CN113484428B - Construction method of peach pit qi-bearing decoction characteristic spectrum - Google Patents
Construction method of peach pit qi-bearing decoction characteristic spectrum Download PDFInfo
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention discloses a construction method of a peach pit qi-bearing soup characteristic map, which comprises the following steps: preparing a mixed reference solution of gallic acid reference substance, catechin reference substance, amygdalin reference substance, epicatechin gallate reference substance, glycyrrhizin reference substance, apiose glycyrrhizin reference substance, cinnamaldehyde reference substance, cinnamic acid reference substance, chrysophanol-1-O-glucoside reference substance, chrysophanol-8-O-glucoside reference substance, emodin-8-O-glucoside reference substance, rhein reference substance, glycyrrhizic acid reference substance, emodin reference substance and chrysophanol reference substance; preparing a sample solution; then the mixture is injected into a liquid chromatograph for testing, and a characteristic spectrum of the peach pit qi-bearing decoction is established. The method provided by the invention has good repeatability, is accurate and reliable, can provide a data basis for the quality control of the peach pit qi-bearing soup, and effectively ensures the stability and controllability of the quality of the peach pit qi-bearing soup product.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicine quality analysis and detection, in particular to a construction method of a peach pit qi-bearing soup characteristic spectrum.
Background
The peach pit qi-bearing decoction comes from Shang Han Lun (typhoid treatise on Zhang Zhongjing) in the Han Dynasty: the disease of the sun is not resolved, and the bladder is heat-accumulated, so that the patients get down from the blood, and the patients get up. The other part is not attacked, and the other part is firstly released; for the patients with frequent exterior syndrome but urgent knots in lower abdomen, it is advisable to treat qi-supporting decoction of peach pit. Fifty peach kernels (peeled tip), four rhubarb, two cassia twig, two liquorice (roasted), and two mirabilite. The five flavors are superior, seven liters of water are boiled to take two liters and half, dregs are removed, internal mirabilite is removed, the internal mirabilite is higher, the internal heat is lower, the internal heat is slightly boiled, and the five flavors are taken after warm taking, and three days are taken after taking. The peach pit qi-bearing decoction is widely applied clinically, is not only limited to symptoms such as lower abdominal distention and pain, dark stool, spontaneous urination, delirium and mania of the accumulation of blood in the sun, but also widely applied to symptoms such as cerebral trauma sequelae, gynaecology dysmenorrhea, amenorrhea, retention of urine in kidney diseases, prostatitis or retention of urine caused by hyperplasia of prostate, and the like. Peach pit qi-bearing soup is not developed into a Chinese patent medicine in China at present, but is developed into a Chinese prescription preparation for wide clinical use in Japan, and the Chinese prescription preparation is sold in a pharmacy as a Chinese prescription medicine and has a strong development value in China.
At present, the research on the peach pit qi-bearing decoction is mainly focused on the aspect of pharmacological research, and the research on the material basis, the extraction process, the multi-index component content measurement and the characteristic spectrum of the decoction is less, and the system is lacking. And less research is conducted on how to measure the consistency of mass-produced preparations and traditional decoction quality.
Disclosure of Invention
The invention aims to solve the technical problem of providing a construction method of the peach pit qi-bearing soup characteristic spectrum, which has good repeatability, is accurate and reliable, can provide a data basis for mass production quality control of the peach pit qi-bearing soup, and ensures the stability and controllability of the quality of the peach pit qi-bearing soup product.
In order to solve the technical problems, the invention provides a construction method of a peach pit qi-bearing soup characteristic map, which comprises the following steps:
(1) Respectively taking gallic acid reference substance, catechin reference substance, amygdalin reference substance, epicatechin gallate reference substance, glycyrrhizin reference substance, apiose glycyrrhizin reference substance, cinnamaldehyde reference substance, cinnamic acid reference substance, chrysophanol-1-O-glucoside reference substance, chrysophanol-8-O-glucoside reference substance, emodin-8-O-glucoside reference substance, rhein reference substance, glycyrrhizic acid reference substance, emodin reference substance, and appropriate amount of chrysophanol reference substance, adding solvent, dissolving or extracting to obtain reference substance solution;
(2) Extracting semen Persicae decoction with solvent to obtain sample solution;
(3) Taking a preset amount of reference substance solution and test substance solution, and injecting into a liquid chromatograph, wherein the liquid chromatograph uses octadecylsilane chemically bonded silica as a filler, methanol as a mobile phase A and phosphoric acid aqueous solution as a mobile phase B for gradient elution, so as to establish a characteristic map of the peach pit qi-supporting soup.
As an improvement of the above technical scheme, the gradient elution is performed according to the following procedure:
0-5 min, mobile phase A from 1% to 8%, mobile phase B from 99% to 92%;
5-21 min, mobile phase A from 8% to 23%, mobile phase B from 92% to 77%;
21-35 min, mobile phase A from 23% -31%, mobile phase B from 77% -69%;
35-45 min, mobile phase A from 31% -55%, mobile phase B from 69% -45%;
45-51 min, mobile phase A from 55% -65%, mobile phase B from 45% -35%;
51-56 min, mobile phase A from 65% -83%, mobile phase B from 35% -17%;
56-65 min, mobile phase A from 83% to 100% and mobile phase B from 17% to 0%.
As an improvement of the technical scheme, in the step (3), respectively sucking 1-3 mu L of each of the reference substance solution and the test substance solution, and injecting into a liquid chromatograph for detection, wherein the liquid chromatograph uses octadecylsilane chemically bonded silica gel as a filler, the column length is 150mm, the inner diameter is 2.1mm, the particle diameter is 1.6 mu m, and the column temperature is 30-38 ℃; the liquid chromatograph takes methanol as a mobile phase A, takes 0.15-0.25% phosphoric acid aqueous solution as a mobile phase B, has the flow rate of 0.25-0.36 mL/min and the detection wavelength of 220-290 nm.
As an improvement of the technical scheme, in the step (3), respectively sucking 1 mu L of each of the reference substance solution and the test substance solution, and injecting the solution into a liquid chromatograph for detection, wherein the liquid chromatograph uses octadecylsilane chemically bonded silica gel as a filler, the column length is 150mm, the inner diameter is 2.1mm, the particle diameter is 1.6 mu m, and the column temperature is 38 ℃; the liquid chromatograph takes methanol as a mobile phase A, takes 0.2% phosphoric acid aqueous solution as a mobile phase B, and has the flow rate of 0.3mL/min.
As an improvement of the technical scheme, in the step (3), when the detection time is 0-30 minutes, the detection wavelength is 220nm; the detection wavelength is 290nm when the detection time is 30-45 minutes, and 260nm when the detection time is 45-65 minutes.
As an improvement of the technical scheme, in the step (1), a gallic acid reference substance, a catechin reference substance, an amygdalin reference substance, an epicatechin gallate reference substance, a glycyrrhizin reference substance, a apiose glycyrrhizin reference substance, a cinnamaldehyde reference substance, a cinnamic acid reference substance, a chrysophanol-1-O-glucoside reference substance, a chrysophanol-8-O-glucoside reference substance, a emodin-8-O-glucoside reference substance, a rhein reference substance, a glycyrrhizic acid reference substance, a emodin reference substance and a chrysophanol reference substance are respectively taken, and methanol is respectively added to prepare a solution containing 15 mug of gallic acid, 100 mug of catechin, 30 mug of amygdalin, 50 mug of epicatechin gallate, 30 mug of glycyrrhizin, 70 mug of apiose glycyrrhizin, 10 mug of cinnamaldehyde, 10 mug of cinnamic acid, 50 mug of chrysophanol-1-O-glucoside, 40 mug of chrysophanol-8-O-glucoside, 50 mug of emodin-8-O-glucoside, 20 mug of rhein, 100 mug of chrysophanic acid and 10 mug of chrysophanol reference substance per 1 ml.
As an improvement of the technical scheme, in the step (2), the extraction solvent is 20-100% methanol, the extraction time is 10-30 min, and the extraction mode is ultrasonic extraction.
As an improvement of the above technical solution, the step (2) includes:
taking 0.15-0.25 g of peach kernel qi-bearing soup freeze-dried powder, precisely weighing, placing into a conical bottle with a plug, adding 10mL of 70% methanol, weighing, respectively taking out, adopting ultrasonic waves with the power of 200-300W and the frequency of 40-50 kHz to treat for 20min, taking out, cooling, weighing again, supplementing the lost weight with 70% methanol, filtering, and taking out the subsequent filtrate to obtain the sample solution.
As an improvement of the technical scheme, the characteristic spectrum of the peach pit qi-bearing soup comprises 15 characteristic peaks; wherein, peak 1 is gallic acid peak, peak 2 is catechin peak, peak 3 is amygdalin peak, peak 4 is epicatechin gallate peak, peak 5 is glycyrrhizin peak, peak 6 is apigenin glycyrrhizin peak, peak 7 is cinnamaldehyde peak, peak 8 is cinnamic acid peak, peak 9 is chrysophanol-1-O-glucoside peak, peak 10 is chrysophanol-8-O-glucoside peak, peak 11 is emodin-8-O-glucoside peak, peak 12 is large Huang Suanfeng, peak 13 is glycyrrhizic acid peak, peak 14 is emodin peak, and peak 15 is chrysophanol peak.
As an improvement of the technical scheme, the peach pit qi-supporting soup comprises the following components in parts by weight: 15 parts of peach kernel, 55.2 parts of rheum officinale, 27.6 parts of cinnamon, 27.6g of liquorice and 27.6g of mirabilite.
As an improvement of the technical scheme, the preparation method of the peach pit qi-supporting soup comprises the following steps: soaking semen Persicae, radix et rhizoma Rhei, cortex Cinnamomi and Glycyrrhrizae radix in 1200-1500 mL water, boiling with strong fire, boiling with slow fire until the liquid medicine is 400-600 mL, filtering, adding Natrii sulfas, and heating and boiling.
As an improvement of the technical scheme, the preparation method of the peach pit qi-supporting soup comprises the following steps: soaking semen Persicae, radix et rhizoma Rhei, cortex Cinnamomi and Glycyrrhrizae radix in 1400mL water, boiling with strong fire, boiling with slow fire until the medicinal liquid is 500mL, filtering, adding Natrii sulfas, and heating and boiling.
The implementation of the invention has the following beneficial effects:
the invention establishes the characteristic spectrum of the peach pit qi-supporting soup, and totally calibrates 22 characteristic peaks, can fully display the characteristics of chemical components of the peach pit qi-supporting soup, has abundant information quantity of the characteristic peaks, and can comprehensively reflect the quality information of the peach pit qi-supporting soup, thereby achieving the purpose of comprehensively and effectively controlling the quality of the peach pit qi-supporting soup product. Meanwhile, the method is stable, accurate and reliable, and quality monitoring of characteristic components of a plurality of medicines is realized.
Drawings
FIG. 1 is a characteristic spectrum of the peach pit qi-supporting decoction according to the invention when the wavelength of 220nm is adopted for measurement;
FIG. 2 is a characteristic spectrum of the peach pit qi-supporting decoction according to the invention when measured at a wavelength of 260 nm;
FIG. 3 is a characteristic spectrum of the peach pit qi-supporting decoction according to the invention when measured with a wavelength of 290nm;
FIG. 4 is a characteristic spectrum of the peach pit qi-supporting decoction according to the invention when acetonitrile-phosphoric acid is used as a mobile phase for measurement;
FIG. 5 is a characteristic spectrum of the peach pit qi-supporting decoction according to the invention when methanol-water is used as a mobile phase for measurement;
FIG. 6 is a characteristic spectrum of the peach pit qi-supporting decoction according to the invention when methanol-phosphoric acid is used as a mobile phase for measurement;
FIG. 7 is a characteristic spectrum of the peach pit qi-supporting decoction according to the invention when different chromatographic columns are used for measurement;
FIG. 8 is a characteristic spectrum of the peach pit qi-supporting soup according to the invention when different column temperatures are used for measurement;
FIG. 9 is a characteristic spectrum of the peach pit qi-bearing soup according to the invention when different flow measurements are used;
FIG. 10 is a characteristic spectrum of peach kernel qi-supporting decoction, peach kernel control medicinal material and peach kernel deficiency negative sample in the specific investigation of the characteristic spectrum of the peach kernel qi-supporting decoction; wherein, peak 1 is gallic acid peak, peak 2 is catechin peak, peak 3 is amygdalin peak, peak 4 is epicatechin gallate peak, peak 5 is glycyrrhizin peak, peak 6 is apigenin glycyrrhizin peak, peak 7 is cinnamaldehyde peak, peak 8 is cinnamic acid peak, peak 9 is chrysophanol-1-O-glucoside peak, peak 10 is chrysophanol-8-O-glucoside peak, peak 11 is emodin-8-O-glucoside peak, peak 12 is large Huang Suanfeng, peak 13 is glycyrrhizic acid peak, peak 14 is emodin peak, and peak 15 is chrysophanol peak;
FIG. 11 is a characteristic spectrum of a peach pit decoction for qi, a rhubarb control drug and a rheum officinale negative sample in the specific investigation of the characteristic spectrum of the peach pit decoction for qi; wherein, peak 1 is gallic acid peak, peak 2 is catechin peak, peak 3 is amygdalin peak, peak 4 is epicatechin gallate peak, peak 5 is glycyrrhizin peak, peak 6 is apigenin glycyrrhizin peak, peak 7 is cinnamaldehyde peak, peak 8 is cinnamic acid peak, peak 9 is chrysophanol-1-O-glucoside peak, peak 10 is chrysophanol-8-O-glucoside peak, peak 11 is emodin-8-O-glucoside peak, peak 12 is large Huang Suanfeng, peak 13 is glycyrrhizic acid peak, peak 14 is emodin peak, and peak 15 is chrysophanol peak.
FIG. 12 is a characteristic spectrum of a peach pit decoction, a cinnamon control drug and a cinnamon deficiency negative sample in the specific investigation of the characteristic spectrum of the peach pit decoction; wherein, peak 1 is gallic acid peak, peak 2 is catechin peak, peak 3 is amygdalin peak, peak 4 is epicatechin gallate peak, peak 5 is glycyrrhizin peak, peak 6 is apigenin glycyrrhizin peak, peak 7 is cinnamaldehyde peak, peak 8 is cinnamic acid peak, peak 9 is chrysophanol-1-O-glucoside peak, peak 10 is chrysophanol-8-O-glucoside peak, peak 11 is emodin-8-O-glucoside peak, peak 12 is large Huang Suanfeng, peak 13 is glycyrrhizic acid peak, peak 14 is emodin peak, and peak 15 is chrysophanol peak;
FIG. 13 is a characteristic spectrum of a peach pit carrier qi decoction, a licorice control drug and a liquorice-deficient negative sample in the specific investigation of the characteristic spectrum of the peach pit carrier qi decoction; wherein, peak 1 is gallic acid peak, peak 2 is catechin peak, peak 3 is amygdalin peak, peak 4 is epicatechin gallate peak, peak 5 is glycyrrhizin peak, peak 6 is apigenin glycyrrhizin peak, peak 7 is cinnamaldehyde peak, peak 8 is cinnamic acid peak, peak 9 is chrysophanol-1-O-glucoside peak, peak 10 is chrysophanol-8-O-glucoside peak, peak 11 is emodin-8-O-glucoside peak, peak 12 is large Huang Suanfeng, peak 13 is glycyrrhizic acid peak, peak 14 is emodin peak, and peak 15 is chrysophanol peak;
fig. 14 is a characteristic spectrum superposition diagram of 20 batches of peach pit qi-bearing soup samples; wherein, peak 1 is gallic acid peak, peak 2 is catechin peak, peak 3 is amygdalin peak, peak 4 is epicatechin gallate peak, peak 5 is glycyrrhizin peak, peak 6 is apigenin glycyrrhizin peak, peak 7 is cinnamaldehyde peak, peak 8 is cinnamic acid peak, peak 9 is chrysophanol-1-O-glucoside peak, peak 10 is chrysophanol-8-O-glucoside peak, peak 11 is emodin-8-O-glucoside peak, peak 12 is large Huang Suanfeng, peak 13 is glycyrrhizic acid peak, peak 14 is emodin peak, and peak 15 is chrysophanol peak;
FIG. 15 is a graph showing a peak superposition of peach pit qi-supporting decoction characteristic spectrum reference chromatograms, wherein peak 1 is a gallic acid peak, peak 2 is a catechin peak, peak 3 is a amygdalin peak, peak 4 is an epicatechin gallate peak, peak 5 is a glycyrrhizin peak, peak 6 is a apigenin glycyrrhizin peak, peak 7 is a cinnamaldehyde peak, peak 8 is a cinnamic acid peak, peak 9 is a chrysophanol-1-O-glucoside peak, peak 10 is a chrysophanol-8-O-glucoside peak, peak 11 is a emodin-8-O-glucoside peak, peak 12 is a large Huang Suanfeng, peak 13 is a glycyrrhizic acid peak, peak 14 is a emodin peak, and peak 15 is a chrysophanol peak;
fig. 16 is a mass spectrum of total ion flow and a liquid chromatogram of the peach pit gas-bearing decoction sample solution.
Detailed Description
The present invention will be described in further detail with reference to the drawings and the detailed description, for the purpose of making the objects, technical solutions and advantages of the present invention more apparent.
The peach pit qi-supporting decoction is from typhoid fever theory and is generally considered to be composed of peach kernels, rheum officinale, cassia twig, honey-fried licorice root and mirabilite in the prior description. Based on ancient book documents, the determined peach pit qi-bearing soup prescription is as follows: 15.0g of peach kernel (mountain peach), 55.2g of rhubarb (palmleaf rhubarb), 27.6g of cinnamon, 27.6g of fried licorice and 27.6g of mirabilite. Wherein, each medicinal material is identified to meet the regulations of the relevant item of the Chinese pharmacopoeia of 2020 edition. It should be noted that Zhang Zhongjing is marked below the traditional Chinese medicine in the typhoid treatise, is closest to the stir-frying method in the modern pharmacopoeia, and stir-frying without adding auxiliary materials in ancient times is divided into two methods of stir-frying yellow and stir-frying charcoal, and by combining the 'Chinese pharmacopoeia' of 2020 edition and referring to the processing standards in other places, the peach kernel is defined as the boiled peach kernel or the mountain peach kernel, the rheum officinale is the rheum palmatum, the cassia twig is the cinnamon and the prepared liquorice is the stir-fried liquorice.
Further, the preparation method of the peach pit qi-bearing decoction traditional decoction based on ancient book literature examination is as follows: taking the four materials except mirabilite, adding 1400mL of water, soaking, boiling with strong fire, keeping boiling with slow fire to about 500mL of liquid medicine, filtering with a screen, adding mirabilite into the filtrate, and heating to slight boiling to obtain the peach pit qi-supporting decoction.
The peach pit qi-bearing soup samples adopted in the invention are all peach pit qi-bearing soup freeze-dried powder, specifically, the peach pit qi-bearing soup is stirred evenly and split-packed in brown penicillin bottles, and the penicillin bottles are transferred into a vacuum circulation pump vacuum freeze dryer for freeze drying, and taken out to obtain the freeze-dried powder, thus obtaining the peach pit qi-bearing soup freeze-dried powder. In addition, in order to comprehensively reflect the quality information of the peach pit qi-bearing soup, the inventor collects not less than 15 batches of each medicinal material at 3 places of production and prepares samples for research.
In order to comprehensively reflect the quality information of the peach pit qi-supporting soup and realize comprehensive and effective control of the quality of the peach pit qi-supporting soup product, the invention provides a method for establishing a characteristic spectrum of the peach pit qi-supporting soup, which is described in detail below:
1 instrument and reagent
The instrument, reagent and reagent information used in the invention are shown in tables 1 to 3:
table 1 instrument information summary table
TABLE 2 summary of reagent information
TABLE 3 control information
TABLE 4 20 batch corresponding physical decoction piece formulation information table
2 chromatographic conditions and preparation of reference solution and test solution
2.1 chromatographic conditions
Chromatographic conditions: octadecylsilane chemically bonded silica is used as filler (column length 150mm, inner diameter 2.1mm, particle diameter 1.6 μm, chromatographic column: waters CORTECS T3 column); methanol is taken as a mobile phase A,0.2% phosphoric acid aqueous solution is taken as a mobile phase B, and gradient elution is carried out according to the regulations in the table; column temperature is 38 ℃; the flow rate is 0.3ml per minute; the sample injection amount is 1 mul, the detection wavelength is 220nm in 0-30 min, 290nm in 30-45 min and 260nm in 45-65 min. The theoretical plate number should be not less than 10000 calculated according to catechin peaks.
TABLE 5 gradient elution table
2.2 preparation of control solution
Precisely weighing appropriate amounts of gallic acid reference, catechin reference, amygdalin reference, epicatechin gallate reference, glycyrrhizin reference, apigenin reference, cinnamaldehyde reference, cinnamic acid reference, chrysophanol-1-O-glucoside reference, chrysophanol-8-O-glucoside reference, emodin-8-O-glucoside reference, rhein reference, glycyrrhizic acid reference, emodin reference, and chrysophanol reference, precisely weighing, and adding methanol to obtain solution containing gallic acid 15 μg, catechin 100 μg, amygdalin 30 μg, epicatechin gallate 50 μg, glycyrrhizin 30 μg, apigenin 70 μg, cinnamaldehyde 10 μg, cinnamic acid 10 μg, chrysophanol-1-O-glucoside 30 μg, chrysophanol-8-O-glucoside 40 μg, emodin-8-O-glucoside 50 μg, rhein 20 μg, rhein 100 μg, and chrysophanol 10 μg per 1 ml.
2.3 preparation of sample solutions
Taking about 0.25g of peach kernel qi-bearing soup freeze-dried powder, grinding, precisely weighing, placing into a conical bottle with a plug, precisely adding 10mL of 70% methanol, performing ultrasonic treatment (power is 250W, frequency is 45 kHz) for 20 minutes, taking out, cooling, weighing again, supplementing the weight of loss with 70% methanol, shaking uniformly, filtering, and taking subsequent filtrate.
2.4 assay
Precisely sucking 1 μl of each of the control solution and the sample solution, and measuring with a liquid chromatograph. The characteristic spectrum of the sample should show chromatographic peaks corresponding to the retention time of the chromatographic peaks of the reference substance. And calculating according to a traditional Chinese medicine chromatographic characteristic spectrum similarity evaluation system, wherein the similarity between the characteristic spectrum of the sample to be tested and the contrast characteristic spectrum is not lower than 0.90. 15 characteristic peaks should be presented in the characteristic spectrum of the sample; wherein, peak 1 is gallic acid peak, peak 2 is catechin peak, peak 3 is amygdalin peak, peak 4 is epicatechin gallate peak, peak 5 is glycyrrhizin peak, peak 6 is apigenin glycyrrhizin peak, peak 7 is cinnamaldehyde peak, peak 8 is cinnamic acid peak, peak 9 is chrysophanol-1-O-glucoside peak, peak 10 is chrysophanol-8-O-glucoside peak, peak 11 is emodin-8-O-glucoside peak, peak 12 is large Huang Suanfeng, peak 13 is glycyrrhizic acid peak, peak 14 is emodin peak, and peak 15 is chrysophanol peak.
3. Determination of chromatographic conditions
3.1 Determination of the optimal absorption wavelength
Examining different absorption wavelengths; the absorption wavelengths are 220nm, 260nm and 290nm respectively. A Waters CORTECS T3 (2.1X105 mm,1.6 μm) column was used; gradient elution was performed with methanol as mobile phase a and 0.2% phosphoric acid as mobile phase B, as specified in table 5; the flow rate is 0.3ml per minute; the sample injection amount is 1 μl; the results are shown in FIGS. 1 to 3.
Experimental results show that through full wavelength scanning, more chromatographic peaks in the chromatogram of the sample have higher response values, but the chromatographic peaks of cinnamaldehyde and cinnamic acid have maximum absorption at 290nm, and factors such as the number, the abundance, the separation degree and the baseline drift condition of each chromatographic peak of the obtained chromatogram under different wavelengths are comprehensively considered, and finally the detection wavelength is 220nm when the test time is 0-30 minutes; when the test time is 30-45 minutes, the detection wavelength is 290nm; when the test time is 45-65 minutes, the detection wavelength is 260nm.
3.2 investigation of different mobile phases
The mobile phase type was examined by selecting acetonitrile-phosphoric acid, methanol-water, methanol-0.1 vol% phosphoric acid aqueous solution, and methanol-0.2 vol% phosphoric acid aqueous solution as mobile phases, respectively, and using the gradient elution procedure as in table 5. A Waters CORTECS T3 (2.1X105 mm,1.6 μm) column was used; the flow rate is 0.3ml per minute; the sample injection amount is 1 μl; the detection wavelength is 220nm when the test time is 0-30 minutes; when the test time is 30-45 minutes, the detection wavelength is 290nm; when the test time is 45-65 minutes, the detection wavelength is 260nm. The results are shown in FIGS. 4 to 6.
As can be seen from the figure, acetonitrile-phosphoric acid is adopted as a mobile phase, the chromatographic components of the sample have earlier peak separation, and the chromatographic peak separation is poor; methanol-water is used as a mobile phase, so that the chromatographic peak is fewer and the separation degree is poor; methanol-phosphoric acid is used as a mobile phase, so that the chromatogram information is rich, the chromatographic peak separation degree is good, the base line is stable, and the peak outlet time is proper. Finally, methanol was used as mobile phase A and 0.2% phosphoric acid aqueous solution was used as mobile phase B, and gradient elution was performed according to the procedure of Table 5.
3.3 investigation of different chromatographic columns
The effect of three columns of Waters CORTECS T3 (1.6 μm,150 mm. Times.2.1 mm), phenomnex Omega 1.6 μm PS C18 100A (1.6 μm,150 mm. Times.2.1 mm) and Waters ACQUITY UPLC HSS T3 (1.8 μm,150 mm. Times.2.1 mm) on the chromatographic behavior of the peach kernel gas bearing soup feature profile was compared, respectively. In addition to chromatographic columns, other test conditions are described in section 2.1. The results are shown in FIG. 7.
Experimental results show that different chromatographic columns have great influence on the peak numbers, peak shapes and peak separation degrees of the characteristic spectrum of the peach kernel support qi decoction, a Waters CORTECS T3 column (1.6 mu m,150mm multiplied by 2.1 mm) is used as the chromatographic column, the peak numbers of the chromatographic peaks are slightly more, the peak shapes are better, and the base line is stable, so that the Waters CORTECS T3 column (1.6 mu m,150mm multiplied by 2.1 mm) is selected as the characteristic spectrum research chromatographic column of the peach kernel support qi decoction.
3.4 investigation of different column temperatures
The influence of the column temperature of 30 ℃, 35 ℃, 38 ℃ and 40 ℃ on the characteristic spectrum of the peach pit qi-bearing soup is examined respectively. In addition to column temperature, other test conditions were as described in section 2.1. The results are shown in FIG. 8.
The experiment result shows that different chromatographic column temperatures have certain influence on the chromatographic behavior of the peach pit qi-supporting decoction characteristic spectrum, and the conditions of chromatographic peak shape, peak separation degree, baseline and the like are comprehensively considered, so that the chromatographic column temperature of the peach pit qi-supporting decoction characteristic spectrum research is determined to be 38 ℃.
3.5 investigation of flow Rate
The effect of the flow rates of 0.25ml per minute, 0.3ml per minute and 0.35ml per minute on the chromatographic behavior of the peach pit qi-bearing decoction characteristic spectrum was examined respectively. In addition to flow rate, other test conditions are as described in section 2.1. The results are shown in FIG. 9.
The results show that different flow rates have certain influence on the chromatographic behavior of the characteristic spectrum of the peach pit qi-supporting decoction, the conditions of chromatographic peak separation degree, column pressure and the like are comprehensively considered, and the flow rate of the mobile phase of the characteristic spectrum of the peach pit qi-supporting decoction is studied and determined to be 0.3ml per minute.
3.6 determination of chromatographic conditions
According to the above experiments, the chromatographic conditions were determined as follows: octadecylsilane chemically bonded silica is used as filler (Waters CORTECS T3 column, column length of 150mm, inner diameter of 2.1mm, particle diameter of 1.6 μm); gradient elution was performed using methanol as mobile phase a and 0.2% phosphoric acid aqueous solution as mobile phase B, as specified in table 5 above; the detection wavelength is 220nm for 0-30 min, 290nm for 30-45 min, 260nm for 45-65 min, and the column temperature is 38 ℃; the flow rate was 0.3ml per minute.
4 investigation of sample solution preparation method
4.1 extraction solvent investigation
The influence of different extraction solvents on the characteristic spectrum of the peach pit qi-bearing decoction is examined respectively, 20% methanol, 50% methanol, 70% methanol and 100% methanol are selected as the extraction solvents, and the characteristic spectrum results of the different extraction solvents are compared.
The preparation method of the sample solution comprises the following steps: taking about 0.25g of peach kernel qi-bearing soup freeze-dried powder preparation (batch number: S9), precisely weighing, placing into a conical bottle with a plug, precisely adding 20% methanol, 50% methanol, 70% methanol and 100% methanol 10ml respectively, weighing, performing ultrasonic treatment (power 250W and frequency 45 kHz) for 20 minutes, taking out, cooling, weighing again, supplementing the lost weight with corresponding extraction solvent, filtering, and taking the subsequent filtrate.
The chromatographic conditions are as follows: gradient elution was performed as specified in Table 5 using a Waters CORTECS T3 column (1.6 μm,150 mm. Times.2.1 mm) as the column, methanol as mobile phase A, and 0.2% phosphoric acid as mobile phase B; column temperature is 38 ℃; the flow rate is 0.3ml per minute; the detection wavelength is 220nm when the test time is 0-30 minutes; when the test time is 30-45 minutes, the detection wavelength is 290nm; when the test time is 45-65 minutes, the detection wavelength is 260nm.
The results of different extraction solvent investigation of the peach pit qi-bearing decoction feature maps are shown in Table 6.
Table 6 table of results of solvent extraction tests on peach pit qi-supporting decoction feature patterns
Experimental results: by comparing the characteristic patterns of the 4 extraction solvents, when the extraction solvent is methanol, the cinnamic aldehyde chromatograph is not obvious, and when 20 percent of methanol, 50 percent of methanol and 70 percent of methanol are used as the extraction solvents, the peak numbers of the characteristic patterns of the peach pit qi-supporting soup are the same, wherein when 70 percent of methanol is used as the extraction solvent, the total peak area/sample weighing amount is larger, and the characteristic patterns of the peach pit qi-supporting soup can be fully represented, so that 70 percent of methanol is used as the extraction solvent of the characteristic patterns of the peach pit qi-supporting soup.
4.2 extraction time investigation
And (3) observing the influence of different extraction times on the characteristic spectrum of the peach pit qi-bearing soup, and selecting three different extraction times of 10 minutes, 20 minutes and 30 minutes.
Taking about 0.25g of peach pit qi-bearing soup freeze-dried powder preparation (batch number: S9), precisely weighing, placing into conical bottles with plugs, precisely adding 10ml of 70% methanol respectively, weighing, respectively taking out, performing ultrasonic treatment (power 250W, frequency 45 kHz) for 10 minutes, 20 minutes and 30 minutes, taking out, cooling, weighing again, supplementing the lost weight with 70% methanol, filtering, and taking out the subsequent filtrate.
Chromatographic conditions: gradient elution was performed as specified in Table 5 using a Waters CORTECS T3 column (1.6 μm,150 mm. Times.2.1 mm) as the column, methanol as mobile phase A, and 0.2% phosphoric acid as mobile phase B; column temperature is 38 ℃; the flow rate is 0.3ml per minute; the detection wavelength is 220nm when the test time is 0-30 minutes; when the test time is 30-45 minutes, the detection wavelength is 290nm; when the test time is 45-65 minutes, the detection wavelength is 260nm.
The results of different extraction solvent investigation of the peach pit qi-bearing decoction feature maps are shown in Table 7.
TABLE 7 peach pit qi-bearing decoction feature map extraction time investigation result table
The results show that: by comparing the influence of different extraction times on the characteristic spectrum of the peach pit qi-bearing decoction, the total peak area/sample weighing of ultrasonic extraction for 10 minutes and 30 minutes is smaller than that of ultrasonic extraction for 20 minutes, and ultrasonic extraction is selected for 20 minutes in order to ensure that the peach pit qi-bearing decoction is completely extracted without loss of components.
4.3 determination of the method for preparing the sample solution
According to the experimental results, the pretreatment method of the peach pit qi-bearing soup characteristic spectrum sample is determined as follows:
taking about 0.25g of peach kernel qi-bearing soup freeze-dried powder, precisely weighing, placing into a conical bottle with a plug, precisely adding 10ml of 70% methanol, performing ultrasonic treatment (power is 250W and frequency is 45 kHz) for 20 minutes, taking out, cooling, weighing again, supplementing the weight of loss with 70% methanol, shaking uniformly, filtering, and taking the subsequent filtrate.
5. Methodological verification
5.1 Investigation of specificity
And respectively taking peach pit qi-supporting soup without each medicine, and preparing according to a preparation method of a sample solution to obtain a negative sample solution without each medicine.
Taking reference medicinal materials of peach kernel, rhubarb, cinnamon and liquorice respectively, and preparing reference medicinal material reference solution of each medicinal material according to a preparation method of the reference medicinal material reference solution of the reference medicinal material.
And (3) taking a proper amount of gallic acid reference substance, catechin reference substance, amygdalin reference substance, epicatechin gallate reference substance, glycyrrhizin reference substance, apigenin reference substance, glycyrrhizic acid reference substance, cinnamaldehyde reference substance, cinnamic acid reference substance, rhein reference substance, emodin reference substance and chrysophanol reference substance, precisely weighing, and respectively adding methanol to prepare a solution containing 15 mug of gallic acid, 100 mug of catechin, 30 mug of amygdalin, 50 mug of epicatechin gallate, 30 mug of glycyrrhizin, 70 mug of apioside, 100 mug of glycyrrhizic acid, 10 mug of cinnamaldehyde, 10 mug of cinnamic acid, 20 mug of rhein, 10 mug of emodin and 10 mug of chrysophanol per 1ml of reference substance, thereby obtaining a reference substance solution of each reference substance.
1 μl of each of the sample solution, the negative sample solution lacking each medicinal material, the reference substance solution of each medicinal material, and the reference substance solution of each reference substance is injected into a liquid chromatograph, and the sample is injected and analyzed under the chromatographic condition of section 2.1, and the results are shown in fig. 10-13.
As can be seen from fig. 10 to 13: 15 common chromatographic peaks of the peach kernel qi-supporting decoction preparation characteristic spectrum are detected, wherein 1 peak (No. 3 peak) is derived from peach kernel: 10 peaks ( peaks 1, 2, 4, 8, 9, 10, 11, 12, 14 and 15) are derived from rheum officinale, 3 peaks ( peaks 2, 7 and 8) are derived from cinnamon, 3 peaks ( peaks 5, 6 and 13) are derived from liquorice, and 0 peak is derived from mirabilite; 1 peak (No. 2 peak, catechin) is common to radix et rhizoma Rhei and cortex Cinnamomi, and 1 peak (No. 8 peak, cinnamic acid) is common to radix et rhizoma Rhei and cortex Cinnamomi; the established characteristic spectrum can reflect all medicinal components in the prescription except for the mineral medicine mirabilite.
In addition, as can be seen from fig. 10 to 13, the sample chromatogram has the same chromatographic peak at the retention time corresponding to the control chromatogram, and the negative is not interfered, thus demonstrating that the method has good specificity.
5.2 integrity inspection
The preparation method comprises the steps of precisely sucking 1 mu l of sample solution of peach kernel qi-bearing decoction freeze-dried powder preparation (batch number: S9), injecting into an ultra-high liquid chromatograph, measuring according to determined chromatographic conditions (section 2.1), maintaining the same gradient, prolonging elution time by one time under the mobile phase proportion of a gradient end point, analyzing a characteristic map, and showing that no hysteresis peak appears, thus the method has good integrity.
5.3 precision investigation
Taking a sample solution of a peach kernel qi-bearing soup freeze-dried powder preparation (batch number: S9), repeatedly sampling for 6 times according to a determined chromatographic condition (section 2.1), taking epicatechin gallate peaks as reference peaks in a sampling volume of 1 mul, calculating the relative retention time and relative peak area of each common characteristic peak, calculating RSD values, and simultaneously adopting traditional Chinese medicine chromatographic characteristic spectrum similarity (2012 edition) evaluation software to calculate the similarity of each characteristic spectrum, wherein the RSD of each common characteristic peak relative retention time is 0.04% -0.39%, the RSD of each common characteristic peak relative peak area is 0.12% -1.36%, and the similarity is 1.000, so that the instrument precision is good, and the experimental results are shown in tables 8-10.
Table 8 results table (relative retention time) for examining precision of peach pit qi-bearing decoction feature pattern
Table 9 precision investigation results table (relative peak area) of peach pit qi-bearing decoction characteristic spectrum
Table 10 similarity analysis results of peach pit qi-supporting decoction characteristic spectrum precision test
5.4 stability investigation
Sample injection is carried out on sample solutions of peach kernel qi-bearing soup freeze-dried powder preparations (batch number: S9) according to determined chromatographic conditions (section 2.1) for 0, 3, 6, 9, 12, 15, 18, 21 and 24 hours, the sample injection volume is 1 mul, epicatechin gallate peaks are taken as reference peaks, the relative retention time and the relative peak area of each common characteristic peak are calculated, RSD values are calculated, meanwhile, the similarity of each characteristic spectrum is calculated by adopting traditional Chinese medicine chromatographic characteristic spectrum similarity (2012 edition) evaluation software, the RSD of each common characteristic peak relative retention time is 0.05% -0.43%, the RSD of the relative peak area is 0.80% -2.96%, the similarity is 1.000, the stability of the sample solutions in 24 hours is good, and the experimental results are shown in tables 11-13.
Table 11 results table (relative retention time) for investigating the stability of the Taohuanqi decoction feature map
Table 12 results table (relative peak area) for investigating stability of peach pit qi-supporting decoction characteristic spectrum
TABLE 13 similarity analysis results of peach pit qi-supporting decoction characteristic spectrum stability test
5.5 repeatability investigation
About 0.25g of the same batch of peach pit qi-bearing soup freeze-dried powder preparation (batch number: S9) is taken, precisely weighed, 6 parts are parallel, a sample solution (section 2.3) is prepared according to a determined method, and 1 μl is injected according to a determined chromatographic condition (section 2.1). The relative retention time and the relative peak area of each common characteristic peak are calculated by taking the epicatechin gallate peak as a reference peak, the RSD value is calculated, meanwhile, the similarity of each characteristic spectrum is calculated by adopting Chinese medicine chromatographic characteristic spectrum similarity (2012 edition) evaluation software, and as a result, the RSD of each common characteristic peak relative retention time is 0.00-0.34%, the RSD of the relative peak area is 1.06-3.95%, and the similarity is 1.000, which indicates that the method has good repeatability. The results of the experiments are shown in tables 14 to 16.
Table 14 peach pit qi-bearing decoction feature map repeatability test results table (relative retention time)
Table 15 peach pit qi-bearing decoction feature map repeatability test results table (relative peak area)
Table 16 similarity analysis results of characteristic spectrum repeatability test of peach pit qi-supporting soup
6 determination of samples of different batches and determination of the Co-Peak
Taking 20 batches of peach pit qi-bearing soup corresponding to the real objects, preparing the test solution according to a determined test solution preparation method (section 2.3), and respectively injecting 1 μl according to a determined chromatographic condition (section 2.1), and measuring. And (3) taking epicatechin gallate peaks as reference peaks, calculating the relative retention time and the relative peak area of each common characteristic peak, calculating RSD values, and simultaneously adopting traditional Chinese medicine chromatographic characteristic spectrum similarity (2012 edition) evaluation software to calculate the similarity of each characteristic spectrum. The results are shown in tables 17 to 19, and the chromatograms are shown in FIG. 14.
The result shows that the relative retention time of the common characteristic peak has the RSD of 0.00-0.19 percent, and the RSD of the relative peak area of the common characteristic peak has the RSD of 0.00-51.70 percent because of certain difference of the content of the corresponding components of the common characteristic peak of each batch of samples, the deviation is larger, and the similarity range of the characteristic spectrum of the 20 batches of nuclear gas-bearing soup preparation is 0.937-0.998 and is larger than 0.9. And analyzing the characteristic spectrum of 20 batches of peach pit qi-bearing decoction preparations to determine 15 common characteristic peaks.
Table 17-20 batch peach pit qi-bearing decoction freeze-dried powder preparation characteristic spectrum similarity table
Table 18 results of relative retention time of characteristic spectrum of 20 batches of peach pit qi-bearing decoction freeze-dried powder preparation
Table 19 results of the relative peak areas of the characteristic maps of the 20 batches of peach pit qi-bearing decoction freeze-dried powder preparation
7 chemical composition test study of characteristic map
And identifying the chemical components contained in the peach pit qi-supporting decoction by adopting high-resolution mass spectrometry according to the chemical components contained in each medicinal flavor in the peach pit qi-supporting decoction.
7.1 high resolution Mass Spectrometry Specification
Mass spectrometry conditions: waters ACQUITY UPLCTM I-Class type ultra-high performance liquid chromatograph; the Wolth ultra-high performance liquid phase time-of-flight high resolution mass spectrometry system (Xex G2-XS QTOF MS) and the data processing system is a UNIFI 1.8 workstation (Waters, manchester, U.K.); waters Xselected HSS T3 (column length 150mm, inner diameter 2.1mm, particle size 2.5 μm). Nitrogen is used as atomizing and taper hole gas of a mass spectrum ion source; electrospray ionization positive ion mode; capillary voltage: 2.0KV; taper hole voltage: 40V; ion source temperature: 120 ℃; desolventizing gas temperature: 450 ℃; desolventizing gas flow rate: 800L/h; scanning time: 0.5s; scanning time interval: 0.02s; mass-to-charge ratio range: 50-1200; data acquisition mode: MSE is collected under ESI (+); correcting a mass axis by using a sodium formate solution, and correcting mass accuracy by using Leucine enkephalin (Leucine-enkephalin) as an internal standard, wherein the gas flow of a taper hole is as follows: 50L/h.
Chromatographic conditions: octadecylsilane chemically bonded silica is used as filler (Waters Xselected HSS T column, column length 150mm, inner diameter 2.1mm, particle diameter 2.5 μm); gradient elution was performed as specified in table 20 with methanol as mobile phase a and aqueous water (containing 2mmol/L ammonium acetate and 0.05% formic acid) as mobile phase B; column temperature is 40 ℃; the flow rate is 0.35ml per minute; the sample injection amount is 1 μl, and the wavelength is 260nm.
Preparation of a control solution: 2.2 items;
preparation of test solution: the same as in item 2.3.
Table 20 chromatographic assignment study gradient elution program table
The identification results are shown in Table 21, and the chromatograms are shown in FIG. 15 and FIG. 16.
Table 21 identification results of characteristic spectrum components of peach pit qi-supporting decoction
In conclusion, the invention establishes the characteristic spectrum of the peach pit qi-supporting soup, the characteristic spectrum can fully display the chemical component characteristics of the peach pit qi-supporting soup, the characteristic peak information quantity is rich, and the quality information of the peach pit qi-supporting soup can be comprehensively reflected, so that the purpose of comprehensively and effectively controlling the quality of the peach pit qi-supporting soup product can be achieved. Meanwhile, the characteristic map construction method provided by the invention has the advantages of good repeatability, accuracy, reliability and good stability.
While the foregoing is directed to the preferred embodiments of the present invention, it will be appreciated by those skilled in the art that changes and modifications may be made without departing from the principles of the invention, such changes and modifications are also intended to be within the scope of the invention.
Claims (10)
1. The construction method of the peach pit qi-bearing soup characteristic spectrum is characterized by comprising the following steps of:
(1) Respectively taking gallic acid reference substance, catechin reference substance, amygdalin reference substance, epicatechin gallate reference substance, glycyrrhizin reference substance, apiose glycyrrhizin reference substance, cinnamaldehyde reference substance, cinnamic acid reference substance, chrysophanol-1-O-glucoside reference substance, chrysophanol-8-O-glucoside reference substance, emodin-8-O-glucoside reference substance, rhein reference substance, glycyrrhizic acid reference substance, emodin reference substance, and appropriate amount of chrysophanol reference substance, adding solvent, dissolving or extracting to obtain reference substance solution;
(2) Extracting semen Persicae decoction with solvent to obtain sample solution; the extraction solvent is 20-70% methanol;
(3) Taking a preset amount of reference substance solution and test substance solution, and injecting into a liquid chromatograph, wherein a chromatographic column of the liquid chromatograph is a Waters CORTECS T3 column, the column length is 150mm, the inner diameter is 2.1mm, and the particle size is 1.6 mu m; the liquid chromatograph uses methanol as a mobile phase A and a 0.15% -0.25% phosphoric acid aqueous solution as a mobile phase B to perform gradient elution, and a characteristic map of the peach pit qi-supporting soup is established;
wherein the gradient elution is performed according to the following procedure:
0-5 min, wherein the mobile phase A is from 1% to 8%, and the mobile phase B is from 99% to 92%;
5-21 min, wherein the mobile phase A is from 8% to 23%, and the mobile phase B is from 92% to 77%;
21-35 min, 23% -31% of mobile phase A and 77% -69% of mobile phase B;
35-45 min, mobile phase A from 31% -55%, mobile phase B from 69% -45%;
45-51 min, wherein the mobile phase A is from 55% to 65%, and the mobile phase B is from 45% to 35%;
51-56 min, mobile phase A from 65% -83%, mobile phase B from 35% -17%;
56-65 min, 83% -100% of mobile phase A and 17% -0% of mobile phase B;
wherein, when the detection time is 0-30 minutes, the detection wavelength is 220nm; the detection wavelength is 290nm when the detection time is 30-45 minutes, and 260nm when the detection time is 45-65 minutes.
2. The method for constructing the peach pit qi-bearing soup feature map according to claim 1, wherein in the step (3), 1-3 μl of each of a reference substance solution and a test substance solution is sucked and injected into a liquid chromatograph for detection; wherein the column temperature of the chromatographic column is 30-38 ℃; the liquid chromatograph takes methanol as a mobile phase A, takes 0.15% -0.25% phosphoric acid aqueous solution as a mobile phase B, and the flow rate is 0.25-0.36 mL/min.
3. The method for constructing a characteristic spectrum of the peach pit qi-bearing decoction according to claim 1, wherein in the step (3), 1 μl of each of a reference substance solution and a test substance solution is sucked and injected into a liquid chromatograph for detection, wherein the column temperature of a chromatographic column of the liquid chromatograph is 38 ℃; the liquid chromatograph takes methanol as a mobile phase A, takes 0.2% phosphoric acid aqueous solution as a mobile phase B, and has the flow rate of 0.3mL/min.
4. The method for constructing a characteristic spectrum of peach pit qi-bearing soup according to claim 1, wherein in the step (1), a gallic acid reference, a catechin reference, an amygdalin reference, an epicatechin gallate reference, a glycyrrhizin reference, a apioside glycyrrhizin reference, a cinnamaldehyde reference, a cinnamic acid reference, a chrysophanol-1-O-glucoside reference, a chrysophanol-8-O-glucoside reference, a emodin-8-O-glucoside reference, a rhein reference, a glycyrrhizic acid reference, a emodin reference and a chrysophanol reference are respectively taken, and methanol is respectively added into each 1mL of a solution containing 15 mug of gallic acid, 100 mug of catechin, 30 mug of amygdalin, 50 mug of epicatechin gallate, 30 mug of glycyrrhizin, 70 mug of apioside glycyrrhizin, 10 mug of cinnamaldehyde, 10 mug of cinnamic acid, 30 mug of chrysophanol-1-O-glucoside, 40 mug of chrysophanol-8-O-glucoside, 50 mug of chrysophanol-50 mug, 10 mug of chrysophanol reference solution, and 10 mug of chrysophanol reference solution are respectively obtained.
5. The method for constructing the peach pit qi-bearing soup feature map according to claim 1, wherein in the step (2), the extraction time is 10-30 min, and the extraction mode is ultrasonic extraction.
6. The method for constructing the peach pit qi-bearing soup feature map as claimed in claim 1, wherein the step (2) comprises:
taking 0.15-0.25 g of peach kernel qi-bearing soup freeze-dried powder, precisely weighing, placing into a conical bottle with a plug, adding 10mL of 70% methanol, weighing, respectively taking out, adopting ultrasonic waves with the power of 200-300W and the frequency of 40-50 kHz to treat for 20min, taking out, cooling, weighing again, supplementing the lost weight with 70% methanol, filtering, and taking out the subsequent filtrate to obtain the sample solution.
7. The method for constructing the peach pit qi-bearing decoction feature map according to claim 1, wherein the peach pit qi-bearing decoction feature map comprises 15 feature peaks; wherein, peak 1 is gallic acid peak, peak 2 is catechin peak, peak 3 is amygdalin peak, peak 4 is epicatechin gallate peak, peak 5 is glycyrrhizin peak, peak 6 is apigenin glycyrrhizin peak, peak 7 is cinnamaldehyde peak, peak 8 is cinnamic acid peak, peak 9 is chrysophanol-1-O-glucoside peak, peak 10 is chrysophanol-8-O-glucoside peak, peak 11 is emodin-8-O-glucoside peak, peak 12 is large Huang Suanfeng, peak 13 is glycyrrhizic acid peak, peak 14 is emodin peak, and peak 15 is chrysophanol peak.
8. The method for constructing the peach pit qi-supporting soup feature map according to claim 1, wherein the peach pit qi-supporting soup is composed of the following components in parts by weight: 15 parts of peach kernel, 55.2 parts of rheum officinale, 27.6 parts of cinnamon, 27.6 parts of liquorice, and 27.6 parts of mirabilite.
9. The method for constructing the peach pit qi-supporting soup feature map according to claim 1, wherein the preparation method of the peach pit qi-supporting soup is as follows: soaking semen Persicae, radix et rhizoma Rhei, cortex Cinnamomi and Glycyrrhrizae radix in 1200-1500 mL of water, boiling with strong fire, boiling with slow fire until the liquid medicine is 400-600 mL, filtering, adding Natrii sulfas, and heating and boiling.
10. The method for constructing the peach pit qi-supporting soup feature map according to claim 9, wherein the preparation method of the peach pit qi-supporting soup is as follows: soaking semen Persicae, radix et rhizoma Rhei, cortex Cinnamomi and Glycyrrhrizae radix in 1400mL water, boiling with strong fire, boiling with slow fire until the medicinal liquid is 500mL, filtering, adding Natrii sulfas, and heating and boiling.
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