RESUMO
Objective To establish an HPLC method for simultaneous determination of uridine, vernine, adenosine, cordycepin, and N6-(2-hydroxyethyl)-adenosine in Cordyceps (Cordyceps militaris, Cordyceps Fungus Powder CS-4, Hirsutella sinensis, and C. sinensis), and determine the characteristic components of C. militaris to provide the scientific basis for quality control of C. militaris and its extract. Methods HPLC was performed on Inertsil ODS-3 column (250 mm × 4.6 mm, 5 μm) with mobile phase A (acetonitrile) and B (water) for gradient elution. The flow rate was 0.8 mL/min, detection wavelength was 260 nm, and column temperature was 30 ℃. Results All mentioned five nucleosides can be detected from C. militaris. However, cordycepin and N6-(2-hydroxyethyl)-adenosine were undetected in the other three Cordyceps species. The sample preparation method of C. militaris has a great effect on the content of nucleosides. The content of uridine, vernine and adenosine was the highest in samples prepared by ultrasonic extraction 180 min. Cordycepin was stable form six sample preparation methods, and N6-(2-hydroxyethyl)-adenosine was unbearable to heat and acid. The contents of cordycepin and N6-(2-hydroxyethyl)-adenosine were the same in four preparation methods. Conclusion This experiment provides a basis for the quality analysis of C. militaris and its extracts. Cordycepin and N6-(2-hydroxyethyl)-adenosine can be used as markers for the quality control of C. militaris and its extracts.
RESUMO
Objective: To establish an HPLC fingerprint method for water-soluble components in Cordyceps sinensis which could provide the basis for authenticity identification and quality evaluation of C. sinensis. Methods: HPLC method was adopted to construct the fingerprint of water soluble components in C. sinensis and its main confused species (C. liangshanensis, C. gunnii and artificial C. militaris). Total 16 common peaks were marked, and 12 common peaks were confirmed by the method with tandem high performance liquid chromatography (HPLC) and quadrupole time-of-flight mass spectrometry (Q-TOF MS). Results: The common mode of HPLC fingerprint for C. sinensis was developed. The method could distinguish between C. sinensis and its confused species, and 12 common peaks were identified, including cytosine, uracil, cytidine, hypoxanthine + guanine, uridine, thymine, adenine, inosine, guanosine, thymidine, adenosine, and cordycepin respectively. Conclusion: This method is simple and easy, providing scientific basis for identification of C. sinensis.
RESUMO
Objective To establish the HPLC fingerprint of cultured Cordyceps militaris. MethodsHPLC Method was used for the determination of C. militaris on Agilent ZOBAX SB-Aq C18 column (250 mm?4.0 mm, 5 ?m), and measuring with methanol 5%—60% in 30 min as elution detective wavelength at 260 nm, injection sample of 1.0 ?L and flow rate of 1.0 mL/min. The systematic optimized design of the chromatographic conditions, such as supersonic extraction, mobile phase, and detective wavelength as well was carried out. Results Fingerprint consisted of 11 common peaks and the result of methodology determination fitted to the related standards. The evaluation of cultured C. militaris by systematically comparing chromatograms with a professional analytical software recommended by National Institute for the Control of Pharmaceutical and Biological Products has been established. Conclusion The method is accurate, simple, and useful for the quality control of cultured C. militaris.