
  4. Cho, J.Y., Kim, A.R., Yoo, E.S., Baik, K.U., and Park, M.H., Immunomodulatory effect of arctigenin, a lignan compound, on tumour necrosis factor-alpha and nitric oxide production, and lymphocyte proliferation, J. Pharm. Pharmacol. 51, 1267-1273 (1999). https://doi.org/10.1211/0022357991777001
  5. Cho, M.K., Park, J.W., Jang, Y.P., Kim, Y.C., and Kim, S. G., Potent inhibition of lipopolysaccharide-inducible nitric oxide synthase expression by dibenzylbutyrolactone lignans through inhibition of $I-{\kappa}B{\alpha}$ phosphorylation and of p65 nuclear translocation in macrophages, Int. Immunopharm. 2, 105-116 (2002). https://doi.org/10.1016/S1567-5769(01)00153-9
  6. Eich, E., Pertz, H., Kaloga, M., Schulz, J., Fesen,M.R., Mazumder, A., and Pommier, Y., (−)-Arctigenin as a Lead Structure for Inhibitors of Human Immunodeficiency Virus Type-1 Integrase, J. Med. Chem. 39, 86-95 (1996). https://doi.org/10.1021/jm950387u
  7. Eklund, P.C., Backman, M.J. Kronberg, L., Smeds, A.I., and Sjoholm, R.E. Identification of lignans by liquid chromatography-electrospray ionization ion-trap mass spectrometry, J. Mass Spectrom. 43, 97-107 (2008).
  8. Guo, H., Liu, A-H., Li, L., and Guo, D-A. Simultaneous determination of 12 major constituents in Forsythia suspensa by high performance liquid chromatography-DAD method. J. Pharm. Biomed. Anal. 43, 1000-1006 (2007). https://doi.org/10.1016/j.jpba.2006.09.033
  9. Guo, H., Liu, A-H., Ye, M., Yang M., and Guo, D-A., Characterization of phenolic compounds in the fruits of Forysthia suspense by highperformance liquid chromatography coupled with electrospray ionization tandem mass spectrometry., Rapid Commun. Mass Spectrom. 21, 715- 729 (2007). https://doi.org/10.1002/rcm.2875
  10. Hikino, H., Constituents and physiological actions of Schizonepeta and Forsythia, J. of Traditional Sino Japanese Medicines. 3, 41-45 (1982).
  11. Hirose, M., Yamaguchi, T., Lin, C., Kimoto, N., Futakuchi, M., Kono, T., Nishibe, S., and Shirai, T., Effects of arctiin on PhIP-induced mammary, colon and pancreatic carcinogenesis in female Sprague- Dawley rats and MeIQx-induced hepatocarcinogenesis in male F344 rats, Cancer Lett. 155, 79-88 (2000). https://doi.org/10.1016/S0304-3835(00)00411-0
  12. Huang, D.M., Guh, J.H., Chueh, S.C., and Teng, C.M., Modulation of anti-adhesion molecule MUC-1 is associated with arctiin-induced growth inhibition in PC-3 cells, Prostate. 59, 260-267 (2004). https://doi.org/10.1002/pros.10364
  13. Huanhuan Qu, Baixue Li, Xu Li, Guangzhong Tu, Juan Lü, and Wenji Sun, Qualitative and quantitative analyses of three bioactive compounds in different parts of Forsythia suspensa by highperformance liquid chromatography-electrospray ionization-mass spectrometry, Microchem. J. 89, 159-164 (2008). https://doi.org/10.1016/j.microc.2008.02.002
  14. Ishida, J., Wang, H.K., Oyama, M., Cosentino, M.L., Hu, C.Q., and Lee, K.H., Anti-AIDS agents 46. Anti-HIV activity of harman, an anti-HIV principle from Symplocos setchuensis, and its derivatives, J. Nat. Prod. 64, 958-960 (2001). https://doi.org/10.1021/np0101189
  15. Ishizuka, O., Kumazawa, N., Ohta, S., Kamogawa, A., and Shinoda, M., Effects of various methanol extracts of crude drugs on experimental subacute and chronic hepatic injury, Yakugaku Zasshi. 112, 174-182 (1992). https://doi.org/10.1248/yakushi1947.112.3_174
  16. Jang, Y.P., Kim, S.R., and Kim, Y.J., Nueroprotective dibenzylbutyrolactone lignans of Torreya nucifera, Planta Med. 67, 470-472 (2001). https://doi.org/10.1055/s-2001-15804
  17. Kang, H.S., Lee, J.Y., and Kim, C.J., Anti-inflammatory activity of arctigenin from Forsythiae Fructus, J. Ethnopharmacol. 116, 305-312 (2008). https://doi.org/10.1016/j.jep.2007.11.030
  18. Kim, S.H., Jang, Y.P., Sung, S.H., Kim, C.J. et al., Hepatoprotective dibenzylbutyrolactone lignans of Torreya nucifera against $CCl_4$-induced toxicity in primary cultured rat hepatocytes, Biol. Pharm. Bull. 26, 1202-1205 (2003). https://doi.org/10.1248/bpb.26.1202
  19. Kimura, Y., Okuda, H., Nishibe, S., and Arichi, S., Effects of Caffeoylglycosides on Arachidonate Metabolism in Leukocytes, Planta Med. 53, 148-153 (1987). https://doi.org/10.1055/s-2006-962658
  20. Kitagawa, S., Nishibe, S., and Baba, H., Studies on the Chinese crude drug "forsythiae fructus". VIII. On isolation of phenylpropanoid glycosides from fruits of Forsythia koreana and their antibacterial activity, Yakugaku Zasshi. 107(4), 274-278 (1987). https://doi.org/10.1248/yakushi1947.107.4_274
  21. Kitagawa, S., Tsukamoto, H., Hisada, S., Nishibe, S., and Studies on the Chinese Crude Drug "Forsythiae Fructus." VII. A New Caffeoyl Glycoside from Forsythia viridissima. Chem. Pharm. Bull. 32, 1209- 1213 (1984). https://doi.org/10.1248/cpb.32.1209
  22. Lee, E.B. and Keum, H.J., Pharmacological Studies on Forsythiae Fructus, Kor. J. Pharmacogn. 19(4), 262-269 (1988).
  23. Lee, J.S., Min, B.S., and Bae, K.W., Effects of Forsythiae fructus on L1210 and HL-60 cells, Yakhak Hoeji. 40, 462-467 (1996).
  24. Lim, H., Lee, J.G., Lee, S.H., Kim, Y.S., and Kim, H.P., Antiinflammatory activity of phylligenin, a lignan from the fruits of Forsythia koreana, and its cellular mechanism of action, J. Ethnopharmacol. 118, 113-117 (2008). https://doi.org/10.1016/j.jep.2008.03.016
  25. Miura, M., Ohta, S., and Kamogawa, A., Shinoda, M, Basic study of assay method of choleretic effect and the screening of crude drugs, Yakugaku Zasshi. 107, 992-1000 (1987). https://doi.org/10.1248/yakushi1947.107.12_992
  26. Namba, T., The Encyclopedia of Wakan Yaku. Hoikusha, Osaka, 1993, pp. 296-298.
  27. Nikaido, T., Ohmoto, T., Kinoshita, T., Sankawa, U., Nishibe, S., and Hisada, S., Inhibition of cyclic AMP phosphodiesterase by lignans. Chem. Pharm. Bull. 29. 3586-3592 (1981). https://doi.org/10.1248/cpb.29.3586
  28. Nishibe, S., Chiba, M., and Hisada, S., Studies on the Chinese crude drug "forsythiae fructus." I. Constituents of Forsythiae fruits on the market, Yakugaku Zasshi. 97, 1134-1137 (1977a). https://doi.org/10.1248/yakushi1947.97.10_1134
  29. Nishibe, S., Chiba, M., and Hisada, S., Studies on the Chinese crude drug "Forsythiae fructus." II. Comparative examination on the lignin glucosides of Forsythia fruits of the original plants listed in the Japanese Pharmacopoeia Ed. IX, Shoyakugaku Zasshi. 31, 131-135 (1977b).
  30. Nishibe, S., Noguchi, Y., Yoshida, A., and Kawamura, T., On the marker compounds and methods for chemical identification of forsythia fruit and plantago seed. Nat. Medicine. Tokyo, Japan, 55(5), 272-275 (2001).
  31. Nishibe, S., Okabe, K., Tsukamoto, H., Sakushima, A., Hisada, S., Baba, H., and Akisada, T. Studies on the Chinese crude drug ''Forsythiae Fructus''. VI. The structure and antibacterial activity of suspensaside isolated from Forsythia suspensa. Chem. Pharm. Bull. 30, 4548-4553 (1982a). https://doi.org/10.1248/cpb.30.4548
  32. Nishibe, S., Okabe, K., Tsukamoto, H., Sakushima, A., and Hisada, S., Studies on the Chinese crude drug Forsythia fructus. V. The structure of forsythiaside isolated from Forsythia suspensa. Chem. Pharm. Bull. 30, 1048-1050 (1982b). https://doi.org/10.1248/cpb.30.1048
  33. Nishibe, S., The plant origins of herbal medicines and their quality evaluation. Yakugaku Zasshi. 122, 363-379 (2002). https://doi.org/10.1248/yakushi.122.363
  34. Noro, Y., Hisata, Y., Okada, K., Kawamara, T., Tanaka, T., and Nishibe, S., Phenylethanoid glycosides in the fruits of Forsythia spp. Shoyakugaku Zasshi. 45, 327-332 (1991).
  35. Ozaki, Y., Rui, J., Tang, Y, and Satake, M., Antiinflammatory effect of Forsythia suspensa Vahl and its active fraction. Biol. Pharm. Bull. 20, 861-864 (2000).
  36. Pharmacopoeia Commission of PRC, Pharmacopoeia of the People's Republic of China, vol.1, Chemical Industry Press, Beijing, 2005, pp.117-118.
  37. Prieto, J.M., Recio, M.C., Ciner, R.M., Manez, S., Giner, E.M., and Rios J.L., Influence of traditional Chinese anti-inflammatory medicinal plants on leukocyte and platelet functions, J. Pharm. Pharmacol. 55, 1275-1282 (2003). https://doi.org/10.1211/0022357021620
  38. Rho, Y.S., Antifungal Studies on Components from the Pericarp of Forsythia viridissima (I), Kor. J. Pharmacog. 6, 143-147 (1975).
  39. Ricci, A., Fiorentino, A., Piccolella, S., Golino, A., Pepi, F., D'Abrosca, B., Letiza, M., and Monaco, P. : Furofranic glycosylated lignans: a gas-phase ion chemistry investigation by tandem mass spectrometry. Rapid Commun. Mass Spectrom. 22, 3382-3392 (2008). https://doi.org/10.1002/rcm.3746
  40. Schinella, G.R., Tournier, H.A., Prieto, J.M., Mordujovich, Rios, J.L., Antioxidant activity of anti-inflammatroy plant extracts. Life Sci. 70(9), 1023-1033 (2003).
  41. Schmidt, T.J., Henmati, S., and Fuss, E., Alfermann, A.W., A Combined HPLC-UV and HPLC-MS method for the identification of lignans and its application to the lignans of Linum usitatissimum L. and L. bienne Mill., Phytochem. Anal. 17, 299-311 (2006). https://doi.org/10.1002/pca.918
  42. Takagi, K., Kimura, M., Harada, M., and Otsuka, Y., Pharmacology of Medicinal Herbs in East Asia. Nanzando, Tokyo, 187-188 (1982).
  43. Takasaki, M., Konoshima, T., Komatsu, K., Tokuda, H., and Nishino, H., Cancer chemopreventive activity of lignans from the aerial part of Saussurea medusa. Cancer Lett. 158, 53-59 (2000). https://doi.org/10.1016/S0304-3835(00)00499-7
  44. Tokar, M. and Klimek, B., The content of lignan glycosides in Forsythia flowers and leaves. Acta Poloniae Pharmaceutica. 61(4), 273-278 (2004).
  45. Vlietinck, A.J., De Bruyne, T., Apers, S., and Pieters, L.A., Plant-derived leading compounds for chemotherapy of human immunodeficiency virus (HIV) infection, Planta Med. 64, 97-109 (1998). https://doi.org/10.1055/s-2006-957384
  46. WHO and Natural Products Research Institute, Seoul National Univ., WHO Regional Publications, Western Pacific Series No.21, MEDICINAL PLANTS IN THE REPUBLIC OF KOREA., World Health Organization, 1998, pp.125. ISBN 9290611200
  47. Woo, W.S. and Lee, E.B., The Screening of Biological Active Plants in Korea Using Isolated Organ Preparations (II) -Anticholinergic and Oxytocic Actions in the Ileum and Uterus, Kor. J. Pharmacog. 10, 27- 30 (1979).
  48. Xinyixueyuan, Dictionary of Traditional Chinese Medicine, Shanghai Scientific & Technologic Press, Shanghai, 292, 1977, pp. 1111-1113
  49. Ye, M., Yan, Y. and Guo, D-A., Characterization of phenolic compounds in the Chinese herbal drug Tu-Si-Zi by liquid chromatography coupled to electrospray ionization mass spectrometry, Rapid Commun. Mass Spectrom. 19, 1469-1484 (2005). https://doi.org/10.1002/rcm.1944