Optimization Of The Wine-Soaking And Roasting Processing Technique And Quality Standards For Cistanche Deserticola Using AHP-CRITIC Entropy Weight Method Combined With Orthogonal Experimental Design
Feb 27, 2025
2.8 Quality Research
2.8.1 Characteristic Identification
After processing 19 batches of Cistanche deserticola decoction pieces using the optimized alcohol-soaking and roasting method, their characteristics were described as follows:
They appear as irregular thick slices with a dark brown or yellowish-brown surface. The cross-sections exhibit dot-like vascular bundles arranged in wavy rings. The texture is relatively hard, with a slight alcohol aroma, a sweet taste, and a slightly bitter aftertaste. Representative samples (S1–S3) are shown in Figure 2.
Figure 2: Samples of three batches of alcohol-soaked and roasted Cistanche deserticola decoction pieces.
CISTANCHE EXTRACT WITH 16% ACTEOSIDE
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2.8.2 Inspection
Following the methods specified in the 2020 edition of the Chinese Pharmacopoeia, moisture content and total ash content were measured in triplicate, and the results are presented in Table 12.
The moisture content of 19 batches of alcohol-soaked and roasted Cistanche deserticola decoction pieces ranged from 5.56% to 9.14%.
The total ash content ranged from 4.64% to 7.63%.
Considering fluctuations in decoction piece quality, the mean values of moisture content and total ash content were increased by 30% to establish the following quality standards:
Moisture content should not exceed 9.56%.
Total ash content should not exceed 8.15%.
2.8.3 Determination of Extractable Matter and Component Content
Following the 2020 edition of the Chinese Pharmacopoeia, the content of extractable matter was determined. The content of active components was measured according to the method described in Section 2.1, with results shown in Table 12.
The extractable matter content ranged from 57.38% to 71.91%, with an average value of 66.48%.
The total content of echinacoside and verbascoside ranged from 0.37% to 1.52%.
Considering fluctuations in decoction piece quality, the average values were reduced by 30% to establish the following quality standards:
Extractable matter content should not be less than 46.54%.
Total content of echinacoside and verbascoside should not be less than 0.59%.
Figure 2 Samples of three batches of wine-soaked roasted Cistanche deserticola slices
Table 12: Determination Results of Moisture Content, Total Ash Content, Extract Content, and Component Content in 19 Batches of Alcohol-Soaked and Roasted Cistanche Decoction Pieces (%)
| Sample | Moisture (%) | Total Ash (%) | Extract (%) | Echinacoside (%) | Cistanoside A (%) | Tubuloside A (%) | Verbascoside (%) | Isoverbascoside (%) | 2′-Acetylverbascoside (%) |
|---|---|---|---|---|---|---|---|---|---|
| S1 | 8.43 | 6.56 | 70.39 | 0.96 | 0.20 | 0.01 | 0.27 | 0.06 | 0.27 |
| S2 | 7.23 | 7.28 | 69.17 | 0.90 | 0.20 | 0.03 | 0.23 | 0.06 | 0.05 |
| S3 | 8.02 | 7.26 | 70.09 | 1.22 | 0.23 | 0.03 | 0.30 | 0.06 | 0.02 |
| S4 | 8.40 | 6.66 | 70.73 | 0.78 | 0.23 | 0.05 | 0.28 | 0.04 | 0.12 |
| S5 | 8.01 | 7.63 | 68.73 | 0.81 | 0.23 | 0.03 | 0.31 | 0.06 | 0.21 |
| S6 | 9.14 | 7.42 | 66.09 | 0.19 | 0.11 | 0.01 | 0.18 | 0.03 | 0.18 |
| S7 | 8.77 | 6.57 | 63.14 | 0.39 | 0.16 | 0.01 | 0.17 | 0.03 | 0.14 |
| S8 | 7.07 | 6.60 | 65.56 | 0.30 | 0.16 | 0.04 | 0.09 | 0.02 | 0.13 |
| S9 | 8.32 | 5.33 | 63.16 | 0.57 | 0.24 | 0.02 | 0.21 | 0.04 | 0.09 |
| S10 | 6.18 | 6.93 | 64.51 | 0.41 | 0.10 | 0.03 | 0.29 | 0.04 | 0.10 |
| S11 | 8.60 | 6.68 | 68.47 | 0.60 | 0.23 | 0.03 | 0.34 | 0.05 | 0.15 |
| S12 | 7.58 | 6.15 | 69.24 | 1.18 | 0.16 | 0.04 | 0.37 | 0.05 | 0.12 |
| S13 | 6.32 | 4.64 | 69.89 | 0.79 | 0.16 | 0.04 | 0.19 | 0.04 | 0.02 |
| S14 | 5.56 | 4.83 | 62.95 | 0.56 | 0.07 | 0.03 | 0.08 | 0.04 | 0.02 |
| S15 | 6.67 | 7.36 | 62.81 | 0.92 | 0.10 | 0.02 | 0.20 | 0.10 | 0.02 |
| S16 | 6.49 | 6.49 | 71.91 | 0.36 | 0.08 | 0.02 | 0.10 | 0.05 | 0.02 |
| S17 | 5.32 | 4.84 | 63.01 | 0.63 | 0.21 | 0.02 | 0.07 | 0.02 | 0.02 |
| S18 | 6.93 | 5.34 | 57.38 | 0.47 | 0.11 | 0.02 | 0.07 | 0.02 | 0.02 |
| S19 | 5.62 | 5.52 | 65.82 | 0.66 | 0.14 | 0.04 | 0.12 | 0.04 |
3 Discussion
Chromatographic Condition Optimization
During the optimization of chromatographic conditions, several mobile phase compositions were compared, including:
Methanol/Acetonitrile - 0.2% Phosphoric Acid Solution
Methanol/Acetonitrile - 0.% Formic Acid Solution
Methanol/Acetonitrile - 0.2% Formic Acid Solution
Methanol/Acetonitrile - 0.5% Formic Acid Solution
It was found that when using Acetonitrile - 0.2% Phosphoric Acid Solution as the mobile phase, the baseline showed slight deviations. When using Methanol - 0.1% Formic Acid Solution, the separation of chromatographic peaks was relatively poor. Ultimately, Acetonitrile - 0.5% Formic Acid Solution was selected as the mobile phase due to its better peak shape and separation performance.
For detection wavelength and column temperature, after considering ease of operation and accuracy, the detection wavelength of 330 nm provided by the Pharmacopoeia was selected for echinacoside and verbascoside, and a column temperature of 30°C was found to offer good absorption for all six components.
Selection of Evaluation Indicators for Alcohol-Soaked and Roasted Cistanche Decoction Pieces
In selecting intrinsic quality evaluation indicators, both comprehensiveness and scientific validity were emphasized. The selected marker components included:
High-content compounds such as echinacoside and verbascoside
Compounds with significant content changes before and after processing, such as isoverbascoside, 2′-acetylverbascoside, and cistanoside A
Biologically active components, such as tubuloside A
For calculating weight coefficients and scores, the AHP-CRITIC comprehensive weighting method was used, which effectively combined subjective and objective factors, reducing subjective bias while ensuring data stability and variability. This method minimizes interference, ensuring scientific and stable results, and enhances the credibility of the multi-indicator comprehensive scoring approach.
Evaluation of Processing Techniques for Alcohol-Soaked and Roasted Cistanche
This study investigated the processing technology of alcohol-soaked and roasted Cistanche using a single-factor experiment to evaluate the effects of:
Type and amount of alcohol
Soaking duration
Roasting temperature
Roasting time
Effect of Different Alcohol Types
A comparison of Huangjiu (yellow rice wine), Mijiu (rice wine), and Baijiu (Chinese distilled liquor) showed that under Huangjiu processing conditions, the six component contents were higher than in Mijiu, and echinacoside and verbascoside were more stable, consistent with previous research findings [7,8].
Effect of Alcohol Quantity
The Pharmacopoeia prescribes a Huangjiu-to-Cistanche ratio of 10:2 to 10:3. However, when using a 10:1 ratio in this study, the echinacoside and verbascoside contents were significantly higher than those obtained using the Pharmacopoeia's alcohol dosage.
Effect of Soaking Time
According to traditional Chinese medicine texts, "soaking for three days and roasting finely" is recommended. Therefore, the soaking time was tested at 6 to 72 hours.
Effect of Roasting Temperature and Time
The term "roasting" (焙, bèi) traditionally refers to baking on a heated ceramic tile, which can reach temperatures up to 200°C. The "Anhui Provincial Processing Standards" specify that roasting should use low heat, so a temperature range of (85±5)°C to (185±5)°C was tested.
Since longer roasting times lead to greater loss of volatile components, roasting times of 20 to 40 minutes were selected, and the optimal parameter ranges for each factor were determined.
Optimization of Processing Parameters Using AHP-CRITIC and Orthogonal Design
Using the AHP-CRITIC entropy weight method, the weight coefficients of each evaluation index in the alcohol-soaked and roasted processing method were determined. Then, orthogonal experimental design was applied to optimize the processing parameters, leading to the final optimized processing method:
100 g of purified Cistanche
10 g of Huangjiu
Soaking for 26 hours
Roasting at (155±5)°C for 30 minutes
Cooling naturally to obtain the final product
The established quality standards for alcohol-soaked and roasted Cistanche decoction pieces were:
Moisture content ≤ 9.56%
Total ash content ≤ 8.15%
Ethanol-soluble extract content ≥ 46.54%
Total echinacoside and verbascoside content ≥ 0.59%
Challenges and Future Research Directions
Experimental results showed significant quality differences among different batches, which may be attributed to variations in the raw Cistanche material.
Currently, several challenges remain in the study of Cistanche processing, such as:
The ideal appearance characteristics of alcohol-processed Cistanche decoction pieces, which should be dark brown, glossy, moderately soft in texture, sweet in taste, and slightly aromatic with alcohol
Differences in texture between alcohol-steamed, alcohol-stewed, and roasted decoction pieces, where steamed and stewed pieces tend to be softer, while roasted pieces are harder
Whether these textural differences correspond to intrinsic quality changes, requiring further research
Challenges and Future Research Directions
Although this study successfully optimized the alcohol-soaked and roasted Cistanche processing method and established quality standards, significant differences were observed among different batches of processed decoction pieces. This variation is likely due to inherent quality differences in the raw Cistanche material, highlighting the need for further standardization in raw material selection and processing techniques.
Currently, several challenges remain in the study of Cistanche processing, such as:
The ideal appearance characteristics of alcohol-processed Cistanche decoction pieces, which should be dark brown, glossy, moderately soft in texture, sweet in taste, and slightly aromatic with alcohol
Differences in texture between alcohol-steamed, alcohol-stewed, and roasted decoction pieces, where steamed and stewed pieces tend to be softer, while roasted pieces are harder
Whether these textural differences correspond to intrinsic quality changes, requiring further research
This study provides a scientific basis for optimizing the alcohol-soaked and roasted Cistanche processing technique, but further investigations are needed to enhance standardization and ensure batch-to-batch consistency.
