| Forsythia suspensa (Thunb.) Vahl belongs to Oleaceae, Forsythia. It falls into perennial deciduous shrub, distributing in China's Henan, Shanxi and Shaanxi provinces. Its main chemical compositions include phillyrin, forsythoside, rutin, oleanolic acid etc. It has the effect of resisting bacteria, diminishing inflammation and allaying fever. It is often used to treat Cold, Scrofula, Urinary infection etc. As one of the traditional Chinese clinical herbs, F. suspense has a relatively high medicinal value.F. suspense from Shangluo area in Shaanxi province is unadulterated medicinal materials. According to textual research, the characteristic geographical location, climate and soil are favorable for F. suspense 's growth. The total area of wild F. suspense and purchasing amount in Shangluo area ranks first in Shaanxi province. Shangluo is one of main areas producing F. suspense in China. Currently there is no international standard for Chinese traditional medicine's production and quality. Thus, quality control and stability of traditional Chinese medicine become key issues in production. The development and application of technology of fingerprints provide an effective method for quality control of traditional Chinese medicine.This paper studies on the F. suspense from Shangluo production base of Medicinal Materials. Adopting HPLC, we determined and compared phillyrin contents in the same part of different F. suspense and different parts of the same F. suspense. Then we established HPLC fingerprints diagram, which provides new methods and theoretical foundation for quality control of F. suspense from Shangluo production base of Medicinal Materials.Results of the experiment:1 .We compared several methods to distill phillyrin in different materials of F. suspense and finally selected ultrasonic distilling method using pure methanol. Compared with the distilling methods mentioned in references, the method we chooseis simple, easy to manipulate and materials are adequately distilled. The processing flow of F. suspense's leaves and fruits is: pick—>-drying at normal temperature—>crush and sieve using 60-hole sieve—>-measure accurately—^ultrasonic for 30min—?distilled liquid;the processing flow of F. suspense's flowers is: pick—>drying at normal temperature—-?crush—>measure accurately—^ultrasonic for 30min—>distilled liquid;2. Adopting HPLC, we determined the content of phillyrin in the same part of different F. suspense: the phillyrin content in the flowers of F. suspensa is 0.80 %±0.34%;the phillyrin content in the leaves of green F. suspensa is 3.05 %±1.21%;the phillyrin content in the leaves of ripe F. suspensa is 2.48 %±1.31%;the phillyrin content in green F. suspensa is 0.28%±0.10%;the phillyrin content in ripe F. suspensa is 0.20%±0.03%. These all accord with 2005 pharmacopoeia edition indicating phillyrin content should not be lower than 0.15 %,-3. Adopting HPLC, we determined the phillyrin content in different parts of the same F. suspensa: the average phillyrin content in the flowers of F. suspensa is 0.68 %;the average phillyrin content in the leaves of green F. suspensa is 3.37%;the average phillyrin content in the leaves of ripe F. suspensa is 2.99%;the average phillyrin content in green F. suspensa is 0.19%;the average phillyrin content in ripe F. suspensa is 0.13%. We thus concluded that the phillyrin content in different parts of the same F. suspensa are: leaves of green F. suspensa > leaves of ripe F. suspensa> flowers of F. suspensa > green F. suspensa > ripe F. suspensa;4. We establish separate HPLC measuring methods for phillyrin content in the leaves, fruits and flowers of F. suspensa. Chromatogram condition is: wavelength 205nm;flowing speed l.Oml/min;mobile phase elutes at A(methanol)-B(0.1% icy acetic acid) grads. ?F. suspensa's leaves: 20%—25%A (0.01—5 min.) , 25%— 30%A (5—15 min) , 30%—40%A (15—40 min) , 40%—50%A (40—65 min) ? ?F. suspensa's fruits and flowers: 15 %—20% A (0.01 — 5 min.) , 20%—25% A(5~20 min) , 25%—40% A (20—32 min) , 40%—42% A (32—50 min) , 42%~50%A (50-60 min) , 50% A (60-65 min);5. Via systematic research of fingerprints of different parts of F. suspensa, weestablished fingerprints research methods for different parts of F. suspensa. Methodology showed: measuring precision of F. suspensa's leaves RSD<2.38%, reappearance RSD<2.73%, stability RSD<4.97%, measuring precision of F. suspensa's fruits and flowesr RSD<2.45%, reappearance RSD< 2.07%, stability RSD<1.70%, all samples are stable within 48h;6. We observed HPLC fingerprints of different parts of 12 batches of F. suspensa samples. The total crests of F. suspensa's leaves, fruits and flowers are 13n 10 and 13 respectively. Compared with the standard chromatogram diagram, in HPLC fingerprints of the F. suspensa's leaves, we ascertained that the 10th crest was phillyrin;in HPLC fingerprints of the F. suspensa's fruits, we ascertained that the 6th crest was phillyrin;in HPLC fingerprints of F. suspensa's flowers, we ascertained that the 1 lth crest was phillyrin. At the same time, we set down parameters of technology of fingerprints in different parts of F. suspensa, using phillyrin as a comparing crest,We calculated relative maintaining time, relative areas and similarities of mutual crests of 12 samples. |
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