Abstract
Purpose: To develop an ultra-performance liquid chromatography-tandem mass spectrometry (UPLCMS/MS) method for the simultaneous determination of 7 major components of Ginkgo leaf (kaempferol, quercetin, isorhamnetin, ginkgolides A, ginkgolides B, ginkgolides C and bilobalide) in dog plasma. Methods: Beagle dog plasma samples were spiked with internal standard (domperidone), acidified with HCl and extracted twice by liquid-liquid extraction using ethyl acetate. Chromatographic separation was achieved on an Acquity UPLC BEH C18 column (100 x 2.1 mm, 1.7 μm) by gradient elution with a run time of 4.0 min. The specificity, linearity, precision, recovery, matrix effect and stability of the method were determined.Results: The method showed high selectivity of the flavonols and terpene lactones in plasma samples. The concentration of the 7 target compounds showed good linear relationship with the peak area ratios of each analyte to internal standard. Lower limit of quantification (LLOQ) was 1.232, 0.240, 0.200, 1.330, 0.960, 0.696, 0.470 ng•mL-1 for kaempferol, quercetin, isorhamnetin, bilobalide, ginkgolides A, ginkgolides B and ginkgolides C, respectively. Recovery of all QC samples ranged from 77.68 to 105.07 %. Matrix effect derived from QC samples was in the range of 85.09 – 113.14 %. The stability of the analytes, calculated as RSD at three concentrations, was < 15 %.Conclusion: The developed method is simple, rapid and sensitive and can be applied to the determination of kaempferol, quercetin, isorhamnetin, ginkgolides A, ginkgolides B, ginkgolides C and bilobalide in dog plasma.Keywords: Ultra-performance liquid chromatography-tandem mass spectrometry, Ginkgo biloba, Beagle dog plasma, Kaempferol, Quercetin, Isorhamnetin, Ginkgolides A, Ginkgolides B, Ginkgolides C, Bilobalide
Highlights
The extract of Ginkgo biloba (Ginkgo, Ginkgoaceae) leaves has been considered as valuable medicine and dietary supplements for more than 40 years
A simple, rapid and sensitive ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the simultaneous determination of the 7 major components Ginkgo Biloba leaves in dog plasma
Selectivity was evaluated by comparing the chromatograms of six individual blank dog plasma samples with those of corresponding standard plasma samples spiked with the analytes and IS, and plasma samples obtained after oral administration of drugs
Summary
The extract of Ginkgo biloba (Ginkgo, Ginkgoaceae) leaves has been considered as valuable medicine and dietary supplements for more than 40 years. Pharmacology experiments showed that it is helpful to the prevention and treatment of cardiovascular diseases [1,2] and cerebral insufficiency [3,4,5,6,7]. Such pharmacological activities are attributed to the flavonoid glycosides and terpene lactones. A simple, rapid and sensitive ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the simultaneous determination of the 7 major components Ginkgo Biloba leaves (kaempferol, quercetin, isorhamnetin, ginkgolides A, ginkgolides B, ginkgolides C and bilobalide) in dog plasma.
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