CN115856122A - Method for simultaneously detecting multiple medicinal components in Yu Huang oral liquid by UPLC-PDA - Google Patents

Abstract

The invention belongs to the technical field of veterinary drug detection, and particularly relates to a method for simultaneously detecting multiple medicinal components in Yu Huang oral liquid by using UPLC-PDA. The invention can simultaneously determine the content of 10 components of berberine hydrochloride, palmatine hydrochloride, baicalin, geniposide, gallic acid, paeoniflorin, rhein, emodin, physcion and chrysophanol in Yu Huang oral liquid, thereby simultaneously detecting 7 medicines of coptis chinensis, phellodendron, scutellaria baicalensis, gardenia, myrobalan, white paeony root and rhubarb.

Description

Method for simultaneously detecting multiple medicinal components in Yu Huang oral liquid by UPLC-PDA

Technical Field

The invention belongs to the technical field of veterinary drug detection, and particularly relates to a method for simultaneously detecting multiple medicinal components in Yu Huang oral liquid by using UPLC-PDA.

Background

The information disclosed in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.

Yu Huang oral liquid is a Chinese veterinary drug prescription preparation, and is collected in 2017 edition of Chinese medicinal roll p184 from veterinary drug national standard, and the prescription comprises: 250g of radix curcumae, 220g of myrobalan, 50g of gardenia, 50g of scutellaria baicalensis, 50g of rheum officinale, 30g of radix paeoniae alba, 50g of golden cypress and 50g of coptis chinensis. In the standard, thin layer chromatography is used for detecting gardenia (geniposide), myrobalan (gallic acid), coptis (berberine hydrochloride) and rhubarb in Yu Huang oral liquid, and high performance liquid chromatography is used for detecting the content of baicalin. It takes about 2-3 days to complete a sample using the standard test method.

By adopting thin-layer chromatography, the influence of environmental temperature and humidity and manual operation on spots is very large, the spots are often unclear, the result is difficult to judge and time is consumed, fei Shiji, the toxicity of a developing agent for chromatography is found to be large in actual inspection work, and the developing agent for chromatography is large in harm to experimenters, such as trichloromethane, ethyl acetate, methanol, acetone, formic acid, benzene, isopropanol, ether, petroleum ether (30-60 ℃), ethyl formate and other various reagents. And the operation is complicated, the time consumption is more, and various effective components can not be detected simultaneously.

In addition, the thin-layer chromatography is complex in preparation in the early stage, multiple steps of pre-saturation, unfolding, airing, checking and checking results and the like are needed, time and labor are wasted, interference caused by factors such as environmental temperature and humidity is large, edge effects and the like exist, thin-layer spots are difficult to distinguish sometimes by methods, and reproducibility is poor.

Disclosure of Invention

In order to solve the defects of the prior art, the invention aims to provide a method for simultaneously detecting multiple medicinal components in Yu Huang oral liquid by using UPLC-PDA. The invention can simultaneously determine the content of 10 components of berberine hydrochloride, palmatine hydrochloride, baicalin, geniposide, gallic acid, paeoniflorin, rhein, emodin, physcion and chrysophanol in Yu Huang oral liquid, thereby simultaneously detecting 7 medicines of coptis chinensis, phellodendron, scutellaria baicalensis, gardenia, myrobalan, white paeony root and rheum officinale.

In order to achieve the purpose, the invention is realized by the following technical scheme:

in a first aspect, the invention provides a method for simultaneously detecting multiple medicinal components in Yu Huang oral liquid by UPLC-PDA, which comprises the following steps:

A. dissolving berberine hydrochloride, palmatine hydrochloride, baicalin, geniposide, gallic acid, paeoniflorin, rhein, emodin, physcion and chrysophanol reference substances in a solvent to respectively prepare reference stock solutions, and mixing the reference stock solutions to obtain a mixed reference stock solution;

B. dissolving the Yuhuang oral liquid sample with a solvent, performing ultrasonic extraction, centrifuging, diluting and filtering to obtain a sample solution to be detected;

C. injecting the mixed contrast stock solution into an UPLC-PDA chromatograph, performing gradient elution and detection, determining the nature by retention time, and drawing a standard curve according to the corresponding relation between the peak area measured by the sample amount and the sample amount;

D. and C, injecting the sample solution to be detected into an UPLC-PDA chromatograph, performing gradient elution and detection under the same conditions as those in the step C, measuring the peak area of each target object, determining the qualitative retention time, quantifying according to the standard curve manufactured in the step C, and calculating the content of the effective components in the Yu Huang oral liquid sample.

The beneficial effects obtained by one or more technical schemes of the invention are as follows:

1. the invention adopts UPLC-PDA method to carry out determination, and uses waters ACQUITY UPLC HSST ₃ The chromatographic column is an optimized chromatographic column, acetonitrile (0.05 percent of phosphoric acid) -0.05 percent of phosphoric acid aqueous solution is subjected to gradient elution, 3D scanning (190 nm-400 nm) is used for detecting by using detection wavelengths of 232nm,238nm,257nm,278nm and 289nm, and 10 main components in Yu Huang oral liquid can be simultaneously detected at one time.

2. According to the determination method, a test sample is extracted by taking (V50% methanol: V0.05% phosphoric acid aqueous solution = 1:1) as an extraction solvent, the toxicity is lower compared with trichloromethane ethyl ether and the like used in a thin layer, the peak area response value of a compound is moderate, the sample is simple and convenient to process, the reagent and the time are saved, the used solvent is low in toxicity, safe and environment-friendly to operate, and the harm to people and the environment is low; and 4 thin layer survey of former standard, the time such as processing development is dried is long, the multiple reagent of adoption: benzene, acetone, ethyl acetate, isopropanol, methyl acetate and the like, wherein the benzene belongs to a class of carcinogens, the used reagent has high toxicity, spots are sometimes unclear, the influence of environment temperature and humidity, personnel operation and the like is large, and the spot result is sometimes difficult to judge.

3. The determination method of the invention uses the high performance liquid chromatograph, presents the result in the form of chromatogram and spectrogram, is convenient and fast, has visual result, is easy to judge, has low detection limit, greatly shortens the detection time, and improves the detection efficiency, the detection sensitivity and the detection effect.

4. According to the method, a sample can be detected in about 2 hours from sample treatment to product judgment on the machine; in the traditional Chinese medicine rolls of the 2017 edition of the national standard of veterinary medicines, during the thin-layer detection, the pretreatment of samples is complicated, some samples need to be heated and refluxed, some samples need to be dried by distillation in a water bath, and some samples need to be shaken and extracted. The method needs to prepare the developing agent, needs multiple steps of presaturation, development, airing and the like, and needs about 2-3 days for one sample.

Drawings

The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification, illustrate exemplary embodiments of the invention and together with the description serve to explain the invention and not to limit the invention.

FIG. 1 is a spectrum diagram of berberine hydrochloride, palmatine hydrochloride, baicalin, geniposide, gallic acid, paeoniflorin, rhein, emodin, physcion and chrysophanol;

FIG. 2 is a chromatogram of the mixed control sample of example 1 at a detection wavelength of 232 nm;

FIG. 3 is a chromatogram of the mixed control sample of example 1 at a detection wavelength of 238 nm;

FIG. 4 is a chromatogram of the mixed control sample of example 1 at a detection wavelength of 257 nm;

FIG. 5 is a chromatogram of the mixed control sample of example 1 at a detection wavelength of 278 nm;

FIG. 6 is a chromatogram of the mixed control sample of example 1 at a detection wavelength of 289 nm;

FIG. 7 is a chromatogram of a 0037 sample in example 1 at a detection wavelength of 232 nm;

FIG. 8 is a chromatogram of a 0037 sample in example 1 at a detection wavelength of 238 nm;

FIG. 9 is a chromatogram of a 0037 sample in example 1 at a detection wavelength of 257 nm;

FIG. 10 is a chromatogram of a 0037 sample in example 1 at a detection wavelength of 278 nm;

FIG. 11 is a chromatogram of a 0037 sample in example 1 at a detection wavelength of 289 nm;

FIG. 12 is a chromatogram of a 1751 sample in example 2 at a detection wavelength of 232 nm;

FIG. 13 is a chromatogram of a 1751 sample in example 2 at a detection wavelength of 238 nm;

FIG. 14 is a chromatogram of the 1751 sample from example 2 at a detection wavelength of 257 nm;

FIG. 15 is a chromatogram of the 1751 sample from example 2 at a detection wavelength of 278 nm;

FIG. 16 is a chromatogram of the 1751 sample from example 2 at a detection wavelength of 289 nm.

Detailed Description

It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.

In a first exemplary embodiment of the present invention, a method for simultaneously detecting multiple pharmaceutical components in Yu Huang oral liquid by using UPLC-PDA comprises the following steps:

A. dissolving berberine hydrochloride, palmatine hydrochloride, baicalin, geniposide, gallic acid, paeoniflorin, rhein, emodin, physcion and chrysophanol reference substances in a solvent to respectively prepare reference stock solutions, and mixing the reference stock solutions to obtain a mixed reference stock solution;

B. dissolving a sample of the Yuhuang oral liquid by using a solvent, performing ultrasonic extraction, centrifuging, diluting and filtering to obtain a sample solution to be detected;

C. injecting the mixed contrast stock solution into an UPLC-PDA chromatograph, performing gradient elution and detection, determining the nature by retention time, and drawing a standard curve according to the corresponding relation between the peak area measured by the sample amount and the sample amount;

D. and C, injecting the sample solution to be detected into an UPLC-PDA chromatograph, performing gradient elution and detection under the same conditions as those in the step C, measuring the peak area of each target object, determining the qualitative retention time, quantifying according to the standard curve manufactured in the step C, and calculating the content of the effective components in the Yu Huang oral liquid sample.

In one or more embodiments of this embodiment, the solvent in step a is one or more of absolute ethanol, methanol, chloroform.

In one or more embodiments of this embodiment, the solvent in step B is methanol, absolute ethanol, 50% aqueous methanol, 50% aqueous ethanol, methanol: 0.05% phosphoric acid = 50.

50% aqueous methanol solution, 50% aqueous ethanol solution, methanol: the concentrations in the mixed solution of 0.05% phosphoric acid =50 were all volume concentrations.

In one or more embodiments of the present embodiment, the ultrasonic extraction time in step B is 15-30min, the centrifugation speed is 10000r/min, and the centrifugation time is 5-20min.

In one or more embodiments of this embodiment, the sample size in step C is 0.5-5 μ L, the mobile phase flow rate is 0.34-0.36mL/min, and the column temperature is 39-41 ℃.

In one or more embodiments of this embodiment, the mobile phase in step C comprises mobile phase a and mobile phase B, the mobile phase a is a 0.05% by volume solution of acetonitrile phosphate, and the mobile phase B is a 0.05% by volume solution of phosphoric acid in water.

In one or more embodiments of this embodiment, the gradient elution in step C comprises the following stages:

0min,8% mobile phase a,92% mobile phase B, initial curve;

0.5min,8% mobile phase a,92% mobile phase B, curve 6;

3min,20% mobile phase A,80% mobile phase B, curve 6;

4.5min,25% mobile phase a,75% mobile phase B, curve 6;

5.5min,28% mobile phase a,72% mobile phase B, curve 6;

8.5min,28% mobile phase a,72% mobile phase B, curve 6;

10min,50% mobile phase a,50% mobile phase B, curve 6;

11min,50% mobile phase a,50% mobile phase B, curve 6;

11.5min,70% mobile phase a,30% mobile phase B, curve 6;

14min,70% mobile phase a,30% mobile phase B, curve 6;

14.5min,8% mobile phase a,92% mobile phase B, curve 6;

15min,8% mobile phase a,92% mobile phase B, curve 6.

In one or more embodiments of this embodiment, step C uses 3D scanning (190-400 nm) with detection wavelengths of one or more of 232nm,238nm,257nm,278nm,289 nm.

In one or more embodiments of this embodiment, step B uses 8% mobile phase a and 92% mobile phase B as dilutions for dilution in admixture with supernatant 1:1 obtained by centrifugation.

In one or more embodiments of the embodiment, the Yu Huang oral liquid sample can simultaneously detect the content of berberine hydrochloride, palmatine hydrochloride, baicalin, geniposide, gallic acid, paeoniflorin, rhein, emodin, physcion and chrysophanol through a chromatogram and a spectrogram, and further simultaneously detect the content of coptis chinensis, phellodendron amurense, scutellaria baicalensis, gardenia, myrobalan, radix paeoniae alba and rhubarb in a Yu Huang oral liquid sample.

In order to make the technical solution of the present invention more clearly understood by those skilled in the art, the technical solution of the present invention will be described in detail below with reference to specific examples and comparative examples.

In the embodiment of the invention:

the source of the reference substance: purchased from units such as China institute for veterinary medicine, china institute for food and drug testing, and China institute for biological product testing.

The source of the test sample is as follows: yellow stagnation oral liquid: 1571 sample, jinan Guangsheng Biotech limited; 0037 sample manufactured by Shandong Parsen pharmaceutical Co., ltd. Is a sample produced by enterprises according to the prescription of pharmacopoeia strictly.

An instrument device: BP211D analytical balance (sidoris, germany); waters AcquisytTM ultra high performance liquid chromatograph (Waters corporation, USA), PDA detector, chromatographic column: waters ACQUITY UPLC HSS T3 chromatographic column (2.1 mm. Times.100mm, 1.8 μm).

Example 1

0037 detection of samples

Dissolving berberine hydrochloride, palmatine hydrochloride, baicalin, geniposide, gallic acid, paeoniflorin, rhein, emodin, physcion and chrysophanol in suitable solvent, and diluting to desired volume to obtain respective control stock solutions; the control stock solutions were mixed as shown in table 1 to obtain a mixed control stock solution. Weighing a proper amount of benzoic acid reference substance, independently dissolving the benzoic acid reference substance by using ethanol to prepare benzoic acid stock solution, and adding 10 mu L of benzoic acid stock solution into 10ml of mixed reference substance solution, wherein the total volume is almost ignored.

TABLE 1 preparation table of 10 kinds of mixed control for Yuhuang oral liquid

2.0mL of a 0037 sample was precisely measured, placed in a 10mL volumetric flask, and mixed with methanol: dissolving a mixed solution of (0.05% phosphoric acid) =50, fixing the volume to a scale, weighing, ultrasonically extracting for 20min, cooling to room temperature, weighing, complementing the weight loss by using a proper solvent, taking out 5mL, centrifuging at 10000rpm/min for 10min, taking a supernatant, diluting with a mixed solution consisting of 8% phosphoric acid acetonitrile solution with the volume concentration of 0.05% and 92% phosphoric acid water solution with the volume concentration of 0.05% by 1:1, filtering, and obtaining a 0037 sample solution for liquid chromatography determination.

The UPLC detection instrument is an ultra-high performance liquid chromatograph, the PDA detector is a photodiode array detector, and the chromatographic column is a Waters ACQUITY UPLC HSST ₃ The chromatographic column has a column temperature of 40 ℃, and the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is a phosphate acetonitrile solution with a volume concentration of 0.05%, and the mobile phase B is a phosphate acetonitrile solution with a volume concentration of 0.05%The flow rate of the phosphoric acid aqueous solution was 0.35mL/min, and the elution procedure was as shown in Table 2. PDA selects 3D scanning (190-400 nm), and simultaneously multi-wavelength detection selects channels of 232nm,238nm,257nm,278nm and 289nm.

TABLE 2 gradient elution procedure

	Flow rate of flow	A％	B％	Curve
					Initial	0.35	8	92	Initial
0.5	0.35	8	92	6
					3	0.35	20	80	6
4.5	0.35	25	75	6
					5.5	0.35	28	72	6
8.5	0.35	28	72	6
					10	0.35	50	50	6
11	0.35	50	50	6
					11.5	0.35	70	30	6
14	0.35	70	30	6
					14.5	0.35	8	92	6
15	0.35	8	92	6

And injecting the mixed control stock solution into an UPLC-PDA chromatograph, performing gradient to reserve time for qualitative determination, and drawing a standard curve according to the corresponding relation between the peak area measured by the sample amount and the sample amount. The results of the test with the mixed control stock solution are shown in table 3.

Table 3 mixed control stock solution chromatographic data

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0037 injecting the sample solution into UPLC-PDA chromatograph, performing gradient elution and detection, measuring peak area of each target object, determining the nature by retention time, quantifying according to standard curve, and calculating the content of effective components in Yu Huang oral liquid sample. 0037 chromatographic data of the sample solutions are shown in table 4.

TABLE 4 0037 chromatographic data for sample solutions

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As shown in fig. 1, the spectra of the various compounds are characterized as follows: wavelength at the peak of gallic acid: 215.8nm, 272.0nm;

the wavelength at the gardenoside peak: 240.2nm;

wavelength at the paeoniflorin peak: 232.3nm;

wavelength at baicalin peak: 277.5nm, 316.3nm;

wave length at wave crest of palmatine hydrochloride: 226.2nm, 273.9nm, 345.5nm;

wave length at the wave crest of berberine hydrochloride: 229.2nm, 264.7nm, 347.9nm;

wavelength at peak of chrysophanol: 225.2nm, 257.9nm, 288.0nm;

wavelength at emodin peak: 222.5nm, 288.6nm;

wavelength at rhein peak: 231.0nm and 258.5nm;

physcion peak wavelength: 223.1nm, 266.5nm and 286.1nm.

Multiple compounds are qualitatively identified by chromatogram retention time and features of the spectrogram.

As shown in fig. 2-6, chromatograms of the mixed control stock solutions obtained under PDA channels with different wavelengths indicate: geniposide paeoniflorin hardly responds at 278nm and 289 nm; paeoniflorin did not respond at 257 nm; all materials have response values at 238nm and 232nm; rhein response at 238nm was much lower than 232 nm. FIGS. 7-11 are chromatograms of 0037 sample solutions obtained in PDA channels with different wavelengths, which, by comparison of the chromatograms, respond relatively better to compounds at 232nm, in order to optimize the wavelength. Therefore, the effective component content was calculated using the chromatographic data of the 0037 sample solution obtained using the 232nm channel, and the results are shown in Table 5.

TABLE 5 0037 chromatographic data of sample solution in 232nm channel and effective component content calculation results

0037 the sample detects 9 compounds, wherein the content of baicalin is 10.8mg/ml at most, the content of gallic acid is 3.1mg/ml, the content of geniposide is 2.8mg/ml and paeoniflorin is 267.3 mu g/ml, rhein 158.862 mu g/ml, emodin 82.33 mu g/ml, berberine hydrochloride 78.326 mu g/ml, palmatine hydrochloride 27.788 mu g/ml, chrysophanol 18.612 mu g/ml, and physcion is not detected. In addition, the preservative benzoic acid of the sample is detected, quantification is not carried out, and the preservative is well separated from other compounds, so that the requirements are met.

Example 2

1751 detection of samples

Unlike example 1, the test sample was changed from the 0037 sample to 1751 sample.

1751 the chromatograms of the sample solution at 232nm,238nm,257nm,278nm and 289nm are shown in FIGS. 12-16, and the data are shown in Table 6. Similar to example 1, geniposide paeoniflorin was almost non-responsive at 278nm and 289 nm; paeoniflorin did not respond at 257 nm; all materials have response values at 238nm and 232nm; rhein response at 238nm was much lower than 232 nm. And through comparison of chromatograms, each compound at 232nm has relatively better response and is optimized for wavelength.

TABLE 6 1751 sample solution chromatography data

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Therefore, the effective component content was calculated using the chromatographic data of the 1571 sample solution obtained through the 232nm channel, and the results are shown in Table 7. 1751, detecting 7 compounds, wherein the baicalin content is 2.2mg/ml at most, the gallic acid 468.276 μ g/ml, the geniposide 279.765 μ g/ml and the paeoniflorin 336.440 μ g/ml are less, the emodin, the berberine hydrochloride and the palmatine hydrochloride are lower in the range of 30-50 μ g/ml, and rhein, chrysophanol and physcion are not detected. The preservative benzoic acid of the sample is detected, quantification is not carried out, the preservative is well separated from other compounds, and the requirements are met.

TABLE 7 1571 chromatographic data of sample solution at 232nm channel and effective component content calculation results

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The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made by using various liquid phases of different brands, chromatographic columns of different brands, different mobile phase compositions and elution modes, etc. within the spirit and principle of the present invention, should be included in the protection scope of the present invention.

Claims (10)

1. A method for simultaneously detecting multiple medicinal components in Yu Huang oral liquid by UPLC-PDA is characterized by comprising the following steps:

A. dissolving berberine hydrochloride, palmatine hydrochloride, baicalin, geniposide, gallic acid, paeoniflorin, rhein, emodin, physcion and chrysophanol reference substances in a solvent to respectively prepare reference stock solutions, and mixing the reference stock solutions to obtain a mixed reference stock solution;

B. dissolving the Yuhuang oral liquid sample with a solvent, performing ultrasonic extraction, centrifuging, diluting and filtering to obtain a sample solution to be detected;

C. injecting the mixed contrast stock solution into an UPLC-PDA chromatograph, performing gradient elution and detection, determining the nature by retention time, and drawing a standard curve according to the corresponding relation between the peak area measured by the sample amount and the sample amount;

D. and C, injecting the sample solution to be detected into an UPLC-PDA chromatograph, performing gradient elution and detection under the same conditions as those in the step C, measuring the peak area of each target object, determining the qualitative retention time, quantifying according to the standard curve manufactured in the step C, and calculating the content of the effective components in the Yu Huang oral liquid sample.

2. The method for simultaneously detecting Yu Huang oral liquids by UPLC-PDA as claimed in claim 1, wherein the solvent in step a is one or more of absolute ethanol, methanol, and chloroform.

3. The method for simultaneously detecting Yu Huang oral liquids by UPLC-PDA as claimed in claim 1, wherein the solvent in step B is methanol, absolute ethanol, 50% aqueous methanol, 50% aqueous ethanol, methanol: 0.05% phosphoric acid = 50.

4. The method for simultaneously detecting Yu Huang oral liquids by UPLC-PDA as claimed in claim 1, wherein ultrasonic extraction time in step B is 15-30min, centrifugation speed is 10000r/min, and centrifugation time is 5-20min.

5. The method for simultaneously detecting Yu Huang oral liquids by UPLC-PDA as claimed in claim 1, wherein the sample amount in step C is 0.5-5 μ L, the mobile phase flow rate is 0.34-0.36mL/min, and the column temperature is 39-41 ℃.

6. The method for simultaneously detecting Yu Huang oral liquids by UPLC-PDA as claimed in claim 1, wherein the mobile phase in step C comprises mobile phase a and mobile phase B, the mobile phase a is 0.05% by volume acetonitrile phosphate solution, and the mobile phase B is 0.05% by volume aqueous phosphoric acid solution.

7. The method for simultaneously detecting Yu Huang oral liquids with UPLC-PDA as claimed in claim 6, wherein the gradient elution in step C comprises the following stages:

0min,8% mobile phase a,92% mobile phase B, initial curve;

0.5min,8% mobile phase a,92% mobile phase B, curve 6;

3min,20% mobile phase A,80% mobile phase B, curve 6;

4.5min,25% mobile phase a,75% mobile phase B, curve 6;

5.5min,28% mobile phase a,72% mobile phase B, curve 6;

8.5min,28% mobile phase a,72% mobile phase B, curve 6;

10min,50% mobile phase a,50% mobile phase B, curve 6;

11min,50% mobile phase a,50% mobile phase B, curve 6;

11.5min,70% mobile phase a,30% mobile phase B, curve 6;

14min,70% mobile phase a,30% mobile phase B, curve 6;

14.5min,8% mobile phase a,92% mobile phase B, curve 6;

15min,8% mobile phase a,92% mobile phase B, curve 6.

8. The method for simultaneously detecting Yu Huang oral liquids using UPLC-PDA as claimed in claim 1 wherein 3D scanning (190-400 nm) is used in step C with detection wavelength of one or more of 232nm,238nm,257nm,278nm,289 nm.

9. The method for simultaneously detecting multiple medicinal components in Yu Huang oral liquid by UPLC-PDA as claimed in claim 6, wherein 8% mobile phase a and 92% mobile phase B are used as diluent in step B, and mixed and diluted with supernatant 1:1 obtained by centrifugation.

10. The method for simultaneously detecting multiple medicinal components in Yu Huang oral liquid by UPLC-PDA as in claims 1-9, wherein Yu Huang oral liquid sample can simultaneously detect berberine hydrochloride, palmatine hydrochloride, baicalin, geniposide, gallic acid, paeoniflorin, rhein, emodin, physcion and chrysophanol content by chromatogram and spectrogram, and further simultaneously detect the composition content of coptis chinensis, phellodendron, scutellaria baicalensis, gardenia, myrobalan, radix paeoniae alba and rhubarb in Yu Huang oral liquid sample.

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