高效液相色谱梯度洗脱法测定不同中国产西红花药材中西红花苷Ⅰ、Ⅱ的含量  

Content determination of crocus Ⅰ and crocus Ⅱ in Crocus sativus L. from various places in China by HPLC with gradient elution

作　　者：李顺旭[1,2] 杨大坚[2] 李荣东[1] 史伟[1,2] 董玮玮[2] 彭江丽[1,2] 

机构地区：[1]湖南中医药大学药学院,长沙410208 [2]深圳市中药药学及分子药理学研究重点实验室,广东深圳518057

出　　处：《中南药学》2010年第12期886-889,共4页Central South Pharmacy

基　　金：香港赛马会慈善信托基金资助项目(编号:JCICM-5-07)

摘　　要：目的建立高效液相色谱梯度洗脱法测定国产西红花中西红花苷Ⅰ和西红花苷Ⅱ的含量。方法采用反相高效液相色谱法,以Agilent ZORBAX StableBond SB-Aq(4.6 mm×250 mm,5μm)柱为色谱柱;流动相为0.01%甲酸(A)-乙腈(B),进行梯度洗脱,0～8 min(75%～70%A),8～10 min(70%～60%A),10～25 min(60%～30%A);流速:1.0 mL.min-1;进样量:5μL;测定波长:440 nm;柱温:30℃。结果西红花苷Ⅰ和西红花苷Ⅱ的保留时间大约分别为5.71 min和8.83 min。以峰面积对进样浓度线性回归,西红花苷Ⅰ回归方程:Y=24.93X-0.254 3,r=1.000,线性范围3.125～200μg.mL-1,西红花苷Ⅱ回归方程:Y=36.11X+8.370,r=1.000,线性范围3.125～200μg.mL-1。西红花苷Ⅰ和西红花苷Ⅱ的回收率分别为101.1%和100.2%,RSD分别为1.7%和1.4%。结论本方法可用于测定不同国产西红花药材中西红花苷Ⅰ、Ⅱ的含量,本方法操作简便,结果准确。Objective To establish an HPLC with gradient elution method to determine the content of crocus I and cro cus II in Crocus sativus L. from various places in China. Methods Chromatographic conditions included column Agi lent ZORBAX StableBond SB-Aq （4. 6 mm×250 mm, 5 um） with 0. 01% formic acid （A） acetonitrile （B） as the mobile phase gradient elution： 0 8 min （75%-70%A）, 8-10 min （70%-60%A）, and 10-25 min （60%-30%A）. The flow rate was 1.0 mL · min- 1, the injection volume was 5 uL, the detection wavelength was 440 nm, and the column temperature was 30 ℃. Results The retention time of crocus I and crocus II was about 5.71 min and 8. 83 min respectively. The regression equation for crocus I was Y= 24.93 X-0. 254 3, r =1. 000, and the linear range was 3. 125-200 ug· mL-1. That for crocus II was Y 36.11X＋8. 370, r= 1. 000, and the linear range was 3. 125-200 ug · mL -1. The average recovery of crocus I and crocus II was 101.1% and 100.2%, RSD was 1.7% and 1.4% respectively. Conclusion The method is accurate and sensitive, which can be used for the content determina- tion of crocus I and crocus II in Crocus sativus L. from various places in China.

关 键 词：西红花苷Ⅰ 西红花苷Ⅱ 西红花 高效液相色谱 梯度洗脱 

分 类 号：R927.2[医药卫生—药学] R284.1