Sand-fried szechwan chinaberry fruit processing technology and quality evaluation method for determining optimal processing technology parameters

Abstract

The invention relates to a sand-fried szechwan chinaberry fruit processing technology and a quality evaluation method for determining optimal processing technology parameters, and belongs to the technical field of traditional Chinese medicine processing and quality control, clean river sand is taken and placed in a pot, medium fire heating is carried out, cut szechwan chinaberry fruit decoction pieces are put into the pot when the temperature is stable, the szechwan chinaberry fruit decoction pieces are fried until the surface is slightly yellow, and the szechwan chinaberry fruit decoction pieces are taken out of the pot, cooled and taken out. An orthogonal design experiment is adopted, the sand frying temperature, the river sand weight multiple, the frying time and the medicinal material granularity are taken as investigation factors, the optimal processing technology of the sand-fried szechwan chinaberry fruit is screened out, and the optimal sand frying technology is subjected to technology verification. According to the processing method, the limitations of non-uniform heating and inconsistent processing degrees of a traditional clear frying method are improved, the defects can be effectively improved by adopting a sand frying method, a uniform processed product is prepared, and the purposes of toxicity reduction and efficiency improvement of processing are achieved. The established HPLC method for measuring the contents of the four medicinal components of the szechwan chinaberry fruit and the fingerprint spectrum method can be used for quickly and integrally evaluating the quality and the processing effect of the szechwan chinaberry fruit processed product.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine processing and quality control, and specifically relates to a sand-fried Toosendan fruit processing technology and a quality evaluation method for establishing optimal processing technology parameters. Background Art

[0002] Toosendan fruit is the dried mature fruit of the plant Toosendan Sieb.et Zucc. of the Meliaceae family. It is also known as Toosendan, Golden Bell, Toosendan Fruit, etc. It is a traditional Chinese medicine for regulating qi. It has the effects of soothing the liver, promoting qi and relieving pain, and killing insects. It is often used for chest and flank, abdominal distension and pain, hernia pain, abdominal pain caused by worms, etc. Toosendan fruit is slightly toxic when raw, but it can reduce toxicity and enhance pharmacological effects after processing, so it is often used after processing.

[0003] Ancient books record that the processing methods of Toosendan fruit include cleaning, cutting, salting, vinegaring, stir-frying, and medicinal juice processing. With the development of traditional Chinese medicine, the processing methods of Toosendan fruit are now mainly cleaning and stir-frying. The processing methods of Toosendan fruit recorded in all editions of the "Chinese Pharmacopoeia" are cleaning and stir-frying. Due to the irregular shape and size of Toosendan fruit itself, the stir-frying method is unevenly heated and the processing effect is poor, which may cause the processed product to fail to achieve the purpose of "reducing toxicity and increasing efficacy", affecting clinical use. In addition, the quantitative index of Toosendan fruit in the "Chinese Pharmacopoeia" 2020 uses high-performance liquid chromatography-mass spectrometry to determine the content of toosendanin, which has high requirements for method operation, detection instruments and technical conditions.

[0004] Among the existing studies, there are few studies on the processing technology of sand-fried Toosendan fruit. There are no clear regulations on the process parameters such as sand-frying temperature, sand multiples, medicinal material particle size and frying temperature. Therefore, establishing the optimal sand-frying process conditions for Toosendan fruit is crucial to achieve the purpose of its processing to reduce toxicity and increase efficacy; establishing a multi-component content determination method and combining it with fingerprint spectrum for comprehensive quality evaluation are of great significance for actual production and quality inspection. Summary of the invention

[0005] The purpose of the present invention is to propose a sand-fried Toosendan fruit processing technology and a quality evaluation method for establishing the best processing technology parameters. Compared with the stir-frying method, the sand-fried method can evenly and effectively control the heating temperature of Toosendan fruit, improve the stability of the processing technology, and more easily obtain a "reduced toxicity and increased efficacy" processed product, and the appearance color of the processed medicinal material is uniform. At the same time, the quality evaluation method can comprehensively, objectively and accurately evaluate the processing technology and quality of sand-fried Toosendan fruit by establishing a multi-index content and fingerprint spectrum evaluation method.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is:

[0007] Firstly, the invention proposes a sand-fried Toosendan fruit processing technology, which comprises the steps of putting river sand into a pot, heating it with medium fire, adding the Toosendan fruit that has been crushed in advance when the temperature is stable, frying the Toosendan fruit until the surface is slightly yellow, taking it out of the pot, cooling it, and taking it out to obtain the product.

[0008] As a preferred technical solution of the present invention, in the processing process, Toosendan seeds are pre-crushed to a medicinal material with a particle size of 1.5 to 2.5 cm, the amount of river sand added is 20 times that of the medicinal material, the frying temperature is 120° C., and the frying time is 6 minutes.

[0009] Secondly, the present invention also proposes a quality evaluation method for determining the optimal processing parameters of sand-fried Toosendan fruit, comprising the following steps:

[0010] (1) Select L9(3 4 ) orthogonal table, select river sand weight multiple 10-20 times, frying temperature 120-180℃, frying time 3-9min and medicinal material particle size 0.5-3.5cm as four investigation factors, each factor has three levels, and conduct an orthogonal test of sand frying processing of Chuanlianzi;

[0011] (2) The contents of the four main active ingredients and the yield of dry extract of Toosendan Fructus fried in sand by the above-mentioned orthogonal test were measured as evaluation indicators;

[0012] (3) Calculating scores for the measurement results of step (2) to obtain a comprehensive score;

[0013] (4) The above comprehensive scoring results were analyzed by variance analysis to obtain the optimal processing parameters of stir-fried Toosendan fruit;

[0014] (5) Conduct process verification on the above-mentioned optimal preparation process.

[0015] Among them, each factor described in step (1) takes three levels, namely, the weight multiples of river sand are 10 times, 15 times, and 20 times, the frying temperature is 120°C, 150°C, and 180°C, the frying time is 3min, 6min, and 9min, and the particle size of the medicinal material is 0-1.5cm, 1.5-2.5cm, and 2.5-3.5cm.

[0016] Among them, the four main active ingredients in step (2) are rutin, quercetin, isotoshurin and toosendan, and the content determination method is as follows:

[0017] ① Preparation of mixed reference solution: Take appropriate amount of rutin, quercetin, isotoshurin and toosendan, dissolve and dilute with methanol to prepare a solution containing 0.001806 mg of rutin, 0.01254 mg of quercetin, 0.19072 mg of isotoshurin and 0.0664 mg of toosendan per 1 mL, to obtain a mixed reference solution;

[0018] ② Preparation of test solution: Weigh 5.0 g of stir-fried Toosendan fruit, place in a round-bottom flask, add 50 mL of 70% ethanol, heat and reflux for extraction for 1 h, and cool; transfer 2.0 mL of the solution into an EP tube, centrifuge at 12000 r / min for 20 min, and filter with a 0.22 μm microporous membrane to obtain the filtrate to obtain the test solution;

[0019] ③Chromatographic conditions:

[0020] High performance liquid chromatography; Shim-pack Scepter C18-120 chromatographic column (2.1×100 mm, 3 μm);

[0021] The mobile phase was acetonitrile (A)-0.05% formic acid in water (B), with a flow rate of 0.3 mL / min;

[0022] The detection wavelength was 210 nm; the column temperature was 30 °C; the injection volume was 2.0 μL;

[0023] The elution procedure is:

[0024] 0-5min, 3%→13%A, 97%→87%B;

[0025] 5-15min, 13%→25%A, 87%→75%B;

[0026] 15-25min, 25%→40%A, 75%→60%B;

[0027] 25-30min, 40%→60%A, 60%→40%B;

[0028] 30-33min, 60% A, 40% B;

[0029] 33-34min, 60%→3%A, 40%→97%B;

[0030] 34-36min, 3%A, 97%B.

[0031] The comprehensive score calculation formula in step (3) is:

[0032] Z=[(Y1 / Y 1max )×0.2090+(Y2 / Y 2max )×0.1826+(Y3 / Y 3max )×0.1839+(Y4 / Y 4max )×0.1959+(Y G /

[0033] Y Gmax )×0.2279]×100%

[0034] Among them, Y1, Y2, Y3 and Y4 represent the contents of rutin, quercetin, isotoshurin and toosendan, respectively. G Indicates the yield of dry extract.

[0035] The optimal processing parameters of sand-fried Toosendan seeds obtained in step (4) are: particle size of medicinal material 1.5-2.5 cm, weight multiple of river sand 20 times, frying temperature 120° C., and frying time 6 min.

[0036] Wherein, in step (5), comparing the fingerprints and the contents of the medicinal components of the raw, stir-fried and sand-fried products of Toosendan Fructus to verify the optimal sand-frying process comprises the following steps:

[0037] ① Establish a fingerprint method, conduct methodological verification to examine precision, stability and repeatability, and then use the same chromatographic conditions to detect the samples to be tested. The test results are compared and evaluated by similarity evaluation software. The results show that the similarity of the fried products is above 0.90, and the similarity of the stir-fried products is above 0.89, and the repeatability is relatively good.

[0038] ② The contents of rutin, quercetin, isotoscherichia coli and toosendan in raw, stir-fried and sand-fried products were tested. The results showed that the content of toosendania coli was in the order of sand-fried product > raw product > stir-fried product, and the content of isotoscherichia coli was in the order of stir-fried product > raw product > stir-fried product. Since the toxicity of isotoscherichia coli is much greater than that of toosendanin, stir-frying reduces the content of isotoscherichia coli, which is in line with the principle of toxicity reduction during processing, while the content of toosendania coli increases slightly, which is in line with the principle of synergistic enhancement during processing; the total content of the four components is in the order of sand-fried product > stir-fried product > raw product.

[0039] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0040] (1) The present invention adopts an orthogonal experimental design, with sand frying temperature, river sand weight multiples, frying time, and medicinal material particle size as the main investigation factors, and the contents of four medicinal ingredients, namely, rutin, quercetin, isotoxin and toosendanin, and the dry extract yield as evaluation indicators. The independence weight method is used to calculate the comprehensive score, and the best processing technology of sand frying Toosendan fruit is selected; since the traditional processing method of stir-frying itself has the limitations of uneven heating and poor processing effect, it may cause the processed product to fail to achieve the purpose of "reducing toxicity and increasing efficacy", affecting clinical use; after changing to the sand frying method, the heating temperature of Toosendan fruit can be evenly and effectively controlled, the processing technology is more stable, it is easier to obtain a processed product with "reducing toxicity and increasing efficacy", and the appearance and color of the processed medicinal material are better.

[0041] (2) The present invention adopts HPLC method to carry out content determination and fingerprint comparison of medicinal components of Toosendan Fructus, selects reasonable chromatographic conditions and chromatographic columns, has good separation effect, high reproducibility and excellent identification effect. The method verification shows that the instrument has good precision, good sample stability and good method repeatability, meets the technical requirements of content determination and fingerprint, and provides a better evaluation method for quality control of Toosendan Fructus. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 This is the appearance of 9 batches of sand-fried products in orthogonal experiments.

[0043] Figure 2 It is the HPLC comparison chart of four mixed reference substances and Toosendan samples (1. rutin; 2. quercetin; 3. isotolin, 4. toosendan).

[0044] Figure 3 It is an overlay of HPLC fingerprints of 14 batches of different processed products of Toosendan Fructus (sand-fried S1-6, lightly stir-fried S7-12, and raw product S13-14). DETAILED DESCRIPTION

[0045] The processing process steps proposed by the present invention are as follows: put clean river sand into a pot, heat it over medium heat, wait for the temperature to stabilize, put in the cut Chuanlianzi medicinal pieces, stir-fry until the surface is slightly yellow, take it out of the pot, let it cool, and take it out. An orthogonal design experiment is adopted, and the sand frying temperature, river sand weight multiples, frying time, and medicinal material particle size are used as investigation factors to screen out the best sand frying Chuanlianzi processing technology, and the best sand frying technology is verified, and the verification results show that it meets the processing principle of reducing toxicity and increasing efficiency.

[0046] The quality control steps proposed in the present invention are as follows: taking an appropriate amount of Toosendan processed product powder, heating and refluxing extraction, centrifugation, and filtering to obtain a test solution; using high performance liquid chromatography (HPLC) C18 chromatographic column and DAD detector to determine the contents of four components such as Toosendanin, and recording the fingerprint spectrum, as a result, the contents of the four components and UPLC spectrum (such as Figure 2 shown).

[0047] The traditional processing method of Toosendan Fructus, stir-frying, has the limitations of uneven heating and inconsistent processing degree. The improved sand stir-frying method of the present invention can effectively improve this disadvantage, prepare a uniform processed product, and achieve the purpose of "reducing toxicity and increasing efficacy" of processing. The established HPLC method for determining the content of four active ingredients in Toosendan Fructus and fingerprint spectrum method can quickly and holistically evaluate the quality and processing effect of Toosendan Fructus processed products.

[0048] The present invention is further described in detail below in conjunction with embodiments and drawings.

[0049] A quality evaluation method for determining the optimal processing parameters of sand-fried Toosendan fruit comprises the following steps:

[0050] (1) Select L9(3 4 ) orthogonal table, selected river sand weight multiples of 10-20 times, frying temperature of 120-180℃, frying time of 3-9min and medicinal material particle size of 0.5-3.5cm as four investigation factors, each factor took three levels, and carried out orthogonal experiment on sand frying of Toosendan fruit.

[0051] Table 1. Orthogonal factor level table

[0052]

[0053] (2) The Fructus Toosendans fried in sand by the above orthogonal test (such as Figure 1 The contents of four main active ingredients, toosendanin, isotosendanin, rutin and quercetin, as well as the yield of dry extract were determined as evaluation indicators.

[0054] The determination methods of the contents of the four main active ingredients, toosendanin, isotosendanin, rutin and quercetin, are as follows:

[0055] ① Preparation of mixed reference solution: Take appropriate amount of each reference substance of rutin, quercetin, isotoshurin and toosendan, dissolve and dilute with methanol to make a solution containing 0.001806 mg of rutin, 0.01254 mg of quercetin, 0.19072 mg of isotoshurin and 0.0664 mg of toosendan per 1 mL to obtain a mixed reference solution.

[0056] ② Preparation of the test solution: Weigh 5.0 g of stir-fried Toosendan seeds, place in a round-bottom flask, add 50 mL of 70% ethanol, heat under reflux for extraction for 1 h, and cool; transfer 2.0 mL of the solution into an EP tube, centrifuge at 12000 r / min for 20 min, and filter with a 0.22 μm microporous filter membrane, take the filtrate, and obtain the test solution.

[0057] ③Chromatographic conditions:

[0058] High performance liquid chromatography; Shim-pack Scepter C18-120 chromatographic column (2.1×100 mm, 3 μm);

[0059] The mobile phase was acetonitrile (A)-0.05% formic acid in water (B), with a flow rate of 0.3 mL / min;

[0060] The detection wavelength was 210 nm; the column temperature was 30 °C; the injection volume was 2.0 μL;

[0061] The elution procedure is:

[0062] 0-5min, 3%→13%A, 97%→87%B;

[0063] 5-15min, 13%→25%A, 87%→75%B;

[0064] 15-25min, 25%→40%A, 75%→60%B;

[0065] 25-30min, 40%→60%A, 60%→40%B;

[0066] 30-33min, 60% A, 40% B;

[0067] 33-34min, 60%→3%A, 40%→97%B;

[0068] 34-36min, 3%A, 97%B.

[0069] Table 2. Orthogonal experimental analysis results

[0070]

[0071] (3) Use the independence weight method to calculate the scores for the measurement results of step (2).

[0072] Table 3. Calculation results of the independence weight method

[0073]

[0074] The comprehensive score calculation formula is:

[0075] Z=[(Y1 / Y 1max )×0.2090+(Y2 / Y 2max )×0.1826+(Y3 / Y 3max )×0.1839+(Y4 / Y 4max )×0.1959+(Y G /

[0076] Y Gmax )×0.2279]×100%

[0077] Table 4. Intuitive analysis table

[0078]

[0079] (4) The variance analysis was performed on the results of the above comprehensive scores to obtain the best optimization process for stir-frying Toosendan seeds in sand.

[0080] Table 5. ANOVA table

[0081]

[0082] From the intuitive analysis results, it can be seen that the order of factors affecting the total content of the four components of Chuanlianzi is: B>D>A>C. The order of the levels of factor B is B3>B1>B2, the order of the levels of factor D is D2>D3>D1, the order of the levels of factor A is A1>A3>A2, and the order of the levels of factor C is C2>C3>C1. At the same time, the results of variance analysis show that the four factors of frying temperature, river sand weight multiples, frying time and medicinal material particle size have no significant effect on the four components and extract yield in Chuanlianzi at the level of α=0.10.

[0083] Therefore, the optimal sand frying process parameters of Toosendan seeds selected by the present invention are B3D2A1C2, that is, Toosendan seeds with a particle size of 1.5 to 2.5 cm are buried and stir-fried with 20 times sand at 120° C. for 6 minutes.

[0084] (5) Process verification of the above-mentioned optimal process:

[0085] Sample preparation and detection: Take 14 portions of the same batch of Chuanlianzi, 50g each. Six of them were fried at 120℃ for 6 minutes using the best sand-frying method, i.e., Chuanlianzi with a particle size of 1.5-2.5cm was buried in 20 times sand. Another six were fried at the same temperature to the same color by the stir-frying method. The remaining two were not processed. The test solution was prepared uniformly according to the above preparation method, and the relevant components were determined according to the above-mentioned chromatographic conditions. The content was calculated, and the similarity of the fingerprints of the two processed products was compared.

[0086] Table 6. Contents of four components in fried Toosendan fruit

[0087]

[0088] Table 7. Contents of four components in stir-fried Toosendan fruit

[0089]

[0090] Table 8. Contents of four components in raw Toosendan fruit

[0091]

[0092] The contents of rutin, quercetin, isotoshulin and toosendan in raw, stir-fried and sand-fried products were tested. The results showed that the content of toosendan was in the order of sand-fried product > raw product > stir-fried product, and the content of isotoshulin was in the order of stir-fried product > raw product > stir-fried product. Since the toxicity of isotoshulin is much greater than that of toosendan, the reduction of isotoshulin content by sand-frying is in line with the principle of toxicity reduction by processing, and the content of toosendan slightly increased, which is in line with the principle of synergistic effect by processing. The total content of the four components was in the order of sand-fried product > stir-fried product > raw product.

[0093] The detection results of the fingerprint spectrum are compared and evaluated by similarity evaluation software (see the similarity comparison chart Figure 3 ), the similarities of the stir-fried products were all above 0.90, and those of the stir-fried products were all above 0.89, with good repeatability.

[0094] The above contents are merely examples and explanations of the concept of the present invention. The technicians in this technical field may make various modifications or additions to the specific embodiments described or replace them in a similar manner. As long as they do not deviate from the concept of the invention or exceed the scope defined by the claims, they should all fall within the protection scope of the present invention.

Claims

1. A process for preparing stir-fried Toosendan fruit, characterized in that: Put river sand in a pot and heat it over medium heat. When the temperature stabilizes, add the pre-cut Toosendan seeds and stir-fry until the surface turns slightly yellow. Remove from the pot, let cool, and take out.

2. The process for preparing the stir-fried Toosendan fruit as claimed in claim 1, characterized in that: Toosendan seeds were pre-cut into medicinal materials with a particle size of 1.5 to 2.5 cm, the amount of river sand added was 20 times that of the medicinal materials, the frying temperature was 120°C, and the frying time was 6 minutes.

3. A quality evaluation method for determining the optimal processing parameters of stir-fried Toosendan fruit, characterized in that: The steps include: (1) Select L9(3 4 ) orthogonal table, select river sand weight multiple 10-20 times, frying temperature 120-180℃, frying time 3-9min and medicinal material particle size 0.5-3.5cm as four investigation factors, each factor has three levels, and conduct an orthogonal test of sand frying processing of Chuanlianzi; (2) The contents of the four main active ingredients and the yield of dry extract of Toosendan Fructus fried in sand by the above-mentioned orthogonal test were measured as evaluation indicators; (3) Calculating scores for the measurement results of step (2) to obtain a comprehensive score; (4) The above comprehensive scoring results were analyzed by variance analysis to obtain the optimal processing parameters of stir-fried Toosendan fruit; (5) Conduct process verification on the above-mentioned optimal preparation process.

4. The quality evaluation method according to claim 3, characterized in that: Each factor in step (1) is set at three levels, namely, the weight multiple of river sand is 10 times, 15 times, and 20 times, the frying temperature is 120°C, 150°C, and 180°C, the frying time is 3min, 6min, and 9min, and the particle size of the medicinal material is 0-1.5cm, 1.5-2.5cm, and 2.5-3.5cm.

5. The quality evaluation method according to claim 3, characterized in that: The four main active ingredients in step (2) are rutin, quercetin, isotoshurin and toosendan, and the content determination method is as follows: ① Preparation of mixed reference solution: Take appropriate amount of rutin, quercetin, isotoshurin and toosendan, dissolve and dilute with methanol to prepare a solution containing 0.001806 mg of rutin, 0.01254 mg of quercetin, 0.19072 mg of isotoshurin and 0.0664 mg of toosendan per 1 mL, to obtain a mixed reference solution; ② Preparation of test solution: Weigh 5.0 g of stir-fried Toosendan fruit, place in a round-bottom flask, add 50 mL of 70% ethanol, heat and reflux for extraction for 1 h, and cool; transfer 2.0 mL of the solution into an EP tube, centrifuge at 12000 r / min for 20 min, and filter with a 0.22 μm microporous membrane to obtain the filtrate to obtain the test solution; ③Chromatographic conditions: High performance liquid chromatography; Shim-pack Scepter C18-120 chromatographic column (2.1×100 mm, 3 μm); The mobile phase was acetonitrile (A)-0.05% formic acid in water (B), with a flow rate of 0.3 mL / min; The detection wavelength was 210 nm; the column temperature was 30 °C; the injection volume was 2.0 μL; The elution procedure is: 0-5min, 3%→13%A, 97%→87%B; 5-15min, 13%→25%A, 87%→75%B; 15-25min, 25%→40%A, 75%→60%B; 25-30min, 40%→60%A, 60%→40%B; 30-33min, 60% A, 40% B; 33-34min, 60%→3%A, 40%→97%B; 34-36min, 3%A, 97%B.

6. The quality evaluation method according to claim 3, characterized in that: The comprehensive score calculation formula in step (3) is: Z=[(Y1 / Y 1max )×0.2090+(Y2 / Y 2max )×0.1826+(Y3 / Y 3max )×0.1839+(Y4 / Y 4max )×0.1959+(Y G / Y Gmax )×0.2279]×100％ Among them, Y1, Y2, Y3 and Y4 represent the contents of rutin, quercetin, isotoshurin and toosendan, respectively. G Indicates the yield of dry extract.

7. The quality evaluation method according to claim 3, characterized in that: The optimal processing parameters of sand-fried Toosendan seeds in step (4) are: particle size of medicinal material 1.5-2.5 cm, weight multiple of river sand 20 times, frying temperature 120° C., and frying time 6 min.

8. The quality evaluation method according to claim 3, characterized in that: In step (5), the fingerprints and the contents of the medicinal components of the raw, stir-fried and sand-fried products of Toosendan Fructus are compared to verify the optimal sand-frying process, which comprises the following steps: ① Establish a fingerprint method, conduct methodological verification to examine precision, stability and repeatability, and then use the same chromatographic conditions to detect the samples to be tested. The test results are compared and evaluated by similarity evaluation software. The results show that the similarity of the fried products is above 0.90, and the similarity of the stir-fried products is above 0.89, and the repeatability is relatively good. ② The contents of rutin, quercetin, isotoscherichia coli and toosendan in raw, stir-fried and sand-fried products were tested. The results showed that the content of toosendania coli was in the order of sand-fried product > raw product > stir-fried product, and the content of isotoscherichia coli was in the order of stir-fried product > raw product > stir-fried product. Since the toxicity of isotoscherichia coli is much greater than that of toosendanin, stir-frying reduces the content of isotoscherichia coli, which is in line with the principle of toxicity reduction during processing, while the content of toosendania coli increases slightly, which is in line with the principle of synergistic enhancement during processing; the total content of the four components is in the order of sand-fried product > stir-fried product > raw product.

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