ABSTRACT
Objective::The processing method of red ginseng was determined by comparing the effects of different steaming time and pressure on the total content of six ginsenosides. Method::The contents of ginsenoside Rg1, Re, Rf, Rb1, Rc and Rb2 were determined by ultra high performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-QqQ-MS/MS). The Waters ACQUITY UPLC BEH C8 column (2.1 mm×100 mm, 1.7 μm) was used. The mobile phase was 0.1% formic acid aqueous solution (A) and 0.1% formic acid acetonitrile solution (B) for gradient elution (0-4 min, 81%-79%A; 4-6.3 min, 79%-75%A; 6.3-6.5 min, 75%-71%A; 6.5-9.5 min, 71%A; 9.5-16.5 min, 71%-68.5%A; 16.5-16.6 min, 68.5%-60%A; 16.6-19 min, 60%-100%A). The flow rate was set at 0.4 mL·min-1 and the column temperature was set at 35 ℃. The mass spectrographic analysis employed electrospray ionization (ESI) and negative ion collection mode with capillary ionization voltage of 2.5 kV, desolvation temperature of 350 ℃, desolvation gas flow of 700 L·h-1 and cone gas flow of 50 L·h-1. Multiple reaction monitoring (MRM) mode was used to collect information, the collection range was m/z 100-1 500, detection was performed by MRM mode at m/z 799.59-637.49 for ginsenoside Rg1, m/z 945.54-475.79 for ginsenoside Re, m/z 799.59-475.49 for ginsenoside Rf, m/z 1 107.59-783.97 for ginsenoside Rb1, m/z 1 077.58-783.96 for ginsenoside Rc, m/z 1 077.75-191.19 for ginsenoside Rb2. Result::When the steaming time was 3 hours, the total mass fraction of six ginsenosides in each sample group was 7.099 8-16.768 5 mg·g-1, and the total amount of the six ginsenosides in atmospheric steaming was 2.5-12.6 times of that in pressurized steaming, which was obviously better than that in pressurized steaming. Conclusion::Under the conditions of this experiment, the best processing method of red ginseng is atmospheric steaming for 3 hours with fresh ginseng.
ABSTRACT
Objective: To study the changes rule of active ingredients content and moisture status during the process of dry (drying/steaming) and rehydration (decoction), which could provide technical support for optimizing the dry/rehydration conditions of Chinese medicine and scientifically determine the end point of the process, and it also provides a new scientific perspective for exploring the differences in fresh/dry/processing of traditional Chinese medicine. Methods: Low-field NMR and imaging techniques (LF-NMR, MRI) were used to determine the change of water with time; The content changes of main composition of ginsenosides in different samples were determined by HPLC. Results: The results of determination of moisture and chemical composition showed that: The fresh ginseng was steamed for 180 min. At this time, the water was saturated, the ginsenosides tended to be stable, and the content of total ginsenosides was high. When fresh ginseng and red ginseng were dried at different temperatures for 12.5 h, they were not completely dried at 40 ℃ hot-air drying; The sun-dried ginseng still contained 3.02% water at 50 ℃ hot-air drying, and the red ginseng has been dried; Both of them have been dried at 60 ℃, but the content of total ginsenosides in ginseng and red ginseng was the highest at 50 ℃. The comprehensive results showed that ginseng and red ginseng were better at 50 ℃ hot-air drying. During rehydration (decocting), the moisture content of the decoction for 60 min was fully saturated and the content of total ginsenosides was higher, better than 30 min and 120 min, which was a better decocting condition. The moisture content and total ginsenosides content of fresh ginseng were higher than those of steaming/drying/decocting ginseng, suggesting that fresh ginseng is great significance for preserving and exerting the basic state of the initial pharmacodynamics of traditional Chinese medicine. Conclusion: In this study, ginseng was used as an example. LF-NMR/MRI and HPLC techniques were used to focus on the changes of moisture and chemical contents during the drying (drying, processing) and rehydration (decocting) of traditional Chinese medicines. It provides a new technical method for the determination of the dry/rehydration end point and the optimization of process conditions for traditional Chinese medicine, and also provides a new scientific basis for the interpretation and exploration of the theory of fresh/drying/processing of traditional Chinese medicine.
ABSTRACT
Objective: To verify the existence of microRNAs (miRNAs) extracted from fresh ginseng decoction. Methods: Fresh ginseng was prepared into decoction according to the conventional method. The miRNA were extracted from the condensed ginseng decoction by plant microRNA extraction kit. Then miRNA were treated by DNase I and subjected to agarose gel electrophoresis and Agilent 2100 bioanalysis. MiR-159 and miR-6135, which were highly expressed in ginseng, were selected and verified by real-time quantitative PCR to detect the expression in the decoction. Results: Ginseng miRNA were successfully extracted from fresh decoction. MiR-159 and miR-6135 were expressed in fresh decoction with lower levels than those of fresh ginseng. Conclusion: miRNAs stably existed after processing, and retained some stability after high-temperature treatment. The findings provide a valuables basis for the further studies on ginseng miRNAs.