HPLC法测定白果中银杏酸的含量

HPLC Determination of Ginkgolic Acids in Fruits of Ginkgo biloba

目的:建立反相高效液相色谱法测定银杏白果中有毒成分银杏酸含量。方法:用LC/DAD/ESI/MS对白果中的银杏酸进行定性鉴定。白果经石油醚索氏提取、浓缩、定容后,直接进样进行HPLC测定。色谱分析条件:色谱柱为Inertsil ODS-2(250 mm×4.6 mm,5μm),流动相为甲醇-3%HAc溶液(92:8),流速1.0 mL·min-1,柱温40℃,紫外检测波长310 nm。结果:白果中含有5种银杏酸:C15:1,C17:2,C15:0和C17:1(存在2种同分异构体),还有1种作者推测为C17:3,其中C17:1和推测为C17:3的银杏酸为首次在白果中发现。该法精密度和重复性较好,其5次测定的RSD分别为0.71%和3.7%。白果中银杏酸含量达200 mg·kg-1左右,大大超过国际限量标准,其食用安全性应引起高度重视。结论:该法简便、快速、准确,适用于白果及其产品中银杏酸的分析,可为白果产品的质量监控和安全性评价提供基础。

Objective: To establish the determination method of toxic components-ginkgolic acids ( GAs) in fruits of Ginkgo biloba by RP - HPLC. Methods:The GAs in the fruits were identified by LC/DAD/ESI/MS. Through enrichment by increasing the sample amount and using petroleum ether Soxhlet extraction, GAs could be determined directly by HPLC. The chromatographic column was Inertsil ODS -2(250 mm×4. 6 mm,5μm) ;the mobile phase was methanol -3% aqueous acetic acid(92:8) ;the flow rate was 1.0 mL·min-1;the column temperature was 40℃ ;the detection wavelength was at 310 nm. Results: There were five kinds of GAs( C15: 1 ,C17:2,C15: 0,C17:1, and tentatively C17:3)in Ginkgo biloba fruits, thereinto GAs C17:1 and tentatively. C17: 3 were discovered in fruits for the first time. The precision and reproducibility of the method were good,and their RSD(n =5) was 0. 71% and 3. 7% , respectively. The content of GAs in fruits was up to 200 mg·kg-1,much higher than the international limited standard. So edible security should still be attached importance to. Conclusion: The method was found to be simple, fast and accurate,and could be used to GAs analysis in Ginkgo biloba fruits and their products. It could provide the basis on products of Ginkgo biloba fruits in quality control and safety evaluation.