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A reversed phase-high performance liquid chromatography-diode array detector (RP-HPLC-DAD) method was developed to determine the amount of gastrodin, the major active component of Grammatophyllum speciosum pseudobulbs. A previous study suggested that the ethanolic extract of G. speciosum had a potential to encourage stemness of keratinocytes. Therefore, in order to determine the quality of G. speciosum ethanolic extract, the quantitative analysis of gastrodin should be validated. The optimized RP-HPLC condition was achieved within 18 min, using a column, Inertsil ODS-3 (4.6 x 150 mm, 5 μm). The mobile phase consisted of a mixture of water: acetonitrile with a gradient from 1:99 to 100:0 in 14 minutes and keep constant at 1:99 for 4 minutes. The flow rate was 1.7 mL/min with the monitored UV wavelength of 220 nm. Gastrodin was eluted at the retention time of 6.5 minutes. The calibration curve of gastrodin at the concentration of 10 100 μg/mL showed good linearity (r = 0.999). In addition, the intraday and interday precisions of gastrodin were 1.05 and 0.38%, respectively. The percentage recovery was within 99.3 101.5%. Average gastrodin contents in 3 samples were 55.52 58.12 mg/g. The results indicate that this method can be utilized to control the quality of G. speciosum extract.

VALIDATED RP-HPLC METHOD FOR QUANTIFICATION OF GASTRODIN IN ETHANOLIC EXTRACT FROM THE PSEUDOBULBS OF Grammatophyllum speciosum Blume