HPLC Characteristic Fingerprint and Chemical Pattern Recognition of Fermentation Mycelium Preparations

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1 * Lichrospher RP-C mm 250 mm 5 μm ml min nm HPLC doi /cpj R284 A HPLC Characteristic Fingerprint and Chemical Pattern Recognition of Fermentation Mycelium Preparations ZHANG Ping 1 2 ZHENG Tian-jiao 2 3 ZHANG Wen-juan 2 JIAO Kun 4 WEI Feng 2* LIU Bin 1 1. Beijing University of Chinese Medicine Beijing China 2. National Institutes for Food and Drug Control Beijing China 3. China Pharmaceutical University Nanjing China 4. Fuyang Institute for Food and Drug Control Fuyang China ABSTRACT OBJECTIVE To establish a method of HPLC characteristic fingerprint annlysis for the quality control of five fermentation mycelium preparations. METHODS The HPLC analysis was carried out on a C 18 column 4. 6 mm 250 mm 5 μm by gradient elution with methanol-water as mobile phase at a folw rate of 1. 0 ml min - 1 the column temperature was set at 25 and the detection wavelength was set at 260 nm. The HPLC characteristic fingerprint of 68 batches of five fermentation mycelium preparations was developed and the components of adenosine uridine guanosine uracil and adenine were identified. Similarity analysisi and hierarchical cluster analysis HCA were applied to study the HPLC characteristic fingerprint and chemical pattern recognition. RE- SULTS The chemical pattern recognition analysis showed that 15 batches of Bailing capsules 18 batches of Jinshuibao capsules and 10 batches of Xinganbao capsules were clustered together respectively indicating that the preparation quality of single enterprise was consistent. The HPLC chromatograms of 15 batches of Zhiling capsules were different and the difference in samples of different enterprises was obvious. CONCLUSION This method is accurate and reliable and it can be used to control the quality of fermentation mycelium preparations. KEY WORDS fermentation mycelium preparation nucleoside HPLC characteristic chromatogram similarity analysis hierarchical cluster analysis Cordyceps sinensis BerK. Sacc Tel * weifeng@ nifdc. org. cn Chin Pharm J 2015 February Vol. 50 No

2 1 Tab. 1 Information of the fermentation mycelium preparations Name Source of standard Sample Strain Sample standard Batch Manufacturer Ningxinbao capsule The Ministry of health standards WS3-B YBZ Xinganbao capsule The Ministry of health standards WS3-B Zhiling capsule The Ministry of health standards WS3-B Bailing capsule Jinshuibao capsule China Pharmacopoeia 2010 edition Cs-C-Q80 China Pharmacopoeia 2010 edition Cs-4 Cordyceps mycelium powder Cephalosporium sinensis Chen. sp. nov WS HD WS B / 10 1 Artificial Cordyceps mycelium Gliocladium roseum powder link Thom Cordyceps Mortierella powder Mortierella SP WS HD Hirsutella sinensis Liu WS Z Z 15 1 Guo Yu-et Zeng Paecilomyces hepiali 18 1 WS 3 -C Z chen Waters 2695 HPLC Lichrospher RP mm 250 mm 5 μm B - A 0 ~ 13 min 0% B 13 ~ 30 min 0 15% B 30 ~ 40 min 15% 60% B ~ 50 min 60% 95% B 1. 0 ml min nm 20 μl % 1 ml μg g 1 70% 50 ml 1. 1 Waters 2695 Waters 2996 PDA 20 min 70% Empower KQ - 250DE 10 ml 10 ml METTLER TOLEDO AE Milli-Q RSD 5 % RSD 1 % Sigma 98. 0% HPLC 18 RSD 5% 7 RSD 1. 5% Chin Pharm J 2015 February Vol. 50 No h

3 HPLC RSD 5 % 5 S RSD 1. 0 % 24 1 h S S S1 S2 S3 S4 S A - 5 HPLC B - C - D - E - F - Fig. 1 The HPLC characteristic chromatograms of the fermentation mycelium preparations and five reference substances A - HPLC characteristic chromatogram of five reference substances B - HPLC characteristic chromatogram of Bailing capsules C - HPLC characteristic chromatogram of Jinshuibao capsules D - HPLC characteristic chromatogram of Ningxinbao capsules E - HPLC characteristic chromatogram of Xinganbao capsules F - HPLC characteristic chromatogram of Zhiling capsules Chin Pharm J 2015 February Vol. 50 No

4 S chempattern ~ ~ ~ ~ ~ ~ Fig. 2 The similarity analysis results of the fermentation mycelium 68 chempattern preparations Fig. 3 The cluster analysis dendrogram of the fermentation mycelium 68 preparations 296 Chin Pharm J 2015 February Vol. 50 No

5 C A B C D E Fig. 4 The similarity distribution of Zhiling Capsules 5 A - B - C - D - E - Fig. 5 The HPLC characteristic chromatograms of the fermentation mycelium preparations A - HPLC characteristic chromatograms of 15 bathches of Bailing capsules B - HPLC characteristic chromatograms of 18 bathches of Jinshuibao capsules C - HPLC characteristic chromatograms of 10 bathches of Ningxinbao capsules D - HPLC characteristic chromatograms of 10 bathches of Xinganbao capsules E - HPLC characteristic chromatograms of 15 bathches of Zhiling capsule Chin Pharm J 2015 February Vol. 50 No

6 4 2 2 sinensis J 3. 3 ry of diabetic rats J PCR - J. Saudi Med REFERENCES 1 Ch. P 2010 VolⅠ 2010 S JIANG Y YAO Y J. Anamorphic Fungi related to Cordyceps sinensis J. Mycosystema ZHANG M H HONG L GUO Q. Determination of the adenosine content of Fermented Cordyceps powder and Jinshui Bao capsule by HPLC J. Chin Tradit Herb Drugs YANG C HOU S Y YU J et al. Ningxinbao capsule in the treatment of arrhythmia J. Chin J New Drugs Clin Remed ZHANG Z XIE Y Z LI S Z et al. Study progress in Cordyceps. J Microbiol KO W S S HSU S L. Compound Cordyceps TCM-700C exhibits potent hepatoprotective capability in animal model J. Fitoterapia CUI H Y. Protective effects of Cordyceps sinensis on kidney inju-. J Med Sci Yanbian Univ 8 GAO B YANG J. Cordyceps sinensis extract suppresses hypoxiainduced proliferation of rat pulmonary artery smooth muscle cells 9 GUAN C H LIU J. Effect of Dongchong Xiacao on pulmonary function and Th1 /Th2 of rat model with chronic obstructive pulmonary disease J. Zhejiang J Integr Tradit Chin West Med ZHANG P XIAO X Y LI Y K et al. Chemical analysis of nucleosides and alkalines in five preparations of fermental Cordyceps J. Chin J Pharm Anal ZHANG P XIAO X Y HUANG W et al. RP-HPLC-UV Determination of ergosterol in five preparations of fermental cordyceps J. Chin J Pharm Anal Luc Besan on doi /cpj Chin Pharm J 2015 February Vol. 50 No

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