Comparison of HPLC fingerprint between enzymatic Calculus bovis and natural Calculus bovis
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- Ιπποκράτης Παπαφιλίππου
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1 HPLC * HPLC - DAD DiamonsilC mm 4. 6 mm 5 μm - -1% 370 nm 1. 0 ml /min 40 R A Comparison of HPLC fingerprint between enzymatic Calculus bovis and natural Calculus bovis LI Ke QI Yong-xiu YU Xiu-ling WANG Fu-gang LI Bing-long GAO Yun-sheng * Dept. of Pharmacy Taishan Medical College Taian Shandong China KEY WORDS natural Calculus bovis calculus bovis cultivated by glucuronidase HPLC fingerprint ABSTRACT AIM To develop an HPLC-PDA fingerprint analysis method of natural Calculus bovis in comparison with Calculus bovis cultivated by glucuronidase. METHODS An HPLC analysis was carried out on a Diamonsil C 18 column 4. 6 mm 250 mm 5 μm with the mobile phase consisting Methanol A -chloroform -1% acetic acid aqueous solution in gradient mode. The detection wavelength was set at 370 nm. The flow rate was 1. 0 ml min - 1 and the analytic time was 40 min. RESULTS Under the above chromatographic conditions the mutual mode of HPLC fingerprints was set up Calculus bovis cultivated by glucuronidase had a similar fingerprint chromatogram to the natral Calculus bovis but there was a little difference in their concent. CONCLUSION The method is in good precision and reproducibility and could be available for the identification of various substitutes of Calculus bovis. Calculus bovis Bostaurus 2-3 domesticus Gmelin % ~ 2% Waters DAD Calculus bovis cultivated by glucuronidase Empower Waters Sarto- rious BT 25S 1 / CBCG ZRQN060 J10LF Tel likegaofeng@ 126. com. * Tel ysgao@ tsmc. edu. cn. 1
2 KQ-500DE AP-01P VACUUM PUMP Autoscience Autoscience HPLC Fig. 2 HPLC chromatogram of natural Calculus bovis 2. 3 in dimethyl sulfoxide DMSO nm nm 4 Bostaurus domesticus Gmelin nm ml nm HPLC 2. 2 Fig. 3 HPLC chromatogram of natural Calculus bovis at 450 nm 10 ml min 6000 r /min 10 min 2 20 μl nm HPLC 1 2 Fig. 4 HPLC chromatogram of natural Calculus bovis ~ 400 nm DAD at 370 nm min 40 min 40 min 1 Fig HPLC HPLC chromatogram of natural Calculus bovis in methanol-chloroform-hydrochloride-h 2 O min HPLC Fig. 5 HPLC chromatogram of natural Calculus bovis in 60 minites 2
3 DiamonsilC mm 4. 6 mm 5 μm A - B -1% C 0 ~ 10 min 65% A 6% B 29% C 11 ~ 30 min 82 % A 15% B 3% C 31 ~ 40 min 65 % A 6% B 29% C 1. 0 ml / min 370 nm 0 ~ 40 min 20 μl Fig. 6 RSD HPLC chromatogram of 10 batches of natural Calculus bovis % 0. 97% h RSD % 1. 34% Fig. 7 Mutual pattern of HPLC chromatogram on natural Calculus bovis RSD % 1. 55% μl HPLC 2004A 0 ~ 40 min HPLC Fig. 8 HPLC chromatogram on 10 batches of Calculus bovis cultivated by glucuronidase and mutual pattern 6 7 of natural Calculus bovis HPLC A % S HPLC 10 3
4 Tab Relative reserving time of 10 batches of natural Calculus bovis samples Tab Relative peak area of 10 batches of natural Calculus bovis samples A - B A - B -1% C A - B -1% C 0 ~ 10 min 65% A 6% B 29% C 11 ~ 30 min82 % A 15% B 3% C 31 ~ 40 min 65 % A 6% B 29% C HPLC HPLC J β- J J HPLC-ELSD J HPLC J J
5 J M HPLC * HPLC Agilent Zorbax Extend-C mm 250 mm 5 μm % 0. 2% 268 nm 0. 7 ml /min R A HPLC fingerprint of coptis-evodia couplet medicines TU Yao-sheng LIU Fa-jin SUN Dong-mei ZHUANG Yu-jian Guangdong Provincial Institute of TCM Guangzhou Guangdong China KEY WORDS coptis-evodia couplet medicines HPLC fingerprints ABSTRACT AIM The HPLC fingerprint of coptis-evodia couplet medicines was carried out to illustrate the effect of the compatibility to chemical compounds and to explain the mechenism of this compatibility. METHODS The chromatographic fingerprints were obtained by measuring the coptis-evodia couplet medicine on an Agilent Zorbax extend-c 18 column 4. 6 mm 250 mm 5 μm with the gradient elution solvent system of acetonitrile-0. 3% phosphoric 0. 2% triethylamine-water. The flow rate was 0. 7 ml /min the column temperature was maintained at 20 and the detection wavelength was set at 268 nm. RESULTS Nineteen co-possessing peaks were selected as the fingerprint peaks and preliminary assignment was also performed by taking berberine peak as the reference peak. CONCLUSION From the couplet medicines angle adding coptis into evodia in stew proves beneficial to the stripping of liposoluble alkaloids come from evodia in proportion Agilent 1200 DAD * Tel tcmgdp@ 163. com 5
HPLC- ESI-MS HPLC-ESI-MS HPLC-ESI-MS HPLC 11 HPLC HPLC-ESI-MS. Asterias rollestoni Bell. LC- MS. Vol.11 No.1
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