CN113640431A

CN113640431A - Detection method of strong loquat syrup

Info

Publication number: CN113640431A
Application number: CN202111110852.2A
Authority: CN
Inventors: 眭荣春; 胡军; 胡庭国; 邓晓军; 肖云
Original assignee: Jiangxi Universe Pharmaceuticals Co ltd
Current assignee: Jiangxi Universe Pharmaceuticals Co ltd
Priority date: 2021-09-23
Filing date: 2021-09-23
Publication date: 2021-11-12
Anticipated expiration: 2041-09-23
Also published as: CN113640431B

Abstract

The invention provides a detection method of strong loquat syrup. The method comprises the steps of diluting and extracting a standard strong loquat dew product, adding a solubilizer to improve the extraction amount of effective components in a medicament, reducing the emulsification phenomenon, selecting the standard strong loquat dew product as a standard spectrogram, and avoiding the phenomenon that illegal merchants pass detection only by adding a single effective substance and reduce the phenomenon of insufficient detection. Before liquid chromatography analysis, a thin-layer chromatography is adopted for coarse analysis, whether a product is qualified or not is preliminarily determined, and then a reasonable eluent and an elution sequence are designed for high performance liquid chromatography analysis. Before liquid chromatography testing, a modified filter membrane is required to be used for filtering the test solution, so that impurities in the test solution are reduced, the detection speed and the detection accuracy are improved, and the service life of a chromatographic column is prolonged.

Description

Detection method of strong loquat syrup

Technical Field

The invention relates to the technical field of medicine quality control, in particular to a detection method of strong loquat syrup.

Background

The strong loquat syrup is prepared from seven traditional Chinese medicines such as loquat leaf, poppy shell, stemona root and cynanchum glaucescens, and is a variety collected in the second volume of the Standard of Ministry of health in the patent of Chinese patent medicine preparations; has the effects of nourishing yin, astringing lung, relieving cough and eliminating phlegm: can be used for treating chronic cough, bronchitis, etc. Wherein the loquat leaves are monarch drug, and the poppy capsule is a main drug for treating cough, and contains alkaloid components such as morphine, codeine, papaverine, protopine, etc. As is known, the efficacy of Chinese patent medicine is not from any single active ingredient, but the result of the combined action of a plurality of effective parts, and the prior quality standard of strong loquat dew only controls the identification and the relative density. The two items can not represent the quality of the strong loquat distillate, and the quality risk exists. The fingerprint can represent the components of the strong loquat dew more comprehensively and can control the quality of the strong loquat dew more strictly.

So far, the existing quality standard of the strong loquat syrup is too simple to effectively control the product quality, and the quality of the strong loquat syrup sold in the market is uneven. Because the preparation contains the narcotic poppy capsule, if the dosage of the preparation is improperly controlled, the preparation is easy to addiction after being taken for a long time. Modern chemical and pharmacological studies have shown that the major active ingredients of poppy shell are alkaloids including papaverine, narcotine, thebaine, morphine and codeine, which are highly addictive ingredients. In order to ensure the safety and effectiveness of the preparation, the content thereof must be controlled. The existing quality standard of the medicine only has chemical reaction identification of alkaloid and flavone, and has poor specificity, low controllability and no content determination item. At present, only reports of measuring the content of morphine, codeine or papaverine in poppy shell in the strong loquat syrup by HPLC or gas chromatography are available in the content measurement research. With the development of modern analysis technology, the liquid chromatography-mass spectrometry technology has gradually become an effective means for quantitative analysis of Chinese medicinal components by virtue of its advantages of high sensitivity, strong specificity, rapidness and high efficiency. Therefore, the invention adopts the standard strong loquat dew solution to obtain a characteristic spectrogram special for the strong loquat dew through liquid chromatography so as to improve the quality control level of the strong loquat dew and ensure the safety of products.

Disclosure of Invention

The invention mainly aims to provide a detection method of strong loquat syrup.

The second purpose of the invention is to provide a fingerprint spectrum of the standard strong loquat dew and a test method thereof.

The invention is realized by the following scheme:

a detection method of strong loquat distillate is provided.

The detection method of the strong loquat syrup comprises the following steps:

first), establishment of standard strong loquat dew product detection method

1) Taking 10 parts of a standard strong loquat dew product, adding a diluent to dilute the product, fixing the volume to 5-6 times of the volume of an original product, heating the product to 50 ℃, preserving heat and standing for 1h for later use;

2) filtering the prepared solution by adopting a microporous molecular membrane, collecting filtrate, adding 30-40 parts of diluent into filter residue, heating the mixture to 35 ℃, keeping the temperature and standing for 30min, filtering again to obtain filtrate a and filter residue, adding 40-45 parts of diluent into the filter residue again, heating the mixture to 45 ℃, keeping the temperature and standing for 45min, filtering to obtain filtrate b and filter residue, and uniformly mixing the filtrate, the filtrate a and the filtrate b for later use;

3) putting the combined filtrate into a rotary evaporation drying machine for evaporation treatment to obtain a residue after rotary evaporation drying; taking 0.5 part of residue, adding 10 parts of ethanol solution, and adjusting the pH value to 5.5-6.0 by 0.001mol/L HCl solution until the residue is completely dissolved to obtain solution a;

4) taking 5ml of the solution a, adding an ethanol solution into the solution a to fix the volume to 500ml, respectively putting 20ml, 20ml and 20ml into a 50ml volumetric flask, and adding ethanol to fix the volume to the scale mark; then, respectively filtering the solution with a microporous filter membrane, putting the solution into a liquid chromatograph for test analysis to obtain data of an absorption peak value within 0-35min, introducing the data into Matlab software for drawing, wherein the time is an abscissa, and the average value of the absorption values of 4 groups of data is an ordinate to obtain a standard curve of the strong loquat dew;

II), detection of the sample

1) Treating the strong loquat dew product to be tested according to the method, then obtaining a corresponding spectrogram through a liquid chromatograph, and judging as a qualified product when the relative peak area of the spectrogram and the standard spectrogram is within 2%. The preparation method of the diluent comprises the following steps:

1) heating 50 parts of isopropanol and 30 parts of toluene to 60 ℃, uniformly mixing to obtain a solution m, and keeping the temperature and standing for later use;

2) the prepared solution m and the solubilizer are mixed according to the volume ratio of (100-: (0.3-0.5), adding 20-30 parts of petroleum ether, heating the solution to 75 ℃ at the speed of 1-2 ℃/min, keeping the temperature, stirring for 3-5h, and cooling to room temperature to obtain the product.

The preparation of the solubilizer comprises the following steps:

a. putting 5-9 parts of tetraethylammonium bromide into a reactor, adding 3-5 parts of tetrabutylammonium hydrogen sulfate, adding a toluene solution to completely dissolve the tetraethylammonium bromide, heating the mixture to 50 ℃ at the speed of 3-5 ℃/min to obtain a solution i, and standing and preserving the temperature for later use;

b. putting 2-4 parts of tetramethylammonium fluoride into a reactor, adding an acetone solution, heating to 90 ℃ at the speed of 1-3 ℃/min, dropwise adding 12-15 parts of the solution i into the reactor at the speed of 15-18 drops/min while stirring, heating the solution to 150 ℃ together, and carrying out heat preservation reaction for 4-5 hours to obtain a reactant a;

c. adding 15-20 parts of ethylene glycol dimethyl ether into the reactant a, heating to 50 ℃ at the speed of 1-3 ℃/min, and carrying out heat preservation reaction for 3-4h to obtain the catalyst.

Wherein, before carrying out liquid chromatography, carry out preliminary analysis through thin-layer chromatography earlier, reduce the waste of detection cost, concrete detection method is:

placing the solution a of the standard solution and the sample solution on a thin-layer chromatography silica gel G plate in a spot plate mode, evaporating the liquid in the thin-layer chromatography silica gel G plate by naturally drying, placing the thin-layer chromatography silica gel G plate in a chromatography solution, obliquely placing the chromatography solution until the chromatography solution climbs to 6/7 areas of the silica gel plate, taking out the chromatography solution and drying the chromatography solution in the air, placing the chromatography solution under an ultraviolet lamp with a wavelength of 256mm to observe whether the positions of the spots are the same, if the positions are the same, carrying out the next analysis, and if the positions are not the same, determining the samples as non-qualified products, and eliminating the samples.

Wherein, the preparation of the chromatographic solution comprises the following steps:

a. putting 15-20 parts of cyclohexane into a reactor, adding 40-45 parts of ethyl acetate solution, heating to 50 ℃, adding 1-3 parts of propylene carbonate, heating to 70 ℃, and carrying out heat preservation reaction for 3 hours to obtain an intermediate product a;

b. adding 30-40 parts of petroleum ether into the intermediate product a, heating to 80 ℃, reacting for 5-6 hours in a heat preservation way, standing and preserving heat to obtain an intermediate product b;

c. and (3) uniformly mixing the intermediate product b and ethanol according to the volume ratio of 7:1 to obtain the chromatography liquid.

Wherein, the elution sequence in the liquid chromatogram of the step 4) is as follows:

0-5 min, 95% of phase A and 5% of phase B;

at 5-12min, phase A is 95-90%, and phase B is 5-10%;

when 12-25min, the phase A is 90-83%, and the phase B is 10-17%;

at 25-31min, the A phase is 83-81%, and the B phase is 17-19%;

31-35min later, the phase A is 81-80%, and the phase B is 19-20%;

wherein, the organic phase adopts methanol solution;

0.45 percent aqueous phosphoric acid solution with the flow rate of 1.0 ml/min; the column temperature is 25-28 ℃, and the detection wavelength is 213 mm;

wherein, 0.2 percent of triethylamine solution is added into the organic phase; the column was Agilent HC-C18250X 4.6mm 5 μm.

Wherein, the step 4) also comprises the modification of the microporous filter membrane, and the specific modification step is as follows:

1) soaking the microporous filter membrane in ethanol solution, heating to 38-42 deg.C at a rate of 1-2 deg.C/min, simultaneously performing microwave vibration, maintaining the temperature and shaking for 2 hr, taking out, washing with methanol solution, and standing;

2) putting the filter membrane prepared in the step into a dichloromethane solution, adding a modifier, heating to 80 ℃ at the speed of 3-5 ℃/min, preserving heat and standing for 4-5h to obtain a primary modified filter membrane;

3) and (3) soaking the preliminary modified filter membrane in a methanol solution, controlling the temperature of the methanol solution to be 0-3 ℃, keeping the temperature, standing for 3-4h, taking out and airing to obtain the modified filter membrane.

The preparation method of the modifier comprises the following steps:

i) putting 1-3 parts of sulfonated polyether ether ketone into a reactor, adding a methanol solution, heating to 42-45 ℃ at the speed of 1-3 ℃/min, then adding 2-4 parts of N, N-dimethylformamide diethyl acetal into the reactor while the reactor is hot, heating to 60 ℃, and carrying out heat preservation reaction for 3-4 hours to obtain a product a;

ii) adding 2-4 parts of 2, 3-glycidoxypropyl trimethoxysilane into the product a, adding a dichloromethane solution into the product a, starting a condensation reflux device, heating to 145 ℃ at the speed of 4-6 ℃/min, and carrying out heat preservation reaction for 4-6h to obtain a product b;

iii) adding 4-6 parts of N, N-dimethoxyamide into the product b, then adding a dimethyl sulfoxide solution, heating to 80-85 ℃, and carrying out heat preservation reaction for 4-5h to obtain a product c;

iv) adding 1-2 parts of 3, 3-diethoxypropionic acid ethyl ester into the product c, then adding a toluene solution, heating to 153-157 ℃ at the speed of 2-4 ℃/min, starting a condensation reflux device during heating, and carrying out heat preservation reaction for 5-6 hours to obtain the product.

The invention has the following advantages:

the method provided by the invention is to dilute and extract the standard strong loquat dew product, extract the effective components in the strong loquat dew and analyze the effective components, and the principle is to extract the effective components in the strong loquat dew; wherein the diluent contains solubilizer, which can improve the extraction rate of effective components in the strong loquat distillate. The effective components of the solubilizer can improve the extraction content of the medicament, reduce the emulsification phenomenon, improve the analysis precision and improve the repeatability of the detection result. The effective components in the solubilizer are suitable for a high performance liquid chromatograph, a standard strong loquat dew product is selected as a standard spectrogram, and the spectrogram obtained by testing various samples is detected and analyzed, so that the phenomenon that illegal merchants pass detection only by adding a single effective substance is avoided, and the phenomenon that the detection is insufficient is reduced.

The effective components in the herbal medicines are extracted by the diluent, after the effective components are obtained, a reasonable elution phase, an elution sequence and elution time need to be designed, meanwhile, the influence of the solubilizer is reduced, and the influence on the analysis result is avoided. Because the methanol solution is selected in the elution process, triethylamine which is an organic substance is added into the methanol solution to adjust the pH value, and the tailing condition of a characteristic peak in liquid chromatogram is reduced; in the same way, a corresponding buffering agent is added into the water phase, the pH value of the whole elution phase is adjusted, and the phenomena of asymmetric peak, broad peak and split peak are reduced. Before liquid chromatography analysis, thin-layer chromatography is adopted for coarse analysis, whether the strong loquat syrup product is qualified or not is preliminarily determined, the resolution efficiency is improved, and meanwhile, the influence effect of the solubilizer is further reduced by preliminarily judging through the thin-layer chromatography.

Because the solubilizer component and the special elution sequence added in the diluent try to reduce the influence of the solubilizer on the experimental result, but the phenomena of broad peak, peak cracking and impurity peak at the blank position still occur when a sample is tested, the sample needs to be filtered before the liquid chromatography test, and the common filter membrane cannot effectively remove impurities in the test solution during the filtration, so that the filler in the chromatographic column can be damaged in the test process, the chromatographic column is blocked, the service life of the chromatographic column is shortened, and the filter membrane before entering the column needs to be modified before the sample solution is detected and analyzed; the influence of the solubilizer on experimental results is further reduced by modifying the filter membrane, the influencing factors in the test solution are filtered, the service life of the chromatographic column is prolonged, and the detection cost is reduced; the modified filter membrane can effectively reduce impurities in the test solution, and improve the detection speed and the detection accuracy; by modifying the filter membrane, the solubilizer in the diluent and the elution speed and sequence, the method can detect the quality problem of the strong loquat syrup quickly and efficiently at low resistance.

Detailed description of the invention

Example 1

A detection method of strong loquat distillate is provided.

The detection method of the strong loquat syrup comprises the following steps:

first), establishment of standard strong loquat dew product detection method

1) Taking 10 parts of a standard strong loquat dew product, adding a diluent to dilute the product, fixing the volume to 5.5 times of the volume of an original product, heating the product to 50 ℃, preserving heat and standing for 1 hour for later use;

2) filtering the prepared solution by adopting a microporous molecular membrane, collecting filtrate, adding 35 parts of diluent into filter residue, heating to 35 ℃, keeping the temperature and standing for 30min, filtering again to obtain filtrate a and filter residue, adding 43 parts of diluent into the filter residue again, heating to 45 ℃, keeping the temperature and standing for 45min, filtering to obtain filtrate b and filter residue, and uniformly mixing the filtrate, the filtrate a and the filtrate b for later use;

3) putting the combined filtrate into a rotary evaporation drying machine for evaporation treatment to obtain a residue after rotary evaporation drying; taking 0.5 part of residue, adding 10 parts of ethanol solution, and adjusting the pH value to 5.8 by 0.001mol/L HCl solution until the residue is completely dissolved to obtain solution a;

II), detection of the sample

1) Treating the strong loquat dew product to be tested according to the method, then obtaining a corresponding spectrogram through a liquid chromatograph, and judging as a qualified product when the relative peak area of the spectrogram and the standard spectrogram is within 2%.

The preparation method of the diluent comprises the following steps:

2) the prepared solution m and a solubilizer are mixed according to a volume ratio of 103: 0.4, adding 25 parts of petroleum ether, heating the solution to 75 ℃ at the speed of 1.5 ℃/min, keeping the temperature, stirring for 4 hours, and cooling to room temperature to obtain the product.

The preparation of the solubilizer comprises the following steps:

a. putting 7 parts of tetraethylammonium bromide into a reactor, adding 4 parts of tetrabutylammonium hydrogen sulfate, adding a toluene solution to completely dissolve the tetraethylammonium bromide, heating the tetrabutylammonium hydrogen sulfate to 50 ℃ at the speed of 4 ℃/min to obtain a solution i, and standing and preserving the temperature for later use;

b. putting 3 parts of tetramethylammonium fluoride into a reactor, adding an acetone solution, heating to 90 ℃ at the speed of 2 ℃/min, dropwise adding i 13 parts of the solution into the reactor at the speed of 17 drops/min while stirring, heating the solution to 150 ℃ together, and keeping the temperature for reaction for 4.5 hours to obtain a reactant a;

c. adding 18 parts of ethylene glycol dimethyl ether into the reactant a, heating to 50 ℃ at the speed of 2 ℃/min, and carrying out heat preservation reaction for 3.5 hours to obtain the catalyst.

a. putting 18 parts of cyclohexane into a reactor, adding 43 parts of ethyl acetate solution, heating to 50 ℃, adding 2 parts of propylene carbonate, heating to 70 ℃, and carrying out heat preservation reaction for 3 hours to obtain an intermediate product a;

b. adding 35 parts of petroleum ether into the intermediate product a, heating to 80 ℃, reacting for 5.5 hours in a heat preservation way, standing and preserving heat to obtain an intermediate product b;

0-5 min, 95% of phase A and 5% of phase B;

at 5-12min, phase A is 95-90%, and phase B is 5-10%;

when 12-25min, the phase A is 90-83%, and the phase B is 10-17%;

at 25-31min, the A phase is 83-81%, and the B phase is 17-19%;

31-35min later, the phase A is 81-80%, and the phase B is 19-20%;

wherein, the organic phase adopts methanol solution;

0.45 percent aqueous phosphoric acid solution with the flow rate of 1.0 ml/min; the column temperature is 26 ℃, and the detection wavelength is 213 mm;

1) soaking the microporous filter membrane in ethanol solution, heating to 40 deg.C at a rate of 1.5 deg.C/min, simultaneously performing microwave vibration, maintaining the temperature and shaking for 2 hr, taking out, washing with methanol solution, and standing;

2) putting the filter membrane prepared in the step into a dichloromethane solution, adding a modifier, heating to 80 ℃ at the speed of 4 ℃/min, preserving heat and standing for 4.5 hours to obtain a primary modified filter membrane;

3) and (3) soaking the preliminary modified filter membrane in a methanol solution, controlling the temperature of the methanol solution to be 1 ℃, keeping the temperature and standing for 3.5h, taking out and airing to obtain the modified filter membrane.

The preparation method of the modifier comprises the following steps:

i) putting 2 parts of sulfonated polyether ether ketone into a reactor, adding a methanol solution, heating to 43 ℃ at the speed of 2 ℃/min, then adding 3 parts of N, N-dimethylformamide diethyl acetal into the reactor while the reactor is hot, heating to 60 ℃, and carrying out heat preservation reaction for 3.5 hours to obtain a product a;

ii) adding 3 parts of 2, 3-glycidoxypropyl trimethoxy silane into the product a, adding a dichloromethane solution into the product a, starting a condensation reflux device, heating to 143 ℃ at the speed of 5 ℃/min, and carrying out heat preservation reaction for 5 hours to obtain a product b;

iii) adding 5 parts of N, N-dimethoxyamide into the product b, then adding a dimethyl sulfoxide solution, heating to 83 ℃, and carrying out heat preservation reaction for 4-5h to obtain a product c;

iv) adding 1.5 parts of 3, 3-diethoxypropionic acid ethyl ester into the product c, then adding a toluene solution, heating to 155 ℃ at the speed of 3 ℃/min, starting a condensation reflux device during heating, and carrying out heat preservation reaction for 5.5 hours to obtain the product.

Example 2

A detection method of strong loquat distillate is provided.

The verification method of the strong loquat syrup comprises the following steps:

first), establishment of standard strong loquat dew product detection method

1) Taking 10 parts of a standard strong loquat dew product, adding a diluent to dilute the product, fixing the volume to 5 times of the volume of an original product, heating the product to 50 ℃, preserving heat and standing for 1 hour for later use;

2) filtering the prepared solution by adopting a microporous molecular membrane, collecting filtrate, adding 40 parts of diluent into filter residue, heating to 35 ℃, keeping the temperature and standing for 30min, filtering again to obtain filtrate a and filter residue, adding 40 parts of diluent into the filter residue again, heating to 45 ℃, keeping the temperature and standing for 45min, filtering to obtain filtrate b and filter residue, and uniformly mixing the filtrate, the filtrate a and the filtrate b for later use;

3) putting the combined filtrate into a rotary evaporation drying machine for evaporation treatment to obtain a residue after rotary evaporation drying; taking 0.5 part of residue, adding 10 parts of ethanol solution, and adjusting the pH value to 5.5 by 0.001mol/L HCl solution until the residue is completely dissolved to obtain solution a;

II), detection of the sample

The preparation method of the diluent comprises the following steps:

2) and (3) mixing the prepared solution m and a solubilizer according to a volume ratio of 100: 0.5, adding 20 parts of petroleum ether, heating the solution to 75 ℃ at the speed of 2 ℃/min, keeping the temperature, stirring for 3 hours, and cooling to room temperature to obtain the product.

The preparation of the solubilizer comprises the following steps:

a. placing 5 parts of tetraethylammonium bromide into a reactor, adding 5 parts of tetrabutylammonium hydrogen sulfate, adding a toluene solution to completely dissolve the tetraethylammonium bromide, heating the tetrabutylammonium hydrogen sulfate to 50 ℃ at the speed of 3 ℃/min to obtain a solution i, and standing and preserving the temperature for later use;

b. putting 4 parts of tetramethylammonium fluoride into a reactor, adding an acetone solution, heating to 90 ℃ at the speed of 1 ℃/min, dropwise adding i 15 parts of the solution into the reactor at the speed of 15 drops/min while stirring, heating the solution to 150 ℃ together, and keeping the temperature for 5 hours to obtain a reactant a;

c. adding 15 parts of ethylene glycol dimethyl ether into the reactant a, heating to 50 ℃ at the speed of 3 ℃/min, and carrying out heat preservation reaction for 3 hours to obtain the catalyst.

a. putting 20 parts of cyclohexane into a reactor, adding 40 parts of ethyl acetate solution, heating to 50 ℃, adding 1 part of propylene carbonate, heating to 70 ℃, and carrying out heat preservation reaction for 3 hours to obtain an intermediate product a;

b. adding 40 parts of petroleum ether into the intermediate product a, heating to 80 ℃, reacting for 5 hours in a heat preservation way, standing and preserving heat to obtain an intermediate product b;

0-5 min, 95% of phase A and 5% of phase B;

at 5-12min, phase A is 95-90%, and phase B is 5-10%;

when 12-25min, the phase A is 90-83%, and the phase B is 10-17%;

at 25-31min, the A phase is 83-81%, and the B phase is 17-19%;

31-35min later, the phase A is 81-80%, and the phase B is 19-20%;

wherein, the organic phase adopts methanol solution;

0.45 percent aqueous phosphoric acid solution with the flow rate of 1.0 ml/min; the column temperature is 25 ℃, and the detection wavelength is 213 mm;

1) soaking the microporous filter membrane in ethanol solution, heating to 42 deg.C at a rate of 1 deg.C/min, simultaneously performing microwave vibration, maintaining the temperature and shaking for 2 hr, taking out, washing with methanol solution, and standing;

2) putting the filter membrane prepared in the step into a dichloromethane solution, adding a modifier, heating to 80 ℃ at the speed of 3 ℃/min, preserving heat and standing for 5 hours to obtain a primary modified filter membrane;

3) and (3) soaking the preliminary modified filter membrane in a methanol solution, controlling the temperature of the methanol solution to be 0 ℃, keeping the temperature and standing for 4 hours, taking out and airing to obtain the modified filter membrane.

The preparation method of the modifier comprises the following steps:

i) putting 3 parts of sulfonated polyether ether ketone into a reactor, adding a methanol solution, heating to 45 ℃ at the speed of 1 ℃/min, then adding 2 parts of N, N-dimethylformamide diethyl acetal into the reactor while the reactor is hot, heating to 60 ℃, and carrying out heat preservation reaction for 4 hours to obtain a product a;

ii) adding 2 parts of 2, 3-glycidoxypropyl trimethoxy silane into the product a, adding a dichloromethane solution into the product a, starting a condensation reflux device, heating to 140 ℃ at the speed of 6 ℃/min, and carrying out heat preservation reaction for 6 hours to obtain a product b;

iii) adding 4 parts of N, N-dimethoxyamide into the product b, then adding a dimethyl sulfoxide solution, heating to 85 ℃, and carrying out heat preservation reaction for 4 hours to obtain a product c;

iv) adding 2 parts of 3, 3-diethoxypropionic acid ethyl ester into the product c, then adding a toluene solution, heating to 157 ℃ at the speed of 2 ℃/min, starting a condensation reflux device during heating, and carrying out heat preservation reaction for 5 hours to obtain the product.

Example 3

A detection method of strong loquat distillate is provided.

first), establishment of standard strong loquat dew product detection method

1) Taking 10 parts of a standard strong loquat dew product, adding a diluent to dilute the product, fixing the volume to 6 times of the volume of an original product, heating the product to 50 ℃, preserving heat and standing for 1h for later use;

2) filtering the prepared solution by adopting a microporous molecular membrane, collecting filtrate, adding 30 parts of diluent into filter residue, heating to 35 ℃, keeping the temperature and standing for 30min, filtering again to obtain filtrate a and filter residue, adding 45 parts of diluent into the filter residue again, heating to 45 ℃, keeping the temperature and standing for 45min, filtering to obtain filtrate b and filter residue, and uniformly mixing the filtrate, the filtrate a and the filtrate b for later use;

3) putting the combined filtrate into a rotary evaporation drying machine for evaporation treatment to obtain a residue after rotary evaporation drying; taking 0.5 part of residue, adding 10 parts of ethanol solution, and adjusting the pH value to 6.0 by 0.001mol/L HCl solution until the residue is completely dissolved to obtain solution a;

II), detection of the sample

The preparation method of the diluent comprises the following steps:

2) the prepared solution m and solubilizer are mixed according to a volume ratio of 105: 0.3, adding 30 parts of petroleum ether, heating the solution to 75 ℃ at the speed of 1 ℃/min, keeping the temperature, stirring for 5 hours, and cooling to room temperature to obtain the product.

The preparation of the solubilizer comprises the following steps:

a. putting 9 parts of tetraethylammonium bromide into a reactor, adding 3 parts of tetrabutylammonium hydrogen sulfate, adding a toluene solution to completely dissolve the tetraethylammonium bromide, heating the mixture to 50 ℃ at the speed of 5 ℃/min to obtain a solution i, and standing and preserving the temperature for later use;

b. putting 2 parts of tetramethylammonium fluoride into a reactor, adding an acetone solution, heating to 90 ℃ at the speed of 3 ℃/min, dropwise adding i 12 parts of the solution into the reactor at the speed of 18 drops/min while stirring, heating the solution to 150 ℃ together, and keeping the temperature for reacting for 4 hours to obtain a reactant a;

c. adding 20 parts of ethylene glycol dimethyl ether into the reactant a, heating to 50 ℃ at the speed of 1 ℃/min, and carrying out heat preservation reaction for 4 hours to obtain the catalyst.

a. putting 15 parts of cyclohexane into a reactor, adding 45 parts of ethyl acetate solution, heating to 50 ℃, adding 3 parts of propylene carbonate, heating to 70 ℃, and carrying out heat preservation reaction for 3 hours to obtain an intermediate product a;

b. adding 30 parts of petroleum ether into the intermediate product a, heating to 80 ℃, reacting for 6 hours in a heat preservation way, standing and preserving heat to obtain an intermediate product b;

0-5 min, 95% of phase A and 5% of phase B;

at 5-12min, phase A is 95-90%, and phase B is 5-10%;

when 12-25min, the phase A is 90-83%, and the phase B is 10-17%;

at 25-31min, the A phase is 83-81%, and the B phase is 17-19%;

31-35min later, the phase A is 81-80%, and the phase B is 19-20%;

wherein, the organic phase adopts methanol solution;

0.45 percent aqueous phosphoric acid solution with the flow rate of 1.0 ml/min; the column temperature is 28 ℃, and the detection wavelength is 213 mm;

1) soaking the microporous filter membrane in ethanol solution, heating to 38 deg.C at a rate of 2 deg.C/min, simultaneously performing microwave vibration, maintaining the temperature and shaking for 2 hr, taking out, washing with methanol solution, and standing;

2) putting the filter membrane prepared in the step into a dichloromethane solution, adding a modifier, heating to 80 ℃ at the speed of 5 ℃/min, preserving heat and standing for 4 hours to obtain a primary modified filter membrane;

3) and (3) soaking the preliminary modified filter membrane in a methanol solution, controlling the temperature of the methanol solution to be 3 ℃, keeping the temperature and standing for 3 hours, taking out and airing to obtain the modified filter membrane.

The preparation method of the modifier comprises the following steps:

i) putting 1 part of sulfonated polyether ether ketone into a reactor, adding a methanol solution, heating to 42 ℃ at the speed of 3 ℃/min, then adding 4 parts of N, N-dimethylformamide diethyl acetal into the reactor while the reactor is hot, heating to 60 ℃, and carrying out heat preservation reaction for 3 hours to obtain a product a;

ii) adding 4 parts of 2, 3-glycidoxypropyl trimethoxy silane into the product a, adding a dichloromethane solution into the product a, starting a condensation reflux device, heating to 145 ℃ at the speed of 4 ℃/min, and carrying out heat preservation reaction for 4 hours to obtain a product b;

iii) adding 6 parts of N, N-dimethoxyamide into the product b, then adding a dimethyl sulfoxide solution, heating to 80 ℃, and carrying out heat preservation reaction for 5 hours to obtain a product c;

iv) adding 1 part of 3, 3-diethoxypropionic acid ethyl ester into the product c, then adding a toluene solution, heating to 153 ℃ at the speed of 4 ℃/min, starting a condensation reflux device during heating, and carrying out heat preservation reaction for 6 hours to obtain the product.

Comparative example 1

A method for testing strong loquat distillate.

Wherein, the establishment step 1) of the standard strong loquat dew product detection method in the strong loquat dew detection method is as follows:

1) taking 10 parts of a standard strong loquat dew product, adding a diluent into the standard strong loquat dew product for dilution, diluting to a constant volume of 5.5 times of the volume of an original product, and standing at normal temperature for 1 hour for later use;

the rest is the same as example 1.

Comparative example 2

A method for testing strong loquat distillate.

Wherein, the establishing step 2) of the standard strong loquat dew product detection method in the strong loquat dew detection method is as follows:

2) filtering the prepared solution by adopting a microporous molecular membrane, collecting filtrate, adding 35 parts of diluent into filter residue, heating to 35 ℃, keeping the temperature and standing for 30min, filtering again to obtain filtrate a and filter residue, adding 43 parts of diluent into the filter residue again, heating to 40 ℃, keeping the temperature and standing for 45min, filtering to obtain filtrate b and filter residue, and uniformly mixing the filtrate, the filtrate a and the filtrate b for later use;

the rest is the same as example 1.

Comparative example 3

A method for testing strong loquat distillate.

2) filtering the prepared solution by adopting a microporous molecular membrane, collecting filtrate, adding 35 parts of diluent into filter residue, heating to 35 ℃, keeping the temperature and standing for 30min, filtering again to obtain filtrate a and filter residue, and uniformly mixing the filtrate a and the filter residue for later use;

the rest is the same as example 1.

Comparative example 4

A method for testing strong loquat distillate.

Wherein, the establishing step 3) of the standard strong loquat dew product detection method in the strong loquat dew detection method is as follows:

3) putting the combined filtrate into a rotary evaporation drying machine for evaporation treatment to obtain a residue after rotary evaporation drying; taking 0.5 part of residue, adding 10 parts of ethanol solution, and adjusting the pH value to 5.3 by 0.001mol/L HCl solution until the residue is completely dissolved to obtain solution a;

the rest is the same as example 1.

Comparative example 5

A method for testing strong loquat distillate.

Wherein the diluent is an n-butanol solution.

The rest is the same as example 1.

Comparative example 6

A method for testing strong loquat distillate.

The preparation method of the diluent comprises the following steps:

2) adding 25 parts of petroleum ether into the prepared solution m, heating the solution to 75 ℃ at the speed of 1.5 ℃/min, keeping the temperature, stirring for 4 hours, and cooling to room temperature to obtain the product.

The rest is the same as example 1.

Comparative example 7

A method for testing strong loquat distillate.

The preparation method of the diluent comprises the following steps:

and uniformly mixing 50 parts of isopropanol, 30 parts of toluene, 0.4 part of solubilizer and 25 parts of petroleum ether to obtain the high-performance composite material.

The rest is the same as example 1.

Comparative example 8

A method for testing strong loquat distillate.

The preparation method of the diluent comprises the following steps:

2) the prepared solution m and a solubilizer are mixed according to a volume ratio of 103: 0.7, adding 25 parts of petroleum ether, heating the solution to 75 ℃ at the speed of 1.5 ℃/min, keeping the temperature, stirring for 4 hours, and cooling to room temperature to obtain the product.

The rest is the same as example 1.

Comparative example 9

A method for testing strong loquat distillate.

Wherein, the preparation of the solubilizer comprises the following steps of a:

a. putting 11 parts of tetraethylammonium bromide into a reactor, adding 4 parts of tetrabutylammonium hydrogen sulfate, adding a toluene solution to completely dissolve the tetraethylammonium bromide, heating the tetrabutylammonium hydrogen sulfate to 50 ℃ at the speed of 4 ℃/min to obtain a solution i, and standing and preserving the temperature for later use;

the rest is the same as example 1.

Comparative example 10

A method for testing strong loquat distillate.

Wherein, the preparation of the solubilizer comprises the following steps of a:

a. putting 7 parts of tetraethylammonium bromide into a reactor, adding 2 parts of tetrabutylammonium hydrogen sulfate, adding a toluene solution to completely dissolve the tetraethylammonium bromide, heating the tetrabutylammonium hydrogen sulfate to 50 ℃ at the speed of 4 ℃/min to obtain a solution i, and standing and preserving the temperature for later use;

the rest is the same as example 1.

Comparative example 11

A method for testing strong loquat distillate.

Wherein, the preparation of the solubilizer comprises the following steps of a:

a. putting 7 parts of tetraethylammonium bromide into a reactor, adding 4 parts of tetrabutylammonium hydrogen sulfate, adding a toluene solution to completely dissolve the tetraethylammonium bromide, heating the mixture to 50 ℃ at the speed of 6 ℃/min to obtain a solution i, and standing and preserving the temperature for later use;

the rest is the same as example 1.

Comparative example 12

A method for testing strong loquat distillate.

Wherein, the preparation of the solubilizer comprises the following steps of:

b. putting 6 parts of tetramethylammonium fluoride into a reactor, adding an acetone solution, heating to 90 ℃ at the speed of 2 ℃/min, dropwise adding i 13 parts of the solution into the reactor at the speed of 17 drops/min while stirring, heating the solution to 150 ℃ together, and keeping the temperature for reaction for 4.5 hours to obtain a reactant a;

the rest is the same as example 1.

Comparative example 13

A method for testing strong loquat distillate.

Wherein, the preparation of the solubilizer comprises the following steps of:

b. putting 3 parts of tetramethylammonium fluoride into a reactor, adding an acetone solution, dropwise adding i 13 parts of the solution into the reactor at a speed of 17 drops/min while stirring, heating to 150 ℃ at a speed of 2 ℃/min, and carrying out heat preservation reaction for 4.5 hours to obtain a reactant a;

the rest is the same as example 1.

Comparative example 14

A method for testing strong loquat distillate.

Wherein, the preparation of the solubilizer comprises the following steps of c:

c. adding 18 parts of ethylene glycol dimethyl ether into the reactant a, heating to 50 ℃ at the speed of 5 ℃/min, and carrying out heat preservation reaction for 3.5 hours to obtain the catalyst.

The rest is the same as example 1.

Comparative example 15

A method for testing strong loquat distillate.

Wherein, the preparation of the solubilizer comprises the following steps:

a. and (2) putting 7 parts of tetraethylammonium bromide into a reactor, adding 4 parts of tetrabutylammonium hydrogen sulfate, 3 parts of tetramethylammonium fluoride and 18 parts of ethylene glycol dimethyl ether into the reactor, adding a toluene solution to completely dissolve the tetrabutylammonium hydrogen sulfate, heating the mixture to 120 ℃ at the speed of 4 ℃/min, and carrying out heat preservation reaction for 3-4 hours to obtain the catalyst.

The rest is the same as example 1.

Comparative example 16

A method for testing strong loquat distillate.

a. putting 22 parts of cyclohexane into a reactor, adding 43 parts of ethyl acetate solution, heating to 50 ℃, adding 2 parts of propylene carbonate, heating to 70 ℃, and carrying out heat preservation reaction for 3 hours to obtain an intermediate product a;

The rest is the same as example 1.

Comparative example 17

A method for testing strong loquat distillate.

c. and (3) uniformly mixing the intermediate product b and ethanol according to the volume ratio of 6:1 to obtain the chromatography liquid.

The rest is the same as example 1.

Comparative example 18

A method for testing strong loquat distillate.

0-5 min, 95% of phase A and 5% of phase B;

5-14min later, the phase A is 95-90%, and the phase B is 5-10%;

at 14-25min, phase A is 90-83%, and phase B is 10-17%;

at 25-31min, the A phase is 83-81%, and the B phase is 17-19%;

31-35min later, the phase A is 81-80%, and the phase B is 19-20%;

wherein, the organic phase adopts methanol solution;

The rest is the same as example 1.

Comparative example 19

A method for testing strong loquat distillate.

0-5 min, 95% of phase A and 5% of phase B;

at 5-12min, phase A is 95-90%, and phase B is 5-10%;

when 12-25min, the phase A is 90-84%, and the phase B is 10-16%;

at 25-31min, the A phase is 84-81%, and the B phase is 16-19%;

31-35min later, the phase A is 81-80%, and the phase B is 19-20%;

wherein, the organic phase adopts methanol solution;

The rest is the same as example 1.

Comparative example 20

A method for testing strong loquat distillate.

0-5 min, 95% of phase A and 5% of phase B;

at 5-12min, phase A is 95-90%, and phase B is 5-10%;

when 12-25min, the phase A is 90-83%, and the phase B is 10-17%;

at 25-31min, the A phase is 83-81%, and the B phase is 17-19%;

31-35min later, the phase A is 81-75%, and the phase B is 19-25%;

wherein, the organic phase adopts methanol solution;

The rest is the same as example 1.

Comparative example 21

A method for testing strong loquat distillate.

0-5 min, 95% of phase A and 5% of phase B;

at 5-12min, phase A is 95-90%, and phase B is 5-10%;

when 12-25min, the phase A is 90-83%, and the phase B is 10-17%;

at 25-31min, the A phase is 83-81%, and the B phase is 17-19%;

31-35min later, the phase A is 81-80%, and the phase B is 19-20%;

wherein, the organic phase adopts methanol solution;

0.45 percent aqueous phosphoric acid solution with the flow rate of 0.5 ml/min; the column temperature is 26 ℃, and the detection wavelength is 213 mm;

The rest is the same as example 1.

Comparative example 22

A method for testing strong loquat distillate.

0-5 min, 95% of phase A and 5% of phase B;

at 5-12min, phase A is 95-90%, and phase B is 5-10%;

when 12-25min, the phase A is 90-83%, and the phase B is 10-17%;

at 25-31min, the A phase is 83-81%, and the B phase is 17-19%;

31-35min later, the phase A is 81-80%, and the phase B is 19-20%;

wherein, the organic phase adopts methanol solution;

0.45 percent aqueous phosphoric acid solution with the flow rate of 1.0 ml/min; the column temperature is 33 ℃, and the detection wavelength is 213 mm;

The rest is the same as example 1.

Comparative example 23

A method for testing strong loquat distillate.

0-5 min, 95% of phase A and 5% of phase B;

at 5-12min, phase A is 95-90%, and phase B is 5-10%;

when 12-25min, the phase A is 90-83%, and the phase B is 10-17%;

at 25-31min, the A phase is 83-81%, and the B phase is 17-19%;

31-35min later, the phase A is 81-80%, and the phase B is 19-20%;

wherein, the organic phase adopts methanol solution;

0.45 percent aqueous phosphoric acid solution with the flow rate of 1.0 ml/min; the column temperature is 26 ℃, and the detection wavelength is 213 mm; the column was Agilent HC-C18250X 4.6mm 5 μm.

The rest is the same as example 1.

Comparative example 24

A method for testing strong loquat distillate.

Wherein, the microporous filter membrane in the step 4) is not modified.

The rest is the same as example 1.

Comparative example 25

A method for testing strong loquat distillate.

Wherein, the step 4) also comprises the modification of the microporous filter membrane, and the step 1) in the specific modification step is as follows:

1) soaking the microporous filter membrane in an ethanol solution, simultaneously carrying out microwave vibration on the microporous filter membrane, vibrating the microporous filter membrane at normal temperature for 2 hours, taking out the microporous filter membrane, washing the microporous filter membrane with a methanol solution, and standing for later use;

the rest is the same as example 1.

Comparative example 26

A method for testing strong loquat distillate.

Wherein, the step 4) also comprises the modification of the microporous filter membrane.

2) putting the filter membrane prepared in the step into a dichloromethane solution, adding a modifier, heating to 75 ℃ at the speed of 4 ℃/min, preserving heat and standing for 4.5 hours to obtain a primary modified filter membrane;

The rest is the same as example 1.

Comparative example 27

A method for testing strong loquat distillate.

3) and (3) soaking the preliminary modified filter membrane in a methanol solution, standing at normal temperature for 3.5h, taking out and airing to obtain the modified filter membrane.

The rest is the same as example 1.

Comparative example 28

A method for testing strong loquat distillate.

The preparation method of the modifier comprises the following steps:

i) putting 5 parts of sulfonated polyether ether ketone into a reactor, adding a methanol solution, heating to 43 ℃ at the speed of 2 ℃/min, then adding 3 parts of N, N-dimethylformamide diethyl acetal into the reactor while the reactor is hot, heating to 60 ℃, and carrying out heat preservation reaction for 3.5 hours to obtain a product a;

The rest is the same as example 1.

Comparative example 29

A method for testing strong loquat distillate.

The preparation method of the modifier comprises the following steps:

i) putting 2 parts of sulfonated polyether ether ketone into a reactor, adding a methanol solution, heating to 43 ℃ at the speed of 5 ℃/min, then adding 3 parts of N, N-dimethylformamide diethyl acetal into the reactor while the reactor is hot, heating to 60 ℃, and carrying out heat preservation reaction for 3.5 hours to obtain a product a;

The rest is the same as example 1.

Comparative example 30

A method for testing strong loquat distillate.

The preparation method of the modifier comprises the following steps:

i) putting 2 parts of sulfonated polyether ether ketone and 3 parts of N, N-dimethylformamide diethyl acetal into a reactor, adding a methanol solution into the reactor, heating to 60 ℃ at the speed of 2 ℃/min, and carrying out heat preservation reaction for 3.5 hours to obtain a product a;

The rest is the same as example 1.

Comparative example 31

A method for testing strong loquat distillate.

The preparation method of the modifier comprises the following steps:

ii) adding 3 parts of 2, 3-glycidoxypropyl trimethoxy silane into the product a, adding a dichloromethane solution into the product a, starting a condensation reflux device, heating to 130 ℃ at the speed of 5 ℃/min, and carrying out heat preservation reaction for 5 hours to obtain a product b;

The rest is the same as example 1.

Comparative example 32

A method for testing strong loquat distillate.

The preparation method of the modifier comprises the following steps:

iii) adding 3 parts of N, N-dimethoxyamide into the product b, then adding a dimethyl sulfoxide solution, heating to 83 ℃, and carrying out heat preservation reaction for 4-5 hours to obtain a product c;

The rest is the same as example 1.

Comparative example 33

A method for testing strong loquat distillate.

The preparation method of the modifier comprises the following steps:

iv) adding 4 parts of 3, 3-diethoxypropionic acid ethyl ester into the product c, then adding a toluene solution, heating to 155 ℃ at the speed of 3 ℃/min, starting a condensation reflux device during heating, and carrying out heat preservation reaction for 5.5 hours to obtain the product.

The rest is the same as example 1.

Comparative example 34

A method for testing strong loquat distillate.

The preparation method of the modifier comprises the following steps:

i) putting 2 parts of sulfonated polyether ether ketone, 3 parts of 2, 3-glycidoxypropyl trimethoxy silane, 5 parts of N, N-dimethoxyamide and 1.5 parts of 3, 3-diethoxypropionic acid ethyl ester into a reactor, adding a methanol solution, heating to 43 ℃ at the speed of 2 ℃/min, adding 3 parts of N, N-dimethylformamide diethyl acetal into the reactor while the temperature is still hot, heating to 60 ℃, and carrying out heat preservation reaction for 8 hours to obtain the sulfonated polyether ether ketone.

The rest is the same as example 1.

Comparative example 35

The detection method in the patent No. CN 106198832B 'production quality control method of strong loquat dew' is adopted to detect the strong loquat dew product.

Experimental example 1 reproducibility of standard fingerprint of standard strong loquat syrup

The high performance liquid phase fingerprint spectrogram of the standard loquat dew obtained by the examples and the comparative examples is subjected to reproducibility analysis test, the relative peak area RSD is detected and analyzed, and the specific test analysis results are shown in Table 1:

TABLE 1 reproducibility of liquid chromatography

As can be seen from the data in table 1, the fingerprints of the standard strong loquat dew obtained in examples 1 to 3 have excellent reproducibility, the relative peak areas are all within 1%, the RSD of the comparative examples is all above 1%, the repeatability of the characteristic fingerprint is poor, and when the RSD data is greater than 3%, the fingerprint cannot be used as the judgment fingerprint of the standard strong loquat dew for detecting samples. Among them, comparative examples 1 to 4 change the process to the process in the test of the standard strong loquat dew sample, and further change the standard curve of the test, so the reproducibility is reduced, but still can be used as the standard judgment curve. Comparative examples 5 to 8 change the diluent or the preparation process and the effective ingredient of the diluent, resulting in a decrease in stability of data, but a part of the test standard curve is still available; comparative examples 9 to 15 change the formulation or preparation method of the solubilizing agent, resulting in change of the effective ingredient entering the liquid chromatography column, so that the reproducibility was lowered; comparative examples 18 to 23 changed the formulation of the eluent or the elution order of the eluent during the detection process, which further led to the decrease of the repeatability of the characteristic fingerprint spectrum; comparative examples 24-34 alter the microporous membrane modification process and thus also have an effect on the RSD of the fingerprint.

Experimental example 2 detection efficiency of strong loquat syrup

90 samples of strong loquat distillate produced in the same batch are detected by spectrograms of the standard strong loquat distillate obtained by the examples and the comparative examples, and 10 interference components distributed by pure effective components in the strong loquat distillate are detected at the same time, wherein the preparation method of the interference component solution is as follows:

precisely weighing 13.74mg of papaverine hydrochloride reference substance, 11.64mg of narcotine reference substance, 15.50mg of thebaine reference substance, 11.57mg of morphine reference substance and 14.52mg of codeine phosphate reference substance, respectively placing in a 20mL measuring flask, adding a proper amount of methanol solution containing 0.5% formic acid, ultrasonically dissolving, fixing the volume to scale, and shaking up to obtain the reference substance stock solution. Precisely measuring 30uL of papaverine reference substance stock solution, 40uL of narcotine reference substance stock solution, 15uL of thebaine reference substance stock solution, 1400uL of morphine reference substance stock solution and 300uL of codeine reference substance stock solution, placing into a 10mL measuring flask, and adding methanol to constant volume to obtain a mixed reference substance solution. Precisely absorbing morphine-D₃codeine-D₃And (3) placing 1.0mL of each internal standard solution into a 20mL measuring flask, and adding methanol to dilute to a scale mark to obtain a mixed internal standard solution.

Precisely measuring 1.0mL of a sample, placing the sample into a 50mL polytetrafluoroethylene centrifuge tube with a plug, adding 200uL of mixed internal standard working solution, adding 5mL of water, shaking uniformly, precisely adding 15.0mL of acetonitrile, vortexing for 1min, adding a QuEChERS AOAC extraction salt bag, rapidly shaking, vortexing for 1min, centrifuging for 5min at 10000r/min, transferring 10mL of supernatant into a 15mL of QuEChERS AOAC dispersed solid phase extraction tube, covering a bottle cap, performing vortex shaking for 2min with force, centrifuging for 5min at 8000r/min, taking supernatant, filtering with a 0.22um organic microporous filter membrane, and taking filtrate to be detected.

The detection efficiency is determined by the detection time and the detection accuracy, wherein the calculation formula of the detection efficiency is

η=(（T_max-T_i）×100%/T_max)*0.55+β*0.45

Wherein η (%) is the detection efficiency;

T_max(min) is the conventional longest length of a single sampleDetecting time;

T_i(min) is the detection time of the ith sample;

β (%) is the accuracy;

the specific addition results are shown in table 2:

TABLE 2 efficiency of the assay

As can be seen from Table 2, the accuracy rates of examples 1-3 are all 100%, and the relative detection time has the largest value in all comparative examples, which indicates that the detection time of examples 1-3 is the shortest in the total detection time, so the detection efficiency is the highest; the prior art is adopted for detection in the comparative example 36, but because an interference group exists, the comparative example 36 can only judge whether the sample is strong loquat dew according to the existing maximum component detection in the sample, and ignores the existence of other tiny components in the strong loquat dew, so that the accuracy is reduced, and the identification capability of a spectrogram is weak. Comparative examples 1 to 4 change the pretreatment process of the strong loquat syrup, and comparative examples 5 to 8 change the effective components in the diluent, and the changes result in the reduction of the extraction rate of the effective components in the product, so the efficiency of detecting the product is reduced; comparative examples 9 to 15 were modified for the preparation of the solubilizing agent in the diluent, wherein comparative examples 9 to 10 and comparative example 12 were modified for the ratio of raw material components in the solubilizing agent, and comparative examples 11 and comparative examples 13 to 15 were modified for the process in the preparation of the solubilizing agent, which resulted in the modification of the effective components in the solubilizing agent, lowering the extraction rate of the solubilizing agent, and thus in the detection process, lowering the detection efficiency; comparative examples 16 to 17 change the composition of the chromatography liquid, resulting in poor chromatography effect, but rather, prolonged detection time, further reducing detection efficiency.

Claims

1. A detection method of strong loquat syrup is characterized by comprising the following steps: the detection method of the strong loquat syrup comprises the following steps:

first), establishment of standard strong loquat dew product detection method

II), detection of the sample

2. The method for detecting loquat leaf extract according to claim 1, wherein the method comprises the following steps: the preparation method of the diluent comprises the following steps:

3. The method for detecting loquat leaf extract according to claim 2, wherein the method comprises the following steps: preparation of the solubilizer:

4. The method for detecting loquat leaf extract according to claim 1, wherein the method comprises the following steps: before liquid chromatography, preliminary analysis is carried out through thin-layer chromatography, waste of detection cost is reduced, and the specific detection method comprises the following steps:

5. The method of claim 4, wherein the method comprises the following steps: the preparation of the chromatographic solution comprises the following steps:

6. The method for detecting loquat leaf extract according to claim 1, wherein the method comprises the following steps: the elution sequence in the liquid chromatography of step 4) is as follows:

0-5 min, 95% of phase A and 5% of phase B;

at 5-12min, phase A is 95-90%, and phase B is 5-10%;

when 12-25min, the phase A is 90-83%, and the phase B is 10-17%;

at 25-31min, the A phase is 83-81%, and the B phase is 17-19%;

31-35min later, the phase A is 81-80%, and the phase B is 19-20%;

wherein, the organic phase adopts methanol solution;

7. The method for detecting loquat leaf extract according to claim 1, wherein the method comprises the following steps: step 4) also comprises the modification of the microporous filter membrane, and the specific modification steps are as follows:

8. The method of claim 7, wherein the method comprises the steps of: the preparation method of the modifier comprises the following steps: