HPLC波长切换法同时测定热毒清锭中3种有效成分含量

目的 建立HPLC波长切换法,同时测定热毒清锭中绿原酸,木犀草苷和延胡索乙素含量.方法 采用流动相：乙腈-0.4%磷酸溶液-0.4%（以三乙胺调至pH 6.5）磷酸溶液系统按梯度洗脱分离测定;检测波长波长绿原酸326nm;木犀草苷350nm;延胡索乙素282nm.色谱柱;Waters Symmertry C18（4.6mm×150mm,5μm）柱.结果 线性范围分别为绿原酸0.0227～0.363μg（r=0.9999）;木犀草苷0.00388μg～0.0621μg（r=0.9998）;延胡索乙素0.274～4.38μg（r=0.9999）,回收率分别为98.6%、98.3%和98.8%.RSD依次为1.1%、1.0%和1.2%.结论 本法应用梯度洗脱和波长切换技术,将热毒清锭中绿原酸,木犀草苷和延胡索乙素很好的分离,并通过波长切换技术,选择待测成分检测波长,减少其他成分的吸收干扰,提高检测灵敏度,专属性,可同时测定热毒清锭中绿原酸,木犀草苷和延胡索乙素含量,从而提升该制剂质量的控制水平.

Simultaneous determination of Chlorogenic acid,Luteolin and Tetrahydropalmatine in Reduqing Ingots by HPLC with the wavelength switching method

OBJECTIVE To develop a HPLC wavelength switching method for simultaneous determination of Chlorogenic acid, Luteolin and Tetrahydropalmatine in Reduqing Ingots. METHODS The mobile phase consisted of acetonitrile,0. 4% phosphoric acid solution and 0.4% phosphoric acid solution （ PH was adjusted to 6. 5 using triethylamine） in gradient mode. The detection wavelength was set at 326nm for chlorogenie acid,350nm for luteolin and 282nrn for tetrahydropalmatine. The column： Waters Symmertry C18 （4.6mm × 150mm, 5μm ） column. RESULTS The linear ranges were 0. 0227 - 0. 363μg （ r = 0. 9999 ） for ehlorogenie acid, 0. 00388 - 0. 0621 μg（ r = 0. 9998 ）for luteolin and 0. 274 - 4.38μg （r = 0. 9999 ） for tetrahydropalmatine. The average recover- ies of the compounds above were 98.6% ,98.3% and 98.8% respectively. The RSDs were 1.1% ,1.0% and 1.2% in sequence. CONCLUSION The method was established to obtain good separation of ehlorogenie acid, luteolin and tetrahydropalmatine using the gradient elution. The wavelength switching technology with selective detection wavelength can reduce the interference absorption of other ingredients and improve the sensitivity and specificity of the detection,which can be used for simultaneous determination of Chlorogenie acid, Luteolin and Tetrahydropalma- tine in reduqing ingots and thereby enhancing the level of guality control of the preparation.