Processing method of pollen typhae charcoal

Abstract

The invention relates to cattail pollen, in particular to a processing method of cattail pollen charcoal. A preparation method of charred pollen typhae is characterized by comprising the following steps: taking a proper amount of pollen typhae, putting the pollen typhae into a proper frying pan or frying equipment, frying while heating, controlling the fire power to enable the frying temperature of the materials to be 190-220 DEG C and the frying time to be 6-11 minutes, taking out the charred pollen typhae when the charred pollen typhae is brown or black brown, and cooling in the air to obtain charred pollen typhae decoction pieces. Wherein the total content of the 5-hydroxymethyl-2-furoic acid and the furoic acid is used as a standard or a quality standard for judging the moderate degree of heating, and the total content of the 5-hydroxymethyl-2-furoic acid and the furoic acid is 0.35 mg / g or above. The pollen typhae charcoal obtained by the method meets the requirements, and is simple and easy to implement and controllable in quality.

Description

Technical Field

[0001] The present invention relates to pollen typhus, and in particular to a method for preparing pollen typhus charcoal. Background Art

[0002] The Chinese medicine Puhuang is the dried pollen of Typha angustifolia L., Typhaorientalis Presl. or plants of the same genus, Typhaaceae. It has the effects of stopping bleeding, removing blood stasis and relieving stranguria. It is used to treat hematemesis, epistaxis, hemoptysis, metrorrhagia, traumatic bleeding, amenorrhea, dysmenorrhea, and painful stranguria.

[0003] The processed products included in the 2020 edition of the Chinese Pharmacopoeia include raw Puhuang (crushed into lumps and sieved) and Puhuang charcoal (clean Puhuang and stir-fry until brown, shaped like Puhuang, with a brown or black brown surface. It has a burnt aroma and tastes slightly bitter and astringent). Puhuang charcoal made by stir-frying raw Puhuang has obvious anti-hemorrhagic effects.

[0004] Puhuang is a pollen-based Chinese medicine with a loose texture. It is very difficult to control the temperature when frying it to charcoal, which can easily lead to "insufficient" or "excessive" phenomena, which in turn leads to different hemostatic effects. "Ben Cao Meng Quan" points out: "In the manufacture of all medicines, it is important to be moderate. If it is insufficient, the efficacy will be difficult to obtain, and if it is excessive, the smell will be lost." It accurately points out the importance of the degree of processing to ensure the efficacy of the medicine.

[0005] The main components of Typhae Pollen are flavonoids, organic acids, alkanes, tannins, polysaccharides, amino acids, etc. Its modern pharmacological effects mainly involve the cardiovascular system, anti-atherosclerosis, improvement of sugar metabolism, regulation of immunity and anti-inflammatory, uterine excitation and anti-tumor effects. However, the current research on the intrinsic chemical components of Typhae Pollen mainly focuses on flavonoid components. The 2020 edition of the Chinese Pharmacopoeia uses isorhamnetin-3-O-neohesperidin and typhaneoglucoside as indicators for quality control of Typhae Pollen. However, the charring of Typhae Pollen is a continuous high-temperature carbonization process involving chemical reactions such as degradation, generation and transformation of multiple compounds. The content of indicator components such as isorhamnetin-3-O-neohesperidin and typhaneoglucoside is greatly reduced, and it is no longer suitable as a quality control indicator for Typhae Pollen charcoal. The control of the heat during the frying process of Typhae Pollen is an important factor in determining the quality of Typhae Pollen charcoal. However, due to the lack of effective and controllable indicators closely related to the cooking heat, the quality of Typhae Pollen charcoal actually produced is not stable.

[0006] Therefore, it is urgent to find specific, objective and accurate indicators for judging the frying temperature control and to obtain a preparation method for high-quality Puhuang charcoal that is suitable for the intrinsic chemical composition of Puhuang charcoal. Summary of the invention

[0007] Purpose of the invention:

[0008] The first invention purpose of the present invention is to provide a method for preparing Puhuang charcoal, which can obtain Puhuang charcoal that meets quality requirements and is simple and easy to operate.

[0009] The second invention purpose of the present invention is to use the sum of 5-hydroxymethyl-2-furoic acid and furoic acid content as an indicator for evaluating the degree of fire and provide a corresponding quantitative standard.

[0011] A preparation method of Puhuang charcoal, characterized in that it is achieved according to the following steps: taking an appropriate amount of Puhuang and placing it in a suitable frying pan or frying equipment, frying while heating, controlling the firepower to make the frying temperature of the material at 190-220°C, frying time 6-11min, taking out when the color of Puhuang charcoal is brown or black brown, and cooling it to obtain Puhuang charcoal slices.

[0012] The method for preparing pollen charcoal is characterized in that the sum of the contents of 5-hydroxymethyl-2-furoic acid and furoic acid is used as a standard or quality standard for judging whether the temperature is "moderate", wherein the sum of the contents of 5-hydroxymethyl-2-furoic acid and furoic acid is above 0.35 mg / g.

[0013] Beneficial Effects

[0014] 1. The present invention determines the best processing technology:

[0015] That is, the best process is determined by adjusting the temperature and time of the processing, that is, controlling the firepower to make the material frying temperature at 190-220°C, the frying time is 6-11 minutes, and taking out the Puhuang charcoal when the color is brown or black brown. The method of obtaining Puhuang charcoal meets the requirements, is simple and easy to operate, and has controllable quality.

[0016] 2. The sum of 5-hydroxymethyl-2-furoic acid and furoic acid contents is used as an indicator for evaluating the degree of fire and provides a corresponding quantitative standard.

[0017] The sum of the contents of 5-hydroxymethyl-2-furoic acid and furoic acid can be used as a temperature standard during the frying process, and can also be used as a quality standard for Puhuang charcoal after processing. The operation is simple and easy.

[0018] The prior art shows that isorhamnetin-3-O-neohesperidin and typhaneoglucoside are indicators for quality control of raw pollen, but they are not applicable to pollen charcoal. The present invention finds that two known compounds (5-hydroxymethyl-2-furoic acid and furoic acid) appear after processing. The compounds are produced after processing and are not inherent in raw pollen. The present invention finds for the first time that they can be used as indicators for controlling the temperature of frying. There are three levels of temperature in the processing of pollen charcoal: "not enough", "moderate" and "too much", and the standard is vague and difficult to quantify. The present invention finds that the overall change trend of 5-hydroxymethyl-2-furoic acid and furoic acid is consistent, and the total content of the two increases with the increase of frying time and temperature, and then shows a downward trend. Therefore, the sum of the contents of 5-hydroxymethyl-2-furoic acid and furoic acid can be used as a quantitative standard for evaluating the temperature. The details are as follows:

[0019] As shown in Table 2, samples P1-P2 correspond to "not enough": the total amount of 5-hydroxymethyl-2-furoic acid and furoic acid is 0.064-0.193 mg / g; samples P3-P4 correspond to "moderate". When fried to the state of sample P3, the average content of 5-hydroxymethyl-2-furoic acid and furoic acid increased by 74.60% and 21.50% respectively, and the total content increased by 96.10%, which may be produced by the decomposition of flavonoid glycosides.

[0020] Samples P3-P4 correspond to "moderate": with the increase of frying time and temperature, that is, when the sample P4 was fried, the average content of 5-hydroxymethyl-2-furoic acid decreased by 25.74%, furoic acid continued to increase, the average content increased by 6.98%, and the total content decreased by 18.42%, which was less than that of 5-hydroxymethyl-2-furoic acid. This shows that in addition to the decomposition of flavonoid glycosides to produce furoic acid during the frying process during this period, 5-hydroxymethyl-2-furoic acid was also degraded to produce furoic acid;

[0021] P5-P6 corresponds to "too much": as the charcoal-roasting process continued, that is, when the sample was roasted to the P5 state, 5-hydroxymethyl-2-furoic acid and furoic acid decreased significantly, and their average contents decreased by 95.44% and 50.87% respectively, and the total content decreased by 80.70%. The degradation degree of 5-hydroxymethyl-2-furoic acid was more serious, and its decrease was 1.88 times that of furoic acid and 1.18 times that of the total content, indicating that while 5-hydroxymethyl-2-furoic acid was degraded to produce furoic acid, furoic acid was also degraded and destroyed. However, due to the continuous increase in charcoal-roasting time and temperature, that is, when the sample was roasted to the P6 state, the two components were severely destroyed, and their content values ​​approached zero.

[0022] After a large number of sample experiments (100 batches), it was confirmed that the actual measured results of the total content of the two components in the "moderate" samples were between 0.372 and 0.858 mg / g, so it is stipulated that the total amount of these two components in Puhuang charcoal must not be less than 0.35 mg / g. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 , samples of pollen typhae fried at different temperatures;

[0024] Figure 2 , Representative HPLC graphs of Puhuang decoction pieces and samples prepared at different cooking temperatures;

[0025] Figure 3 , separation process of chemical components of pollen charcoal;

[0026] Figure 4 , Dynamic changes of the contents of two components in Puhuang charcoal samples with different frying temperatures. A is 5-hydroxymethyl-2-furoic acid; B is furoic acid; C is the sum of the contents of the two components (sum);

[0027] Figure 5 , bar chart of the content of two components in Puhuang charcoal samples fried at different temperatures. DETAILED DESCRIPTION

[0028] Example 1 Preparation of dynamic samples of pollen charcoal with different frying temperatures

[0029] 1. Sample

[0030] In this experiment, a total of 6 batches of commercially available Typha aurea samples were collected, and detailed information is shown in Table 1.

[0031] Table 1 Information of commercially available pollen charcoal samples

[0032]

[0033] 2. Preparation method

[0034] Preparation equipment: Demashi IH-BC-3500A induction cooker.

[0035] According to the charcoal roasting method under the general rule 0213 of the 2020 edition of the "Chinese Pharmacopoeia", samples were taken from the same batch of Puhuang decoction pieces according to the color changes of the pieces during the frying process, and the material temperature (℃) at each sampling was measured using an infrared thermometer. Six groups of Puhuang charcoal samples with different frying temperatures were taken from each batch, cooled, and numbered P1 to P6 respectively; a total of 4 batches were fried, numbered C1 to C4, and each batch included 6 groups of Puhuang charcoal samples with different frying temperatures.

[0036] The details are as follows:

[0037] Investigation of processing technology: 500 g of Puhuang medicinal material was put into a frying pan, heated (power 1000W), and stir-fried manually. The processing temperature and color change of Puhuang were detected in real time. Samples were taken at different times, and the frying time, material temperature, and properties of Puhuang charcoal were recorded.

[0038] 3. Results:

[0039] Table 2 Processing technology exploration and results

[0040]

[0041] As shown in Table 2 and Figure 1 As shown in the figure, the best processing technology of Puhuang charcoal is: take an appropriate amount of Puhuang and put it in a frying pan or frying equipment, heat it, and fry it while heating. Control the firepower to make the frying temperature of the material between 190 and 220℃, and the frying time is 6 to 11 minutes. When the color of Puhuang charcoal is brown or black brown, take it out and let it cool to obtain Puhuang charcoal slices.

[0042] Example 2 Quality Standard Investigation of the Typha Pollen Carbon Obtained in Example 1

[0043] 1. Materials

[0044] 1.1 Instrument

[0045] Thermo Ultimate 3000 liquid chromatograph (DAD detector); Bruker DRX-500MHz nuclear magnetic resonance spectrometer; Waters 2535 semi-preparative liquid chromatograph; Agilent 1260 high performance liquid chromatograph; column chromatography silica gel (80-100, 100-200 mesh); GF254 silica gel plate (100mm×50mm); C18 MB100-40 / 75 (FUJI SIL YSIACHEMICAL LTD); Sephadex LH-20; preparative reverse phase chromatography column (250mm×20mm, 10μm) (Daisogel, Japan);

[0046] 1.2 Reagents

[0047] Methanol, acetonitrile (chromatographic grade); formic acid (chromatographic grade).

[0048] 1.3 Samples

[0049] In this experiment, a total of 6 batches of commercially available Puhuang charcoal samples were collected, as shown in Table 1; 4 batches of homemade Puhuang charcoal dynamic samples with different frying temperatures were prepared, as shown in Table 2, samples P1-P6.

[0050] 2. Detection methods and results

[0051] 2.1 Chromatographic conditions

[0052] Agilent ZORBA X SB-Aq column (250×4.6mm, 5μm), mobile phase methanol (B)-0.1% formic acid water (A), gradient elution (0~30min, 1%→3%B); flow rate 1.0mL·min -1 , column temperature 30°C, detection wavelength 254nm, injection volume 10μL.

[0053] 2.2 Preparation of test solution and reference solution

[0054] 2.2.1 Preparation of test solution

[0055] Take 0.5 g of each sample powder, weigh it accurately, put it in a stoppered conical flask, add 50 mL of 70% methanol, seal it and shake it well, weigh the mass, reflux and extract for 1 hour, cool and replenish it, take the supernatant and filter it through a 0.22 μm microporous filter membrane to obtain the product.

[0056] 2.2.2 Preparation of reference solution

[0057] Accurately weigh appropriate amounts of reference substances 5-hydroxymethyl-2-furoic acid and furoic acid, and accurately add methanol to prepare mixed reference substance solutions with concentrations of 0.152 mg / mL and 0.156 mg / mL, respectively.

[0058] 2.3 HPLC analysis of Puhuang charcoal samples at different frying temperatures

[0059] Take samples of Puhuang charcoal fried at different temperatures, prepare the test solution according to "2.2", inject and analyze according to the chromatographic conditions of "2.1", and record the chromatogram.

[0060] result:

[0061] like Figure 2 As shown, peak 1 began to appear in the P1 sample, and peak 1 and peak 2 appeared in the P2 sample. These two components were not found in the raw product. It was speculated that these two components were unknown compounds newly generated after the charcoal was fried on Puhuang, so they were separated and identified.

[0062] 2.4 Confirmation of unknown compounds

[0063] 2.4.1 Extraction and separation

[0064] Accurately weigh 1 kg of "moderate" pollen charcoal (C39) sample, add 6 L of 1:1 (v / v) chloroform / methanol mixed solvent, extract at 30 °C with ultrasound for 4 hours, filter out the extract, repeat the extraction 3 times, combine the extracts, and concentrate under reduced pressure to obtain 146.1 g of total extract.

[0065] The extract was dissolved in 50°C warm water, extracted with petroleum ether and ethyl acetate three times respectively, concentrated and dried under reduced pressure to obtain 52g of petroleum ether phase extract, 16g of ethyl acetate phase extract and 76g of aqueous phase extract.

[0066] The aqueous extract was subjected to normal phase silica gel column chromatography, reverse phase column chromatography, gel chromatography and the like to obtain compound 1 and compound 2 (separation process see Figure 3 shown).

[0067] After separation, the final weight of compound 1 was 8 mg, and the weight of compound 2 was 10 mg.

[0068] 2.4.2 Structural identification

[0069] Compound 1: white powder, 1 H NMR (500MHz, CD3OD) δ: 7.13 (1H, d, J = 3.0Hz), 6.45, (1H, d, J = 3.0Hz), 4.56 (2H, s). The above data are basically consistent with those reported in the literature (TOSHIRO MATSUI, AKIHIRO KUDO, SHINJITOKUDA, KIYOSHI MATSUMOTO, AND HIROKAZU HOSOYAMA, J. Agric. Food Chem. 2010, 58, 10876-10879), so the compound was identified as 5-hydroxymethyl-2-furoic acid (alias: 5-hydroxymethyl-2-furancarboxylic acid).

[0070]

[0071] Compound 2: white powder, 1 H NMR (500 MHz, CD3OD) δ: 7.70 (1H, d, J = 1.0 Hz), 7.18 (1H, d, J = 3.0 Hz), 6.57 (1H, dd, J = 1.0, 3.0 Hz). The above data are consistent with the literature report (He Chengjun, Peng Cheng, Dai Ou, Yang Lian, Liu Juan, Guo Li, Xiong Liang, Liu Zhaohua. Chinese Herbal Medicine, 2014, 45 (21): 3048–

[0072] 3052), so the compound was identified as furoic acid (also known as 2-furancarboxylic acid).

[0073]

[0074] 2.5 Determination of the contents of two components in Puhuang charcoal samples cooked at different temperatures

[0075] 2.5.1 Methodological investigation

[0076] 2.5.1.1 Investigation of linear relationship

[0077] Accurately pipette an appropriate amount of 5-hydroxymethyl-2-furoic acid and furoic acid reference stock solutions, and dilute them into reference solutions of different mass concentrations by multiple dilution method. Perform the determination according to the chromatographic conditions under 2.1. Perform linear regression with the peak area as the ordinate (y) and the mass concentration as the abscissa (x). Simultaneously perform the determination of the detection limit (S / N=3) and the quantification limit (S / N=10), see Table 3.

[0078] Table 3 Regression equation, linear range, detection limit and quantification limit of the two components

[0079]

[0080] 2.5.1.2 Precision investigation

[0081] Inject the mixed reference solution of "2.2.2" 6 times according to the chromatographic conditions of "2.1", record the peak areas of the two components to be tested, and calculate the RSD. The RSD of the peak areas of the two components to be tested are 0.78% and 0.94% respectively, indicating that the precision of the instrument is good.

[0082] 2.5.1.3 Repeatability study

[0083] Take 6 portions of Typha charcoal (P3) powder, prepare the test solution according to "2.3.1", inject according to the chromatographic conditions of "2.1", record the peak areas of the two components to be tested, calculate the content and RSD. The RSD of the content of the two components to be tested is 0.92% and 0.70%, respectively, indicating that the method has good repeatability.

[0084] 2.5.1.4 Stability study

[0085] The test solution of Typha charcoal (P3) powder was prepared according to "2.3.1". The samples were injected according to the chromatographic conditions of "2.1" at 0, 2, 4, 8, 12, 16, 20, and 24 hours after preparation, and the peak areas of the two components to be tested were recorded and the RSD was calculated. The peak area RSDs of the two components to be tested within 24 hours were 0.62% and 1.08%, respectively, indicating that the test solution was stable within 24 hours.

[0086] 2.5.1.5 Investigation of sample recovery rate

[0087] Take 9 portions of known content of Typha charcoal (P3) powder, each portion is about 0.25g, accurately weighed, placed in a stoppered conical flask, three portions as a group, three groups of samples were accurately added with equal amounts of reference solution at 1:0.8, 1:1, 1:1.2, prepare the test solution according to "2.3.1", inject according to the chromatographic conditions of "2.1", record the peak area of ​​the two components to be tested, and calculate the recovery rate and RSD of each component to be tested. The average recovery rate of the two components to be tested was 99.93% to 103.8%, and the RSD was 0.44% to 1.9%, indicating that the accuracy of the method is good.

[0088] 2.5.2 Sample content determination

[0089] (1) Prepare the test solution of the samples prepared at different frying temperatures according to "2.2.1", inject the samples according to the chromatographic conditions in "2.1", obtain the peak area of ​​the component to be tested, and calculate the sample content (based on the dry product).

[0090] result:

[0091] As shown in Table 4 and Figure 4As shown in the figure, the overall change trend of 5-hydroxymethyl-2-furoic acid and furoic acid is consistent. The content increases with the increase of frying time and temperature, and then decreases. Among them, when the sample is fried to the P3 state, the average content of 5-hydroxymethyl-2-furoic acid and furoic acid increases by 74.60% and 21.50%, respectively, and the total content increases by 96.10%, which may be produced by the decomposition of flavonoid glycosides; with the increase of frying time and temperature, that is, when the sample is fried to the P4 state, the average content of 5-hydroxymethyl-2-furoic acid decreases by 25.74%, furoic acid continues to increase, the average content increases by 6.98%, and the total content decreases by 18.42%, which is less than 5-hydroxymethyl-2-furoic acid, indicating that in addition to the decomposition of flavonoid glycosides to produce furoic acid during the frying process during this period, 5-hydroxymethyl-2 -Furfuric acid is also degraded to generate furoic acid; as the charcoal roasting process continues, that is, when the sample is roasted to the P5 state, 5-hydroxymethyl-2-furfuric acid and furoic acid decrease significantly, and their average contents decrease by 95.44% and 50.87% respectively, and the total content decreases by 80.70%. The degradation degree of 5-hydroxymethyl-2-furfuric acid is more serious, and its decrease is 1.88 times that of furoic acid and 1.18 times that of the total content, indicating that while 5-hydroxymethyl-2-furfuric acid is degraded to generate furoic acid, furoic acid is also degraded and destroyed. However, due to the continuous increase in charcoal roasting time and temperature, that is, when the sample is roasted to the P6 state, the two components are severely destroyed, and their content values ​​approach zero.

[0092] From the above analysis, we can see that these two components can be used to judge the temperature of Puhuang charcoal, and it is more reasonable to use the sum of the contents of the two components as the index for judging the temperature of Puhuang charcoal, which avoids the influence of the conversion of 5-hydroxymethyl-2-furoic acid to furoic acid on the judgment of temperature.

[0093] Table 4 Determination results of the contents of two components in Puhuang charcoal samples with different frying temperatures (mg / g, n=4)

[0094]

[0095] (2) In order to verify the above conclusions, 100 batches of Puhuang charcoal samples with different frying temperatures were prepared according to the method described in Example 1. According to the description of Puhuang in the 2020 edition of the Chinese Pharmacopoeia, "This product is shaped like Puhuang, with a brown or dark brown surface. It has a burnt aroma and tastes slightly bitter and astringent." These 100 batches of samples included Puhuang charcoal samples with three specifications of frying temperatures: "not enough", "moderate", and "excessive". Among them, there were 14 batches of "not enough" samples, 37 batches of "moderate" samples, and 49 batches of "excessive" samples. The test solutions of the 100 batches of samples were prepared according to "2.2.1" and injected according to the chromatographic conditions of "2.1" to obtain the peak area of ​​the component to be tested, and the sample content (calculated as dry product) was calculated.

[0096] Results: As shown in Table 5 and Figure 5As shown in the results, the contents of the two components in the three specifications of Puhuang charcoal, "not enough", "moderate" and "excessive", showed significant differences (P < 0.001). The total content of the two components in 37 batches of "moderate" samples was between 0.372 and 0.858 mg / g. Therefore, it is temporarily stipulated that the total amount of these two components in Puhuang charcoal should not be less than 0.35 mg / g. All 37 batches of “moderate” samples met the requirements, while samples that were below or above the limit did not meet the requirements, indicating that this limit value can be used to judge the degree of fire of Puhuang charcoal.

[0097] Table 5 Determination results of the contents of two components in Puhuang charcoal samples at different frying temperatures (mg / g)

[0098]

[0099] (3) In order to further verify the rationality of the limit value, this experiment collected 6 batches of commercially available pollen charcoal samples, prepared the test solution according to "2.2.1", injected the sample according to the chromatographic conditions of "2.1", obtained the peak area of ​​the component to be tested, and calculated the sample content (based on the dry product). The results are shown in Table 6.

[0100] Table 6 Determination results of the contents of two components in commercially available pollen charcoal samples (mg / g, n=2)

[0101]

[0102] The results showed that the S4 sample (0.32 mg / g) among the 6 batches of commercially available Puhuang charcoal samples was below the limit value. By comparing with the properties specified under Puhuang charcoal in the 2020 edition of the "Chinese Pharmacopoeia", it was analyzed that the commercially available sample was "overcooked", which also verified the rationality of this limit value.

Claims

1. A method for preparing pollen charcoal, characterized in that: The method is achieved by the following steps: 500 g of Puhuang charcoal is placed in a suitable frying pan or frying equipment, frying while heating, controlling the firepower so that the frying temperature of the material is between 190 and 220°C, and the frying time is 6 to 11 minutes. When the color of Puhuang charcoal is brown or dark brown, take it out and let it cool to obtain Puhuang charcoal slices.

2. The method for preparing Typhae Pollen charcoal according to claim 1, characterized in that: The sum of the contents of 5-hydroxymethyl-2-furoic acid and furoic acid is used as the standard or quality standard for judging whether the cooking temperature is "moderate", and the sum of the contents of 5-hydroxymethyl-2-furoic acid and furoic acid is above 0.35 mg / g.

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