A Chinese herbal medicine fermentation state detection method based on infrared temperature measurement technology

Abstract

The application provides a Chinese herbal medicine fermentation state detection method based on infrared temperature measurement technology, and belongs to the technical field of Chinese herbal medicine fermentation. The Chinese herbal medicine is subjected to solid fermentation by using a biological enzyme compound, the temperature inside the fermented Chinese herbal medicine is detected by using infrared temperature measurement technology, and the total sugar and reducing sugar contents of the fermented Chinese herbal medicine are detected; the internal temperature of the fermented Chinese herbal medicine is taken as the horizontal coordinate x, and the total sugar and reducing sugar contents of the fermented Chinese herbal medicine are taken as the vertical coordinates y, respectively, so that a linear prediction model of the total sugar and reducing sugar contents of the fermented Chinese herbal medicine is constructed; and the total sugar and reducing sugar contents of the fermented Chinese herbal medicine are predicted according to the internal temperature of the fermented Chinese herbal medicine and the linear prediction model. The application can detect the temperature of Chinese herbal medicine solid fermentation in real time, and then can predict the total sugar and reducing sugar contents of the Chinese herbal medicine subjected to solid fermentation by using the fermentation temperature, so as to evaluate the fermentation degree and state of the Chinese herbal medicine, and has great significance for the development of the Chinese herbal medicine industry.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of Chinese herbal medicine fermentation, and particularly relates to a Chinese herbal medicine fermentation state detection method based on infrared temperature measurement technology. BACKGROUND

[0002] In recent years, the research on traditional Chinese medicine, especially Chinese herbal medicine, has developed rapidly, resulting in an increasing generation of by-products of Chinese herbal medicine year by year. However, the existing technology is insufficient in the development and utilization of Chinese herbal medicine by-products, and there is a serious waste of resources. Chinese herbal medicine is mainly plant-derived medicinal materials, and the effective components mainly exist in the cytoplasm of cells. The fibrous structure in the cell wall of plants hinders the extraction of effective active components, resulting in a waste of resources due to the unsatisfactory drug efficacy. Fermentation technology is an effective method for improving the content and efficacy of effective components of Chinese herbal medicine, reducing toxicity, reducing side effects, and producing new active components. However, there are problems such as uncontrollable fermentation process, easy contamination of bacteria, inability to evaluate the fermentation degree of Chinese herbal medicine in real time, and large fluctuations under the influence of fermentation conditions.

[0003] In addition, modern fermentation technology incorporates microbiology, bioengineering, pharmacology, and bioinformatics, and liquid fermentation can detect fermentation parameters such as temperature, humidity, pH, and bacterial growth in real time, which greatly improves the quality and safety of fermentation products. However, compared with liquid fermentation, solid fermentation of Chinese herbal medicine is more common. The non-flowing nature of solid fermentation materials makes it difficult to detect the above fermentation parameters in real time.

[0004] Currently, infrared temperature measurement technology has been applied in many medical fields, such as temperature examination and tumor examination. However, there is no existing technology reporting its application in the field of Chinese herbal medicine fermentation. Therefore, it is of great significance for the development of Chinese herbal medicine industry to provide a method for real-time detection of the fermentation state and degree of solid fermentation of Chinese herbal medicine by-products using infrared temperature measurement technology. SUMMARY

[0005] Therefore, the purpose of the present application is to provide a method for real-time determination of the temperature inside the solid fermentation of Chinese herbal medicine using infrared temperature measurement technology, and prediction of the fermentation state and degree of Chinese herbal medicine based on the internal temperature of the fermented Chinese herbal medicine.

[0006] In order to achieve the above-mentioned purpose of the application, the present application provides the following technical solutions:

[0007] The application discloses a Chinese herbal medicine fermentation state detection method based on infrared temperature measurement technology.

[0008] Preferably, the solid fermentation is that the Chinese herbal medicine by-products are dried, crushed through a 2-5 mesh sieve, mixed with water at a mass ratio of 10:6-15, and fermented by adding a biological enzyme compound with a mass percentage of 0.4-5.5%.

[0009] Preferably, the Chinese herbal medicine includes any one or more of forsythia suspensa, honeysuckle, fried bitter apricot kernels, isatis root, hawksbeard, houttuynia cordata, rhodiola rosea and licorice.

[0010] Preferably, the Chinese herbal medicine by-products include residues generated in the extraction and decoction process of traditional Chinese medicine preparations and / or non-medicinal parts of Chinese herbal medicines.

[0011] Preferably, the biological enzyme compound is composed of cellulase, xylanase and protease at a weight ratio of 1-3:1:1.

[0012] Preferably, the cellulase is 60,000 U / g, the xylanase is 60,000 U / g, and the protease is 60,000 U / g.

[0013] Preferably, the fermentation temperature is 40-50 DEG C.

[0014] Preferably, the infrared temperature measurement is that a sample fermentation bag of the fermented Chinese herbal medicine is taken out, cooled for 10-30 min, and then photographed and measured in temperature by using an instrument connected with an infrared thermal imaging device.

[0015] Preferably, the temperature inside the fermented Chinese herbal medicine is detected by using the infrared temperature measurement technology every 1-2 days.

[0016] Preferably, when the fermented Chinese herbal medicine is fermented at 50 DEG C by adding 5.0% of the biological enzyme compound, the total sugar prediction model is y=7.648x+60.643, R 2 =0.9923; the reduction sugar prediction model is y=-1.8228x+165.15, R 2 =0.9983; when the fermented Chinese herbal medicine is fermented at 50 DEG C by adding 0.5% of the biological enzyme compound, the total sugar prediction model is y=3.8452x+230.11, R 2was 0.8968; the predicted model of the reducing sugar was y = -0.7078x + 87.4, R 2 was 0.9976.

[0017] Compared with the prior art, the present application has the following beneficial effects:

[0018] The present application provides a Chinese herbal medicine fermentation state detection method based on infrared temperature measurement technology. In the present application, when a biological enzyme compound is used to perform solid fermentation on Chinese herbal medicine, the infrared temperature measurement technology is used to measure the temperature inside the Chinese herbal medicine solid fermentation, and the total sugar and reducing sugar contents of the fermented Chinese herbal medicine are detected at the same time. A linear prediction model of the total sugar and reducing sugar contents of the fermented Chinese herbal medicine is constructed. Then, according to the internal temperature of the fermented Chinese herbal medicine and the constructed linear prediction model, the total sugar and reducing sugar contents of the fermented Chinese herbal medicine can be predicted. The present application can evaluate the fermentation degree and state of Chinese herbal medicine by detecting the temperature of Chinese herbal medicine solid fermentation in real time and predicting the total sugar and reducing sugar contents of the Chinese herbal medicine solid fermentation by using the fermentation temperature. The present application has great significance for the development of the Chinese herbal medicine industry. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 : Temperature changes of Aspergillus niger (An), Bacillus subtilis (Bs) and biological enzyme compound (En) in the fermentation of Chinese herbal medicine samples under different temperatures and different addition amounts;

[0020] Figure 2 : Changes of viable bacteria, crude fiber, total sugar and reducing sugar in the fermentation of Chinese herbal medicine samples by Aspergillus niger (An), Bacillus subtilis (Bs) and biological enzyme compound (En) under different temperatures and different addition amounts;

[0021] Figure 3 : Correlation analysis between infrared temperature and viable bacteria (CFU), crude fiber (CF), total sugar (TS) and reducing sugar (RS) in the fermentation samples based on Aspergillus niger (An), Bacillus subtilis (Bs) and biological enzyme compound (En);

[0022] Figure 4 : Correlation analysis between infrared temperature and total sugar (TS), reducing sugar (RS) in the fermentation samples by biological enzyme compound (En). DETAILED DESCRIPTION

[0023] The application provides a Chinese herbal medicine fermentation state detection method based on infrared temperature measurement technology, which comprises the following steps: solid fermentation of Chinese herbal medicine by using a biological enzyme compound, detection of the internal temperature of the fermented Chinese herbal medicine by using infrared temperature measurement technology, and detection of the total sugar and reducing sugar contents of the fermented Chinese herbal medicine; taking the internal temperature of the fermented Chinese herbal medicine as the horizontal coordinate x, and taking the total sugar and reducing sugar contents of the fermented Chinese herbal medicine as the vertical coordinate y, a linear prediction model of the total sugar and reducing sugar contents of the fermented Chinese herbal medicine is constructed; and the total sugar and reducing sugar contents of the fermented Chinese herbal medicine are predicted according to the internal temperature of the fermented Chinese herbal medicine and the linear prediction model.

[0024] In the application, the solid fermentation is that the Chinese herbal medicine by-products are dried, crushed through a 2-5 mesh sieve, mixed with water at a mass ratio of 10:6-15, and fermented by adding a biological enzyme compound with a mass percentage of 0.4-5.5%. Preferably, the solid fermentation is that the Chinese herbal medicine by-products are dried, crushed through a 4 mesh sieve, mixed with water at a mass ratio of 10:8-12, and fermented by adding a biological enzyme compound with a mass percentage of 0.5-5%. As an implementable mode, the Chinese herbal medicine by-products are dried at 45 DEG C for 6-12h, preferably 8-10h.

[0025] In the research of the application, it is found that if the addition ratio of the biological enzyme compound is too low, the Chinese herbal medicine is not completely decomposed, which easily causes resource waste, and the linear relationship between the internal temperature of the fermented Chinese herbal medicine and the total sugar and reducing sugar contents of the fermented Chinese herbal medicine is poor; if the addition ratio of the biological enzyme compound is too high, the production cost is increased. In the application, the addition ratio of the biological enzyme compound can fully decompose the Chinese herbal medicine by-products and extract beneficial components in the Chinese herbal medicine by-products, so that resource waste is avoided. Meanwhile, by using the biological enzyme compound and the addition ratio provided in the application, the internal temperature of the fermented Chinese herbal medicine and the total sugar and reducing sugar contents of the fermented Chinese herbal medicine present a good linear relationship in the fermentation process of the Chinese herbal medicine.

[0026] In the present application, the biological enzyme complex is composed of cellulase, xylanase and protease in a weight ratio of 1-3:1:1, preferably the biological enzyme complex is composed of cellulase, xylanase and protease in a weight ratio of 1:1:1. The cellulase is 60,000 U / g, the xylanase is 60,000 U / g, and the protease is 60,000 U / g. In the fermentation of Chinese herbal medicine, microorganisms or enzyme preparations are often used to ferment Chinese herbal medicine. In the research, it is found that when fungi or bacteria are added to ferment Chinese herbal medicine, the internal temperature of the fermented Chinese herbal medicine does not have a good linear relationship with the number of living bacteria, humidity, temperature, crude fiber, total sugar and reducing sugar content and other fermentation parameters in the fermentation, and thus the fermentation state and degree of Chinese herbal medicine solid fermentation cannot be predicted according to the internal temperature of the fermented Chinese herbal medicine. In the present application, the biological enzyme complex provided by the present application is used for solid fermentation of Chinese herbal medicine. In the fermentation process, the internal temperature of the fermented Chinese herbal medicine has a good linear relationship with the total sugar and reducing sugar content of the fermented Chinese herbal medicine, and thus the linear prediction model of the total sugar and reducing sugar content of the fermented Chinese herbal medicine can be constructed, and the total sugar and reducing sugar content of the fermented Chinese herbal medicine can be predicted according to the internal temperature of the fermented Chinese herbal medicine and the linear prediction model.

[0027] In the present application, the Chinese herbal medicine includes any one or several of forsythia, honeysuckle, fried bitter apricot kernel, isatis root, rhizoma tatarinowii, fishy-smelling grass, rhodiola and licorice.

[0028] In the present application, the Chinese herbal medicine by-product includes the residues produced in the extraction and decoction process of traditional Chinese medicine preparation, and / or the non-medicinal parts of Chinese herbal medicine.

[0029] In the present application, the Chinese herbal medicine is a commonly used Chinese herbal medicine in traditional Chinese medicine, which is used in large quantities in daily life, and the amount of related by-products of Chinese herbal medicine is large. The Chinese herbal medicine is mainly a plant-derived medicinal material. After the effective components in the medicinal material are extracted and utilized, the by-products still contain unutilized medicinal components. After further processing, the by-products can be used as additives of commercial feed to promote animal growth.

[0030] In the present application, the fermentation temperature is 40-50℃.

[0031] In the present application, the infrared temperature measurement is to take out the sample fermentation bag of the fermented Chinese herbal medicine, cool for 10-30 min, preferably cool for 20 min, and then use an instrument connected with an infrared thermal imaging device to take a photo and measure the temperature. As an implementable way, the infrared thermal imaging device in the present application is CompactPro, SeeThermal, USA.

[0032] In the present application, the temperature inside the fermented Chinese herbal medicine is detected by infrared temperature measurement technology every 1-2 days.

[0033] In the present application, the fermented Chinese herbal medicine is added with 5.0% of the biological enzyme compound by mass percentage, and when fermented at 50℃, the total sugar prediction model is y=7.648x+60.643, R 2 =0.9923; the prediction model of the reducing sugar is y=-1.8228x+165.15, R 2 =0.9983; the fermented Chinese herbal medicine is added with 0.5% of the biological enzyme compound by mass percentage, and when fermented at 50℃, the total sugar prediction model is y=3.8452x+230.11, R 2 =0.8968; the prediction model of the reducing sugar is y=-0.7078x+87.4, R 2 =0.9976. The linear prediction model of the total sugar and the reducing sugar content in the fermented Chinese herbal medicine constructed in the present application can accurately predict the total sugar and the reducing sugar content in the fermented Chinese herbal medicine by using the internal temperature of the fermented Chinese herbal medicine, and further can evaluate the fermentation degree and the fermentation state of the Chinese herbal medicine.

[0034] Unless otherwise specified, the reagents involved in the present application are all commercially available products, and can be obtained through stable commercial channels.

[0035] The technical solutions provided by the present application will be described in detail below in combination with the embodiments, but they should not be understood as limiting the scope of protection of the present application.

[0036] Example 1

[0037] The residues and non-medicinal parts of Chinese herbal medicine produced in the processing of forsythia, honeysuckle, fried bitter apricot kernel, isatis root, mao kudou, fishy-smelling grass, red sage and licorice root were dried at 45℃ for 8h;

[0038] The dried Chinese herbal medicine by-products were crushed through a 4-mesh sieve, mixed with water at a mass ratio of 10:8, added with 5.0% of the biological enzyme compound by mass percentage, and then fermented at 50℃;

[0039] The biological enzyme compound is composed of cellulase, xylanase and protease at a weight ratio of 1:1:1; the cellulase in the biological enzyme compound is 60,000 U / g, the xylanase is 60,000 U / g, and the protease is 60,000 U / g;

[0040] The internal temperature of the fermented Chinese herbal medicine was detected once every 1-2 days by using infrared temperature measurement technology; when measuring the temperature, the sample fermentation bag was taken out, cooled for 30 minutes, and then photographed by using the mobile phone of the infrared thermal imaging equipment (Compact Pro, SeeThermal, USA);

[0041] The temperature inside the fermented Chinese herbal medicine is detected by infrared temperature measurement technology at the 1st day, the 4th day and the 7th day of fermentation, and the total sugar and reducing sugar contents of the fermented Chinese herbal medicine are detected at the same time; the temperature inside the fermented Chinese herbal medicine is taken as the horizontal coordinate x, and the total sugar and reducing sugar contents of the fermented Chinese herbal medicine are taken as the vertical coordinate y respectively, so as to construct a linear prediction model of the total sugar and reducing sugar contents in the fermented Chinese herbal medicine.

[0042] Example 2

[0043] The residues and non-medicinal parts of Chinese herbal medicine produced in the extraction and decoction process of forsythia suspense, honeysuckle, fried bitter apricot kernel, isatis root, rhizoma tatarinowii, yuxingcao, rhodiola and licorice are dried at 45℃ for 8h;

[0044] The dried Chinese herbal medicine by-products are crushed through a 4-mesh sieve, mixed with water at a mass ratio of 10:8, and added with a biological enzyme complex at a mass percentage of 0.5%, and then subjected to fermentation treatment at 50℃;

[0045] The biological enzyme complex is composed of cellulase, xylanase and protease at a weight ratio of 1:1:1; the cellulase in the biological enzyme complex is 60,000 U / g, the xylanase is 60,000 U / g, and the protease is 60,000 U / g;

[0046] The temperature inside the fermented Chinese herbal medicine is detected by infrared temperature measurement technology every 1-2 days; when the temperature is measured, the sample fermentation bag is taken out, cooled for 30 minutes, and then photographed by a mobile phone connected with an infrared thermal imaging device (Compact Pro, SeeThermal, USA);

[0047] The temperature inside the fermented Chinese herbal medicine is detected by infrared temperature measurement technology at the 1st day, the 4th day and the 7th day of fermentation, and the total sugar and reducing sugar contents of the fermented Chinese herbal medicine are detected at the same time; the temperature inside the fermented Chinese herbal medicine is taken as the horizontal coordinate x, and the total sugar and reducing sugar contents of the fermented Chinese herbal medicine are taken as the vertical coordinate y respectively, so as to construct a linear prediction model of the total sugar and reducing sugar contents in the fermented Chinese herbal medicine.

[0048] Example 3

[0049] The difference between this example and Example 1 is that the fermentation is carried out at 40℃.

[0050] Example 4

[0051] The difference between this example and Example 2 is that the fermentation is carried out at 40℃.

[0052] Example 5

[0053] The residues produced in the extraction and decoction process of forsythia suspense, honeysuckle and fried bitter apricot kernel are dried at 45℃ for 10h;

[0054] The dried Chinese herbal medicine by-products are crushed through a 5-mesh sieve, mixed with water at a mass ratio of 10:12, and added with a biological enzyme compound at a mass percentage of 3.5%, and then subjected to fermentation treatment at 45°C;

[0055] The biological enzyme compound is composed of cellulase, xylanase and protease at a weight ratio of 1:1:1, wherein the cellulase is 60,000 U / g, the xylanase is 60,000 U / g, and the protease is 60,000 U / g in the biological enzyme compound;

[0056] The internal temperature of the fermented Chinese herbal medicine is detected every 1-2 days by using infrared temperature measurement technology; when the temperature is measured, the sample fermentation bag is taken out, cooled for 30 minutes, and then photographed by using a mobile phone of an infrared thermal imaging device (Compact Pro, SeeThermal, USA);

[0057] The internal temperature of the fermented Chinese herbal medicine is detected every 1-2 days by using infrared temperature measurement technology; when the temperature is measured, the sample fermentation bag is taken out, cooled for 30 minutes, and then photographed by using a mobile phone of an infrared thermal imaging device (Compact Pro, SeeThermal, USA);

[0058] Comparative Example 1

[0059] The residues and non-medicinal parts of Chinese herbal medicines produced in the extraction and decoction process of forsythia, honeysuckle, fried bitter apricot kernels, isatis root, mao kudou, fishy-smelling grass, red sage and licorice are dried at 45°C for 8h;

[0060] The dried Chinese herbal medicine by-products are crushed through a 4-mesh sieve, mixed with water at a mass ratio of 10:8, and added with Aspergillus niger at a mass percentage of 3.5%, and then subjected to fermentation treatment at 30°C; wherein the effective viable count of the Aspergillus niger is 10 6 CFU / g;

[0061] The internal temperature of the fermented Chinese herbal medicine is detected every 1-2 days by using infrared temperature measurement technology; when the temperature is measured, the sample fermentation bag is taken out, cooled for 30 minutes, and then photographed by using a mobile phone of an infrared thermal imaging device (Compact Pro, SeeThermal, USA).

[0062] Comparative Example 2

[0063] The residues and non-medicinal parts of Chinese herbal medicines produced in the extraction and decoction process of forsythia, honeysuckle, fried bitter apricot kernels, isatis root, mao kudou, fishy-smelling grass, red sage and licorice are dried at 45°C for 8h;

[0064] The dried Chinese herbal medicine by-products are crushed through a 4-mesh sieve, mixed with water at a mass ratio of 10:8, and added with Bacillus subtilis at a mass percentage of 3.5% for fermentation treatment at 30°C; wherein the effective viable count of the Bacillus subtilis is 10 6 CFU / g.

[0065] Every 1-2 days, the internal temperature of the fermented Chinese herbal medicine is detected by using infrared temperature measurement technology; when the temperature is measured, the sample fermentation bag is taken out, cooled for 30 minutes, and then photographed by using a mobile phone of an infrared thermal imaging device (CompactPro, SeeThermal, USA).

[0066] Comparative Example 3

[0067] The residues and non-medicinal parts of Chinese herbal medicine produced by extraction and decoction during the processing of Forsythia, honeysuckle, fried bitter almonds, isatis root, maojunzhu, yuxingcao, red sage and licorice are dried at 45°C for 8h;

[0068] The dried Chinese herbal medicine by-products are crushed through a 4-mesh sieve, mixed with water at a mass ratio of 10:8, and added with Bacillus subtilis at a mass percentage of 3.5% for fermentation treatment at 40°C; wherein the effective viable count of the Bacillus subtilis is 10 6 CFU / g.

[0069] Every 1-2 days, the internal temperature of the fermented Chinese herbal medicine is detected by using infrared temperature measurement technology; when the temperature is measured, the sample fermentation bag is taken out, cooled for 30 minutes, and then photographed by using a mobile phone of an infrared thermal imaging device (CompactPro, SeeThermal, USA).

[0070] Example 6

[0071] In this example, the internal temperature changes of the fermented Chinese herbal medicine samples under different fermentation treatment conditions of Chinese herbal medicine are compared

[0072] The internal temperatures of the fermented Chinese herbal medicine samples of Examples 1-4 and Comparative Examples 1-3 are measured on the 0th day, the 1st day, the 2nd day and the 4th day of fermentation, and the results are shown in Figure 1 .

[0073] Under the fermentation condition of 30℃, the temperature of the second day of fermentation of the Chinese herbal medicine using Aspergillus niger and Bacillus subtilis showed the highest, and the temperature difference of the two was not large overall. In the fermentation treatment of Bacillus subtilis, the temperature of the Chinese herbal medicine fermented at 40℃ was significantly higher than that of the Chinese herbal medicine fermented at 30℃ on the 1st, 2nd and 4th day of fermentation. In the comparison of Bacillus subtilis and biological enzyme complex, the temperature difference of the two treatments was not large under the condition of 40℃. Under the condition of the same temperature and different addition amount, the temperature difference of the fermentation samples of the biological enzyme complex was not large. Overall, compared with the treatment of Aspergillus niger fermented Chinese herbal medicine, the higher the internal temperature of the Chinese herbal medicine sample fermented at a relatively high temperature by Bacillus subtilis and enzyme preparation.

[0074] Example 7

[0075] This example compares the changes of viable cell count, crude fiber, total sugar and reducing sugar of the fermented Chinese herbal medicine samples under different fermentation treatment conditions of Chinese herbal medicine

[0076] On the 1st, 2nd and 4th day of fermentation, the samples of Examples 1-4 and Comparative Examples 1-3 were collected and analyzed for viable cell count, crude fiber, total sugar and reducing sugar content, and the results are shown in Table 1. Figure 1

[0077] (1) Viable cell count detection

[0078] 1g of sample was added to 9mL of sterilized distilled water, mixed thoroughly, and then diluted by 10 times. The sample was cultured in LB agar medium at 30℃ for 24 hours, and the viable cell count was determined.

[0079] (2) Crude fiber detection

[0080] Prepare 0.13mol / L sulfuric acid solution, 0.23mol / L potassium hydroxide solution and 0.5mol / L hydrochloric acid solution. Crush the sample to 10-40 mesh. Put the sieved sample into a glass or metal container and dry it in a forced air drying oven at 105℃ for 4 hours. Put the dried sample into a drying dish and cool it for at least 30 minutes. Weigh 1g of sample, label the filter bag, and then put the sample into the filter bag. Seal the filter bag and soak it in acetone to remove the fat in the sample. Pour enough 0.5mol / L hydrochloric acid solution into the glass beaker and place it in the fume hood for 20 minutes. After pouring out the hydrochloric acid solution, stand for 10 minutes. Follow the standard procedure of the fiber tester to acid-alkali digest the fermented Chinese herbal medicine sample. Soak the sample in acetone and hydrochloric acid solution in turn, and then evenly put all the filter bags into a forced air drying oven at 105℃ for 4 hours. Cool for 30 minutes, then put the filter bags into a crucible, and then put the crucible into a muffle furnace at 550℃ for 4 hours. Cool and weigh to calculate.

[0081] (3) Total sugar detection​

[0082] Take 1 g sample in a 100 mL conical flask, add 10 mL hydrochloric acid (6 mol / L) solution and 15 mL distilled water, and cook in a 100°C water bath for 30 min, stirring while cooking. Take out the conical flask and, after the liquid cools, centrifuge at 4000 rpm for 10 min. Take the supernatant, pour 20 mL distilled water into the centrifuge tube and centrifuge, and mix the two supernatants and make up to 100 mL. Take 1 ml of the total sugar extract, add 2 mL of DNS solution, and place in a 100°C water bath for 10 min. After cooling, add 9 mL of distilled water, mix well, take 200 uL of the solution into a 96-well plate, and measure the absorbance at 540 nm. Calculate and analyze.

[0083] (4) Reducing sugar detection

[0084] Take 1 g sample in a 100 mL conical flask, add 10 mL hydrochloric acid (6 mol / L) solution and 15 mL distilled water, and cook in a 100°C water bath for 30 min, stirring while cooking. Take out the conical flask and, after the liquid cools, centrifuge at 4000 rpm for 10 min. Take the supernatant, pour 20 mL distilled water into the centrifuge tube and centrifuge, and mix the two supernatants and make up to 100 mL. Take 1 ml of the total sugar extract, add 2 mL of DNS solution, and place in a 100°C water bath for 10 min. After cooling, add 9 mL of distilled water, mix well, take 200 uL of the solution into a 96-well plate, and measure the absorbance at 540 nm. Calculate and analyze.

[0085] The results show that, in Comparative Example 1, when Aspergillus niger is used to ferment Chinese herbal medicine at 30°C, the microbial count is maintained at 10 7 CFU / g, and decreases to 10 3 CFU / g on the 7th day; in Comparative Examples 2 and 3, when Bacillus subtilis is used to ferment Chinese herbal medicine at 30°C and 40°C, the opposite results are obtained. In Examples 1-4, when the biological enzyme complex is used to ferment Chinese herbal medicine, the viable count shows a trend of first increasing and then decreasing, but at 50°C and with 5.0% biological enzyme complex addition, the viable count shows a decreasing trend with the fermentation days. Crude fiber and plant polysaccharides are degraded to produce reducing sugar. Under the conditions of biological enzyme complex 5.0% / 40°C, 0.5% / 40°C and 5.0% / 50°C, crude fiber and polysaccharides show a gradual downward trend with the progress of the treatment time. Reducing sugar is a good indicator of fiber degradation ability. Compared with Aspergillus niger and Bacillus subtilis, the use of biological enzyme complex to ferment Chinese herbal medicine results in higher reducing sugar content; among them, the effect of high-dose and high-temperature treatment on reducing sugar production is more prominent. Overall, with the decrease of crude fiber and total sugar content, the reducing sugar content increases.

[0086] Example 8

[0087] The present embodiment compares the correlation of infrared temperature measurement of fermented Chinese herbal medicine samples under different fermentation conditions of Chinese herbal medicine with viable cell number, crude fiber, total sugar and reducing sugar

[0088] The internal temperature of fermented Chinese herbal medicine of Examples 1-4 and Comparative Examples 1-3 was measured on the 1st, 2nd and 4th day of fermentation, and the viable cell number (CFU), crude fiber (CF), total sugar (TS) and reducing sugar (RS) content in the fermented Chinese herbal medicine were also measured at the same time. The correlation of infrared temperature measurement of fermented Chinese herbal medicine samples with viable cell number, crude fiber, total sugar and reducing sugar was analyzed, and the results are shown in Figure 3

[0089] The results show that, as a means of measuring the internal temperature of fermented Chinese herbal medicine, thermal imaging shows different correlations between measured temperature and other results under different treatment methods of Chinese herbal medicine. In Comparative Example 1, using Aspergillus niger under fermentation conditions at 30°C (An30°C), the measured temperature showed a negative correlation with CFU, CF, TS and RS, and a significant negative correlation with crude fiber. Similarly, under fermentation conditions at 30°C (Bs30°C), in Comparative Example 2, using Bacillus subtilis to ferment the sample, the measured temperature showed a positive correlation with CF and a negative correlation with TS.

[0090] In Examples 1-4, using a biological enzyme complex under the same temperature and different additive dosages, the measured temperature showed a positive correlation with TS. Under the same additive dosage and different temperatures, the measured temperature also showed a positive correlation with TS. In Example 1, adding 5.0% biological complex enzyme under fermentation conditions at 50°C, the measured temperature showed a significant negative correlation with RS.

[0091] Overall, the correlation analysis between the measured temperature of fermented Chinese herbal medicine samples and different fermentation indicators shows different trends due to different treatment methods, fermentation temperatures and additive proportions. However, in relation to the fermentation treatment method using a biological enzyme complex, the measured temperature shows a negative correlation with reducing sugar in the fermented sample. Generally, the higher the thermal imaging temperature in the sample, the more intense the biological reaction, the more sufficient the degradation of crude fiber, and the more total sugar and reducing sugar produced. The increase in reducing sugar promotes the growth of microorganisms, leading to an increase in viable cell number and a decrease in reducing sugar content. Based on the positive correlation between temperature and total sugar content and the negative correlation between temperature and reducing sugar content under enzymatic treatment, infrared temperature measurement can be used to assist in measuring the content of polysaccharides and reducing sugars in fermented Chinese herbal medicine samples under enzymatic conditions.

[0092] Example 9

[0093] ​The embodiment is a linear prediction model of total sugar and reducing sugar content in the fermented Chinese herbal medicine constructed according to the embodiments 1 and 2, and the result is shown in Figure 4 .

[0094] As shown in Figure 4 A and 4B, under the condition of fermenting Chinese herbal medicine at 50℃ and adding 0.5% bio-enzyme complex in the embodiment 2, the infrared temperature measurement is used to predict the content of total sugar and reducing sugar in the fermentation product, wherein the prediction R 2 =0.8968 of total sugar, the internal temperature x of the fermented Chinese herbal medicine and the content y of total sugar show a positive linear relationship, and the prediction model is y=3.8452x+230.11; the prediction R 2 =0.9976 of reducing sugar, the internal temperature x of the fermented Chinese herbal medicine and the content y of reducing sugar show a negative linear relationship, and the prediction model is y=-0.7078x+87.4.

[0095] As shown in Figure 4 C and 4D, under the condition of fermenting Chinese herbal medicine at 50℃ and adding 5.0% bio-enzyme complex in the embodiment 1, the infrared temperature measurement has a good prediction effect on the content of total sugar and reducing sugar in the fermentation product, wherein the content y of total sugar shows a positive linear relationship, the prediction model is y=7.648x+60.643, and R 2 is 0.9923; the content y of reducing sugar shows a negative linear relationship, the prediction model is y=-1.8228x+165.15, and R 2 is 0.9983.

[0096] It can be seen that when the bio-enzyme complex is used to ferment Chinese herbal medicine, under the condition of sufficient enzymolysis, the fermentation temperature can well predict the content of total sugar and reducing sugar in the fermentation product.

[0097] The above only describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the principles of the present application, several improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A method for detecting the fermentation state of traditional Chinese medicine based on infrared thermometry, characterized in that, Traditional Chinese medicine was fermented in a solid state using a bio-enzyme complex. The internal temperature of the fermenting herbs was detected using infrared thermometry, and the total sugar and reducing sugar content of the herbs was also measured. A linear prediction model for the total sugar and reducing sugar content of the fermenting herbs was constructed, with the internal temperature of the herbs as the x-axis and the total sugar and reducing sugar content as the y-axis. The total sugar and reducing sugar content of the fermenting herbs was then predicted based on the internal temperature and the linear prediction model. The solid-state fermentation involves drying and pulverizing the by-products of traditional Chinese medicine, passing them through a 2-5 mesh sieve, mixing them with water at a mass ratio of 10:6-15, and adding 0.4-5.5% of a bio-enzyme complex by mass for fermentation. The bioenzyme complex is composed of cellulase, xylanase, and protease in a weight ratio of 1-3:1:1; the cellulase concentration is 60,000 U / g, the xylanase concentration is 60,000 U / g, and the protease concentration is 60,000 U / g. The fermentation temperature is 40-50℃; The infrared temperature measurement involves taking out the fermentation bag of the fermented Chinese herbal medicine sample, cooling it for 10-30 minutes, and then taking a picture and measuring the temperature using an instrument connected to an infrared thermal imaging device. The fermented herbal medicine consisted of a bio-enzyme complex with 5.0% by mass added. When fermented at 50°C, the total sugar prediction model was y = 7.648x + 60.643, R0 2 The value is 0.9923; the prediction model for the reducing sugar is y = -1.8228x + 165.15, R0. 2 The value is 0.9983; the fermented herbal medicine is made by adding 0.5% by mass of a bio-enzyme complex, and when fermented at 50℃, the total sugar prediction model is y = 3.8452x + 230.11, R0 2 The value is 0.8968; the prediction model for the reducing sugar is y = -0.7078x + 87.4, R0 2 It is 0.9976.

2. The detection method according to claim 1, characterized in that, The Chinese herbal medicines include any one or more of the following: forsythia, honeysuckle, stir-fried bitter almond, isatis root, Dryopteris crassirhizoma, houttuynia cordata, rhodiola rosea, and licorice.

3. The detection method according to claim 1, characterized in that, The by-products of traditional Chinese medicine include the dregs produced during the extraction and decoction of traditional Chinese medicine preparations, and / or the non-medicinal parts of traditional Chinese medicine.

4. The detection method according to claim 1, characterized in that, The internal temperature of the fermented Chinese herbal medicine is detected every 1-2 days using infrared thermometry.

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