NL2026922A - Method for optimizing radix aconiti lateralis processing process - Google Patents
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Abstract
The present invention provides a method for optimizing a radix Aconiti lateralis processing method, relating to the technical field of traditional Chinese medicine processing. Thelnethod.for‘optimizing'a.radiz:Aconiti lateralis19rocessing method includes carrying out experimental design on to—be—optimized factors through a Box—Behenken design method to obtain. processed. products with different factors and different parameters; performing liquid chromatography—mass spectrometry detection on the processed products to obtain respective fingerprints; after common peaks of the fingerprints are screened, analyzing main causes of areas of the common peaks to construct a comprehensive evaluation function. of processed. radix Aconiti lateralis products; calculating a score of a comprehensive evaluation function of each.group according to the function, constructing an equation between the to—be—optimized factors and the score, and predicting an optimal process parameter according to the equation. The method.for optimizing a radix.Aconiti lateralis processingnethodprovidaïbythepresentinventionissuitable for optimizing factors in various processingInethods, has wide universality and is easy to operate.
Description
TECHNICAL FIELD The present invention relates to the technical field of traditional Chinese medicine processing, and in particular to a method for optimizing a radix Aconiti lateralis processing process.
BACKGROUD Radix Aconiti lateralis 1s a processed product of the subroot of Acormitum Carmichaeli Debx. The radix Aconiti lateralishas rigidand drymedicinal properties, the medicinal effect moves around inside the body and does not stick to a certain part. The radix Aconiti lateralis can help the heart yang to dredge the channels and collaterals, replenish spleen yang to help function in transportation and invigorate the kidney Yang to benefit the fire. The radix Aconiti lateralis is a main drug that warms the interior to reinforce yang and has effects of restoring vang to stem counterflow, tonifying fire and helping yang, dispelling cold and relieving pain. Because of toxicity, the radix Aconiti lateralis is mostly processed and then used as a medicine.
A Box-Behnken design is of a response curve design type and does not contain an embedded factor or fractional factorial design. The Box-Behnken design has a processing combination located at the midpoint of the edge of a test space, and requires at least three factors, which can effectively estimate first-order and second-order coefficients.
Fingerprint refers to a spectrogram that is obtained under fixed test conditions after proper treatment of a substance and can indicate chemical characteristics of the substance, and has characteristics of integrity and fuzziness. Liquid chromatography-mass spectrometry is a powerful tool for qualitative and quantitative analysis of chemical components in mixtures due to its advantages of exclusiveness, rapidness,
sensitiveness, and the like. At present, a fingerprint technology, a chemometric analysis method and chromatography-mass spectrometry have been widely used in the identification and quality evaluation of traditional Chinese medicines.
Principal component analysis (PCA) is a statistical method. Through orthogonal transformation, a group of variables that may be correlated are converted into a group of linearly uncorrelated variables, and the converted group of variables is called principal components.
At present, commercially available processed radix Aconiti lateralis products include salted radix Aconiti lateralis, giant typhonium rhizome, radix Aconiti lateralis having outer skin removed and longitudinally into two pieces, yellow radix Aconiti lateralis, black prepared lateral root of aconite, and the like. The processed radix Aconiti lateralis products are processed according to ancient methods in the place of origin, and undergo complicated procedures such as soaking in brine or saline water, decoction, blanching and steaming. However, the radix Aconiti lateralis processed through the methods has more medicinal property loss, properties and flavors are lost, and the effectiveness is greatly reduced. For this reason, the toxicity and efficacy and other quality of the processed radix Aconiti lateralis products need to be comprehensively evaluated in this field. However, at present, this field lacks a simple and effective method for optimizing specific parameters of a radix Aconiti lateralis processing method, which makes the improvement of the radix Aconiti lateralis processing method stagnate.
SUMMARY In order to overcome the defect that the prior art lacks a simple and effective method for optimizing a radix Aconiti lateralis processing method, the present invention provides a method for optimizing a radix Aconiti lateralis processing process, which can synthesize the advantages and disadvantages of a plurality of performances and evaluate processing conditions for the best comprehensive quality.
In order to achieve the above invention objective, the present invention provides the following technical solution: The present invention provides a method for optimizing a radix Aconiti lateralis processing process, including the Following steps: (1): selecting factors in at least three to-be-optimized radix Aconiti lateralisprocessingmethods, designing response surface tests on various factors at different levels by a Box-Behenken experimental design method, and respectively processing the radix Aconiti lateralis according to the designed factors and levels to obtain processed radix Aconiti lateralis products with different factors and different levels; (2): performing high performance liquid chromatography-mass spectrometry detection on each of the processed radix Aconiti lateralis products obtained in step (1) to obtain fingerprints of each of the processed radix Aconiti lateralis products; {3): selecting one of the fingerprints of the processed radix Aconiti lateralis products obtained in step (2) as a reference spectrum, performing full peak matching by using a multipoint correction method, screening common peaks of the fingerprints of all the processed radix Aconiti lateralis products, analyzing main causes of areas of the common peaks, and calculating a weight coefficient of the area of each of the common peaks, thereby constructing a comprehensive evaluation function for obtaining to-be-optimized factors; (4): according to the comprehensive evaluation function obtained in step (3), substituting the data of different factors and different levels designed in step (1) into calculation to obtain comprehensive evaluation scores of processed radix Aconiti lateralis products with different factors and different levels; and (5) performing multiple regression fitting and binomial analysis on each of the comprehensive evaluation scores obtained in step (4), constructing an equation between the comprehensive evaluation score and the to-be-optimized factors, and determining optimal factor parameters according to the equation.
Preferably, the radix Aconiti lateralis processing method includes a yin radix Aconiti lateralis processing method.
Preferably, the yin radix Aconiti lateralis processing method includes mixing salted radix Aconiti lateralis with ginger juice, carrying out covered moistening, steaming the mixture, and drying to obtain a product.
Preferably, the factors include the relative dosage, processing time and processing temperature of the to-be-processed radix Aconiti lateralis.
Preferably, when the radix Aconiti lateralis processing method is the yin radix Aconiti lateralis processing method, the factors include the mass ratio of the ginger juice to the to-be-processed radix Aconiti lateralis, covered moistening time and steaming time.
Preferably, the optimal factor parameters obtained after the optimization of the factors according to steps (1) to (5) are as follows: the mass ratio of the salted radix Aconiti lateralis to the ginger juice is 1:0.1665, the covered moistening time is 12 h, and the steaming time is 9.45 h.
Compared with the prior art, the present invention has the following beneficial effects: The present invention provides a method for optimizing a radix Aconiti lateralis processing method. The method includes carrying out experimental design on to-be-optimized factors through a Box-Behenken design method to obtain processed products with different factors and different parameters; performing liquid chromatography-mass spectrometry detection on the processed products to obtain respective fingerprints; after common peaks of the fingerprints are screened, analyzing main causes of areas of the common peaks to construct a comprehensive evaluation function of processed radix Aconiti lateralis products; calculating a score of a comprehensive evaluation function of each group according to the function, constructing an equation between the to-be-optimized factors and the score, and predicting an optimal process parameter according to the equation.
1. The method for optimizing a radix Aconiti lateralis processing method provided by the present invention is suitable 5 for optimizing factors in various processing methods, has wide universality and 1s easy to operate.
2. The method for optimizing a radix Aconiti lateralis processing method provided by the present invention is a comprehensive evaluation of the quality of processed radix Aconiti lateralis products, and an effective optimization method is provided for improving the guality of the processed radix Aconiti lateralis products.
BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is an overlay chart of liquid chromatography-mass spectrometry fingerprints when a vin radix Aconiti lateralis processing method is optimized; FIG. 2 is a diagram of ovarian morphology observation results; and FIG. 3 is a diagram of testicular morphology observation results.
DESCRIPTION OF THE EMBODIMENTS The present invention provides a method for optimizing a radix Aconiti lateralis processing process, including the following steps.
(1): Select factors in at least three to-be-optimized radix Aconiti lateralis processing methods, design response surface tests on various factors at different levels by a Box-Behenken experimental design method, and respectively process the radix Aconiti lateralis according to the designed factors and levels to obtain processed radix Aconiti lateralis products with different factors and different levels.
In the present invention, the central combination design of the Box-Behnken is a synthesis of statistical design experimental techniques, certain data is obtained by using the Box-Behnken experimental design and experiments, multivariate equations of higher degree are adopted to fit a functional relationship between factors and effect values, and optimal process parameters are solved through analysis of regression ecduations, so that a statistical analysis method for solving the multivariable problem is implemented. Compared with orthogonal and uniform designs, results obtained by the method are visual and convenient to analyze, and the experimental accuracy is improved at the same time.
In the present invention, the radix Aconiti lateralis processing method includes but is not limited to a vin radix Aconiti lateralis processing method. The radix Aconiti lateralis processing method provided by the present invention includes a method for directly processing radix Aconiti lateralis, and also includes a method for further processing processed radix Aconiti lateralis. Yin radix Aconiti lateralis is also called white prepared lateral root of aconite. The yin radix Aconiti lateralis processing method includes mixing salted radix Aconiti lateralis with ginger juice, carrying out covered moistening, steaming the mixture, and drying to obtain a product.
In the present invention, the factors preferably include the relative dosage, processing time and processing temperature of the to-be-processed radix Aconiti lateralis. More preferably, when the radix Aconiti lateralis processing method is the yin radix Aconiti lateralis processing method, the factors include the mass ratio of the ginger juice to the to-be-processed radix Aconiti lateralis, covered moistening time and steaming time.
(2): Perform high performance liquid chromatography-mass spectrometry detection on each of the processed radix Aconiti lateralis products obtained in step (1) to obtain fingerprints of each of the processed radix Aconiti lateralis products.
(3): Select one of the fingerprints of the processed radix Aconiti lateralis products obtained in step (2) as a reference spectrum, perform full peak matching by using a multipoint correction method, screen common peaks of the fingerprints of all the processed radix Aconiti lateralis products, analyze main causes of areas of the common peaks, and calculate a weight coefficient of the area of each of the common peaks, thereby constructing a comprehensive evaluation function for obtaining to-be-optimized factors.
(4): According to the comprehensive evaluation function obtained in step (3), substitute the data of different factors and different levels designed in step (1) into calculation to obtain comprehensive evaluation scores of processed radix Aconiti lateralis products with different factors and different levels.
As shown in the embodiment of the present invention, the comprehensive evaluation function {comprehensive score (CC)) of the yin radix Aconiti lateralis processing method on the mass ratio of the ginger juice to salted radix Aconiti lateralis, covered moistening time and steaming time is as follows: CC=0.146665 P1+0.092501 P2+0.208911 P3+0.207245 P4+0.037561 P5+0.121143 P6+0.0697 P7+0.116274 P8, where Pl to P8 refer to peak areas of the first to eighth common peaks.
(5): Perform multiple regression fitting and binomial analysis on each of the comprehensive evaluation scores obtained in step (4), construct an equation between the comprehensive evaluation score and the to-be-optimized factors, and determine optimal factor parameters according to the equation. In the present invention, the optimal parameter has the highest comprehensive score.
As shown in the embodiment of the present invention, the equation between the mass ratio of the ginger juice to the salted radix Aconiti lateralis, covered moistening time and steaming time in the yin radix Aconiti lateralis processing method and the comprehensive evaluation score is: CC=6.67-0.22A+0.17B+0.23C+0.63AB~-1.10AC+0.12BC~1.02A%+
0.15B2-0.66C2, In the embodiment shown in the present invention, the optimized optimal factor parameters are as follows: the mass ratio of the salted radix Aconiti lateralis to the ginger juice is 1:0.1665, the covered moistening time is 12 h, and the steaming time is 2.45 h.
That is, the yin radix Aconiti lateralis processing method optimized according to the optimization method according to the present invention is as follows: The salted radix Aconiti lateralis and the ginger juice are evenly mixed according to a mass ratio of 1:0.1665, subjected to covered moistening for 12 h and then steamed for 9.54 h, the steamed radix Aconiti lateralis is aired and cut or planed into straight slices, and the straight slices are dried at a low temperature and sieved to remove dust to obtain the yin radix Aconiti lateralis.
The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but the embodiments cannot be construed as limiting the protection scope of the present invention.
Embodiment 1 A Box-Behnken design arrangement test was carried out with Design Expert 8.06 software. For factor investigation, it was selected that the mass ratio of salted radix Aconiti lateralis to ginger juice was 1:(0.1-0.3), the covered moistening time was 12-36 h, and the steaming time was 6-10 h. A response surface test (see Table 1) with 3 factors and 3 levels and 17 test points was designed. Processed radix Aconiti lateralis products were prepared and base peak graphs were measured using a liquid chromatograph/mass spectrometer, and graph data was imported into Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System Software (Version 2012.130723) for processing. Full peak matching was carried out using an El sample as reference spectrum by using a multipoint correction method, 8 common chromatogram peaks were determined, and a liquid chromatography-mass spectrometry fingerprint overlay chart of yin radix Aconiti lateralis of 17 batches of yin radix Aconiti lateralis was established (see FIG. 1).
Table 1 experimental design and results
A/S B/h Std Run (Ginger {Covered “B Comprehensive score (CC) (Steaming (Label) (Sequence) usage moistening time) (Comprehensive score) amount) time) 2 1 25 24 8 6.60585 6 2 25 24 8 6.319192 3 25 24 3 6.0574 14 4 25 36 10 6.765552 7 5 25 24 8 7.357424 16 6 25 12 10 6.295182 3 7 15 24 10 6.472946 13 8 35 12 8 4.79861 4 9 25 36 6 5.772497 1 10 35 24 6 5.694962 15 11 25 24 3 6.99873 9 12 15 12 8 6.346419 8 13 35 24 10 3.665539 12 14 35 36 3 6.51921 15 36 8 5.530611 17 16 15 24 6 4.114475 11 17 25 12 6 5.797127
PeakView V1.2 software was used to derive original spectral data of peak areas of 8 common peaks in the fingerprints of 17 batches of yin radix Aconiti lateralisto forman 8 x 17-order data matrix (see Table 2). The data matrix was imported in SPSS V19.0 analysis software to carry out principal component analysis, calculate the weight coefficient of the area of each peak, and construct a comprehensive evaluation function of yin radix Aconiti lateralis with different process parameters: CC=0.146665P1+0.092501P2+0.208911P3+0.207245P4+0. 037561 P5+0.121143P6+0.0697P7+0.116274P8. Table 2 Areas of common peaks of liquid chromatography-mass spectrometry fingerprints
Peak area (%) Pl P2 p3 P4 P5 P6 P7 P8
Fl 3.509 1.652 8.132 2.195 1.829 2.151 6,742 1.669 F2 2.862 1.507 8.105 2.188 1.927 1.76 6,446 1.791 F3 2.774 1.439 7.213 1.814 2.267 2.656 7,32 1.358 F4 3.254 1.619 8.361 2.903 2.174 2.427 5.245 1.635 F5 3.685 1.714 9.493 2.722 1.766 1.955 7.356 1.602 F6 2.591 1.231 7.635 1.875 2.576 2.731 8.248 1.334 F7 2.847 1.111 8.603 2.524 1.864 2.197 6.189 1.114 F8 2.143 1.5 6.144 1.679 1.365 1.196 4.759 1.363 F9 3.172 1.491 6.539 1.784 1.891 2.098 7.31 1.373 F10 2.774 1.345 6.939 1.882 1.603 1.884 6.664 1.416 Fil 3.426 1.527 9.04 2.306 1.91 2.326 7.984 1.042 F12 3.304 1.398 7.717 2.173 1.312 2.195 5.528 2.475 F13 1.456 0.404 5.441 1.347 1.589 0.784 2.679 0.933 F14 3.338 1.793 7.586 1.621 2.907 2.525 9.527 0.939 F15 3.388 1.613 6.22 1.387 2.37 2.141 7.571 0.727 F16 2.402 1.301 4.501 1.003 1.425 1.675 5.272 1.128 F17 2.934 1.455 6.505 2.268 1.364 1.847 7.434 1.38
The comprehensive evaluation score of each batch was calculated. The higher the score, the better the quality of the batch of samples. Finally, Design Expert V8.0.6 software was used to carry out multiple regression fitting and binomial analysis on comprehensive score data of each group of index components, and the equation of the comprehensive score of the yin radix Aconiti lateralis processing process and ginger usage dosage (A), covered moistening time (B) and steaming time (C) was established: CC=6.67-0.22A+0.17B+0.23C+0.63AB-1.10AC+0.12BC-1.02A2+0 .15B2~0.66C2. An optimal processing process was predicted.
According to the optimal processing process, the ginger usage dosage (A) was 16.65%, covered moistening time (B) was 12 h, and steaming time (C) was 9.54 h.
A method for preparing optimized yin radix Aconiti lateralis includes taking salted radix Aconiti lateralis which has been bleached to remove salt and dried, adding ginger juice with a mass percent of 16.65%, uniformly stirring and performing covered moistening for 12 h, steaming for 9.54 h after covered moistening, spreading and airing on a bamboo sieve after steaming, cooling and then cutting or planing into straight slices, drying the straight slices at low temperature, and sieving to remove dust.
Embodiment 2 Pharmacological experiments were conducted on yin radix Aconiti lateralis prepared by using the preparation method optimized in Embodiment 1.
Experimental grouping: SPF SD rats weighing about 180-220 g were divided into 10 groups with 8 rats in each group, which were divided into a blank female group, a blank male group, a model female group, a model male group, a yin radix Aconiti lateralis female group, a yin radix Aconiti lateralis male group, a yang radix Aconiti lateralis female group, a yang radix Aconiti lateralis male group, a raw radix Aconiti lateralis female group and a raw radix Aconiti lateralis male group.
A method of modeling: Animals in the blank groups were subjected to intragastric administration with normal saline every day and fed conventionally. Rats of the model groups and drug administration groups were given rhubarb decoction at a dosage of 1 ml1/100 g body weight (with a drug concentration of 1 g/ml) once a day for 7 d. (Exhaustive swimming once a day causes overstrain on the basis of intragastric administration of rhubarb), and then 1 g of raw rhubarb and 1 g of raw coptidis rhizoma per 2 g (crude drug) /8 ml of water suspension were administered.
On the 8th day, administration therapy was started. The blank groups and the model groups were subjected to intragastric administration with the same amount of distilled water, and the drug administration group was subjected to intragastric administration with radix Aconiti lateralis at a dosage of 12 g/kg for seven consecutive days.
Sample treatment and detection: The sample treatment and detection were performed 24 h after the last administration; after animals were anesthetized with chloral hydrate, testicular tissue and ovarian tissue were taken immediately and stored in formalin solution for HE staining section observation.
Inspection results:
1. Ovarian morphology observation (See FIG. 2) Follicle growth was active in the blank groups. Follicles at all levels could be seen, and granulosa cells hadmany layers.
The number of follicular granulosa cells at all levels in the model groups decreased significantly.
The number of follicular cells increased in the raw radix Aconiti lateralis group.
The number of follicular cells and corpus luteum increased in the yang radix Aconiti lateralis group.
The number of follicular cells and corpus luteum significantly increased in the yin radix Aconiti lateralis group.
2. Testicular morphology observation (See FIG. 3) Interstitial cells in the blank groups were evenly distributed. The interstitial cells had 5-8 layers.
The model group was loosely arranged with 2-3 layers and had vacuoles.
The yang radix Aconiti lateralis group was orderly arranged with 5-6 layers.
The yin radix Aconiti lateralis group had a chaotic morphology with 2-3 layers.
According to the above results, yin radix Aconiti lateralis and yang radix Aconiti lateralis had good improvement effects.
The above mentioned are only preferred embodiments of the present invention, and it should be noted that those skilled in the art can also make several improvements and retouchings without departing fromthe principles of the present invention.
These improvements and retouchings should also be considered as falling within the protection scope of the present invention.
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| ZHANG HUI ET AL: "Development of Global Chemical Profiling for Quality Assessment of Ganoderma Species by ChemPattern Software", JOURNAL OF ANALYTICAL METHODS IN CHEMISTRY, vol. 2018, 1 January 2018 (2018-01-01), pages 1 - 12, XP055785706, ISSN: 2090-8865, Retrieved from the Internet <URL:http://downloads.hindawi.com/journals/jamc/2018/1675721.xml> DOI: 10.1155/2018/1675721 * |
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