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目的:建立HPLC法同时测定丹参沼泽红假单胞菌转化液中丹参素、原儿茶醛、迷迭香酸、丹酚酸B和丹酚酸A 5种成分的含量,并和丹参对照液进行比较。方法:采用HPLC法,色谱柱为岛津Inert Sustain C18柱(4.6mm×250 mm,5μm);甲醇-0.1%磷酸水梯度洗脱;检测波长为287 nm,流速为0.8 m L·min-1,柱温为26℃,进样量为25μL。结果:丹参素、原儿茶醛、迷迭香酸、丹酚酸B和丹酚酸A分别在20~1000μg·m L-1,5~200μg·m L-1,5~200μg·m L-1,5~1000μg·m L-1,10~200μg·m L-1范围内呈良好的线性关系(r>0.9997)。加样回收率分别为101.31%,101.13%,98.81%,103.47%,99.06%。结论:该方法准确可靠,具有良好的精密度、重复性、稳定性和回收率,可以用于丹参沼泽红假单胞菌转化液中5个酚酸类成分含量的同时测定,为丹参沼泽红假单胞菌转化液的进一步研究奠定基础。
OBJECTIVE: To establish a HPLC method for the simultaneous determination of 5 components of Danshensu, protocatechuic aldehyde, rosmarinic acid, salvianolic acid B and salvianolic acid A in the Salvia spp. Compare. Methods: The HPLC method was performed on a Shimadzu Inert Sustain C18 column (4.6 mm × 250 mm, 5 μm) with a gradient of methanol-0.1% phosphoric acid. The detection wavelength was 287 nm and the flow rate was 0.8 m L · min -1 The column temperature was 26 ℃, the injection volume was 25μL. Results: Danshensu, protocatechuic aldehyde, rosmarinic acid, salvianolic acid B and salvianolic acid A were in the range of 20 ~ 1000μg · m L-1, 5 ~ 200μg · m L-1, 5 ~ 200μg · m L -1.5 ~ 1000μg · m L-1 and 10 ~ 200μg · m L-1, respectively (r> 0.9997). The recoveries of samples were 101.31%, 101.13%, 98.81%, 103.47%, 99.06% respectively. Conclusion: The method is accurate and reliable with good precision, repeatability, stability and recovery. It can be used for the simultaneous determination of five phenolic acids in the Salmonella Rhodopseudomonas aeruginosa transformation solution, Pseudomonas conversion liquid to lay the foundation for further research.