CN110538235A - Bitter orange baked with bran while being fresh in producing area and processing method and application thereof - Google Patents

Abstract

The invention provides a processing method of bitter orange baked with bran when the bitter orange is fresh in a producing area, which relates to the technical field of traditional Chinese medicine processing and comprises the following steps: collecting immature fruits of sour oranges in a production place, removing pulp, cutting peel into thin threads with the thickness of 0.2-0.3 cm, uniformly mixing the thin threads with bran coat while the fruits are fresh, drying the mixture at 160 ℃ for 2 hours, taking out the mixture, and screening off the bran coat to obtain the dried bitter oranges with the fresh bran in the production place; the dosage of the bran coat is 10kg per 100kg of immature fruit of Citrus aurantium L. The production area fresh cutting technology is adopted, repeated drying and water treatment processes are reduced, and manpower is saved; the utilization rate of rice secondary product bran coat is effectively improved; compared with a bran frying process, the bran drying technology can realize controllable heat transfer efficiency in the processing process of the fructus aurantii, and the quality of the finished product of the decoction pieces is more uniform. The effective components such as naringin, hesperidin, neohesperidin, synephrine, D-limonene, gamma-terpinene and the like in the bitter orange baked in the production place while the bitter orange is fresh bran have higher content, and the effect of relieving epigastric distention is better.

Description

Bitter orange baked with bran while being fresh in producing area and processing method and application thereof

Technical Field

The invention belongs to the technical field of traditional Chinese medicine processing, and particularly relates to bitter orange baked with bran at the producing area and a processing method and application thereof.

Background

Fructus Aurantii is dried immature fruit of Citrus aurantium L.of Rutaceae and its cultivar, has effects of regulating qi-flowing, relieving epigastric distention, activating stagnancy and relieving flatulence, and can be used for treating chest and hypochondrium stagnation of qi, fullness and pain, dyspepsia, etc. 2015, Chinese pharmacopoeia, 2008, Jiangxi province, and bran-fried fructus Aurantii. The traditional Chinese medicine theory considers that the bitter orange has stronger dryness and is easy to damage body fluid, and the bitter orange can play a role in reducing dryness and enhancing efficiency after being processed by bran and chaff, thereby being more suitable for clinical application. In the processing process of the traditional fructus aurantii decoction pieces, on one hand, the effective ingredients in the traditional fructus aurantii decoction pieces are lost due to repeated soaking and drying of the medicinal materials in the processes of producing area processing and softening and cutting the medicinal materials; on the other hand, when the batch size of the large-scale production is large, the traditional stir-frying technology is easy to cause uneven heating of the decoction pieces, and the stir-frying machine is more difficult to accurately control the temperature than the oven, so that the quality of the stir-frying decoction pieces of the bitter orange among different batches and among the same batch is difficult to be uniform and controllable.

Disclosure of Invention

The invention aims to provide a processing method of bitter orange baked with bran in a producing area, which improves the contents of Q-Marker (naringin, neohesperidin, hesperidin and synephrine) and main effective ingredients in alcohol extract, volatile oil and aromatic water in the processed bitter orange, and improves the treatment effect of the bitter orange on relieving epigastric distention.

In order to realize the purpose, the invention provides a processing method of bran-baked bitter oranges, which comprises the following steps:

After immature fruits of sour oranges are harvested in a production place, removing pulp, cutting fresh peel into thin threads with the thickness of 0.2-0.3 cm, uniformly mixing the thin threads with bran coat, and drying for 2 hours at 160 ℃; the dosage of the bran coat is 10kg per 100kg of immature fruit of Citrus aurantium L.

And after the baking is finished, screening off the bran coat, and then cooling.

The invention also provides the bran-baked bitter orange prepared by the processing method.

The invention also provides application of the bran-baked fructus aurantii in preparing the bran-baked fructus aurantii decoction pieces for improving the alcohol-soluble effective component content of the fructus aurantii.

Preferably, the alcohol-soluble effective components of the fructus aurantii comprise naringin, neohesperidin, hesperidin, synephrine, D-limonene, gamma-terpinene and the like.

The invention also provides application of the bran-baked fructus aurantii in preparation of bran-baked fructus aurantii decoction pieces for improving the volatile component content of fructus aurantii.

Preferably, the volatile components include D-limonene and gamma-terpinene.

The invention provides a processing method of bran-dried fructus aurantii in a production place while the fructus aurantii is fresh, which utilizes HPLC, UHPLC-Q-TOF/MS and GC-MS component analysis and detection technologies to detect components of 4 processed varieties of fructus aurantii, including raw fructus aurantii, bran-fried fructus aurantii, bran-dried fructus aurantii and bran-fried fructus aurantii, and analyzes the influence of different processing methods on the main effective components, alcohol extract, volatile oil and the types and contents of compounds in aromatic water by combining mathematical statistics methods such as PCA and the like. After comprehensive weighted scoring is carried out by taking 4 components of naringin, neohesperidin, hesperidin and synephrine as indexes, the comprehensive scoring of the bran-dried fructus aurantii in the processed product of the fructus aurantii is the highest, and the processed product is the initially screened optimal decoction piece variety; by identifying the chemical component types in different decoction pieces of fructus aurantii and analyzing the difference of 14 chemical markers with physiological activity among different processed products, the method finds that the bran-dried fructus aurantii has 13 component contents increased and only 1 component content decreased, the component substance basis is superior to other processed products, and the comprehensive weighting scoring is carried out by taking the 14 chemical markers as indexes, so that the bran-dried fructus aurantii is also the best processed product, and the efficacy of the bran-dried fructus aurantii is superior to that of the other processed products; by detecting the types of the volatile components of the fructus aurantii, the D-limonene and the gamma-terpinene are 2 components with the highest content in the volatile oil and the aromatic water, and are the main drug effect substances in the volatile components of the fructus aurantii. The relative contents of D-limonene and gamma-terpinene in different processed products are compared, and the highest content of the components in the bran-baked fructus aurantii is found. The influence of different processed products of fructus aurantii on the gastric residue and intestinal propulsion of a rat with poor Functional Digestion (FD) is compared to discover that the optimal treatment effect of the bran-dried fructus aurantii on the FD rat is achieved when the production place is fresh, and the process for drying the fructus aurantii when the production place is fresh proves that the content of the effective ingredients of the medicine can be improved and the curative effect can be enhanced.

Drawings

FIG. 1 is a HPLC result chart for measuring the contents of naringin, neohesperidin and hesperidin in fructus Aurantii, wherein the letter A is a reference substance and the letter B is a test substance; the number 1 is naringin, 2 is hesperidin, and 3 is neohesperidin;

FIG. 2 is a chart of HPLC results of synephrine content measurement in fructus Aurantii, wherein the letter A is a reference substance and the letter B is a test substance; the number 1 is synephrine.

FIG. 3 is a UPLC-Q-TOF/MS total ion flow diagram of fresh bran-dried fructus Aurantii in positive ion mode in a producing area;

FIG. 4 is a total ion flow diagram of volatile oil of fructus Aurantii by baking with bran in a fresh place;

FIG. 5 is a total ion flow diagram of aromatic water from roasted bitter orange in producing area while fresh bran is being used.

Detailed Description

The invention provides a processing method of bran-dried bitter orange in a producing area, which comprises the following steps: after immature fruits of sour oranges are harvested in a production place, removing pulp, cutting fresh peel into thin threads with the thickness of 0.2-0.3 cm, uniformly mixing the thin threads with bran coat, and drying for 2 hours at 160 ℃; the dosage of the bran coat is 10kg per 100kg of immature fruit of Citrus aurantium L.

The lime is collected from Jiangxi Xingan farmers and is identified as a fresh immature fruit of Citrus aurantium L.

In the processing method of the invention, after the baking is finished, the method preferably further comprises screening off the bran coat and then cooling.

The invention also provides the bran-baked bitter orange prepared by the processing method.

The invention also provides application of the fresh bran-dried fructus aurantii in producing the bran-dried fructus aurantii decoction pieces for improving the alcohol-soluble effective component content of the fructus aurantii.

The alcohol-soluble effective components of the fructus aurantii preferably comprise naringin, neohesperidin, hesperidin and synephrine.

The invention also provides application of the bran-dried fructus aurantii in preparing the bran-dried fructus aurantii decoction pieces for improving the volatile component content of the fructus aurantii in the fresh producing area.

The volatile components of the present invention preferably include D-limonene and gamma terpinene.

The present invention provides a method for processing and application of bran-baked fructus aurantii with fresh producing area, which is described in detail with reference to the following examples, but they should not be construed as limiting the scope of the present invention.

Example 1

Cutting fresh fructus Citri Junoris collected from orchard of Xingan farmer in Jiangxi into two halves, removing fruit pulp, cutting into 0.3cm filament, adding appropriate amount of testa oryzae, mixing, oven drying at 160 deg.C for 2 hr, taking out, sieving to remove testa oryzae, and cooling. 10kg of bran is added per 100kg of lime.

Example 2

Cutting fresh fructus Citri Junoris collected from orchard of Xingan farmer in Jiangxi into two halves, removing pulp, cutting into 0.2cm thin pieces, adding appropriate amount of testa oryzae, mixing, oven drying at 160 deg.C for 2 hr, taking out, sieving to remove testa oryzae, and cooling. 10kg of bran is added per 100kg of lime.

Experiment 1: the Q-Marker content of the fructus aurantii is tested on the fresh bran-dried fructus aurantii in the producing areas prepared in the examples 1 and 2

1 laboratory apparatus

UltiMate 3000 liquid chromatograph (Dionex, usa, containing a PDA-3000 diode array uv detector, Chromeleon workstation); raytek Raynger ST20 infrared thermometer (Raytai, USA), electric heat blast drying cabinet (Shanghai Bingfeng industry Co., Ltd.), KQ-500E type ultrasonic cleaner (Kunshan ultrasonic instruments Co., Ltd.), one hundred thousand electronic analytical balance (Mettler-Tooliduo instruments Shanghai Co., Ltd.), Chinese herbal medicine crusher (Tianjin Tester instruments Co., Ltd.), SHZ-D (III) circulating water type vacuum pump (Steve City Waals instruments Co., Ltd.).

2 reagent

Naringin, hesperidin, neohesperidin control (wakki biotechnology limited, Sichuan, lots wkq16051305, wkq15123105, wkq16041804, respectively), synephrine control (national center for engineering research on the manufacture of solid Chinese medicinal preparations, lot BCTG-0709), methanol, acetonitrile [ chromatographically pure, Germany Merck biotechnology (China) limited ], water was Waha-Ha purified water, and other reagents were analytically pure.

3 preparation of control drug

Raw fructus Aurantii: is prepared according to the processing method under the item of 2015 edition of 'Chinese pharmacopoeia' bitter orange. Removing impurities, cleaning, moistening, slicing, drying, and sieving to remove pulp and core.

Stir-frying fructus aurantii with bran: is prepared according to the processing method under the item of 2015 edition of Chinese pharmacopoeia of bran stir-frying bitter orange. Heating a pot, uniformly scattering a certain amount of wheat bran, heating with medium fire, putting the trifoliate orange shell slices into the pot after the wheat bran is smoked, continuously stirring, taking out the trifoliate orange shell slices when the trifoliate orange shell slices are fried to be light yellow, sieving to remove the wheat bran, and cooling. 10kg of wheat bran is used per 100kg of Zhi Hu tablets.

Stir-frying fructus aurantii with bran: the traditional Chinese medicine is prepared according to a processing method under the item of ' bran-fried bitter orange ' in 2008 edition ' processing standard of traditional Chinese medicine decoction pieces in Jiangxi province. Heating a pot, uniformly scattering a certain amount of bran coat, heating with medium fire, putting the hovenia dulcis thunb slices into the pot after the cigarettes are smoked, continuously stirring, taking out the hovenia dulcis thunb slices when the hovenia dulcis thunb slices are fried to be light yellow, sieving to remove the bran coat, and cooling. 10kg of bran coat is used per 100kg of trifoliate orange shell tablets.

4 preparation of test solutions

Precisely weighing 0.1g each of crude fructus Aurantii, bran-parched fructus Aurantii, and bran-parched fructus Aurantii decoction pieces powder (sieved with No. 3 sieve), placing into a conical flask with a plug, adding 50mL each of methanol, weighing, heating and refluxing for 1.5h, cooling to room temperature, weighing again, adding methanol to balance the weight, shaking, and filtering. Precisely measuring 10mL of the subsequent filtrate, respectively placing in 25mL volumetric flasks, adding methanol to scale, shaking, and filtering with 0.45 μm microporous membrane.

5 methods and results

5.1 content determination of naringin, hesperidin and neohesperidin

5.1.1 chromatographic conditions

Inertsil/WndaSil C18 chromatographic column (250mm × 4.6mm, 5 μm), mobile phase acetonitrile-water (20: 80, pH adjusted by phosphoric acid to 3), flow rate of 1.0mL min-1, column temperature of 30 deg.C, detection wavelength of 283nm, sample injection amount of 10 μ L, and as shown in FIG. 1, naringin, hesperidin and neohesperidin have good separation degree and no interference from other impurities.

5.1.2 preparation of control solutions

Precisely weighing appropriate amount of naringin, hesperidin and neohesperidin, dissolving in methanol, diluting to scale, shaking, and making into reference solution with mass concentration of 2.00, 0.20, 2.00 g.L-1 as stock solution. Precisely measuring 2mL of the reference stock solution in sequence, placing in the same 50mL measuring flask, adding methanol to constant volume to scale, and obtaining mixed reference solution.

5.1.3 Linear relationship investigation

Precisely sucking 1, 5, 10, 15 and 20L of the control solution, measuring according to the chromatographic conditions, and regressing the sample amount by peak area to obtain regression equations of naringin, hesperidin and neohesperidin, wherein the regression equations are Y-23.912X +1.0440 (r-0.9992), Y-25.317X-0.0635 (r-0.9993), Y-26.511X +0.3054 (r-0.9995), and the linear ranges are 0.080-1.600, 0.008-0.160 and 0.080-1.600 mug in sequence.

5.1.4 precision test

And respectively taking 20 mu L of each reference substance solution of naringin, hesperidin and neohesperidin, continuously sampling for 5 times according to the chromatographic conditions, and calculating RSD of the peak areas of the naringin, the hesperidin and the neohesperidin to be 0.5%, 0.4% and 0.6%, respectively, thereby indicating that the precision of the instrument is good.

5.1.5 stability test

The same crude bitter orange decoction pieces are taken, the sample solution prepared in the way is respectively measured for 0 hour, 2 hours, 4 hours, 8 hours and 12 hours according to the chromatographic conditions, and the results show that the RSD of the peak areas of the naringin, the hesperidin and the neohesperidin are respectively 1.2%, 1.5% and 1.9%, which indicates that the sample solution has better stability in 12 hours.

5.1.6 repeatability test

Accurately weighing 6 parts of raw bitter orange powder, each part being 0.5g, measuring the sample solution prepared according to the method according to the chromatographic conditions, and calculating RSD of the peak areas of naringin, hesperidin and neohesperidin to be 1.7%, 1.8% and 2.3% respectively.

5.1.7 sample recovery test

Precisely weighing about 0.1g of fructus Aurantii sample powder (sieved by a third sieve) with known content, and 6 parts in total, precisely adding equivalent diluted solutions of naringin, hesperidin and neohesperidin stock solutions, respectively, preparing a sample solution according to the method, measuring according to the chromatographic conditions, and calculating the recovery rate, wherein the results are shown in Table 1.

TABLE 1 production area naringin, hesperidin and neohesperidin content in freshly bran-baked bitter orange

5.1.8 sample determination

Respectively taking raw fructus aurantii, bran-fried fructus aurantii and fresh bran-dried fructus aurantii decoction piece powder (sieved by a No. three sieve) of a production place, precisely weighing, precisely sucking a reference substance solution and a test substance solution according to the preparation method of the test substance solution and the chromatographic conditions, respectively injecting the reference substance solution and the test substance solution into a high performance liquid chromatograph, measuring peak areas, and calculating the contents of naringin, hesperidin and neohesperidin in each sample according to a regression equation, wherein the results are shown in a table 2.

TABLE 2 naringin, hesperidin and neohesperidin content in different processed products of bitter orange (n ═ 3)

5.2 content determination of synephrine

5.2.1 chromatographic conditions

Inertsil/Wondasil C18 column (250 mm. times.4.6 mm, 5 μm), mobile phase acetonitrile: water (0.1% sodium lauryl sulfate and 0.1% phosphoric acid) ═ 32: 68, the flow rate is 1.0 mL/min < -1 >, the column temperature is 30 ℃, the detection wavelength is 224nm, the sample injection amount is 10 mu L, and the chromatogram is shown in figure 2.

5.2.2 preparation of control solutions

Precisely weighing 1.14mg of synephrine reference substance, placing the synephrine reference substance in a10 mL volumetric flask, adding 50% ethanol, ultrasonically dissolving, fixing the volume to a scale, and shaking up to prepare a reference substance stock solution with the mass concentration of 0.114 g.L < -1 >. Taking 0.1, 0.5, 1.0, 1.5 and 2mL of synephrine reference substance stock solution, respectively placing the synephrine reference substance stock solution in a10 mL volumetric flask, adding 50% ethanol for dilution and constant volume to scale, shaking up, filtering with a 0.45-micrometer microporous filter membrane, and taking subsequent filtrate to obtain the synephrine reference substance solution with the concentrations of 0.00114, 0.0057, 0.0114, 0.0171 and 0.0228 mg/mL < -1 >.

5.2.3 Linear relationship investigation

Precisely sucking different concentrations of synephrine reference substance solution by 10 mu L respectively, injecting into a liquid chromatograph, measuring according to chromatographic conditions under 1.2.4.1, and regressing the sample amount by peak area to obtain the reference substance synephrine with a regression equation of y being 816.22x +1.3571(r being 0.9993), which shows that the linear relation of the synephrine in the range of 0.0114-0.228 mu g is good.

5.2.4 precision test

And taking 10 mu L of synephrine reference substance solution, carrying out continuous sample injection for 5 times according to the chromatographic condition, and calculating the RSD of the peak area to be 0.4 percent, which indicates that the precision of the instrument is good.

5.2.5 stability test

Taking the same bran-baked fructus aurantii decoction pieces, preparing a test solution according to the method, and respectively measuring the test solution for 0 h, 2h, 4 h, 8h and 12h according to the chromatographic conditions, wherein the result shows that the RSD of the synephrine peak area is respectively 2.6%, which indicates that the test solution has better stability in 12 h.

5.2.6 repeatability test

Accurately weighing 6 parts of bran-dried bitter orange powder, wherein each part is 0.5g, preparing a test solution according to the method, and calculating the RSD of the synephrine peak area to be 2.1% according to the chromatographic condition.

5.2.7 sample recovery test

Precisely weighing about 0.1g of bran-baked fructus aurantii sample with known content, respectively precisely adding equivalent synephrine reference substance solution, preparing test sample solution according to the method, measuring according to the chromatographic conditions, and calculating recovery rate, wherein the result is shown in Table 3.

TABLE 3 determination of synephrine content in fresh bran-baked bitter orange in producing area

5.2.8 sample determination

Respectively taking 0.1g of each raw fructus aurantii, bran-fried fructus aurantii, place-of-origin fresh bran-dried fructus aurantii and bran-fried fructus aurantii powder (sieved by a No. three sieve), precisely weighing, precisely sucking a reference substance solution and a test substance solution according to the preparation method of the test substance solution and the chromatographic conditions, respectively injecting the reference substance solution and the test substance solution into a high performance liquid chromatograph, measuring peak areas, and calculating the content of synephrine in each sample according to a regression equation, wherein the results are shown in Table 4.

TABLE 4 content of synephrine in different processed products of fructus Aurantii (n ═ 3)

5.3 comparison of the contents of the main effective components in different processed products

By adopting a multi-index comprehensive weighting scoring method, the contents of naringin, neohesperidin, hesperidin, synephrine and other components in the bitter orange decoction piece variety are comprehensively scored. Naringin and neohesperidin are used as the index of the control of the fructus aurantii decoction pieces in Chinese pharmacopoeia, the weight coefficients are set to be 0.3, the hesperidin and synephrine are used as the main active ingredients of the fructus aurantii, the weight coefficient is set to be 0.2, the higher the content of the above ingredients is, the better the quality of the processed variety is prompted, so that the sample with the highest content is taken as 100 minutes, if the naringin content in the bran-baked fructus aurantii is the highest and 100 minutes, the naringin content score in the raw fructus aurantii is 8.45/9.23 multiplied by 100 which is 91.55, and the calculation methods of the hesperidin, neohesperidin and synephrine content scores are the same. The above components are added according to weight to obtain the comprehensive score of the main component content of the decoction piece variety, for example, the total score of the fresh bran-baked fructus Aurantii in producing area is 8.45/8.45 × 100 × 0.3+6.42/6.84 × 100 × 0.3+1.21/1.32 × 100 × 0.2+0.32/0.32 × 100 × 0.2 ═ 96.48, and the result is shown in table 5.

TABLE 5 comprehensive score of the content of main effective components of different processed fructus Aurantii products (n ═ 3)

According to the experiments, the method for detecting the contents of the main effective components of naringin, hesperidin, neohesperidin and synephrine in the fructus aurantii is established by adopting the high performance liquid chromatography, is stable and reliable, has good reproducibility and can be used for detecting the contents of the components in the fructus aurantii. Through comprehensive weighting analysis, the bran-dried fructus aurantii has the highest comprehensive score, and the traditional fructus aurantii product has the highest comprehensive score based on the crude fructus aurantii decoction pieces, so that the effective components in the fructus aurantii can be damaged to a certain extent by different processing methods, and the effective components such as flavone, alkaloid and the like can be prevented from excessive loss by fresh cutting of the production place.

Experiment 2: comprehensively analyzing chemical components in fresh bran-baked fructus aurantii in producing area

Chemical components in an alcohol extract of fructus aurantii, fructus aurantii stir-baked with bran, fructus aurantii baked with bran while being fresh in producing place and fructus aurantii stir-baked with bran are detected by adopting a UHPLC-Q-TOF/MS method, and Principal Component Analysis (PCA) is carried out by using SIMCA-P13.0 statistical software.

2.1 materials of the experiment

2.1.1 Experimental instruments

triple TOF (TM) 5600 type LC-MS (AB SCIEX, equipped with DuoSprayTM ion source, 30A type liquid chromatography system), AE 240 type one hundred thousandth electronic analytical balance (Metler-Torledo instruments Shanghai Co., Ltd.), FA1004B type electronic balance (Shanghai precision instruments Ltd.), TGL-16B type high-speed refrigerated centrifuge (Shanghai Tingtang scientific instruments factory), GZX-9076MBE type electrothermal blowing dry box (FW Temminck medical equipment factory Co., Ltd.), KQ-250E type medical ultrasonic cleaner (Kunshan ultrasonic instruments Ltd.), and type 135 type Chinese medicine pulverizer (Temminck, Tianjin).

2.1.2 herbs

The same method as the method for preparing fructus aurantii decoction pieces in experiment 1.

2.1.3 reagents and reagents

Naringin, hesperidin, neohesperidin control (the lot numbers of wkq16051305, wkq15123105, wkq16041804, respectively, the mass fractions of which are all more than or equal to 98.0% in the national engineering research center of the manufacturing technology of solid Chinese medicinal preparations), synephrine control (the lot number of BCTG-0709, the mass fraction of which is more than or equal to 98.0%), hesperetin, narirutin, umbelliferone, nobiletin, auraptene control (the purity is not less than 98.0% by the laboratory self-made method of 1H-NMR, 13C-NMR, ESIMS identification, HPLC peak area normalization method), methanol, acetonitrile, formic acid [ mass spectrometric purity, Germany Merke Biotech (China) Limited ], water is ultrapure water, and other reagents are analytically pure.

2.2 methods and results

2.2.1 preparation of test solutions

Precisely weighing 50mg of different processed product powder (sieved by a third sieve) of fructus Aurantii, respectively precisely adding 5mL of methanol, placing in an ultrasonic cleaning machine for ultrasonic extraction at 50 ℃ for 30min, taking out, centrifuging at 4000r min-1 for 15min, taking supernatant, and filtering with a 0.45 μm microporous membrane.

2.2.2 preparation of control solutions

In order to accurately identify the main components in the fructus aurantii, 5mg of hesperidin, naringin, neohesperidin, synephrine, hesperetin, narirutin, umbelliferone, nobiletin and auraptene reference substances are respectively and precisely weighed, dissolved in methanol and respectively dissolved in a10 mL volumetric flask to obtain the reference substance stock solutions. Sucking appropriate amount of each control stock solution, and diluting with methanol to obtain each control solution. The control solution was centrifuged at 4000 r.min-1 for 15min and filtered through a 0.45 μm microporous membrane for UPLC-QTOF-MS analysis.

2.2.3 chromatographic conditions

An ACQUITY UPLC BEH C18 column (2.1 mm. times.100 mm, 1.7 μm); the mobile phase is 0.1 percent formic acid water (A) -0.1 percent formic acid acetonitrile (B), gradient elution is carried out (0-10 min, 95-65 percent A, 10-18 min, 65-25 percent A, 18-21 min, 25-0 percent A, 21-24 min, 0 percent A, 24-24.1 min, 0-95 percent A, 24.1-28 min, 95 percent A), the flow rate is 0.4 mL/min-1; the column temperature is 40 ℃; the sample size was 2. mu.L.

2.2.4 Mass Spectrometry conditions

ESI ion source, data collected in positive ion mode. The flow rate of the dry gas N2 is 10 L.min < -1 >, the temperature of the dry gas is 325 ℃, the temperature of the protective gas is 350 ℃, the flow rate of the protective gas is 10 L.min < -1 >, and the pressure of the atomizer is 275.8 kPa. The nozzle voltage is 500V, the capillary voltage is 4kV, and the scanning range m/z is 50-1200 in the positive ion mode.

2.2.5 data Collection and component identification

Respectively taking 8 batches of crude fructus Aurantii, bran-fried fructus Aurantii, bran-dried fructus Aurantii, and bran-fried fructus Aurantii decoction pieces, and preparing a test sample under the condition of 2.2.1 items; collecting hesperidin, naringin, neohesperidin, synephrine, hesperetin, narirutin, umbelliferone, nobiletin and auraptene reference solutions prepared according to 2.2.2 items, respectively, and detecting the test sample and the reference solution according to chromatographic conditions of 2.2.3 and 2.2.4 items.

Combining the retention time and mass spectrum information of 9 reference substances such as naringin and the like, finally identifying 55 compounds from fructus aurantii, as shown in fig. 3, wherein the labels of the peaks correspond to the numbers in table 6 one by one, and the detailed information of the names, retention time, molecular formulas, errors, ion fragments and the like of the compounds obtained through identification is shown in table 6.

TABLE 6 identification of chemical component information in different pieces of fructus Aurantii by UHPLC-Q-TOF/MS

After the fructus aurantii is stir-fried by bran and bran, no new components are found, the fresh cut fructus aurantii does not cause the quality change of the components compared with the traditional cut fructus aurantii, and the influence of different processing methods on the content of compounds in the fructus aurantii is not completely the same. After stir-frying with bran, the peak area of 7 compounds is obviously increased (CI is more than 1.5); the peak area of 6 compounds is obviously reduced (CI < 0.5); after bran baking, the peak areas of 11 compounds are obviously increased, and the peak areas of 2 compounds are obviously reduced; after being fried by bran, the peak areas of 5 compounds are obviously increased, and the peak areas of 3 compounds are obviously reduced. The area of the ion peak of each processed product of fructus Aurantii in positive ion mode and the index of change before and after processing are shown in Table 7.

TABLE 7 area of ion peak of each processed product of fructus Aurantii in positive ion mode and its index of change before and after processing

2.2.6 partial least squares-discriminant analysis (PLS-DA) of UHPLC-Q-TOF/MS data

The method is characterized in that partial least squares-discriminant analysis (PLS-DA) is adopted to analyze importance projections (VIP) of variables, the VIP can comprehensively summarize the importance of a variable in a model to an X matrix and a Y matrix, the greater the VIP value, the more important the variable is to the model, and the variable with the VIP >1 is generally taken as an important marker of the model. Constructing PLS-DA (R2X [1] ═ 0.629, R2X [2] ═ 0.0961) models of different processed products of fructus aurantii, and screening 14 VIP & gt 1 components as quality markers, wherein the components are respectively as follows: hesperetin (VIP ═ 3.33781), poncirin (VIP ═ 3.3014), narirutin (VIP ═ 2.64193), naringin (VIP ═ 2.64193), hesperidin (VIP ═ 2.56968), neohesperidin (VIP ═ 2.56968), hesperetin (VIP ═ 2.51306), naringenin (VIP ═ 2.2463), deacetyl rhododendron (VIP ═ 2.23414), 5-desmethylnobiletin (VIP ═ 1.67091), hesperetin 5-O-glucoside (VIP ═ 1.60005), hesperetin 7-O-glucoside (VIP ═ 1.60005), neonorth american eriodictyol (VIP ═ 1.50899), and 4 ', 5' -dihydropsoralen (VIP ═ 1.09209).

2.2.7 comparison of the material bases of different processed products

And (3) carrying out comprehensive scoring on the peak areas of 14 chemical markers in different processed products of the bitter orange by adopting a comprehensive weighting scoring method, wherein the component weight coefficient of VIP >2 is set to be 8%, the component weight coefficient of VIP >1 is set to be 5.6%, the higher the peak area of the component is, the better the material base is, and therefore the sample with the highest peak area is taken as 100. The total score of the main component content of the decoction piece variety is obtained by adding 14 chemical markers according to the weight, and is shown in Table 8.

TABLE 8 comprehensive score of chemical marker content of different processed products of fructus Aurantii

The fructus aurantii is cut in a production place and then baked by bran, the component types with obviously increased content in the fructus aurantii are obviously higher than those of the traditional fried and prepared fructus aurantii, and the component types with obviously reduced content do not show obvious rules, so that the situation that the fructus aurantii is baked by bran in the production place is more beneficial to improving the content of chemical components in the fructus aurantii is prompted; after being processed, hesperidin and hesperetin both increase to different degrees, and the content of the bran-dried bitter orange is the highest; the bran (bran) is fried to reduce the neohesperidin content to different degrees, and the bran is baked to increase the neohesperidin content; after being stir-fried with bran, the fructus aurantii has 8 components with increased content and 6 components with reduced content; after the bran is dried, the content of 13 components is increased, and the content of 1 component is reduced; after the bran is fried, the content of 9 ingredients is increased, and the content of 5 ingredients is reduced. By taking the peak areas of 14 chemical markers as evaluation indexes and carrying out multi-index weighting scoring, the highest content of the components in the bran-baked bitter orange is found, and the bran-baked bitter orange is a dominant processed variety of the bitter orange.

Experiment 3: GC-MS combined PCA technology for analyzing volatile oil and chemical components in aromatic water of different processed products of fructus aurantii

3.1 Experimental materials

3.1.1 Experimental instruments

7890B-5977A type gas chromatograph-mass spectrometer (Agilent technologies, Inc., USA), DB-5 quartz capillary column (0.25 μm × 250 μm × 30.0m), ZNHW type intelligent constant temperature electric heating jacket (Consortium of Wahua instruments, Inc., Hippocampus Co., Ltd.), FA1004B type electronic balance (Shanghai precision science instruments, Inc.), FW135 type Chinese medicine grinder (Tester instruments, Inc., Tianjin), Raytek Raynger ST20 infrared thermometer (Retai, USA), and two-stage temperature-regulating electric frying pan (home appliance industry, Zhanjiang, Guangdong province).

3.1.2 herbs

The preparation method of decoction pieces in experiment 1 is the same.

3.1.3 reagents and reagents

Ether (Guangdong Gangxi scientific Co., Ltd.), anhydrous sodium sulfate (Tianjin Di Bo chemical Co., Ltd.).

3.2 methods and results

3.2.1 extraction of essential oils

According to the extraction method of volatile oil in XD (extended X date in the appendix of the national formulary) of 2015 edition, taking a proper amount of raw or prepared pieces of fructus aurantii, crushing, sieving by a No. 3 sieve, precisely weighing 50g of each powder of different processed products of fructus aurantii, placing the powder in a 1L round-bottomed flask, adding 600mL of water and a plurality of crushed ceramic pieces, soaking for 30min, placing the round-bottomed flask in an electric heating jacket, connecting a volatile oil extraction device, heating and extracting for 8h, taking out an upper oil layer in distillate of a volatile oil extractor, namely the volatile oil of the fructus aurantii, adding anhydrous sodium sulfate, standing overnight, adding a proper amount of diethyl ether to prepare a1 g-L-1 sample solution, and. The fructus Aurantii crude volatile oil is yellow transparent oil, and has deep yellow color and special strong fragrance after being processed by bran.

3.2.2 extraction of aromatic Water

Extracting aromatic water by steam distillation, collecting lower water layer of distillate of volatile oil extractor, adding 2 times of diethyl ether, oscillating, extracting for 3 times, mixing extractive solutions, recovering diethyl ether under reduced pressure, adding anhydrous sodium sulfate, standing overnight, and filtering with 0.22 μm microporous membrane to obtain volatile oil. The obtained raw or product aromatic water of fructus Aurantii is clear or semi-clear liquid water, and has special fragrance similar to fructus Aurantii volatile oil.

3.2.3 gas chromatography conditions

The sample port temperature is 260 ℃, the transmission line temperature is 250 ℃, the carrier gas is helium, the flow rate is 1.0mL min < -1 >, the split ratio is 10:1, and the sample injection amount is 1 mu L; the temperature programming is finished after the initial column temperature is 70 ℃, the temperature is raised to 150 ℃ at 2 ℃ and min < -1 >, the temperature is kept for 2min, the temperature is raised to 240 ℃ at 6 ℃ and min < -1 >, the temperature is kept for 3min, the temperature is raised to 300 ℃ at 25 ℃ and min < -1 >, and the temperature is kept for 2 min.

3.2.4 Mass Spectrometry conditions

The ionization mode is electron bombardment ionization source (EI), electron collision energy is 70eV, ion source temperature is 230 ℃, acceleration voltage is 34.6V, resolution is 2500, multiplier voltage is 1.388kV, quadrupole rod temperature is 150 ℃, m/z is 10-650, scanning number is 4.45 times/s, total ion flow graph is recorded, mass spectrogram is searched and identified by NIST08 standard mass spectrogram library, and relative content of each component is calculated by peak area normalization method.

fructus Aurantii volatile oil (figure 4, by relative percentage, contains camphene 0.03%, alpha-phellandrene 0.09%, alpha-terpinene 0.53%, D-limonene 66.75%, m-cymene 3.17%, gamma-terpinene 16.98%, alpha-methyl-alpha- [ 4-methyl-3-pentenyl ] ethylene oxide methanol 0.07%, iso-terpinene 1.09%, cis-linalool oxide 0.19%, linalool 1.72%, (-) -4-terpineol 0.56%, alpha-terpineol 0.57%, (1R, 5R) -rel-carveol 0.03%, thymol 0.13%, trans-sesquiterpene caryophyllene 0.09%, A-copal 0.01%, goldenrain 0.01%, 1, 2, 3, 4, 4a, 5, 6, 8 a-octahydro-7-methyl-4-methylene-1- (1-methylethyl) 0.04%, bicyclo germacadiene 0.08%, (1S, 8aR) -1-isopropyl-4, 7-dimethyl-1, 2, 3, 5, 6, 8 a-hexahydronaphthalene 0.16%, steviol 0.06%, alpha-pymetrol 0.03%, palmitic acid 0.01%, (1S) - (-) -alpha pinene 2.47%, (1S) - (-) -beta-pinene 2.47%, 3-carene 0.01%, L-limonene 0.18%, 5-methyl-2- (1-methylvinyl) -4-hexen-1-ol acetate 0.01%, bicyclo [3.1.0] -4-methyl-1-isopropylhexane didehydro derivative 0.6%, bicyclo [3.1.0] -4-methyl-1-isopropyl-2-hexene 2.29%, bicyclo [4.2.0.] -6, 7-dimethyloctane 0.01%, (+) -limonene oxide 0.02%, 1-methyl-4-prop-1-en-2-yl-cyclohex-2-en-1-ol 0.03%, 7, 7-dimethyl-2-methylene-bicyclo [2.2.1] heptane 0.02%, cyclopropanecarboxamidine hydrochloride 0.02%, carveol 10.06%, (+) -4-carene 0.02%, carvone 0.05%, (Z) -beta-farnesene 0.22%, [ S- (E, E) ] -1-methyl-5-methylene-8- (1-methylethyl) -1, 6-cyclodecadiene 1.28%, camphene 0.01%, alpha-farnesene 0.01%, 1-vinyl-1-methyl-4-prop-2-ylidene-2-prop-1-en-2-ylcyclohexane 0.01%, limonene oxide 20.01%, gamma-apiene 0.03%, gamma-cedrene 0.01%, (-) -a-coumalene 0.01%) and aromatic water (fig. 5, in relative percentages, comprising in total: (1R) - (+) -alpha pinene 0.68%, beta pinene 0.93%, beta myrcene 1.41%, alpha terpinene 0.45%, D-limonene 79.60%, gamma terpinene 12.13%, 9-oxo-pivaloyl-N-acetyl colchicol 0.08%, (+) -4-carene 0.46%, [ S- (E, E) ] -1-methyl-5-methylene-8- (1-methylethyl) -1, 6-cyclodecadiene 0.15%, p-cymene 1.9%, pentacosane 0.08%, octacosane 0.07%, iodooctadecane 0.08%, 9-octyl-heptadecane 0.04%, 7, 9-di-tert-butyl-1-oxaspiro [4.5] -6, 9-decadiene-2, 0.11 percent of 8-diketone, 0.48 percent of 2, 2' -methylene bis- (4-methyl-6-tert-butylphenol), 0.05 percent of 3- (3, 5-di-tert-butyl-4-hydroxyphenyl) methyl propionate, 0.11 percent of precocene II, 0.06 percent of docosane and 4.16 percent of 4-cholesten-3-ketone-semicarbazone), wherein 65 components are identified in the crude volatile oil, wherein D-limonene (66.75%) and gamma-terpinene (16.98%) together account for 83.73% of the volatile oil component of the raw product, and 42 components in the raw product perfume are totally identified, wherein the total amount of the ingredients and the volatile oil is 10, the other 32 ingredients are only detected in aromatic water, and the content of D-limonene (79.60%) and gamma-terpinene (12.13%) in the aromatic water is increased to 91.73%; in addition, bran-fried bitter orange aromatic water ((1R) - (+) - α pinene 0.7%, β -myrcene 2.89%, α -phellandrene 0.03%, α -terpinene 0.12%, D-limonene 76.11%, γ -terpinene 15.39%, terpinolene 1.14%, cis-linalool oxide 0.19%, linalool 0.89%, (-) -4-terpineol 0.36%, α -terpineol 0.41%, (1R, 5R) -rel-carveol 0.08%, 5-isopropenyl-2-methylenecyclohexanol 0.04%, dextrocarvone 0.02%, trans-caryophyllene 0.01%, trans- β -farnesene 0.04%, methyl palmitate 0.01%, cis- Α, Α -5-trimethyl-5-vinyltetrahydrofuran-2-methanol 0.36%, 3.11% of 1, 3, 8-P-menthatriene, 0.06% of 2- (4-methylphenyl) propan-2-ol, 0.01% of n-eicosane, 0.08% of 7, 7-dimethyl-2-methylene-bicyclo [2.2.1] heptane, 0.05% of [ S- (E, E) ] -1-methyl-5-methylene-8- (1-methylethyl) -1, 6-cyclodecadiene, 0.08% of P-mint-1 (7), 1.1% of 3-diene, 0.02% of pentacosane, 0.04% of hexadecane, 0.07% of dibutylhydroxytoluene, 0.03% of heptacosane, 0.03% of octacosane, 0.01% of iodooctadecane, 0.04% of 9-octyl-heptadecane, 0.04% of heneicosane, 7, 9-di-tert-butyl-1-oxaspiro [4.5] -6, 9-decadiene-2, 8-dione 0.02%, 2-methylnonadecane 0.01%, octadecane 0.03%, tetracosane 0.04%, heptadecane 0.02%, 2-bromododecane 0.02%, 1-methyl-4- (1-methylvinyl) benzene 0.04%, bis (1-methyleneethyl) cyclobutene 0.03%, 2, 3, 4, 6-tetramethylphenol 0.01%, 1-iodotridecane 0.06%, pentadecane 0.02%, 5-butyl-nonane 0.01%, methyl 3- (3, 5-di-tert-butyl-4-hydroxyphenyl) propionate 0.03%, 1-butyl-2-isobutyl phthalate 0.01%, 8-methylheptadecane 0.02%, hexacosane 0.01%), wherein 48 components were identified in total with the volatile oil (2-methyl-5- (1-methylethyl) -bicyclo [3.1.0] -2-hexene 0.47%, camphene 0.03%, β -pinene 2.42%, α -phellandrene 0.07%, α -terpinene 0.21%, D-limonene 66.05%, γ -terpinene 15.79%, terpinolene 1.02%, cis-linalool oxide 0.17%, linalool 2.17%, (-) -4-terpineol 0.62%, α -terpineol 0.88%, (1R, 5S) -rel-carveol 0.13%, (1R, 5R) -rel-carveol 0.17%, 4-vinyl-4-methyl-1-prop-2-yl-3-prop-1-en-2-yl-cyclohexene 0.02%, D-carvone 0.08%, trans-caryophyllene 0.15%, trans- β -farnesene 0.25%, a-copal 0.04%, 1.42% of piperylene, (1S, 8aR) -1-isopropyl-4, 7-dimethyl-1, 2, 3, 5, 6, 8 a-hexahydronaphthalene 0.14%, 1, 2, 3, 5, 6, 8 a-hexahydro-4, 7-dimethyl-1- (1-methylethyl) naphthalene 0.01%, elemene 0.07%, eucalyptol 0.16%, steviol 0.08%, alpha-petalol 0.06%, phytone 0.01%, palmitic acid 0.27%, 1S) - (-) -alpha pinene 2.07%, 1S) - (-) -beta-pinene 2.06%, cis-alpha, alpha-5-trimethyl-5-vinyl tetrahydrofuran-2-methanol 0.41%, 2-methyl-1-phenylpropylene 0.02%, 0.02% of 3-methylcyclohexanol, 0.03% of 1, 3, 8-P-menthatriene, 0.01% of 3-carene, 0.04% of 3-methyl-trans-3 a, 4, 7, 7 a-tetrahydroindene, 0.03% of 1-ethyl-4-methyl-2 (1H) -pyridone, 0.02% of 1, 4-dimethyl-4-vinylcyclohexene, 0.01% of isobornyl acetate, 0.03% of citral, 0.02% of decanal, 0.04% of 2- (4-methylphenyl) propan-2-ol, 0.03% of nerol, 0.02% of carvone, 3.35% of o-isopropylbenzene, 0.24% of L-limonene, 0.02% of 5-methyl-2- (1-methylvinyl) -4-hexen-1-ol acetate, 0.01% of neoisolongifolene, 0.01% of 1, 6-diene-3-alcohol humulene, 0.03% of myrcene, 0.1% of (S) -6-vinyl-6-methyl-1- (1-methylethyl) -3- (1-methylethylidene) -cyclohexene, 0.02% of epoxy isomyrcene, 0.02% of myristic acid, 0.01% of 14-methyl pentadecanoic acid methyl ester, 0.01% of 2-octyl cyclopropane octanal, 0.01% of ethofumesate-2-one, 0.02% of (Z, Z) -9, 15-octadecadienoic acid methyl ester, 0.03% of linoleic acid, 0.01% of n-eicosane and 0.04% of nerolidol, wherein 17 components are the same; identifying 20 components in fresh bran-baked fructus Aurantii aromatic water in producing area, wherein the number of the components is 5; 14 of the bran-fried bitter orange aromatic water (2-methyl-5- (1-methylethyl) -bicyclo [3.1.0] -2-hexene 0.09%, (1R) - (+) -alpha pinene 0.39%, beta-myrcene 1.82%, alpha-phellandrene 0.07%, D-limonene 70.58%, gamma-terpinene 14.59%, terpinene 0.66%, 3-carene 0.04%, o-isopropylbenzene 1.87%, (+) -4-carene 0.55%, p-mint-1 (7), 3-diene 0.71%, tetratetradecane 0.05%, triundecane 0.05%, docosane 0.05%), with which volatile oil (2-methyl-5- (1-methylethyl) -bicyclo [3.1.0] -2-hexene 0.52%, beta-myrcene 2.35%, 0.08 percent of alpha-phellandrene, 0.5 percent of alpha-terpinene, 64.04 percent of D-limonene, 14.69 percent of gamma-terpinene, 0.88 percent of terpinene, 0.14 percent of cis-linalool oxide, 1.92 percent of linalool, 0.06 percent of trans-menthyl-2, 8-diene-1-ol, 0.88 percent of (-) -4-terpineol, 0.86 percent of alpha-terpineol, (1R, 5R) -rel-carveol, 0.23 percent of 4-vinyl-4-methyl-1-propyl-2-yl-3-propyl-1-ene-2-yl-cyclohexene, 0.02 percent of dextrocarvone, 0.12 percent of (Z) -3, 7-dimethyl-2, 6-octadiene-1-ol acetate, 0.23 percent of trans-caryophyllene, trans-beta-farnesene 0.24%, valencene 0.01%, 1, 2, 3, 4, 4a, 5, 6, 8 a-octahydro-7-methyl-4-methylene-1- (1-methylethyl) naphthalene 0.05%, piperylene 1.75%, 7, 11-dimethyl-3-methylene-1, 6, 10-dodecanetriene 0.01%, (1S, 8aR) -1-isopropyl-4, 7-dimethyl-1, 2, 3, 5, 6, 8 a-hexahydronaphthalene 0.18%, eucalyptol 0.18%, 3-methyl-4- (2, 6, 6-trimethyl-2-cyclohexen-1-yl) -3-buten-2-ol 0.08%, ALPHA-cumidinol 0.01%, palmitic acid 0.01%, 9-oxo-pivaloyl-N-acetyl colchicol 0.36%, (1S) - (-) -alpha pinene 2.01%, (1S) - (-) -beta-pinene 2.5%, 1, 3, 8-P-menthene 0.04%, citral 0.04%, trans-Z, alpha-bisabolene epoxide, phyllylene compound 0.01%, nerolidol 0.03%, 1-methyl-4-prop-1-en-2-yl-cyclohex-2-en-1-ol 0.05%, 1-vinyl-1-methyl-4-prop-2-ylidene-2-prop-1-en-2-ylcyclohexane 0.11%, 0.02% of 2, 2-dimethyl-3-methylenebicyclo [2.2.1] heptane, 0.02% of o-mentha-1 (7), 0.01% of 8-dien-3-ol, 1.75% of p-cymene, 0.31% of linalool oxide, 0.01% of dihydrolinalool, (2R, 5R) -2-methyl-5-propan-2-ylbicyclo [3.1.0] hexan-2-ol, 0.02% of 1-methyl-4- (1-methylvinyl) cyclohexanol, 0.03% of 1-hydroxymethyl-2-methyl-1-cyclohexene, 0.01% of 3-methyl-2-butenal, 0.06% of cis-citral, 20.13% of carveol, 0.03% of 3-methyl-6- (1-methylethylidene) cyclohex-2-ene-1-one, geranyl acetate 0.02%, 2-isopropyl-5-methyl-9-methylenebicyclodecan-1-ene 0.01%, ALPHA-humulene 0.02%, thujaene 0.02%, guaialene 0.06%, 2- (1-hydroxycycloheptyl) furan 0.03%, 1, 7, 7-trimethyl-2-vinylbicyclo [2, 2, 1] pyrimidin-2-ene 0.06%, 8-hydroxycycloiso-longifolene 0.02%, caryophyllin 0.01%, 1, 2, 3, 4, 4a, 5, 6, 8 a-octahydro-4 a, 8-dimethyl-2- (2-propenyl) -1-naphthol 0.01%, 2-hexenal 0.01%, 4- (4-ethylcyclohexyl) -1-pentyl-cyclohexene 0.01%) 5 of the same components.

As can be seen from tables 9 and 10, the volatile components mainly include alcohols, ketones, esters, acids, alkenes, aldehydes, phenols, hydrocarbons, and other compounds, and the volatile oil and the aromatic water both contain alkenes as their main components. The volatile oil component contains the most alkene compounds with the highest relative content of 90.12-93.09%; secondly, alcohol compounds and hydrocarbon compounds with relative contents of 2.98-5.56% and 1.87-4.01% respectively; the types of esters, ketones and acids are less, and the relative contents are respectively 0.01-0.31%, 0.04-0.17% and 0.00-0.32%; the types of phenols, aldehydes and other compounds are few, and the relative content is low, namely 0.00-0.13%, 0.00-0.13% and 0.00-0.03%. Compared with the volatile oil, the types of the alkene compounds in the aromatic water component are reduced, but the relative content is still the highest, and the relative content is 81.85-98.03%; although the types of the hydrocarbons are obviously increased, the relative contents of the hydrocarbons are not obviously different; the acid and aldehyde are single in type and low in relative content, and the relative content of the acid and aldehyde is only 0.26 percent and 0.06 percent in honey bran bitter orange.

TABLE 9 amount of volatile components in different processed products of fructus Aurantii

TABLE 10 relative content of volatile components in different processed products of fructus Aurantii

In volatile oil, 63 components in the fructus aurantii stir-fried with bran are identified, and 34 new components are added compared with the raw product; identifying 48 components of the fresh bran-dried fructus aurantii in the production place, and adding 24 components compared with the raw decoction pieces; 61 components are identified from the stir-fried fructus aurantii with bran, and 33 new components are added compared with the raw product. In the aromatic water group, 48 components of the fructus aurantii stir-fried with bran are identified, and 24 new components are added compared with the raw product; identifying 20 components of the fresh bran-baked fructus Aurantii in the producing area, and adding 10 new components compared with the raw product; 14 components are identified from the stir-fried fructus aurantii with bran, and 7 newly added components are compared with the raw product. After processing, the types of compounds in the volatile oil are changed greatly, but the relative content difference of various compounds is small, only alcohol compounds are obviously reduced, and other types of components are not changed obviously; in the aromatic water group, the types of compounds and the relative contents of various components are obviously changed, for example, the types and the relative contents of main components of alcohols and alkene hydrocarbon are obviously increased or reduced in the processed products, and the regular change is not seen.

Experiment 4: comparison of the action of different processed products of fructus Aurantii in promoting gastrointestinal motility

4.1 Experimental materials

4.1.1 Experimental instruments

RE52CS rotary evaporator (Shanghai Yangrong Biochemical instruments factory), FA1004B electronic balance (Shanghai precision scientific instruments Co., Ltd.), HH-4 digital display constant temperature water bath (China Hua electric appliances Co., Ltd.), and metabolism cage. ZNHW type intelligent constant temperature electric heating jacket (ZNhw type intelligent instrument responsibility Co., Ltd.), FW135 type traditional Chinese medicine grinder (Tester instruments Co., Ltd., Tianjin), Raytek Raynger ST20 infrared temperature measuring instrument (Raytai Co., USA), and two-grade temperature regulating electric frying pan (household electrical appliance industry Co., Zhanjiang, Guangdong).

4.1.2 Experimental drugs

The experimental medicinal materials are prepared by the same method as the decoction pieces prepared in the experiment 1; domperidone tablet (Xian Yang Sen pharmaceutical Co., Ltd., national drug standard H10910003).

4.1.3 Experimental animals

SD rats, each half male and female, with a weight of 200 + -20 g, were bred in the animal testing center of Nanchang university under the barrier environment of the animal testing center of Jiangxi university of traditional Chinese medicine, under the qualification number SCXK (gan) 2016-.

4.1.4 medicine extract: weighing appropriate amount of processed fructus Aurantii, adding 10 times of water, decocting for 30min, repeating twice, mixing filtrates, and concentrating to obtain medicinal liquid with concentration of 2 g.mL < -1 >; the domperidone tablet is crushed and dissolved by distilled water to prepare a solution of 0.2 g.L < -1 >.

4.1.5 nutritional semi-pastes: and (2) putting 20g of sodium carboxymethylcellulose into 500mL of water, heating and ultrasonically treating the mixture until the sodium carboxymethylcellulose is completely dissolved, adding 16g of milk powder, 8g of starch, 8g of cane sugar and 2g of activated carbon powder, and uniformly stirring the mixture to obtain the sodium carboxymethylcellulose powder.

4.2 methods and results

4.2.1 grouping and Molding

70 SD rats are averagely divided into 7 groups, namely a blank group (A), a raw Zhi shell group (B), a bran fried Zhi shell group (C), a bran baked Zhi shell group (D) with fresh bran at the producing area, a bran fried Zhi shell group (E), an FD model group (F) and a domperidone positive group (G), wherein each group comprises 10 rats. Except the blank group, the FD rat model was replicated for 14 consecutive days by tail-biting rage, diet loss and hydrochloric acid drinking.

4.2.2 methods of administration

After the molding is successful, the rats of each administration group are administered the corresponding medicine in the morning every day since the 15 th day. The raw fructus aurantii, the fructus aurantii stir-fried with bran, the fresh fructus aurantii baked in the producing area and the bran-fried trifoliate group rats are irrigated with the extracting solution of the raw fructus aurantii product according to the dose of 5 g.kg < -1 >, the positive control group is administrated with the domperidone solution according to 10 mL.kg < -1 >, and the blank group and the model group rats are irrigated with the normal saline according to the dose of 10 mL.kg < -1 > for 14 days.

4.2.3 Effect on gastric remnant intestinal propulsion in FD rats

On the 28 th day of the experiment, the patient is fasted for 12h after the last administration, all experimental animals are gavaged with 2mL of nutritional semi-paste, the stomach pouch and the small intestine are taken after anesthesia, and the gastric emptying rate and the small intestine propulsion rate are calculated, and the results are shown in Table 11.

The ratio (%) of small intestine advancing rate (charcoal advancing distance)/total small intestine length × 100% gastric residual rate (total stomach weight-net stomach weight)/weight of semisolid paste × 100%

TABLE 11 gastric residual rate and intestinal transit rate (n ═ 10) for each group of rats

Note that compared with the blank group, 1) P is less than 0.05, 2) P is less than 0.01; comparing with model group 3) P < 0.05, 4) P < 0.01

As can be seen from Table 11, the gastric residual rate and intestinal propulsion rate of the FD model group rats were significantly different from those of the blank group, indicating that the modeling was successful. When the administration dosage is the same, the difference of the fresh bran-baked bitter orange in the producing area is the smallest compared with that of a blank group, and the difference is not significant. The pharmacological research results are consistent with the early-stage chemical component research, and the results prove that the method for baking the fructus aurantii after being cut fresh in the producing area by the bran coat can improve the content of the effective components of the fructus aurantii decoction pieces and enhance the curative effect of relieving epigastric distention.

Experiment 5: parameter comparison of processing method of bran-dried fructus aurantii with fresh producing area

The rice bran addition amount, the baking temperature, the baking time and the slice thickness are set as parameters for comparison, the mass fractions of naringin, hesperidin and neohesperidin and the mass concentration of MTL (methyl thiazolyl tetrazolium) in blood plasma are taken as comprehensive evaluation indexes for comparison, and the collected data are shown in Table 12.

TABLE 12 comparison of processing parameters of fresh bran-baked fructus Aurantii in producing area

The processing is carried out according to the method, the processing technology is reasonable, and the prepared bitter orange decoction pieces baked by bran in the production place have stable quality.

In conclusion, the invention provides the fresh-bran-dried fructus aurantii in the production place, the processing method and the application thereof, the fresh-bran-dried fructus aurantii in the production place is also the best processed variety, and the effect of relieving swelling in the middle and relieving swelling is superior to that of other processed varieties; through earlier stage research, hesperidin, naringin, neohesperidin and synephrine can be used as a Q-Marker of the bitter orange and are the main drug-effect substances in the bitter orange; through UPLC-Q-TOF/MS combined with PCA analysis, it is known that 14 components such as hesperetin can be used as a quality marker of fructus aurantii; by detecting the types of the volatile components of the fructus aurantii, the D-limonene and the gamma-terpinene are 2 components with the highest content in the volatile oil and the aromatic water, and are the main drug effect substances in the volatile components of the fructus aurantii. By comparing the contents of the components in different processed products, the highest content of the bitter orange in the fresh bran baked bitter orange in the production area is found.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims (7)

1. A processing method of bran-dried bitter orange in a producing area is characterized by comprising the following steps: after immature fruits of sour oranges are harvested in a production place, removing fruit pulp, cutting fresh peel into thin threads with the thickness of 0.2-0.3 cm, uniformly mixing the thin threads with bran coat, and drying for 2 hours at the temperature of 160 ℃; the dosage of the bran coat is 10kg per 100kg of immature fruit of Citrus aurantium L.

2. The processing method of claim 1, further comprising screening off the bran coat after the baking is completed, and cooling the bran coat.

3. The dried fructus aurantii with bran in the fresh producing area prepared by the processing method of claim 1 or 2.

4. The use of the fresh bran-baked fructus Aurantii as claimed in claim 3 in preparing bran-baked fructus Aurantii decoction pieces for increasing the alcohol-soluble effective component content of fructus Aurantii.

5. The use of claim 4, wherein the alcohol-soluble effective ingredients of fructus Aurantii include naringin, neohesperidin, hesperidin and synephrine.

6. The use of the bran-baked fructus Aurantii as claimed in claim 3 in the preparation of bran-baked fructus Aurantii decoction pieces for increasing the volatile component content of fructus Aurantii.

7. The use according to claim 6, wherein the volatile components comprise D-limonene and gamma-terpinene.