Based on the results from comparing the effects of steaming and polishing times and processing adjuvants, we could draw the conclusion that the overall chemical composition experienced a great change from three to five cycles of steaming and drying, and five cycles was a simple processing method of Shu Dihuang which obeyed the ancient processing theory.
These results also suggested that the degree of processing must be taken into account when preparing Shu Dihuangtan. Three different pyrolysis and combustion stages were observed: drying and dehydration, pyrolysis and combustion, carbonization and combustion. Since the pyrolysis and combustion characteristics were used to simulate the preparation process of Shu Dihuangtan, we focused more on the pyrolysis and combustion stage.
In the pyrolysis and combustion stagef, two obvious thermal weight loss phases and two thermal weight loss rate peaks appeared for Shu Dihuang with different steaming and drying cycles and adjuvants. Moreover, the intensities of the two thermal weight loss rate peaks were 6. Moreover, the pyrolysis-combustion characteristics of Shu Dihuang processed with no adjuvant were similar. The initial temperature and final temperature in the two phases were The intensities of the thermal weight loss rate peaks were 7.
Along with the increase of steaming and polishing cycles, both the mean intensity of the two thermal weight loss rate peaks and the temperature interval initial and final temperature decreased in the pyrolysis and combustion stage. However, overall, the differences of thermal weight loss and thermal weight loss rate were very small from SW5 to SW9.
Also, the peak temperature of thermal weight loss rate of SW5 was similar to SW6-SW9 Figure 5 , demonstrating that the trends for Shu Dihuang processed with or without adjuvant were similar, so we hold the opinion that five cycles represent a relatively good processing method.
For Shu Dihuang processed with the same steaming and polishing cycles, the pyrolysis-combustion characteristics were also different because of the processing adjuvants with yellow rice wine or not. The intensity of the thermal weight loss rate peak was less for the Shu Dihuang processed with yellow rice wine compared to the Shu Dihuang processed without adjuvants at the same steaming and polishing cycle.
Meanwhile, the Shu Dihuang processed with yellow rice wine spanned a smaller temperature interval during its thermal weight loss. To summarize, the degree of processing and adjuvant are important conditions in the carbonization process for preparing Shu Dihuangtan. The intensity of thermal weight loss rate peak near Moreover, the corresponding temperature of the thermal weight loss rate peak in the second stage of pyrolysis-combustion near For the similarities between any two samples, CS3 and CS9 were relatively higher.
Meanwhile, the relative similarity between CS3 and CSP as well as the sample with several cycles of steaming and drying generally nine cycles in the ancient theory were both obviously high. The global chemical composition of Shu Dihuangtan was affected by both the degree of processing and adjuvant.
Acetonitrile chromatographic grade was purchased from Beijing Bailingwei Tech Co. Beijing, China. Standard reagents: acteoside, catalpol, 5-hydroxymethylfurfural, rehmaionoside A and rehmaionoside D production batch numbers , , , and , respectively were purchased from Chengdu Pufei De Biotech Co.
Chengdu, China. They were confirmed as the roots of Rehmannia glutinosa Libosch. By Prof. Sheng Dihuang was placed in a steamer and heated for 6 h.
This steaming and polishing step was repeated nine times on nine samples which were separated after each cycle. These were the sample batch processed without adjuvants. For the sample batches processed with adjuvant, yellow rice wine was used as the processing adjuvant, and the ratio between the yellow wine and herb was The other conditions were kept the same as above.
The samples were labeled from SW1 to SW9. According to Chinese National Pharmacopoeia , Sheng Dihuang was steamed until the herb turned black. This sample was labeled as SP. The sample was then exposed to sunlight until the surface was slightly dried. After that, the sample was cut into thick slices or pieces and dried completely. A portion of each sample 1. Then the sample was extracted with an ultrasonic extraction device for 1.
An aliquot 25 mL of the filtrate was dried using a water-bath. The residue was dissolved with deionized water and extracted successively with petroleum ether, chloroform, ethyl acetate, and n -butyl alcohol. Reference solutions standards containing catalpa alcohol The mobile phase consisted of 0. The gradient program was developed as shown in Table 8. The flow rate was kept at 1. All the tests below were carried out on Shu Dihuang prepared as described in Section 3.
The precision was determined by repeating HPLC injections of the same sample six times per day. Six independent samples prepared as described in Section 3. The stability was assessed by measuring a single sample solution stored at room temperature at 0, 2, 4, 8, 16 and 24 h.
AIA format files for further processing. A reference sample was selected stochastically from the most representative samples among the middle of the analytical sequence to generate a template.
Then, all of the samples were overlaid based on the template, and common peaks were aligned to the ones in template. The relative retention time RRT and relative peak area RPA were calculated simultaneously and were exported as an excel file for further statistical analysis. The experiments were repeated twice. In summary, the degree of processing steaming and polishing times and processing adjuvant yellow rice wine had a great influence on the overall chemical composition of Shu Dihuangtan.
The optimum processing method of Shu Dihuang was steaming and polishing for five cycles, which followed the ancient processing theory. Since the quality of Shu Dihuang is closely related to the quality of Shu Dihuangtan, the degree of processing and use or not of processing adjuvant of Shu Dihuang are very important for Shu Dihuangtan preparation.
As for the reason why the processed of Shu Dihuang is steaming and drying for several cycles generally nine times in the ancient theory, we need to further combine these results with the pharmacology of Shu Dihuangtan to confirm the reasons. M and M. All authors contributed to the experiments and approved the final manuscript. National Center for Biotechnology Information , U. Journal List Molecules v. Published online Jul Find articles by Xianglong Meng.
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