RESUMO
Objective: By HPLC, to explore the differences of adjuvants for the changes of chemical components of Rehmanniae Radix Praeparata during process by steamed for nine times and shined for nine times. Methods: HPLC was conducted on a Hypersil GOLD aQ C18 column (250 mm×4.6 mm, 5 μm), flow rate was 1 mL/min, and the column temperature was 25℃. With the mobile phase of acetonitrile and 0.1% phosphoric acid water (1:99), the detection wavelength was 210 nm (catalpol), 203 nm (rehmannioside A, rehmannioside D, and leonuride), acetonitrile and 0.1% phosphoric acid water (3:97 and 4:96); 334 nm (actecosode), acetonitrile and 0.1% phosphoric acid water (16:84), and 284 nm (5-HMF), acetonitrile and 0.1% phosphoric acid water (11:89). Sugars was conducted on an Accurasil NH2 column (250 mm×4.6 mm, 5 μm) with the mobile phase of acetonitrile and 0.1% phosphoric acid water (72:28), flow rate was 1 mL/min, the column temperature was 25℃. The detector was ELSD, Alltech2000ES, drift tube temperature was 93℃, and the gas flow rate was 2.6 L/min. The statistic analysis was carried out by SPSS 20.0 and the paired sample t-test was carried out for 12 kinds of chemical composition in Rehmanniae Radix Praeparata obtained by steamed for nine times and shined for nine times and wine-steamed for nine times and shined for nine times. Results: With the increase of steamed times, the contents of catalpol, leonuride, actecosode, stachyose, sucrose, and raffinose in Rehmanniae Radix Praeparata obtained by two ways are all decreased; While the contents of rehmannioside A and rehmannioside D were slightly increased; the contents of 5-HMF, fructose, glucose, and mannose trisaccharide were increased. Conclusion: The established method conforms to the validation requirement of the methodology. In the process of Rehmanniae Radix Praeparata with steamed for nine times and shined for nine times, processing adjuvant has a marked impact on the quality of the processed product. And the related substances of Rehmanniae Radix Praeparata obtained by the two ways have a larger fluctuation in times 3,4, and 6 of steamed and shined processes, and the reason for this still needs to be further studied.