ABSTRACT
OBJECTIVE：To establish a method for the simultan eous determination of the contents of 12 flavonoids in Quzhiqiao. METHODS ：HPLC method was adopted. The determination was performed on Agilent Extend C 18 column with mobile phase consisted of 0.1％ formic acid-acetonitrile （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 35 ℃. The detection wavelength was set at 330 nm，and sample size was 10 μL. The contents of 12 components（such as eriocitrin，narirutin，naringin，naringenin，hesperidin，neohesperidin，hesperide hydrate ，luteolin，hesperide，nobiletin，hesperetin and hesperidolactone ）in 10 batches of Quzhiqiao from different collection places were determined. RESULTS ：The linear range of eriocitrin，narirutin，naringin，naringenin，hesperidin，neohesperidin，hesperide hydrate ，luteolin，hesperide，nobiletin，hesperetin and hesperidolactone were 1.65-16.51，4.50-45.02，35.41-354.12，4.11-41.12，2.29-22.86，34.96-349.56，1.42-14.15，1.50-15.04， 1.83-18.28，1.51-15.08，1.61-16.12，1.28-12.84 μg/mL，respectively（all r＞0.999 7）. The detection limits were 0.165 1，0.450 2， 3.541 2，0.411 2，0.228 6，3.495 6，0.141 5，0.150 4，0.182 8，0.150 8，0.161 2，0.128 4 μg/mL，respectively. The limits of quantitation were 0.547 8，1.487 4，11.663 3，1.360 3，0.758 3，11.594 9，0.466 3，0.497 1，0.601 2，0.499 9，0.532 3，0.424 6 μg/mL，respectively. RSDs of precision （n＝6），reproducibility（n＝6）and stability （24 h，n＝7）tests were all lower than 3％. The average recoveries were 99.50％，99.61％，98.18％，98.85％，98.48％，98.50％，98.25％，99.91％，103.13％，98.82％， 98.44％ ， 100.29％ （RSD＝1.49％ -2.38％ ， n＝6）. The contents of the above 12 components in 10 batches of samples from different collection places were 1.995 5-2.648 8，4.317 7- 5.005 1，33.215 5-34.054 6，3.140 4-3.471 5，3.221 2-3.748 8， 42.746 6-44.026 6，0.202 7-0.239 4，0.191 2-0.208 8，0.080 3- 0.097 9，0.291 9-0.307 1，0.119 9-0.149 1，0.082 7-0.089 8 mg/g. CONC LUSIONS：The method is accurate ，reliable，simple and efficient，which can be used to simultaneous determination of the contents of 12 flavonoids in Quzhiqiao ，and to provide reference for the establishment of quality control standards of Quzhiqiao.
ABSTRACT
OBJECTIVE: To determine the mass scores of naringin and neohesperidin in Fructus aurantii and Citrus chang-shan-huyou with their processed products and evaluate the quality of Fructus aurantii and Citrus changshan-huyou with their pro-cessed products. METHODS:According to the requirements of Chinese Pharmacopoeia(2010 edition)and Zhejiang Province Tradi-tional Chinese Medicine Preparation Standards (2005 edition),the moisture and ash of F. aurantii and C. changshan-huyou with their processed products were detected. And the contents of naringin and neohesperidin were determined. The ZORBAX SB-C18 column was used with the mobile phase of acetonitrile-water(20∶80,V/V)at the flow rate of 1.0 ml/min. The detection wave-length was set at 283 nm,and the column temperature was 40℃.The samples size was 10μl. RESULTS:The moisture of F. au-rantii and C. changshan-huyou was decreased after processing with no obvious change for ash. The contents of naringin and neohes-peridin were decreased,significantly for F. aurantii,and all consistent with the requirements of Chinese Pharmacopoeia(2010 edi-tion)except F. aurantii. The linear range was 0.028 45-0.284 5μg(r=0.999 7)for naringin and 0.085 9-0.858 6μg(r=0.999 6)for neohesperidin;the RSDs of precision,stability and reproducibility tests were no more than 1.36% and the average recovery was re-spectively 96.45%-100.43%(RSD=1.45%,n=6) and 98.36%-102.00%(RSD=1.26%,n=6). CONCLUSIONS: There was no significant difference in the inspection and determination re-sults in F. aurantii and C. changshan-huyou. It is suggested to adjust the limitation of content determination in the Chinese Pharmacopoeia(2010 edition)and processed standards.
ABSTRACT
Objective To clarify the influence on component and pharmacological action of Astragalus polysaccharides (APS) as complementary therapeutic agents prepared by different extraction and purification techniques. Methods Components of APS prepared by different extraction and purification techniques were analyzed, and these APS were used for synergy and attenuation of chemotherapy, radiotherapy treatment with H22 liver cancer and Lewis lung cancer of tumor-bearing mice, and also used for the regulation of immune function to immunosuppression mice. Results Experimental data were analyzed by means of statistical method to get pharmaco-result: A3 (extracted by microwave assistance and purified by membrane separation) > A4 (extracted by refluxing and purified by membrane separation) > A1 (extracted by refluxing and no purification)≈ A2 (extracted by microwave assistance and no purification). There were no significant differences on pharmacodynamic action between A1 and A2. However, compared with A1 and A2,it was worth noting that A3 and A4 exhibited good pharmacodynamic action. Then A3-in and A4-in, the samples in dialyzer after dialysis, were separated and purified to get homogeneous APS, which were the principal constituents of APS in dialyzer, with the molecular weight (Mw) of 7669 and 14 142 determined by HPGPC, respectively. The average Mw of APS outside of the dialyzer, A3-out was 3102 and A4-out 3256, which were the main compositions of A3 and A4, accounted for 79.63% and 53.92%, respectively. Conclusion APS with Mw about 5000 Da exhibit better antitumor effect and immunological activity. Refluxing, microwave assistance extractions, and membrane enrichment techniques bring different cases on Mw distribution, components and pharmacodynamic action, and obviously exhibit relationship among component, Mw distribution, and pharmacological action.