阅读说明：本技术 一种蜜炙桔梗饮片的炮制工艺及其质量标准研究 (Processing technology of honey-fried platycodon grandiflorum decoction pieces and quality standard research of honey-fried platycodon grandiflorum decoction pieces ) 是由 于秀玲 韩风雨 上官同强 邢界红 于 2019-11-28 设计创作，主要内容包括：本发明公开了一种蜜炙桔梗饮片的炮制工艺及其质量标准,重点对其炮制方法进行了研究,在此基础上对蜜炙桔梗饮片的质量控制方法采用了水分、灰分、浸出物、指纹图谱及含量测定的方法。本发明中蜜炙桔梗饮片的质量控制方法的制定填补了蜜炙桔梗饮片质量控制的空白,提高和保证饮片质量稳定,使桔梗临床用药更加合理,保证和提高临床用药疗效。(The invention discloses a processing technology of honey-fried platycodon grandiflorum decoction pieces and a quality standard thereof, which focuses on researching the processing method, and on the basis, a quality control method of the honey-fried platycodon grandiflorum decoction pieces adopts methods of measuring moisture, ash content, extract, fingerprint and content. The formulation of the quality control method of the honey-fried platycodon grandiflorum decoction pieces fills the blank of quality control of the honey-fried platycodon grandiflorum decoction pieces, improves and ensures the quality stability of the decoction pieces, ensures more reasonable clinical medication of platycodon grandiflorum, and ensures and improves the curative effect of the clinical medication.)

1. A processing technology of honey-fried platycodon grandiflorum decoction pieces and quality standard research thereof are characterized by comprising the following steps:

(1) the processing technology of honey-fried platycodon grandiflorum comprises the following steps: adding refined honey 25kg into 100kg decoction pieces, stirring, moistening for 30min, frying in 130 deg.C (bottom temperature) for 17 min;

(2) the characteristics are as follows: the product is in the shape of ellipse or irregular thick tablet, has yellowish brown surface, slightly focal spots, and honey fragrance; the section skin part is yellow white and narrower; the formed layer has obvious ring lines and is brown; the wood part is wide and has more cracks;

(3) and (3) identification: the product is cross-cut: the cells of the cork sometimes remain, and the cork without the peel has a cork layer which contains calcium oxalate prismatic crystals, the inner layer of the cork is narrow, the phloem mammary vessels are scattered, the mammary vessels are slightly thick, the inner layer contains fine granular yellowish-brown substances, the cambium forms a ring, and xylem vessels are scattered singly or a plurality of the xylem vessels are gathered together and arranged in a radial shape. Parenchymal cells contain inulin; secondly, the product is sliced, packed into slices by glycerol absorption, and observed under a microscope, and the fan-shaped or round-like synanthrin crystal can be seen; ③ taking 1g of platycodon grandiflorum powder, adding 30ml of 7% sulfuric acid ethanol-water (1: 3) mixed solution, heating and refluxing for 3 hours, cooling, shaking and extracting for 2 times by using trichloromethane, 20ml each time, combining trichloromethane liquid, adding water and washing for 2 times, 30ml each time, discarding washing liquid, dehydrating the trichloromethane liquid by using anhydrous sodium sulfate, filtering, evaporating filtrate to dryness, and adding 1ml of methanol to residues to dissolve residues to obtain a test solution; another control medicinal material 1g is prepared, and the control medicinal solution is prepared by the same method. Sucking 10ul of each of the two solutions, respectively dropping on the same silica gel H thin layer plate, developing with chloroform-ethyl ether (3: 2) as developing agent, taking out, air drying, soaking the plate in 10% sulfuric acid ethanol solution, heating at 105 deg.C until the spots are clearly developed (about 3min), and displaying the spots with the same color in the chromatogram of the test solution at the position corresponding to the chromatogram of the control solution;

(4) examination of

The water content of the platycodon grandiflorum raw drink tablets does not exceed 12.0 percent, and the platycodon grandiflorum raw drink tablets are measured by referring to a second method (drying method) of 0832 water content measurement method in accordance with the general rules of the four departments of 2015 in the pharmacopoeia of China;

the ash content of the total ash content platycodon grandiflorum raw drink tablets does not exceed 5.0%, and the determination is carried out according to a total ash determination method (a general rule 2302 of the national pharmacopoeia 2015 edition);

(5) extract of plant

Hot dipping method under alcohol soluble extract determination method (general rule 2201) with ethanol as solvent, wherein the content of ethanol should not be less than 43.0%;

(6) determination of content

Precisely weighing 4g of test sample powder, placing the test sample powder into a 50mL centrifuge tube, adding 40mL of methanol, carrying out ultrasonic extraction for 50min, centrifuging for 10min, and extracting supernatant; repeating for three times, and mixing the supernatants; heating and concentrating the supernatant to 15-20 mL, cooling, adding 50mL of diethyl ether, shaking, and removing the supernatant after the supernatant is clarified; adding methanol (20, 10 and 5mL) into the precipitate in times, heating to dissolve the precipitate, cooling, filtering, combining methanol solutions, heating and concentrating to 15-20 mL, cooling, adding diethyl ether 50mL, clarifying, removing supernatant, adding methanol (20, 10 and 5mL) into the precipitate in times, heating to dissolve the precipitate, cooling, filtering, placing the filtrate in an evaporation dish dried to constant weight, evaporating in water bath until the weight is constant, drying at 105 ℃ to constant weight, and calculating to obtain the compound preparation; the total saponin content of the product is not less than 5.4%;

② measuring by high performance liquid chromatography (general rule 0512)

Chromatographic conditions and system adaptability experiment: octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile-water (30: 70) is used as a mobile phase; detecting by an ultraviolet detector; the number of theoretical plates is not less than 3000 calculated according to platycodin D peak;

preparing reference substance solution by precisely weighing appropriate amount of platycodin D reference substance, and adding methanol to obtain solution containing 0.5mg per 1 ml;

preparing test solution by dissolving the powder (sieved with No. two sieve) 0.5g in 25ml methanol, ultrasonic extracting for 30min, shaking, and filtering;

the determination method comprises precisely sucking 20 μ L of reference solution and 20 μ L of test solution, respectively, injecting into liquid chromatograph, determining, and calculating with logarithmic equation by external standard two-point method;

the product contains platycodin D (C57H92O28) not less than 0.10% calculated on dried product;

(7) finger print

Chromatographic conditions YMC-Hydrosphere C18 column (250X 4.6mm, 5 μm); mobile phase water (a) -acetonitrile (B), gradient elution (B): 0-40 min, 15% -25%; 25-30% of 40-80 min; the flow rate is 0.8mL min < -1 >; the column temperature is 35 ℃; the wavelength is 210 nm; the sample introduction amount is 10 mu l, and automatic sample introduction is carried out;

preparing a sample, precisely weighing about 2g of powder (sieved by a second sieve), precisely adding 50% methanol 50ml, weighing, carrying out ultrasonic treatment (power is 250W and frequency is 40kHz) for 30 minutes, cooling, weighing again, complementing the loss reduction weight with 50% methanol, shaking up, filtering, precisely weighing 25ml of subsequent filtrate, drying by evaporation on a water bath, adding 20ml of water into residues, dissolving by slight heating, extracting by shaking with water-saturated n-butyl alcohol for 3 times, 20ml each time, combining n-butyl alcohol solutions, washing with 50ml of ammonia test solution, discarding ammonia solution, washing with 50ml of n-butyl alcohol-saturated water, discarding water solution, evaporating n-butyl alcohol solution, adding methanol into residues to a constant volume of 5ml, and filtering with a 0.22 mu filter membrane for later use before analysis;

(8) the application and dosage are as follows: 3-10 g;

(9) packaging and storing: selecting aluminum foil for non-vacuum packaging, and placing in a ventilated and dry place for moth proofing.

2. The processing technology and quality standard research of the honey-fried platycodon grandiflorum decoction pieces according to claim 1, wherein the content of platycodin D in the obtained honey-fried platycodon grandiflorum decoction pieces is more than 0.20%, and is superior to the market honey-fried platycodon grandiflorum decoction pieces.

Technical Field

The invention belongs to the field of traditional Chinese medicine decoction pieces, and particularly relates to a processing technology of honey-fried platycodon grandiflorum decoction pieces and quality standard research thereof.

Background

Jie Geng is recorded in Shen nong Ben Cao Jing, listed as the inferior product. Susong of the stomach: "it is present at present. Big root like little finger, yellowish white. Growing seedlings in spring, and leaving stems higher than feet. The leaves are similar to apricot leaves and have an oval shape, the four leaves are opposite and are raw, and the tea can be cooked when the tea is tender. The small flowers in summer are purple and have a color similar to morning glory and fructification after autumn. August is collected in August, and if there is no heart, it is called shepherd's purse. The compendium of materia Medica also has records: the root of this herb is firm and straight, so called Jie Geng. Platycodon grandiflorum shepherd's purse is one kind of the Chinese medicinal herbs with sweet taste and bitter taste, so the Chinese medicinal herbs are named as shepherd's purse in the root channel, are also called as sweet platycodon grandiflorum and basically conform to the platycodon grandiflorum used nowadays.

Platycodon grandiflorum is neutral in nature, bitter and pungent in flavor and enters lung meridian; has effects of dispersing lung qi, relieving sore throat, eliminating phlegm, and expelling pus. The traditional Chinese medicine composition can be clinically used for treating cough with excessive phlegm, chest distress, smooth throat and other symptoms, and belongs to the clinical common traditional Chinese medicine. The modern clinical application includes unprocessed and processed, while the processed one is mainly applied to honey products, which is considered to enhance the action of moistening lung to stop cough after being processed with honey and is mostly used for cough caused by lung yin deficiency. However, research reports on the processing technology and quality standard of honey-fried platycodon grandiflorum decoction pieces are few, and honey-fried platycodon grandiflorum decoction pieces are relevant standards in the industry. In order to solve the problems, the processing technology and the quality standard of honey-fried platycodon grandiflorum decoction pieces are urgently needed to improve and ensure the stable quality of the decoction pieces, ensure more reasonable clinical medication of platycodon grandiflorum, ensure and improve the curative effect of the clinical medication, and adapt to the technical requirements of the Chinese herbal pieces on the implementation of production approval code system.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide a processing technology of honey-fried platycodon grandiflorum decoction pieces and a quality standard research thereof, so as to improve and ensure the stable quality of the decoction pieces, ensure that the clinical medication of platycodon grandiflorum is more reasonable, and ensure and improve the curative effect of the clinical medication.

The purpose of the invention is realized by the following technical scheme:

in order to achieve the purpose, the invention adopts the following technical scheme:

1. processing technology of honey-fried platycodon grandiflorum

Adding refined honey 25kg into 100kg decoction pieces, stirring, moistening for 30min, placing into a pan, and parching at 130 deg.C (bottom temperature) for 17 min.

2. Traits

The product is in the form of oval or irregular thick tablet. The surface is yellowish brown, has a little focal spot and has honey fragrance. The section skin part is yellow white and narrower; the formed layer has obvious ring lines and is brown; the wood part is wide and has more cracks.

3. Authentication

(1) The cross section of the product is: cork cells sometimes remain, and those without the outer skin have cork layers, and the cells contain calcium oxalate spherulites. The inner layer of the plug is narrow. The phloem milk ducts are scattered, the walls of the milk ducts are slightly thick, and fine granular yellowish-brown substances are contained in the phloem milk ducts. The cambium is looped. The xylem vessels are scattered individually or several gathered together and arranged radially. The parenchyma cells contain inulin.

(2) Slicing the product, loading into slices with glycerol, and observing under microscope to obtain sector or round inulin crystals.

(3) Taking 1g of platycodon grandiflorum powder, adding 30ml of 7% sulfuric acid ethanol-water (1: 3) mixed solution, heating and refluxing for 3 hours, cooling, extracting for 20ml each time by shaking with chloroform, combining chloroform solutions, washing for 2 times with water, 30ml each time, discarding a washing solution, dehydrating the chloroform solution with anhydrous sodium sulfate, filtering, evaporating filtrate to dryness, and adding 1ml of methanol to the residue to dissolve the residue to obtain a sample solution. Another control medicinal material 1g is prepared, and the control medicinal solution is prepared by the same method. Sucking 10ul of each of the two solutions, respectively dropping on the same silica gel H thin layer plate, developing with chloroform-ethyl ether (3: 2) as developing agent, taking out, air drying, soaking the plate in 10% ethanol sulfate solution, heating at 105 deg.C until the spots are clear (about 3min), and displaying the spots with the same color in the chromatogram of the test solution at the position corresponding to the chromatogram of the control solution.

4. Examination of

The water content of the platycodon grandiflorum raw drink tablets does not exceed 12.0 percent, and the platycodon grandiflorum raw drink tablets are measured by referring to the second method (drying method) of 0832 water content measurement method in the fourth general regulation of 2015 pharmacopoeia.

The ash content of the total ash content of the platycodon root raw drinking tablet is not more than 5.0 percent, and the measurement is carried out according to a total ash content measuring method (a general rule 2302 of the national pharmacopoeia 2015).

5. Extract of plant

Ethanol is used as solvent, and the content of ethanol is not less than 43.0% by hot dipping method under alcohol-soluble extract measuring method (general rule 2201).

6. Determination of content

(1) Precisely weighing 4g of test sample powder, placing in a 50mL centrifuge tube, adding 40mL of methanol, ultrasonically extracting for 50min, centrifuging for 10min, and extracting supernatant. Repeating for three times, and mixing the supernatants. Heating and concentrating the supernatant to 15-20 mL, cooling, adding 50mL of diethyl ether, shaking, clarifying and then discarding the supernatant. Methanol (20, 10, 5mL) was added to the precipitate in portions, and the precipitate was dissolved by heating and allowed to cool. Filtering, combining methanol solutions, heating and concentrating to 15-20 mL, cooling, adding 50mL of diethyl ether, clarifying, removing supernatant, adding methanol (20, 10, 5mL) into precipitate, and heating to dissolve the precipitate. Cooling, filtering, placing the filtrate in evaporating dish, drying in water bath, drying at 105 deg.C to constant weight, and calculating.

The total saponin content of the product should not be less than 5.4%.

(2) Measuring by high performance liquid chromatography (general rule 0512).

Octadecylsilane chemically bonded silica is used as a filler in chromatographic conditions and system adaptability experiments; acetonitrile-water (30: 70) is used as a mobile phase; and detecting by an ultraviolet detector. The number of theoretical plates is not less than 3000 calculated according to platycodin D peak.

Preparation of reference solution A proper amount of platycodin D as reference is precisely weighed and added with methanol to prepare a solution containing 0.5mg per 1 ml.

Preparation of test solution about 0.5g of the powder (sieved by a second sieve) is dissolved in 25ml of methanol, ultrasonic extraction is carried out for 30min, shaking up is carried out, and filtration is carried out, thus obtaining the test solution.

The determination method comprises precisely sucking 20 μ L of reference solution and 20 μ L of test solution, respectively, injecting into liquid chromatograph, determining, and calculating with external standard two-point method logarithmic equation.

The product contains platycodin D (C57H92O28) not less than 0.10% calculated on dried product.

7. Finger print

Chromatographic conditions YMC-Hydrosphere C18 column (250X 4.6mm, 5 μm); mobile phase water (a) -acetonitrile (B), gradient elution (B): 0-40 min, 15% -25%; 25-30% of 40-80 min; the flow rate is 0.8mL min < -1 >; the column temperature is 35 ℃; the wavelength is 210 nm; the sample volume is 10 mul, and automatic sample injection is carried out.

Preparation of sample about 2g of the product powder (passing through a second sieve) is taken, precisely weighed, 50ml of 50% methanol is precisely added, the weight is weighed, ultrasonic treatment (power 250W and frequency 40kHz) is carried out for 30 minutes, cooling is carried out, the weight is weighed again, the loss weight is compensated by 50% methanol, shaking is carried out uniformly, filtration is carried out, 25ml of filtrate is precisely taken, the filtrate is placed on a water bath for evaporation, 20ml of water is added into residue for dissolving by slight heating, shaking extraction is carried out for 3 times by using water saturated n-butyl alcohol, 20ml of water is added each time, n-butyl alcohol solution is combined, 50ml of ammonia test solution is used for washing, ammonia solution is discarded, 50ml of water saturated by n-butyl alcohol is used for washing, water solution is discarded, n-butyl alcohol solution is evaporated to dryness. The sample was passed through a 0.22 μ filter before analysis.

8. The application and dosage are as follows: 3-10 g.

9. Packaging and storing: selecting aluminum foil for non-vacuum packaging, and placing in a ventilated and dry place for moth proofing.

The invention has the following advantages:

the invention standardizes the processing technology of the honey-fried platycodon grandiflorum, and the obtained honey-fried decoction pieces have low water content, luster, bright color, crisp and uniform heating and do not have scorching and burning points. Through research and formulation of the quality standard of honey-fried platycodon grandiflorum decoction pieces, the blank of the quality standard of honey-fried platycodon grandiflorum decoction pieces is filled, the quality stability of the decoction pieces is improved and guaranteed, the clinical medication of platycodon grandiflorum is more reasonable, and the curative effect of the clinical medication is guaranteed and improved.

Drawings

Fig. 1 is a number 1-7 honey-fried platycodon grandiflorum decoction piece thin-layer chromatogram, wherein the number from left to right is 0 reference medicinal materials, 1 Anhui 20180111, 2 Henan 20170720, 3 Jilin Changchun 20170910, 4 Shandong Zibo 20180220, 5 Gansu Chengxian county 20170930, 6 Jiangxi 20180410 and 7 Hebei 20170930 respectively;

fig. 2 is a number of thin-layer chromatograms of honey-fried platycodon grandiflorum decoction pieces numbered 8-17, wherein the numbers from left to right are 0 reference medicinal materials, 8 Chifeng 20170810, 9 Shaanxi 20180110, 10 Sichuan 20170504, 11 Anhui 20180310, 12 Henan 20170808, 13 Jilin Changchun 20180101, 14 Sichuan 20180404, 15 Gansu Chengxian county 20170120, 16 Anhui milli states 20170101, and 17 Hunan market 20180205;

FIG. 3 is a thin-layer chromatogram of honey-fried radix Platycodi decoction pieces numbered 18-27, wherein the number from left to right is 0 control medicinal material, 18 Anhui 2017090619 Hebei 2018011020 Anhui 2017051021 Anhui 2017081022 Anhui 2017102023 Hebei Anhui 2018010924 Hebei 2018040125 Hebei Anhui 2017093026 Hebei Anhui 2017031027 Anhui 20171201

FIG. 4 is a thin-layer chromatogram of honey-fried radix Platycodi decoction pieces numbered 28-37, wherein the number from left to right is 0 of control medicinal material 28 Anhui 2018021029 Hebei 2017120930 Hebei 2017111231 Hebei 2018022032 Shandong 2017102833 Shanxi 2017101134 Shanxi 2018011035 Chifeng 2017111836 Chifeng 2017110137 Chifeng 20171012

FIG. 5 is a thin-layer chromatogram of 38-41 honey-fried radix Platycodi decoction pieces, which is numbered from left to right respectively as 0 control medicinal material 38 Nemeng 2017110139 Shaanxi 2017032040 Sichuan 2017091041 Hebei 20171226

Detailed Description

The following detailed description of the embodiments of the present invention is provided for the purpose of illustration and not limitation, and should not be construed as limiting the scope of the invention.

Comparative example 1 study on processing technique of honey-fried platycodon grandiflorum decoction pieces

1. Design of experiments

The experiment selects honey adding amount (A), frying temperature (B) and frying time (C) of every 100kg of decoction pieces as expedition factors, takes alcohol extract content, total saponin and platycodin D as the optimal frying process expedition indexes of honey-fried platycodon grandiflorum decoction pieces, respectively selects 3 levels through preliminary experiments, adopts an L9(34) table, arranges experimental preparation samples according to an orthogonal table, takes 100g of each group of platycodon grandiflorum raw decoction pieces, uniformly mixes the decoction pieces and quantitative refined honey (the honey-water ratio is 1: 2.5) according to the experimental design, puts the decoction pieces into a frying container after frying and frying are finished after being moistened for 30min, takes out and cools, and puts into a closed container for later use.

TABLE 1 orthogonal design of processing technology research of honey-fried platycodon grandiflorum decoction pieces

2. Evaluation of honey-fried platycodon grandiflorum decoction piece process

The optimal processing technology of honey-fried platycodon grandiflorum decoction pieces is inspected by adopting a weighted comprehensive evaluation method, wherein the weighting coefficient of platycodin D is 0.5, the weighting coefficient of alcohol-soluble extract is 0.25, and the weighting coefficient of total saponin is 0.25. The total mass score is 50 (actual amount of platycodin D/highest amount of platycodin D) +25 (actual amount of alcohol-soluble extract/highest amount of alcohol-soluble extract) +25 (actual amount of total saponins/highest amount of total saponins).

3. Results of orthogonal design experiments

Selecting SPSS20.0 software to process data, and performing comprehensive evaluation on the result by using a comprehensive weighting and scoring method, wherein the content weight coefficients of platycodin D, alcohol extract and total saponin are 0.5, 0.25 and 0.25 respectively. The experimental results are shown in Table 2, and the optimal processing technology is obtained by analyzing the honey adding amount of 25kg per 100kg decoction pieces and frying at 160 ℃ for 17 min.

TABLE 2 experiment results of processing technique optimization of honey-fried platycodon grandiflorum decoction pieces

4. Best process verification test

The optimal process for honey-fried platycodon grandiflorum decoction pieces is determined as follows: adding refined honey 25kg (Mel-water ratio of 1: 2.5) into 100kg decoction pieces, mixing, moistening for 30min, parching at 160 deg.C for 17min, taking out, cooling, and placing in a sealed container.

The optimum technological parameters of the honey-fried platycodon grandiflorum decoction pieces are subjected to 3 times of parallel tests, the content results and the comprehensive scores of platycodon grandiflorum saponin D, alcohol-soluble extract and total saponin are respectively measured, and the RSD value is 0.96%, so that the process is proved to be feasible.

Example 1 thin layer identification

1. Preparation of test solution

Pulverizing all radix Platycodi decoction pieces, sieving with No. 2 sieve, collecting radix Platycodi powder 1g, adding 70% ethanol sulfate-water (1: 3) mixture 30ml, heating and refluxing for 3 hr, cooling, shaking with chloroform for 2 times (20 ml each time), mixing chloroform solutions, washing with water for 2 times (30 ml each time), discarding washing solution, dehydrating chloroform solution with anhydrous sodium sulfate, filtering, evaporating filtrate to dryness, and adding methanol 1ml to dissolve residue.

2. Preparation of control solutions

Collecting radix Platycodi reference material 1g, and making reference material solution with the same method as "1".

3. Conditions of thin layer chromatography

And (2) performing thin-layer chromatography test, sucking 10 μ l of each of the two solutions, respectively dropping on the same H silica gel thin-layer plate, dropping on a strip with a sample width of 6mm and an origin point 10mm away from the bottom edge, developing with chloroform-ethyl ether (3: 2) as developing agent, taking out, air drying, spraying with 10% sulfuric acid ethanol solution, and heating at 105 deg.C until the spots are clearly developed. Spots of the same color appear on the chromatogram of the test solution at the positions corresponding to those on the chromatogram of the control solution.

4. Results of the experiment

Thin layer chromatograms of honey-fried radix Platycodi decoction pieces in each batch are shown in figure 1, figure 2, figure 3, figure 4, and figure 5.

5. Conclusion

The experiment shows that the honey platycodon grandiflorum decoction pieces are prepared into 41 batches, wherein 8 batches of Anhui, 1 batch of Anhui Bozhou, 4 batches of Chifeng, 2 batches of Gansu Chengxian county, 6 batches of Hebei, 3 batches of Hebei Anguo, 2 batches of Henan, 1 batch of Hunan market, 2 batches of Jilin Changchun, 1 batch of Jiangxi, 1 batch of inner Mongolia, 1 batch of Shandong catabo, 2 batches of Shandong, 4 batches of Shaanxi and 3 batches of Sichuan. According to the related thin-layer chromatograms, most spots of the thin-layer chromatograms of the platycodon grandiflorum in all areas of Anhui and Hebei are clear and complete, and the individual areas are very fuzzy, which shows that the platycodon grandiflorum in the area has good quality on the whole, but poor quality stability; the definition difference of thin-layer chromatograms in 4 areas of inner Mongolia, Gansu, Jilin and Shaanxi is great, which shows that the platycodon grandiflorum in the area has good quality and poor quality, and the quality stability is low; the spots of the thin-layer chromatogram map in the Sichuan area are clear and the definition is basically consistent, which shows that the platycodon grandiflorum in the Sichuan area has superior quality and high quality stability; the thin-layer chromatograms in 2 areas of Henan and Shandong have common performance but consistent spot definition, which shows that the platycodon grandiflorum in the 2 areas has common quality but high quality stability; the definition of the thin-layer chromatograms in 2 areas of Hunan and Jiangxi is poor, which indicates that the 2 areas have poor platycodon grandiflorum quality.

Example 2 determination of Total Saponin content

Precisely weighing 4g of test sample powder, placing in a 50mL centrifuge tube, adding 40mL of methanol, ultrasonically extracting for 50min, centrifuging at the rotation speed of 5000r min-1 at room temperature for 10min, and extracting supernatant. Repeating for three times, and mixing the supernatants. And (3) putting the combined supernatant into an electric heating constant-temperature water bath pot, heating and concentrating to 15-20 mL, cooling, adding 50mL of diethyl ether, shaking, and removing the supernatant after the mixture is clarified. Then, methanol (20, 10, 5mL) was added to the precipitate in portions, and the precipitate was dissolved by heating and allowed to cool. Filtering, combining methanol solutions, heating and concentrating to 15-20 mL, cooling, adding 50mL of diethyl ether, shaking the bottle body, removing supernatant after the bottle body is clarified, precipitating, adding methanol (20, 10 and 5mL) in times, and heating to dissolve the precipitate. After cooling, the mixture is filtered, and the filtrate is placed in an evaporating dish which is dried to constant weight. The evaporating dish was evaporated to dryness in an electric constant temperature water bath and dried to constant mass in an electric hot air drying oven at 105 ℃. The total platycodin mass is the difference between the mass of the platycodon grandiflorum before and after evaporation. The content range of the total saponins of the honey-fried platycodon grandiflorum decoction pieces is measured to be 1.66-17.19%.

Analyzing the total saponin content of 41 batches of honey-fried platycodon grandiflorum decoction pieces by using SPSS20.0 software, and performing statistical analysis on the total saponin content of 41 batches of honey-fried platycodon grandiflorum decoction pieces, wherein the average value is 7.20%.

TABLE 3 Total saponins content of honey-fried radix Platycodi decoction pieces

Example 3 determination of platycodin D content

1. Preparation of control solutions

Accurately weighing appropriate amount of platycodin D reference substance, placing in a volumetric flask, and adding methanol to obtain 0.5mg/mL solution. 2. Preparation of test solution

Precisely weighing 0.5g of test sample, placing in a 25mL volumetric flask, adding methanol to the near-scale line, and ultrasonically extracting for 30 min. After cooling, the volume is determined by methanol, shaking up, filtering the subsequent filtrate by a 0.22 μm microporous filter membrane, and storing the filtrate in a sample bottle for later use.

3. Chromatographic conditions

A chromatographic column: waters C18 column (4.6 mm. times.250 mm, 5 μm); mobile phase: acetonitrile-sodium dihydrogen phosphate (pH 3.0, 24: 76 with phosphoric acid); the column temperature is 35 ℃; the flow rate is 0.7 mL/min; the detection wavelength is 210 nm; the amount of the sample was 20. mu.L.

4. Preparation of the Standard Curve

Precisely sucking 2, 4, 8, 12, 16 and 20 mu L of platycodin D reference substance solution, carrying out sample injection measurement according to the chromatographic condition under the item 3, recording the peak area, and carrying out linear regression by taking the peak area x as the ordinate and the reference substance content y as the abscissa. The regression equation of the obtained platycodin D reference substance is that y is 0.0169x-0.0078, which shows that the linear relation of the content of the platycodin D is good within the range of 5.72-0.143 mg. The saponin D regression equation, correlation coefficient and linear range are shown in Table 4.

TABLE 4 results of analysis by linear regression investigation

5. Methodology investigation

5.1 precision test

Injecting 20 μ L of the control sample under item "1", continuously injecting 6 times under chromatographic condition of item 6.3, calculating to obtain RSD value of saponin D of 0.49%, which indicates that the instrument has good precision under the chromatographic condition.

5.2 repeatability test

6 parts of the same platycodon grandiflorum decoction pieces are weighed, sample solutions to be tested are prepared under 6.2 items respectively, sample injection is carried out under the chromatographic condition under the item 3, the average content is 0.1069% and the RSD value is 1.88%, and the experiment has good repeatability under the condition.

5.3 stability test

Sampling 20 mu L of the same platycodon grandiflorum decoction piece test solution for 0 h, 6 h, 12 h, 18 h, 24 h and 48h respectively under the chromatographic condition of item 3, calculating the peak area of platycodin D to obtain the RSD value of 0.83%, which indicates that the platycodon grandiflorum raw decoction piece processed product test solution has better stability for 48h under the condition.

5.4 sample application recovery test

Precisely weighing 1g each of 6 parts of a sample (platycodin D1.164 mg. g-1) with known platycodin D content, preparing a sample solution according to the method under the item 2, adding a proper amount of a saponin D reference substance, measuring the content according to the chromatographic condition under the item 3, and calculating to obtain the average sample addition recovery rate of the platycodin D of 96.17 percent and the RSD value of 1.64 percent.

5.5 assay

Preparing 41 batches of radix Platycodi decoction pieces into test solution, measuring according to chromatographic conditions under specified conditions, measuring peak area, and calculating corresponding content by substituting into corresponding standard curve. The content range of the saponin D of the honey-fried platycodon grandiflorum decoction pieces is measured to be 0.07-0.21%.

Analyzing the content of platycodin D in the rest 40 batches of the secondary platycodon grandiflorum decoction pieces by using SPSS20.0 software, and determining that the group of data has no abnormal value, and the average value is 0.14%.

Platycodin D content in honey-fried platycodon grandiflorum decoction pieces of table 541 batches

Example 4 product quality verification

The three batches of raw medicinal materials planted in a self-owned base are processed by adopting a preferred process, various index components are detected, and the comparison of market product results shows that the honey-fried decoction pieces obtained by the processing process have high quality and stable content.