摘要
目的：建立高效液相色谱法同时测定清热解毒颗粒中绿原酸、栀子苷、木犀草苷、连翘苷、黄芩苷、木犀草素、黄芩素、汉黄芩素共8个成分的含量。方法：采用Phenomenex Luna 5μ-C18色谱柱（4.6 mm×250 mm,5μm）,以甲醇（A）-0.1%磷酸水溶液（B）为流动相,梯度洗脱,流速1 m L·min^-1,检测波长326 nm（0~26 min,测定绿原酸）、238 nm（26~35 min,测定栀子苷）、350 nm（35~44 min,测定木犀草苷）、230 nm（44~48 min,测定连翘苷）、276 nm（48~53 min,测定黄芩苷）、350nm（53~60 min,测定木犀草素）、276 nm（60~85 min,测定黄芩素、汉黄芩素）。结果：各待测组分分离度良好;绿原酸、栀子苷、木犀草苷、连翘苷、黄芩苷、木犀草素、黄芩素、汉黄芩素8个成分的进样量分别在0.041 3~0.413μg（r=0.999 6）,0.034 0~0.340μg（r=0.999 9）,0.037 1~0.371μg（r=0.999 5）,0.026 4~0.264μg（r=0.999 3）,0.196 0~1.960μg（r=0.999 8）,0.010 8~0.108μg（r=0.999 2）,0.021 2~0.212μg（r=0.999 2）,0.005 7~0.057μg（r=0.999 0）与峰面积呈良好的线性关系;加样回收率（n=6）分别为98.4%（RSD=1.6%）、99.2%（RSD=1.4%）、98.5%（RSD=1.7%）、100.7%（RSD=2.0%）、99.0%（RSD=0.9%）、98.8%（RSD=1.9%）、98.1%（RSD=1.8%）、99.9%（RSD=1.3%）。经HPLC测定,3批样品中8个成分的含量分别为绿原酸0.326~0.775 mg·g^-1,栀子苷0.167~0.581 mg·g^-1,木犀草苷0.093~0.216 mg·g^-1,连翘苷0.107~0.304 mg·g^-1,黄芩苷1.574~3.810 mg·g^-1,木犀草素0.092~0.114 mg·g^-1,黄芩素0.175~0.625 mg·g^-1,汉黄芩素0.042~0.188 mg·g^-1。结论：经方法学验证,本方法简单、快速、灵敏、准确,可用于清热解毒颗粒中绿原酸、栀子苷、木犀草苷、连翘苷、黄芩苷、木犀草素、黄芩素、汉黄芩素8个成分的含量测定。
Objective： To develop an HPLC method for simultaneous determination of chlorogenic acid,geniposide,cynaroside,phillyrin,baicalin,luteolin,baicalein and wogonin in Qingrejiedu granules. Methods： The chromatographic separation was carried out on a Phenomenex Luna 5μ-C18column（ 4. 6 mm × 250 mm,5 μm） with a mobile phase consisting of methanol（ A）- 0. 1% phosphoric acid solution（ B） in a gradient mode at a flow rate of 1 m L·min^-1. The UV detection wavelength was set at 326 nm for chlorogenic acid in the first 26 min,238 nm for geniposide during 26- 35 min,350 nm for cynaroside during 35- 44 min,230 nm for phillyrin during 44- 48 min,276 nm for baicalin during 48- 53 min,350 nm for luteolin during 53- 60 min,276 nm for baicalein and wogonin between60 and 85 min. Results： Excellent chromatographic separation was achieved and the ranges for linear correlation of chlorogenic acid,geniposide,cynaroside,phillyrin,baicalin,luteolin,baicalein and wogonin were 0. 041 3- 0. 413μg（ r = 0. 999 6）,0. 034 0- 0. 340 μg（ r = 0. 999 9）,0. 037 1- 0. 371 μg（ r = 0. 999 5）,0. 026 4- 0. 264 μg（ r =0. 999 3）,0. 196 0- 1. 960 μg（ r = 0. 999 8）,0. 010 8- 0. 108 μg（ r = 0. 999 2）,0. 021 2- 0. 212 μg（ r = 0. 999 2）and 0. 005 7- 0. 057 μg（ r = 0. 999 0）,respectively. The average recoveries（ n = 6） of the eight components were98. 4%（ RSD = 1. 6%）,99. 2%（ RSD = 1. 4%）,98. 5%（ RSD = 1. 7%）,100. 7%（ RSD = 2. 0%）,99. 0%（ RSD = 0. 9%）,98. 8%（ RSD = 1. 9%）,98. 1%（ RSD = 1. 8%） and 99. 9%（ RSD = 1. 3%）. The eight ingredient contents in 3 batches of samples were： 0. 326- 0. 775 mg·g^-1（ chlorogenic acid）,0. 167- 0. 581 mg·g^-1（ geniposide）,0. 093- 0. 216 mg·g^-1（ cynaroside）,0. 107- 0. 304 mg·g^-1（ phillyrin）,1. 574- 3. 810 mg·g^-1（ baicalin）,0. 092- 0. 114 mg·g^-1（ luteolin）,0. 175- 0. 625 mg·g^-1（ baicalein） and 0. 042- 0. 188 mg·g^-1（ wogonin） by HPLC detection. Conclusion： The established method is simple,rapid,sensitive and accurate,which can be applied to determine chlorogenic acid,geniposide,cynaroside,phillyrin,baicalin,luteolin,baicalein and wogonin in Qingrejiedu granules.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2015年第9期1601-1605,共5页
Chinese Journal of Pharmaceutical Analysis
关键词
清热解毒颗粒
绿原酸
栀子苷
木犀草苷
连翘苷
黄芩苷
木犀草素
黄芩素
汉黄芩素
高效液相色谱
Qingrejiedu granules
chlorogenic acid
geniposide
cynaroside
phillyrin
baicalin
luteolin
baicalein
wogonin
high performance liquid chromatography
