CN115902052B - HPLC (high Performance liquid chromatography) characteristic spectrum detection method for Russian cherry and preparation thereof - Google Patents

HPLC (high Performance liquid chromatography) characteristic spectrum detection method for Russian cherry and preparation thereof Download PDF

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CN115902052B
CN115902052B CN202310223753.8A CN202310223753A CN115902052B CN 115902052 B CN115902052 B CN 115902052B CN 202310223753 A CN202310223753 A CN 202310223753A CN 115902052 B CN115902052 B CN 115902052B
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罗枫
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Sichuan Zhijiacheng Biotechnology Co ltd
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Abstract

The invention belongs to the field of medicine analysis, and particularly discloses an HPLC (high Performance liquid chromatography) characteristic spectrum detection method of Russian cherry and a preparation thereof, which comprises the following steps: 1) Preparing a reference substance solution; 2) Preparing a sample solution; 3) And respectively sucking the reference substance solution and the sample solution, injecting into a high performance liquid chromatograph, and recording chromatograms. According to the HPLC characteristic spectrum detection method for the Russian ginseng fruits and the preparation thereof, 8 characteristic peaks can be calibrated through the characteristic spectrum established under specific chromatographic conditions, particularly, the chromatographic peaks of the organic acid components of the lycium barbarum acid and the L-malic acid which are special in the Russian ginseng fruits can be identified, the quality control of the Russian ginseng fruits and the preparation thereof is realized through the calibration of the chemical components which are more special, the clinical curative effect of the Russian ginseng fruits is ensured, and the practical popularization and application values are realized.

Description

HPLC (high Performance liquid chromatography) characteristic spectrum detection method for Russian cherry and preparation thereof
Technical Field
The invention relates to the field of medicine analysis, in particular to an HPLC (high Performance liquid chromatography) characteristic spectrum detection method for Russian cherry and a preparation thereof.
Background
The Russian cherry is derived from Malus asiatica of Malus of RosaceaeMalus toringoides(Red.) Hughes or Malus hupehensisMalus transitoria(Batal.) Schneid. The dried mature fruit is a common medicinal material in the folk of Ganmai Tibetan, and has long medicinal and edible history, and has the main effects of clearing lung-heat, eliminating (resolving) phlegm, invigorating stomach, lowering blood pressure, etc. In order to facilitate clinical use and improve patient compliance, russian cherry formula granules are prepared at present.
The traditional Chinese medicine formula particles are different from decoction pieces and traditional decoction in existence form, the preparation process is different, and the quality control requirement cannot be met by only carrying out quality evaluation on a certain component in the aspect of quality control; meanwhile, the correlation of the components between the decoction pieces and the traditional Chinese medicine formula particles cannot be evaluated, so that a characteristic map is required to be added in the quality control of the traditional Chinese medicine formula particles specified by the national bureau for integrally evaluating the quality of the formula particles and comprehensively analyzing the decoction pieces. Patent CN 109060985A discloses an HPLC fingerprint of russian fruits, which can separate out 11 characteristic peaks and identify 6 characteristic components of chlorogenic acid, hyperin, isoquercitrin, quercitrin, phlorizin and quercetin. Proved by experiments, the method can not separate important characteristic components L-malic acid and matrimony vine acid from Russian cherry. Because the total acid content in the russian fruits reaches more than 5%, L-malic acid and medlar acid are unique organic acid components in the russian fruits, it is necessary to develop an HPLC fingerprint detection method of the russian fruit formula particles with good separation degree and stronger specificity.
Disclosure of Invention
In order to solve the problems, the invention provides an HPLC characteristic spectrum detection method for Russian cherry and a preparation thereof, which comprises the following steps:
1) Preparation of a control solution: dissolving reference substances of Lycii acid and L-malic acid in water to obtain reference substance solution;
2) Sample solution preparation: extracting a sample to be detected with water, filtering the extracting solution, and obtaining filtrate to obtain a sample solution;
3) Respectively sucking the reference substance solution and the sample solution, injecting into a high performance liquid chromatograph, and recording chromatograms; the chromatographic conditions were as follows:
chromatographic column: octadecylsilane chemically bonded silica is used as a filler; mobile phase: acetonitrile is taken as a mobile phase A, and a 0.1% phosphoric acid solution is taken as a mobile phase B; gradient elution procedure 0-18 min,1% →3% A; 18-18.5 min,3% -4% A; 18.5-25 min, 4%. Fwdarw. 7%A; 25-30 min,7% -15% A; 30-35 min,15% -31% A.
Further, each 1ml of the reference solution in the step 1) contains 0.1-0.5 mg of medlar acid and 0.1-1 mg of L-malic acid.
Further, in the step 2), the mass-volume ratio of the sample to be detected to water is 0.5-1 g: 5-50 ml; the water is distilled water.
Further, the sample to be detected is Russian fruit medicinal material, russian fruit standard decoction or Russian fruit formula particles.
Further, the extraction in the step 2) is ultrasonic extraction, the power is 500W, the frequency is 40kHz, and the time is 30-60 min.
Further, the chromatographic column in the chromatographic condition in step 3) is: sunFire C18 column, inertsil ODS-3C 18 column or Kromasil C18 column, the column length is 250mm, the inner diameter is 4.6mm, the particle size is 5 mu m, and the column temperature is 25-35 DEG CSample injection amount is 5-25 mu L, flow rate is 0.2-0.7 mL ∙ min -1 The detection wavelength is 200-220 nm.
Further, in the step 3), the column temperature is 30 ℃, the sample injection amount is 10 mu L, and the flow rate is 0.5mL ∙ min in the chromatographic condition -1 The detection wavelength is 210nm.
Further, the chromatogram of the sample solution should show 8 characteristic peaks, wherein 2 peaks should correspond to the retention time of the corresponding reference peak of the reference sample, the peak corresponding to the reference peak of the medlar acid is S peak, the relative retention time of the rest peaks and S peak is calculated, the relative retention time should be within ±10% of the specified value, and the specified value is: peak 1:0.343, peak 2:0.413, peak 3:0.534, peak 6:1.173, peak 7:1.487, peak 8:1.800.
according to the HPLC characteristic spectrum detection method for the Russian ginseng fruits and the preparation thereof, 8 characteristic peaks can be calibrated through the characteristic spectrum established under specific chromatographic conditions, particularly, the chromatographic peaks of the organic acid components of the lycium barbarum acid and the L-malic acid which are special in the Russian ginseng fruits can be identified, the quality control of the Russian ginseng fruits and the preparation thereof is realized through the calibration of the chemical components which are more special, the clinical curative effect of the Russian ginseng fruits is ensured, and the practical popularization and application values are realized.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Drawings
FIG. 1 is a chromatogram for examining the durability of a column (A: sunFire C18 column; B: kromasil C18 column; C: inertsil ODS-3C 18 column);
FIG. 2 3 sample lot profile (S1: PFG001 profile, S2: PFG002 profile, S3: PFG003 profile, R: control profile);
FIG. 3 HPLC chromatogram (A: control profile of Russian crab (Malus asiatica) formula granule; B: HPLC chromatogram of mixed control solution; peak 4:L-malic acid; peak 5: matrimony vine acid);
fig. 4 Russian color fruit pieces, standard decoction, formula particle matching chromatogram.
Detailed Description
The reagents, and equipment used in the embodiments of the invention are all known products and are obtained by purchasing commercially available products. Wherein the Russian crab (Crabapple) formula granule is Crabapple of RosaceaeMalus toringoides(red.) the dried ripe fruit of Hughes is formulated into granule with light brown to reddish brown shape according to the main quality index of standard decoction; slight smell, sour and sweet taste. The specific preparation method comprises the following steps: decocting Russian crabapple decoction pieces 2000g in water, filtering, concentrating the filtrate into fluid extract (dry extract yield 25% -45%), adding appropriate amount of adjuvant, drying (or drying, pulverizing), adding appropriate amount of adjuvant, mixing, granulating, and making 1000 g.
Example 1 feature profile detection of Russian cherry formula particles of the invention
1) Preparation of control solution
Taking reference substances of the medlar acid and the L-malic acid, precisely weighing, and adding distilled water to prepare mixed solutions with the concentration of 0.25mg and 0.5mg in each 1ml respectively as reference substance solutions.
2) Preparation of test solutions
Grinding Russian cherry formula granule 0.5g, placing into a conical flask with a plug, adding distilled water 15ml, performing ultrasonic treatment (power 500W, frequency 40 kHz) for 30min, shaking, filtering, and collecting filtrate.
3) Respectively precisely sucking 10 μl of reference solution and 10 μl of sample solution, injecting into liquid chromatograph, and recording chromatogram. The chromatographic conditions were as follows:
chromatographic column: sunFire C18 column, specification 4.6X1250 mm,5μm; detection wavelength: 210nm; column temperature 30℃and flow rate 0.5mL ∙ min -1 The method comprises the steps of carrying out a first treatment on the surface of the Mobile phase: acetonitrile as mobile phaseA, taking a 0.1% phosphoric acid solution as a mobile phase B; gradient elution procedure: 0-18 min,1% → 3% A; 18-18.5 min,3% -4% A; 18.5-25 min, 4%. Fwdarw. 7%A; 25-30 min,7% -15% A; 30-35 min,15% -31% A.
4) Analytical chromatograms
8 characteristic peaks should be present in the chromatogram of the test solution, 2 of which should correspond to the retention times of the corresponding reference peaks of the control, respectively. The peak corresponding to the reference peak of the medlar acid is an S peak, and the relative retention time of the peak 1, the peak 2, the peak 3, the peak 6, the peak 7, the peak 8 and the S peak is calculated, wherein the relative retention time is within +/-10% of a specified value, and the specified value is: 0.343 (Peak 1), 0.413 (Peak 2), 0.534 (Peak 3), 1.173 (Peak 6), 1.487 (Peak 7), 1.800 (Peak 8).
Example 2 detection of the characteristic Property spectrum of Russian Cryptotaenia
1) Preparation of control solution
Taking reference substances of the medlar acid and the L-malic acid, precisely weighing, and adding distilled water to prepare mixed solutions with the concentration of 0.25mg and 0.5mg in each 1ml respectively as reference substance solutions.
2) Preparation of test solutions
Grinding Russian rue fruit powder 1g, placing into a conical flask with a plug, adding distilled water 20ml, performing ultrasonic treatment (power 500W, frequency 40 kHz) for 45min, shaking, filtering, and collecting the filtrate.
3) Respectively precisely sucking 10 μl of reference solution and 10 μl of sample solution, injecting into liquid chromatograph, and recording chromatogram. The chromatographic conditions were as follows:
chromatographic column: inertsil ODS-3C 18 column, specification 4.6X1250 mm,5μm; detection wavelength: 210nm; column temperature 30℃and flow rate 0.5mL ∙ min -1 The method comprises the steps of carrying out a first treatment on the surface of the Mobile phase: acetonitrile is taken as a mobile phase A, and a 0.1% phosphoric acid solution is taken as a mobile phase B; gradient elution procedure: 0-18 min,1% → 3% A; 18-18.5 min,3% -4% A; 18.5-25 min, 4%. Fwdarw. 7%A; 25-30 min,7% -15% A; 30-35 min,15% -31% A.
4) Analytical chromatograms
8 characteristic peaks should be present in the chromatogram of the test solution, 2 of which should correspond to the retention times of the corresponding reference peaks of the control, respectively. The peak corresponding to the reference peak of the medlar acid is an S peak, and the relative retention time of the peak 1, the peak 2, the peak 3, the peak 6, the peak 7, the peak 8 and the S peak is calculated, wherein the relative retention time is within +/-10% of a specified value, and the specified value is: 0.343 (Peak 1), 0.413 (Peak 2), 0.534 (Peak 3), 1.173 (Peak 6), 1.487 (Peak 7), 1.800 (Peak 8).
Example 3 feature map detection of the Russian color fruit Standard decoction of the invention
1) Preparation of reference solutions
Taking reference substances of the medlar acid and the L-malic acid, precisely weighing, and adding distilled water to prepare mixed solutions with the concentration of 0.25mg and 0.5mg in each 1ml respectively as reference substance solutions.
2) Preparation of test solutions
Grinding 0.5g of lyophilized powder of Russian cherry standard decoction, placing into a conical flask with a plug, adding distilled water 20ml, performing ultrasonic treatment (power 500W, frequency 40 kHz) for 30min, shaking, filtering, and collecting the filtrate.
3) Respectively precisely sucking 10 μl of reference solution and 10 μl of sample solution, injecting into liquid chromatograph, and recording chromatogram. The chromatographic conditions were as follows:
chromatographic column: inertsil ODS-3C 18 column, specification 4.6X1250 mm,5μm; detection wavelength: 210nm; column temperature 30℃and flow rate 0.5mL ∙ min -1 The method comprises the steps of carrying out a first treatment on the surface of the Mobile phase: acetonitrile is taken as a mobile phase A, and a 0.1% phosphoric acid solution is taken as a mobile phase B; gradient elution procedure: 0-18 min,1% → 3% A; 18-18.5 min,3% -4% A; 18.5-25 min, 4%. Fwdarw. 7%A; 25-30 min,7% -15% A; 30-35 min,15% -31% A.
4) Analytical chromatograms
8 characteristic peaks should be present in the chromatogram of the test solution, 2 of which should correspond to the retention times of the corresponding reference peaks of the control, respectively. The peak corresponding to the reference peak of the medlar acid is an S peak, and the relative retention time of the peak 1, the peak 2, the peak 3, the peak 6, the peak 7, the peak 8 and the S peak is calculated, wherein the relative retention time is within +/-10% of a specified value, and the specified value is: 0.343 (Peak 1), 0.413 (Peak 2), 0.534 (Peak 3), 1.173 (Peak 6), 1.487 (Peak 7), 1.800 (Peak 8).
The beneficial effects of the invention are illustrated by experimental examples below:
experimental example 1 Russian color fruit formula particle fingerprint study
1. Chromatographic conditions
The invention determines the characteristic spectrum condition of the traditional Chinese medicine formula granule on the basis of the early test as follows, and detects according to the condition.
Chromatography conditions and system applicability test octadecylsilane chemically bonded silica is used as filler (column length is 250mm, inner diameter is 4.6mm, particle size is 5 μm), acetonitrile is used as mobile phase A, 0.1% phosphoric acid solution is used as mobile phase B, and gradient elution is carried out according to the specifications in the following table; the detection wavelength is 210nm; the flow rate is 0.5ml per minute; the column temperature was 30 ℃.
Figure SMS_1
2. Sample preparation method investigation
On the basis of the early test, distilled water is determined to be used as a solvent, and other extraction condition investigation is carried out.
2.1 extraction time investigation
Grinding Russian crabapple (Malus asiatica) prescription granule 0.5g, placing into conical flask with plug, adding 75% methanol 25ml, respectively ultrasonic treating for 10min, 20min, 30min, and 40min, shaking, filtering, and collecting filtrate. Sample was taken under chromatographic conditions under item "1", and the results are shown in Table 1.
Table 1 investigation of extraction time
Figure SMS_2
As shown in the table, as the extraction time increased, the peak areas of the chromatographic peaks increased, the extraction time was 30min and 40min, and the peak areas of the chromatographic peaks were not greatly different, and the peak areas of the respective chromatographic peaks tended to be stable, so that the optimal extraction time was determined to be 30min.
2.2 sample weighing investigation
Taking 0.1g, 0.3g, 0.5g, 0.7g and 1.0g of Russian crabapple (malus spectabilis) prescription particles, grinding, putting into a conical flask with a plug, respectively adding 20ml of distilled water, respectively carrying out ultrasonic treatment (with the power of 250W and the frequency of 40 kHz) for 30min, taking out, cooling, shaking uniformly, filtering, taking out a subsequent filtrate, and carrying out sample injection according to chromatographic conditions under the item "1", wherein the result is shown in Table 2.
Table 2 sample weighing investigation
Figure SMS_3
As shown in the table, as the sample weighing amount increases, the peak area of each chromatographic peak is increased proportionally, and when the sample weighing amount is 1g, the peak area of the medlar acid peak is overlarge, so that the whole chromatographic chart is not attractive; when the sample weighing amount is 0.3g, the area of partial peak to peak is too small; when the sample weighing amount is 0.5g, the peak area of each peak is moderate, and the total chromatographic peak area is maximum, so that the 0.5g is determined as the optimal sample weighing amount.
Solvent volume investigation
Taking 0.5g of Russian crabapple (malus halliana) prescription granule, grinding, placing into a conical flask with a plug, respectively adding distilled water 5ml, distilled water 10ml, distilled water 15ml and distilled water 20ml, respectively performing ultrasonic treatment (power 250W and frequency 40 kHz) for 30min, taking out, cooling, shaking uniformly, filtering, taking out the subsequent filtrate, and performing sample injection according to chromatographic conditions under the item "1", wherein the result is shown in Table 3.
TABLE 3 solvent volume investigation
Figure SMS_4
As is clear from the table, the peak areas of the respective chromatographic peaks decrease proportionally with the increase of the volume of the solvent, but the number of chromatographic peaks does not change, and when the solvent is 10ml, the peak areas are partially too large, and the optimum volume of the solvent is 15ml by integrating the respective peak conditions.
2.4 preparation method of sample
Grinding Russian crabapple (Malus asiatica) formula granule 0.5g, placing into conical flask with plug, adding distilled water 15ml, ultrasonic treating (power 500W, frequency 40 kHz) for 30min, shaking, filtering, and collecting filtrate.
3. Methodology investigation
3.1 repeatability investigation
Taking 6 parts of Russian crabapple (malus halliana) formula particles, preparing samples according to the item "2.4", continuously sampling according to the chromatographic condition "1", and recording chromatograms, wherein the chromatograms are shown in tables 4-5; the relative retention time and the relative peak area RSD value of each characteristic peak are calculated, and the results are respectively 0.08% -0.32% and 0.02% -0.14%, which shows that the method repeatability is good.
TABLE 4 relative retention time of repeatability
Figure SMS_5
TABLE 5 repeatability versus retained peak area
Figure SMS_6
3.2 Precision investigation
Taking Russian crabapple (malus halliana) formula particle samples, preparing samples according to the item of 2.4, continuously sampling for 6 times according to the chromatographic condition of 1, and recording chromatograms, wherein the chromatograms are shown in tables 6-7; the relative retention time and the relative peak area RSD value of each characteristic peak are calculated, and the results are respectively 0.05% -0.07% and 1.40% -2.90%, so that the instrument precision is good.
TABLE 6 precision investigation of relative retention time
Figure SMS_7
TABLE 7 precision investigation of relative peak areas
Figure SMS_8
3.3 stability investigation
Taking Russian crabapple (malus halliana) formula particle samples, preparing samples according to the item of 1, respectively sampling samples at (0, 2, 4, 8, 12 and 24 and h), and recording chromatograms, wherein the chromatograms are shown in tables 8-9; and calculating the relative retention time and the RSD value of the relative peak area of each characteristic peak, wherein the results are respectively 0.16% -0.43% and 1.21% -2.86%, which shows that the sample solution is stable within 24 hours.
Table 8 stability investigation relative retention time
Figure SMS_9
TABLE 9 stability investigation of relative peak areas
Figure SMS_10
3.4 column durability
Taking the same sample solution, taking chromatographic conditions under the item "1", respectively detecting with SunFire C18 column, inertsil ODS-3C 18 column and Kromasil C18 column, recording the chromatogram, and observing the peak-to-peak condition of the chromatogram. The experimental result shows that each chromatographic peak has good separation degree on different chromatographic columns and symmetrical peak shape, and the chromatographic column has good durability by the method, and the result is shown in figure 1.
4. Establishment of characteristic spectrum
3 batches of Russian cherry formula particles obtained in pilot plant test are prepared according to the term "2.4", and are detected according to the chromatographic condition under the term "1", and the chromatogram is recorded. Determining 8 common chromatographic peaks according to a sample chromatogram result, identifying 2 characteristic peaks which are L-malic acid and matrimony vine acid respectively, wherein peak 5 (matrimony vine acid) has stable peak shape and good separation degree, and selecting the peak as a reference peak (S peak); and matching chromatograms of three Russian cherry prescription granule samples of PFG001, PFG002 and PFG003 with chromatographic peaks in a control characteristic map by adopting a 2012A edition of a traditional Chinese medicine chromatographic fingerprint similarity evaluation system to establish an HPLC characteristic map of the Russian cherry (malus halliana) prescription granule, wherein the HPLC characteristic map is shown in figures 2-3. And (3) injection: the comparison characteristic spectrum is obtained by analyzing characteristic spectrum of 3 Russian russian fruit formula particles by adopting a traditional Chinese medicine chromatographic characteristic spectrum similarity evaluation system (2012A edition), selecting 8 chromatographic methods with good repeatability and separation degree as common peaks, taking a No. 5 peak (medlar acid) with moderate retention time and stable peak output as a reference (S), and generating the comparison characteristic spectrum by adopting a multi-point correction mode and an average method.
5. Similarity analysis
And (3) carrying out similarity analysis on the chromatograms of 3 Russian russia rupestris formula particle samples by adopting traditional Chinese medicine chromatographic fingerprint similarity evaluation software, calculating relative retention time and relative peak area, and finally determining the relative retention time of 8 common peaks. And carrying out correlation analysis on the decoction pieces, the standard decoction and the formula particles, wherein the results are shown in tables 10-12 and fig. 4.
TABLE 10 3 characterization of Russian color fruit formula particle profile versus retention time
Figure SMS_11
TABLE 11 characterization of Russian color fruit formula particle characteristics versus peak area
Figure SMS_12
Table 12 results of similarity evaluation of different samples
Figure SMS_13
The results show that the similarity of the HPLC characteristic patterns of the 3 Russian crab (malus asiatica) formula granules is above 0.900, which indicates that the preparation process of the formula granules is stable. The peak corresponding to the reference peak of the medlar acid is taken as an S peak, the relative retention time of the rest peaks and the S peak is 0.343 (peak 1), 0.413 (peak 2), 0.534 (peak 3), 1.173 (peak 6), 1.487 (peak 7) and 1.800 (peak 8), and the relative retention time of each peak is within the range of +/-10% of a specified value. The characteristic spectrum is adopted to analyze the similarity of the raw materials of the formula particle, the standard decoction and the sample of the formula particle, and the result shows that the raw materials, the standard decoction and the sample of the formula particle have the same characteristic peak, the similarity is more than 0.95, the mass transfer is stable, and the preparation method of the formula particle is suitable.

Claims (5)

1. An HPLC characteristic spectrum detection method for Russian rue fruits and preparations thereof is characterized in that: it comprises the following steps:
1) Preparation of a control solution: dissolving reference substances of Lycii acid and L-malic acid in water to obtain reference substance solution;
2) Sample solution preparation: extracting a sample to be detected with water, filtering the extracting solution, and obtaining filtrate to obtain a sample solution;
3) Respectively sucking the reference substance solution and the sample solution, injecting into a high performance liquid chromatograph, and recording chromatograms; the chromatographic conditions were as follows:
chromatographic column: gunfire C18 column, inertsil ODS-3C 18 column or Kromasil C18 column, the column length is 250mm, the inner diameter is 4.6mm, and the grain diameter is 5 mu m; mobile phase: acetonitrile is taken as a mobile phase A, and a 0.1% phosphoric acid solution is taken as a mobile phase B; gradient elution procedure 0-18 min,1% →3% A; 18-18.5 min,3% -4% A; 18.5-25 min, 4%. Fwdarw. 7%A; 25-30 min,7% -15% A; 30-35 min,15% -31% A, column temperature of 30 ℃, sample injection amount of 10 mu L and flow rate of 0.5mL ∙ min -1 Detection wavelength is 210nm;
8 characteristic peaks should be presented in the chromatogram of the sample solution, wherein 2 peaks should correspond to the retention time of the corresponding reference peaks of the reference substance, the peak corresponding to the reference peak of the medlar acid is S peak, the relative retention time of the rest peaks and S peak is calculated, the relative retention time is within + -10% of the specified value, and the specified value is: peak 1:0.343, peak 2:0.413, peak 3:0.534, peak 6:1.173, peak 7:1.487, peak 8:1.800.
2. the HPLC profile detection method of claim 1, wherein: each 1ml of the reference substance solution in the step 1) contains 0.1-0.5 mg of medlar acid and 0.1-1 mg of L-malic acid.
3. The HPLC profile detection method of claim 1, wherein: and 2), the mass volume ratio of the sample to be detected to water is 0.5-1 g: 5-50 ml; the water is distilled water.
4. A method of HPLC profile detection according to claim 1 or claim 3, wherein: the sample to be detected is Russian cherry medicinal material, russian cherry standard decoction or Russian cherry formula granule.
5. The HPLC profile detection method of claim 1, wherein: the step 2) is ultrasonic extraction, the power is 500W, the frequency is 40kHz, and the time is 30-60 min.
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CN110407897A (en) * 2018-04-28 2019-11-05 四川智佳成生物科技有限公司 The extracting method and detection method of fructus lycii acid in Russia color fruit

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