Studies On Content Determination Of Rhynchophylline And Hplc Fingerprint In U.Macrophylla Wall.

Objective:To build the methods of rhynchophylline content determination and fingerprint of U. macrophylla Wall. and evaluate the quality of U. macrophylla Wall. from different places, different parts and different collecting times. scientifically for controlling the quality of U. macrophylla Wall. and providing test basis for utilization of U. macrophylla Wall. resources.Methods:(1) The HPLC was used to determine rhynchophylline content in U. macrophylla Wall. from different places, different parts and different collecting times.Chromatographic conditions:chromatographic column was Gemini C18(4.6 mm X 250 mm,5μm), mobile phase was methanol- 0.002 mol/L triethylomine (pH value 7.5, adjusted with acetic acid)(64 : 36), flow rate was 1.0ml/min, detective wavelength was 254nm and injection volume was 20μL.Preparation conditions:madicinal materials was weighed 0.2g, then ultrasonic extracted with 15ml 75%methanol lasted for 40 minutes, taked the continued filtrate through 0.45μm microporous membrane. (2) The HPLC was used to determine fingerprint in U. macrophylla Wall. from different places, different parts and different collecting times.Chromatographic conditions:chromatographic column was Gemini C18(4.6 mm×250 mm,5μm), mobile phase was methanol- 0.002 mol/L triethylomine (pH value 7.5, adjusted with acetic acid)(gradient elution), flow rate was 1.0ml/min, detective wavelength was 254nm,column temperature was 30℃and injection volume was 20μL.Preparation conditions:the madicinal materials was weighed 0.2g, ultrasonic extracted with 15mL 75% ethanol lasted for 40 minutes, taked the continued filtrate through 0.45μm microporous membrane.Results:(1) There was a differences in different places, the content of rhynchophylline between 0.143%～0.642%, average rhynchophylline content in branch with hook, hook without branch, branch without hook and stem were similar, which were 0.386%,0.375%,0.393% and 0.351%,leaves was the highest,which was 0.483%, the content of rhynchophylline in different collecting times has some differencs.(2) Similarity in different place were 0.909～0.998;similarity in different parts were that: branch with hook(0.909～0.998), hook without branch(0.883～0.997), branch without hook(0.871～0.999), stem(0.877～0.998) and leaves(0.917～0.992); similarity in different collecting times was 0.975～0.997. Confirmed the main technical parameters, such as six superior peaks and total relative area.Conclution:(1)the methods of rhynchophylline content determination and fingerprint of U. macrophylla Wall. was builded. It showed that the methods of the content determination and fingerprint detection were stable, reliable, repeatable and specific by methodology test. It can be applied to control the quality of U. macrophylla Wall..(2) Rhynchophylline can be detected in different places, and the content was high, but it had some differences between places, U. macrophylla Wall. may be the dominant variety on uncaria; Besides leaves had higher content of rhynchophylline, other parts were similar, and there was a value for development and utilization, the content of rhynchophylline from different collecting times has some differences,but it was indiscipline,which to show that it was reasonable to collect in all years.(3) The standard fingerprint of U. macrophylla Wall. from different places, different parts and different collecting times was builded. Total of the 7 common peaks are recognized as characters of the chromatogram with the rhynchophylline peak as the reference peak.Generally speaking, their similarity were high. The results indicate that the consistency of the fingerprints of U. macrophylla Wall.in Guangxi was good. From six superior peaks and total relative area, it showed that main chemical components which contents were high had little differences, most of them were similar, and the quality was stable.