摘要
用国产高速逆流色谱 (HSCCC)分离纯化中草药———丹参 ,选用正己烷 乙醇 水体系 ,固定相保留率达到78 8%。采用分步洗脱 ,3个产地丹参各分离得到 12个洗脱组分 ,经高效液相色谱仪和紫外光谱仪检测证明 3张HSCCC洗脱图谱中对应洗脱峰为同一组分。HSCCC洗脱图谱不包含非共有峰 ,并且对应洗脱峰保留时间的相对标准偏差RSD <3% ,符合国家标准关于制订指纹图谱方法学考察资料的技术参数。因此 ,HSCCC作为制订指纹图谱的方法之一具有可行性。
In an attempt to apply high-speed counter-current chromatography HSCCC for TCM fingerprints, the separation and purification of the Chinese medicinal plant Salvia miltiorrhiza Bunge of different localities was realized using the technique. The equipments used include a HSCCC (TBE-300) of Shenzhen Tauto Biotech containing three connected preparative coils (diameter of tube=2 6mm, total volume=300mL) and a 20mL sample loop and a HPLC from Shimadzu of Japan with a Ultrasphere C18 column (150×4 6mm ID, 5μm) and a 20μL sample loop. Salvia miltiorrhiza Bunge samples from 3 locations were separated by HSCCC in a Step-wise elution program with solvent systems A (hexane:ethanol:water=10:5 5:4 5) and B (hexane:ethanol:water=10:7:3) at a speed of 900 r/min and a flow-rate of 2mL/min. All the 12 peak fractions were eluted within 13 hours. The contents of each component varied greatly in different samples, which confirmed previous observation that the locations and climates have a great impact on the TCM quality and also indicated a quality control system is necessary to safeguard the quality of the herb. The retention times of the 12 peak fractions from crude extracts of the samples were collected by HPLC and the absorption spectrums of the corresponding peaks were identified. The 12 components of the three crude samples were readily distinguishable and can be used as fingerprints of S. miltiorrhiza Bunge. The relative standard deviation of the HSCCC retention times was less than 3%, which satisfies the requirement of the national standard reference index. The components 7, 8 and 11 from the standards were identified to be crypototanshinone, tanshinone I and tanshinone II A respectively. This study demonstrates that if it is possible to apply HSCCC for TCM fingerprinting, especially with samples of high viscosity and highly absorptive components. The precision and the run time of fingerprinting can be further improved if larger volume and a temperature control system is used. With these and other improvements, HSCCC is expected to play an important role in TCM development.
出处
《生物工程学报》
CAS
CSCD
北大核心
2003年第6期740-744,共5页
Chinese Journal of Biotechnology
关键词
高速逆流色谱
中药
指纹图谱
丹参
high-speed counter-current chromatography (HSCCC), fingerprints of Chinese medicinal plant, Salvia miltiorrhiza Bunge
