Simultaneous determination of adenine, uridine and adenosine in cordyceps sinensis and its substitutes by LC/ESI-MS

Abstract

A simple, sensitive and reproducible high performance liquid chromatography-mass spectrometry coupled with electrospray ionization method for simultaneous separation and determination of adenine, adenosine and uridine was developed. The analytical column is a 2.0 mm × 150 mm Shimadzu VP-ODS column and volume fraction of the mobile phase is 86.5% water, 12.0% methanol and 1.5% formic acid. 2-chloroadenosine was used as internal standard. Selective ion monitoring mode and selective ion monitoring ions at ratio of mass to electric charge of 136 for adenine, 268 for adenosine and 267 for uridine were chosen for quantitative analysis of the three active components. The results show that the regression equations and linear range are Y=0.062X+0.005 and 2.0 – 140.0 µg·mL⁻¹ for adenine, Y=0.049X+0.004 and 4.0–115.0 µg · mL⁻¹ for uridine, Y=0.154X+0.014 and 1.0–125.0 µm · mL⁻¹ for adenosine. The limits of detection are 0.6 µg·mL⁻¹ for adenine, 1.0 µg·mL⁻¹ for uridine and 0.2 µg·mL⁻¹ for adenosine. The recoveries of the three constituents are from 96.6% to 103.2%.

Keywords

high performance liquid chromatography / electrospray ionization interface / mass spectrometry / cordyceps sinensis / nucleoside

References

[1] ZhuJ S, HalpernG M, JonesK. The scientific rediscovery of an ancient Chinese herbal medicine: cordyceps sinensis (part I) [J]. J Altern Complement Med, 1998, 4(3): 289-303

[2] ZhuJ S, HalpernG M, JonesK. The scientific rediscovery of a precious ancient Chinese herbal regimen: cordyceps sinensis (part II) [J]. J Altern Complement Med, 1998, 4(4): 429-457

[3] Institute of Materia Medica of Chinese Academy of Medical SciencesAdvanced Study of Chinese Traditional Herbal Medicine [M], 1995, Beijing, Beijing University of Medical Sciences and Union Medical College Press(in Chinese)

[4] ZhengHu-zhan, DongZe-hong, SheJingModern Study of Traditional Chinese Medicine [M], 1999, Beijing, Xue Yuan Press(in Chinese)

[5] WuHong-zeng, DongXing-hong, MaDe-en, et al.. Comparison study of bioactive components in cordyceps sinensis and its substitutes [J]. Chinese Traditional and Herbal Drugs, 2000, 31(8): 576-578(in Chinese)

[6] LiSao-ping, LiPing, JiHui, et al.. The contents and their change of nucleosides from natural cordyceps sinensis and cultured cordyceps mycelia [J]. Acta Pharmaceutica Sinica, 2001, 36(6): 436-439(in Chinese)

[7] JiangHong, LiuKe, MengShu, et al.. Chemical constituents of the dry sorophore of cordyceps sinensis [J]. Acta Pharmaceutica Sinica, 2000, 35(9): 663-668(in Chinese)

[8] LiXue-qin, BaoTian-tong, WangYan. Determination of mannitol in Dongchongxiacao (cordyceps sinensis) by colorimetric method [J]. Chinese Traditional and Herbal Drugs, 1999, 30(1): 19-21(in Chinese)

[9] LiXue-qin, WangYan, BaoTian-tong. RP-HPLC determination of adenosine in Fermented Cordyceps [J]. China Journal of Chinese Materia Media, 1999, 24(1): 12-14(in Chinese)

[10] ChuXi-ning, ZhangQiang, ChenQiao-mei, et al.. HPLC determination of free uridine, adenine and adenosine in Isariasp [J]. Journal of Pharmaceutical Analysis, 1996, 16(4): 249-251(in Chinese)

[11] GuoCheng, ZhuJie, ZhangXun, et al.. The determination of adenosine and cordycepin in artificial cordyceps [J]. China Journal of Chinese Materia Media, 1998, 23(4): 236-237(in Chinese)

[12] LeungA K, GongF, LiangY Z, et al.. Analysis of the water soluble constituents of cordyceps sinensis with heuristic evolving latent projections [J]. Analytical Letters, 2000, 33(15): 3 195-3 211

[13] YangGen-liang, LiHai-ying, LiuHai-yan, et al.. Determination of adenine and adenosine in cordyceps inensis (Berk.) sacc and artificial cordyceps by capillary zone electrophoresis [J]. Chinese Journal of Analytical Chemistry, 2002, 30(9): 1081-1084(in Chinese)

[14] LiShao-ping, LiPing, DongTing-xia, et al.. Antioxidation activity of different types of natural cordyceps sinensis and cultured cordyceps mycelia [J]. Phytomedicine, 2001, 8(3): 207-212

[15] LiShao-ping, LiPing. Determination of nucleosides in natural cordyceps sinensis and cultured cordyceps mycelia by capillary electrophoresis [J]. Electrophoresis, 2001, 22: 144-150