阅读说明：本技术 一种有柄石韦药材及其饮片、标准汤剂、配方颗粒的特征图谱、构建方法和检测方法 (Characteristic spectrum, construction method and detection method of pyrrosia pedunculata medicinal material, decoction pieces, standard decoction and formula granules thereof ) 是由 周厚成 胡昌江 辜新月 陈玉梅 宋媛 叶志萍 周维 于 2020-10-26 设计创作，主要内容包括：本发明公开了一种有柄石韦药材及其饮片、标准汤剂、配方颗粒的特征图谱、构建方法和检测方法,属于中药检测技术领域,通过高效液相色谱法建立有柄石韦药材及其饮片、标准汤剂、配方颗粒的特征图谱,包括其相应的供试品的溶液制备,对照品的选取和对照溶液的制备进行的研究,得到相应的特征图谱,可以快速判断是否是有柄石韦的药材或饮片或标准汤剂或配方颗粒,同时填补了现有技术中药材有柄石韦质量控制的空白,特别是在有柄石韦药材至有柄石韦配方颗粒成品的整个工艺过程中,质量的整体评价和控制的空白,为之后中药-有柄石韦的研究提供有效研究基础。(The invention discloses a pyrrosia pedunculata medicinal material, a characteristic map of decoction pieces, standard decoction and formula granules of the pyrrosia pedunculata medicinal material, a construction method and a detection method, belonging to the technical field of traditional Chinese medicine detection, establishing characteristic maps of pyrrosia petiolata medicinal material and decoction pieces, standard decoction and formula granules thereof by high performance liquid chromatography, including the corresponding solution preparation of a test sample, the research on the selection of the reference substance and the preparation of the reference solution obtains a corresponding characteristic spectrum, can quickly judge whether the pyrrosia petiolata is a medicinal material or a decoction piece or a standard decoction or a formula granule, and simultaneously fills the blank of the quality control of the pyrrosia petiolata in the prior art, particularly, in the whole process from the pyrrosia peduncularis medicinal material to the pyrrosia peduncularis formula granule finished product, the blank of the overall evaluation and control of the quality provides an effective research basis for the later research of the traditional Chinese medicine pyrrosia peduncularis.)

1. A method for constructing a characteristic spectrum of pyrrosia pedunculata medicinal material, decoction pieces thereof, standard decoction and formula granules is characterized by comprising the following steps:

a. preparation of control solutions: dissolving reference substances of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C in solvent to obtain reference solution;

b. preparation of a test solution: respectively preparing test solution of pyrrosia petiolata medicinal material, decoction pieces, standard decoction and formula granules thereof;

c. and injecting the reference substance solution and the test solution into a high performance liquid chromatograph for detection, and selecting common peaks from the characteristic spectrum of the test solution by taking the characteristic spectrum of the reference substance solution as a reference spectrum to construct the characteristic spectrum of the pyrrosia lingua medicinal material, the decoction pieces, the standard decoction and the formula granules of the pyrrosia lingua medicinal material.

2. The method for constructing the characteristic spectrum of pyrrosia petiolata medicinal material, decoction pieces thereof, standard decoction thereof and formula granules according to claim 1, wherein in the step a, the preparation method of the reference solution comprises the following steps: taking neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C as reference substances, respectively adding 50% methanol, and making into solutions each containing 40 μ g of reference substance per 1 ml.

3. The method for constructing the characteristic spectrum of the pyrrosia pedunculata medicinal material and the decoction pieces, the standard decoction and the formula granules thereof as claimed in claim 2, wherein in the step b, the preparation method of the pyrrosia pedunculata medicinal material test solution and the pyrrosia pedunculata decoction piece test solution comprises the following steps: taking pyrrosia peduncularis medicinal material or pyrrosia peduncularis decoction piece powder, sieving by a second sieve, precisely weighing 0.5g, adding 15ml of water, decocting for 30 minutes, adding 10ml of methanol, sealing, carrying out ultrasonic treatment for 10 minutes, weighing again, complementing the lost weight with methanol, shaking up, filtering, and taking the subsequent filtrate to obtain the pyrrosia peduncularis medicinal material test solution or pyrrosia peduncularis decoction piece test solution.

4. The method for constructing the characteristic spectrum of the pyrrosia petiolata medicinal material, decoction pieces, standard decoction and formula granules of the pyrrosia petiolata medicinal material according to claim 2, wherein in the step b, the method for preparing the test solution of the pyrrosia petiolata standard decoction comprises the following steps: precisely weighing 0.3g of freeze-dried powder of the product, precisely adding 20ml of 50% methanol, sealing, performing ultrasonic treatment for 30 minutes, weighing again, supplementing the lost weight with 50% methanol, shaking up, filtering, and taking the subsequent filtrate to obtain the test solution of the pyrrosia petiolata standard decoction.

5. The method for constructing the characteristic spectrum of pyrrosia petiolata medicinal material, decoction pieces thereof, standard decoction thereof and formula granules according to claim 2, wherein in the step b, the method for preparing the test solution of pyrrosia petiolata formula granules comprises the following steps: taking a proper amount of the granules, accurately weighing about 0.3g after grinding, accurately adding 20ml of 50% methanol, sealing, carrying out ultrasonic treatment for 30 minutes, weighing again, complementing the lost weight with 50% methanol, shaking up, filtering, and taking the subsequent filtrate to obtain a test solution of the pyrrosia petiolata formula granules.

6. The method for constructing the characteristic spectrum of pyrrosia pedunculata medicinal material, decoction pieces thereof, standard decoction thereof and granules according to any one of claims 3 to 5, wherein the detection conditions of the high performance liquid chromatography meet the following requirements:

a chromatographic column: octadecylsilane chemically bonded silica is used as a filler, the column length is 250mm, the inner diameter is 4.6mm, and the particle size is 5 mu m;

column temperature: 30 ℃;

flow rate: 1.0 ml/min;

sample introduction amount: 10 mu l of the mixture;

the detection wavelength is as follows: 330 nm;

the mobile phase A is acetonitrile, the mobile phase B is 0.5 percent phosphoric acid water solution, and gradient elution is carried out.

7. The method for constructing the characteristic spectrum of pyrrosia petiolata medicinal material, decoction pieces thereof, standard decoction thereof and formula granules according to claim 6, wherein the gradient elution process comprises the following steps:

0-5 min, 5-8% of mobile phase A, 95-92% of mobile phase B,

5-20 min, 8-11% of mobile phase A, 92-89% of mobile phase B,

20-25 min, 11% of mobile phase A and 89% of mobile phase B,

25-28 min, 11-15% of mobile phase A, 89-85% of mobile phase B,

28-38 min, 15-19% of mobile phase A, 85-81% of mobile phase B,

38-50 min, 19-20% of mobile phase A, 81-80% of mobile phase B,

50-55 min, 20-28% of mobile phase A, 80-72% of mobile phase B,

55-60 min, 28-30% of mobile phase A, 72-70% of mobile phase B,

60-65 min, the mobile phase A is 30%, and the mobile phase B is 70%.

8. A characteristic spectrum of pyrrosia petiolata medicinal material, decoction pieces, standard decoction and formula granules thereof is characterized in that: comprises characteristic spectrums of pyrrosia pedunculata medicinal materials, decoction pieces, standard decoction and formula granules obtained by adopting a construction method of the characteristic spectrums of pyrrosia pedunculata medicinal materials, the decoction pieces, the standard decoction and the formula granules.

9. The characteristic spectrum of pyrrosia petiolata medicinal material, decoction pieces, standard decoction and formula granules of the pyrrosia petiolata medicinal material according to claim 8 comprises 7 common peaks, wherein the 7 common peaks comprise characteristic peaks of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C.

10. The identification method of pyrrosia pedunculata medicinal material, decoction pieces thereof, standard decoction and formula granules is characterized by comprising the following steps of:

a. taking pyrrosia pedunculata medicinal material or decoction pieces thereof or standard decoction or formula granules thereof to be identified, and respectively preparing sample solutions of the pyrrosia pedunculata medicinal material or the decoction pieces thereof or the standard decoction or the formula granules thereof;

b. respectively carrying out high performance liquid chromatography detection on the sample solutions prepared in the step a to obtain corresponding characteristic chromatograms to be identified;

c. and c, comparing the characteristic chromatogram to be identified obtained in the step b with the corresponding characteristic chromatogram constructed by the method in claim 1, and identifying the chromatogram as qualified if the characteristic chromatogram to be identified is consistent with the corresponding characteristic chromatogram constructed by the method.

Technical Field

The invention belongs to the technical field of traditional Chinese medicine detection, and particularly relates to a pyrrosia pedunculata medicinal material, a characteristic spectrum of decoction pieces, standard decoction and formula granules of the pyrrosia pedunculata medicinal material, a construction method and a detection method.

Background

Pyrrosia peduncularis (Christ) Ching is one of three basic sources of Pyrrosia peduncularis in Chinese pharmacopoeia.

At present, under the item of pyrrosia lingua of the first edition of China pharmacopoeia 2020, a method for measuring the content of a single component of chlorogenic acid by adopting a high performance liquid chromatography is only adopted, and the characteristic spectrum control of the chlorogenic acid is not carried out. Other literature reports establish the fingerprint of pyrrosia lingua medicinal materials and formula granules, such as: HPLC method for determining contents of chlorogenic acid and luteolin in pyrrosia peduncularis and pyrrosia huabei and fingerprint analysis of different extracts [ J ]. chinese wild plant resources, 2015, 34 (1): 19-21. Such as: pyrrosia lingua formula particle HPLC-DAD characteristic spectrum research [ J ] proceedings of Hippocampus science and technology university, 2017, 38 (1): 39-45, etc.

The fingerprint spectrum research of different pyrrosia lingua based on different basic sources reported in the existing literature is only directed at medicinal materials, the report of standard decoction of the medicinal materials is not found, the basic sources are not distinguished in the characteristic spectrum research of pyrrosia lingua formula granules, and the peak recognition of the characteristic spectrum is less.

Therefore, the existing detection method is difficult to effectively compare and analyze the difference and the change of characteristic maps of pyrrosia peduncularis medicinal materials, decoction pieces, standard decoction and formula granules, the transmission condition of the medicinal effect substance basis of pyrrosia peduncularis from the medicinal materials to the formula granules is difficult to deeply know, and the process from pyrrosia peduncularis medicinal materials to finished pyrrosia peduncularis formula granules is difficult to integrally evaluate and control.

Disclosure of Invention

The invention aims to provide a characteristic spectrum, a construction method and a detection method of pyrrosia peduncularis medicinal material and decoction pieces, standard decoction and formula granules thereof, which can solve the problem that the process from pyrrosia peduncularis medicinal material to pyrrosia peduncularis formula granule finished product is difficult to integrally evaluate and control in the prior art so as to fill the blank in the prior art, and the pyrrosia peduncularis quality detection in production is comprehensive and reliable, and meanwhile, the detection time is short, so that the pyrrosia peduncularis preparation method is convenient for large-scale popularization and application.

The purpose of the invention is realized by the following technical scheme:

a method for constructing a characteristic spectrum of pyrrosia pedunculata medicinal material, decoction pieces thereof, standard decoction thereof and formula granules comprises the following steps:

a. preparation of control solutions: dissolving reference substances of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C in solvent to obtain reference solution;

b. preparation of a test solution: respectively preparing test solution of pyrrosia petiolata medicinal material, decoction pieces, standard decoction and formula granules thereof;

c. and injecting the reference substance solution and the test solution into a high performance liquid chromatograph for detection, and selecting common peaks from the characteristic spectrum of the test solution by taking the characteristic spectrum of the reference substance solution as a reference spectrum to construct the characteristic spectrum of the pyrrosia lingua medicinal material, the decoction pieces, the standard decoction and the formula granules of the pyrrosia lingua medicinal material.

Further, in the step a, the preparation method of the reference solution comprises: taking neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C as reference substances, respectively adding 50% methanol, and making into solutions each containing 40 μ g of reference substance per 1 ml.

Further, in step b, the preparation method of the pyrrosia petiolata medicinal material test solution comprises the following steps: taking the powder of the product, sieving the powder by a second sieve, precisely weighing 0.5g, adding 15ml of water, decocting for 30 minutes, adding 10ml of methanol, sealing, carrying out ultrasonic treatment for 10 minutes, weighing again, complementing the weight loss by methanol, shaking up, filtering, and taking the subsequent filtrate to obtain a test solution of the pyrrosia petiolata medicinal material.

Further, in the step b, the preparation method of the pyrrosia petiolata decoction piece test sample solution comprises the following steps: taking the powder, sieving by a second sieve, precisely weighing 0.5g, adding 15ml of water, decocting for 30 minutes, adding 10ml of methanol, sealing, weighing, carrying out ultrasonic treatment for 10 minutes, weighing again, complementing the weight loss by methanol, shaking uniformly, filtering, and taking the subsequent filtrate to obtain a pyrrosia petiolata decoction piece test sample solution.

Further, in step b, the preparation method of the pyrrosia petiolata standard decoction test sample solution comprises the following steps: precisely weighing 0.3g of freeze-dried powder of the product, precisely adding 20ml of 50% methanol, sealing, performing ultrasonic treatment for 30 minutes, weighing again, supplementing the lost weight with 50% methanol, shaking up, filtering, and taking the subsequent filtrate to obtain the test solution of the pyrrosia petiolata standard decoction.

Further, in step b, the preparation method of the test solution of pyrrosia petiolata formula granules comprises the following steps: taking a proper amount of the granules, accurately weighing about 0.3g after grinding, accurately adding 20ml of 50% methanol, sealing, carrying out ultrasonic treatment for 30 minutes, weighing again, complementing the lost weight with 50% methanol, shaking up, filtering, and taking the subsequent filtrate to obtain a test solution of the pyrrosia petiolata formula granules.

The detection conditions of the high performance liquid chromatography meet the following conditions:

a chromatographic column: octadecylsilane chemically bonded silica is used as a filler, the column length is 250mm, the inner diameter is 4.6mm, and the particle size is 5 mu m; column temperature: 30 ℃; flow rate: 1.0 ml/min; sample introduction amount: 10 mu l of the mixture; the detection wavelength is as follows: 330 nm; the mobile phase A is acetonitrile, the mobile phase B is 0.5 percent phosphoric acid water solution, and gradient elution is carried out.

The gradient elution process is as follows:

0-5 min, 5-8% of mobile phase A, 95-92% of mobile phase B,

5-20 min, 8-11% of mobile phase A, 92-89% of mobile phase B,

20-25 min, 11% of mobile phase A and 89% of mobile phase B,

25-28 min, 11-15% of mobile phase A, 89-85% of mobile phase B,

28-38 min, 15-19% of mobile phase A, 85-81% of mobile phase B,

38-50 min, 19-20% of mobile phase A, 81-80% of mobile phase B,

50-55 min, 20-28% of mobile phase A, 80-72% of mobile phase B,

55-60 min, 28-30% of mobile phase A, 72-70% of mobile phase B,

60-65 min, the mobile phase A is 30%, and the mobile phase B is 70%.

A characteristic spectrum of pyrrosia pedunculata medicinal material, decoction pieces thereof, standard decoction and formula granules comprises the characteristic spectrum of pyrrosia pedunculata medicinal material, decoction pieces thereof, standard decoction and formula granules obtained by adopting a construction method of the characteristic spectrum of pyrrosia pedunculata medicinal material, the decoction pieces thereof, the standard decoction and the formula granules.

The characteristic spectrum comprises 7 common peaks, and the 7 common peaks comprise characteristic peaks of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C.

An identification method of pyrrosia pedunculata medicinal material, decoction pieces thereof, standard decoction and formula granules comprises the following steps:

a. taking pyrrosia pedunculata medicinal material or decoction pieces thereof or standard decoction or formula granules thereof to be identified, and respectively preparing sample solutions of the pyrrosia pedunculata medicinal material or the decoction pieces thereof or the standard decoction or the formula granules thereof;

b. respectively carrying out high performance liquid chromatography detection on the sample solutions prepared in the step a to obtain corresponding characteristic chromatograms to be identified;

c. and c, comparing the characteristic chromatogram to be identified obtained in the step b with the corresponding characteristic chromatogram constructed by the method in claim 1, and identifying the chromatogram as qualified if the characteristic chromatogram to be identified is consistent with the corresponding characteristic chromatogram constructed by the method.

The beneficial effects of this technical scheme are as follows:

(1) the invention provides a characteristic spectrum detection method suitable for pyrrosia pedunculata medicinal materials, decoction pieces, standard decoction and formula granules, 7 common peaks are determined, chemical components of the 7 common peaks are identified, and the determined characteristic peaks are respectively neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C which have more chromatographic peaks than the chromatographic peaks identified in the prior art. The constructed HPLC characteristic spectrum has the advantages that the chromatographic peak is well separated, the characteristic spectrum information is rich, the chromatographic peak shape is good, the main chemical component information of pyrrosia petiolata can be comprehensively reflected, and the detection method has good stability and repeatability and is convenient for large-scale popularization and application.

(2) The detection method for the pyrrosia pedunculata medicinal material, the medicinal slices thereof, the standard decoction and the formula granules can integrally control the characteristic components in the pyrrosia pedunculata medicinal material, the medicinal slices thereof, the standard decoction and the formula granules, and ensure the stability of the integral quality of the pyrrosia pedunculata medicinal material, the medicinal slices thereof, the standard decoction and the formula granules.

(3) The invention fills the blank of quality control of the traditional Chinese medicinal material pyrrosia petiolata in the prior art, particularly the blank of quality overall evaluation and control in the whole process from pyrrosia petiolata medicinal materials to pyrrosia petiolata formula granule finished products, and provides an effective research basis for the later research of the traditional Chinese medicine pyrrosia petiolata.

Drawings

The foregoing and following detailed description of the invention will be apparent when read in conjunction with the following drawings, in which:

FIG. 1 is a characteristic diagram for the absorption wavelength investigation in the characteristic diagram creation process in example 4.

Fig. 2 is a characteristic map for column temperature investigation in the characteristic map establishing process in example 5.

FIG. 3 is a feature map for flow rate investigation during feature map creation in example 6.

FIG. 4 is a characteristic spectrum of example 7, which is examined by the method for extracting Podostigma longituba medicinal material.

FIG. 5 is a characteristic diagram of a pyrrosia pedunculata medicinal material subjected to solvent extraction investigation.

FIG. 6 is a characteristic spectrum of pyrrosia pedunculata medicinal material during extraction time investigation.

FIG. 7 is a characteristic diagram of the sample extraction and sample amount investigation during the extraction of pyrrosia pedunculosa medicinal material.

FIG. 8 is a chromatogram peak identification chart when standard decoction with pyrrosia petiolata medicinal material is detected.

Fig. 9 is a characteristic spectrum observed by different chromatographs in the process of establishing the characteristic spectrum of the pyrrosia peduncularis medicinal material.

Fig. 10 is a characteristic diagram for the durability examination of the chromatographic column during the characteristic diagram establishment process of pyrrosia peduncularis.

FIG. 11 is a comparison characteristic spectrum of pyrrosia pedunculata standard decoction.

Fig. 12 is a control profile of pyrrosia pedunculata formulation granules.

FIG. 13 is a control characteristic spectrum of Postoperation folium Pyrrosiae.

FIG. 14 is a control characteristic spectrum of pyrrosia pedunculata decoction pieces.

FIG. 15 is a comparison graph of comparison characteristic spectra of pyrrosia lingua medicinal materials, decoction pieces, standard decoction and formula granules in the invention (Peak 1: neochlorogenic acid, Peak 2 (S): chlorogenic acid, Peak 3: cryptochlorogenic acid, Peak 4: caffeic acid, Peak 5: isochlorogenic acid B, Peak 6: isochlorogenic acid A, Peak 7: isochlorogenic acid C).

Detailed Description

The invention provides a pyrrosia pedunculata medicinal material, a characteristic spectrum, a construction method and a detection method of decoction pieces, standard decoction and formula granules of pyrrosia pedunculata medicinal material, and aims to make the purposes, technical schemes and effects of the invention clearer and more clear and definite, and the invention is further described in detail below. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

The present invention is further illustrated by the following specific examples.

The experimental instruments and materials involved in the examples are as follows:

high performance liquid chromatograph: shimadzu LC-20AD high performance liquid chromatograph, Agilent 1260 high performance liquid chromatograph, Waters e2695 high performance liquid chromatograph;

an electronic balance: ME204E/02, MS205D Mm, XP26 (Mettler-Tollido instruments, Inc.);

an ultra-pure water machine: cell type 1810A (Shanghai Mohler scientific instruments, Inc.);

an ultrasonic cleaner: model KQ600DB (600W, 40 KHz; ultrasonic instruments, Inc. of Kunshan);

a chromatographic column: agilent 5 TC-C18250 × 4.6mm, Kromasil 100-5-C184.6 × 250mm, InertStain C185 μm 4.6 × 250mm, Inertsil ODS-33 μm 4.6 × 250 mm;

reagent: acetonitrile and phosphoric acid are chromatographically pure, water is ultrapure water, and other reagents are analytically pure;

neochlorogenic acid (Vockqi Biotech Co., Ltd., Sichuan, lot number wkq18030107, content 98.0%); chlorogenic acid (China institute for food and drug assay, batch No. 110753-201817, content in 96.8%); cryptochlorogenic acid (China institute for food and drug testing, lot number: 17061401, content in 97.8%); caffeic acid (China institute for testing and testing food and drug; batch No. 110885-; isochlorogenic acid B (Vickqi Biotech, Sichuan, Inc., lot number wkq17060705, 98.0%); isochlorogenic acid A (Dowman Stokes Biotech Ltd, lot number: MUST-18032601, content 98.0%); isochlorogenic acid C (Vickqi Biotech, Sichuan, Inc., lot number wkq17120111, 98.0%);

the pyrrosia petiolata medicinal material batch number is as follows: XLS201808882, XLS201808883, XLS201808884, XLS201808885, XLS201808886, XLS201808887, XLS201808888, XLS201808889, XLS201808890, XLS201808891, XLS201903139, XLS201903140, XLS201903141, XLS201903142, XLS201903143, XLS201903144, XLS201903145, XLS201903146, XLS201903147, XLS 201903148;

the batch number of pyrrosia pedunculata decoction pieces: SW181210, SW181211, SW181212, SW181213, SW181214, SW181215, SW181216, SW181217, SW181218, SW181219, SW190301, SW190302, SW190303, SW190304, SW190305, SW190306, SW190307, SW190308, SW190309, SW 190310;

the freeze-dried powder batch of standard pyrrosia petiolata decoction is as follows: SWBT181210, SWBT181211, SWBT181212, SWBT181213, SWBT181214, SWBT181215, SWBT181216, SWBT181217, SWBT181218, SWBT181219, SWBT190301, SWBT190302, SWBT190303, SWBT190304, SWBT190305, SWBT190306, SWBT190307, SWBT190308, SWBT190309, SWBT 190310;

postophyllia tenore formula granule batch number: SY1907003, SY1907001, SY 1907002.

Example 1

The embodiment relates to a pyrrosia petiolata medicinal material and a test sample of medicinal slices, pyrrosia petiolata standard decoction and pyrrosia petiolata formula granular preparation, and the specific preparation process comprises the following steps:

the preparation method of the pyrrosia petiolata medicinal material test solution comprises the following steps: taking the powder of the product, sieving the powder by a second sieve (the inner diameter of a sieve pore is 850 mu m +/-29 mu m, 24 meshes), about 0.5g, precisely weighing, placing the powder in a conical flask with a plug, adding 15ml of water, weighing, decocting for 30 minutes, cooling, weighing again, supplementing the lost weight with water, precisely adding 10ml of methanol, sealing the plug, weighing, carrying out ultrasonic treatment (the power is 600W, the frequency is 40 kHz) for 10 minutes, cooling, weighing again, supplementing the lost weight with methanol, shaking uniformly, filtering, and taking the subsequent filtrate to obtain the product.

The preparation method of the pyrrosia petiolata decoction piece test sample solution comprises the following steps: weighing about 0.5g of the powder (sieved by a second sieve), precisely weighing, placing in a conical flask with a plug, adding 15ml of water, weighing, decocting for 30 minutes, cooling, weighing again, supplementing the lost weight with water, precisely adding 10ml of methanol, sealing, weighing, ultrasonically treating (power 600W, frequency 40 kHz) for 10 minutes, cooling, weighing again, supplementing the lost weight with methanol, shaking uniformly, filtering, and taking the subsequent filtrate.

The preparation method of the pyrrosia petiolata standard decoction test solution comprises the following steps: taking about 0.3g of freeze-dried powder of pyrrosia petiolata standard decoction, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of 50% methanol respectively, sealing the plug, weighing, ultrasonically treating for 30 minutes, cooling, weighing again, supplementing the lost weight with 50% methanol, shaking uniformly, filtering, and taking the subsequent filtrate.

The preparation method of the test solution of pyrrosia petiolata formula granules comprises the following steps: taking a proper amount of the product, grinding, precisely weighing about 0.3g, placing into a conical flask with a plug, precisely adding 20ml of 50% methanol, sealing the plug, weighing, performing ultrasonic treatment for 30 minutes, cooling, weighing again, supplementing the lost weight with 50% methanol, shaking uniformly, filtering, and taking the subsequent filtrate.

Example 2

The embodiment relates to establishment of a fingerprint spectrum obtained by pyrrosia petiolata medicinal material and decoction pieces thereof, pyrrosia petiolata standard decoction and pyrrosia petiolata formula granular preparation, and specifically comprises the following steps:

a. preparation of control solutions: dissolving reference substances of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C in solvent to obtain reference solution;

b. preparation of a test solution: respectively preparing test solution of pyrrosia petiolata medicinal material, decoction pieces, standard decoction and formula granules thereof;

c. and (3) detection: and injecting the reference substance solution and the test solution into a high performance liquid chromatograph for detection, and selecting common peaks from the characteristic spectrum of the test solution by taking the characteristic spectrum of the reference substance solution as a reference spectrum to construct the characteristic spectrum of the pyrrosia lingua medicinal material, the decoction pieces, the standard decoction and the formula granules of the pyrrosia lingua medicinal material.

The detection conditions of the high performance liquid chromatography meet the following conditions: a chromatographic column: octadecylsilane chemically bonded silica is used as a filler, the column length is 250mm, the inner diameter is 4.6mm, and the particle size is 5 mu m; column temperature: 30 ℃; flow rate: 1.0 ml/min; sample introduction amount: 10 mu l of the mixture; the detection wavelength is as follows: 330 nm; the mobile phase A is acetonitrile, the mobile phase B is 0.5 percent phosphoric acid water solution, and gradient elution is carried out. The number of theoretical plates is not less than 3000 calculated according to chlorogenic acid peak.

The gradient elution process comprises the following steps:

0-5 min, 5-8% of mobile phase A, 95-92% of mobile phase B,

5-20 min, 8-11% of mobile phase A, 92-89% of mobile phase B,

20-25 min, 11% of mobile phase A and 89% of mobile phase B,

25-28 min, 11-15% of mobile phase A, 89-85% of mobile phase B,

28-38 min, 15-19% of mobile phase A, 85-81% of mobile phase B,

38-50 min, 19-20% of mobile phase A, 81-80% of mobile phase B,

50-55 min, 20-28% of mobile phase A, 80-72% of mobile phase B,

55-60 min, 28-30% of mobile phase A, 72-70% of mobile phase B,

60-65 min, the mobile phase A is 30%, and the mobile phase B is 70%.

d. And c, taking the characteristic map of the reference substance solution in the step c as a reference map, selecting common peaks from the characteristic maps of the test substance solution, and constructing the characteristic maps of pyrrosia lingua medicinal materials, decoction pieces, standard decoction and formula granules of the pyrrosia lingua medicinal materials.

The characteristic spectrum constructed by pyrrosia petiolata medicinal material has 7 characteristic peaks, wherein the peak corresponding to the chlorogenic acid reference substance is an S peak. The relative retention time of each characteristic peak to the S peak is calculated and should be within ± 10% of the specified value. The specified values are: 0.633 (peak 1), 1.067 (peak 3), 1.117 (peak 4), 2.058 (peak 5), 2.156 (peak 6), 2.354 (peak 7).

The chromatogram of the sample of the constructed pyrrosia petiolata decoction pieces should present 7 characteristic peaks, the peak corresponding to the chlorogenic acid reference substance is an S peak, and the relative retention time of each characteristic peak and the S peak is calculated and should be within +/-10% of a specified value. The specified values are: 0.633 peak 1), 1.066 (peak 3), 1.117 (peak 4), 2.057 (peak 5), 2.154 (peak 6), 2.352 (peak 7).

The chromatogram of the test sample of the constructed pyrrosia petiolata standard decoction should present 7 characteristic peaks, the peak corresponding to the chlorogenic acid reference substance is the S peak, the relative retention time of the S peak of each characteristic peak is calculated, and the relative retention time is within +/-10% of the specified value. The specified values are: 0.633 (peak 1), 1.066 (peak 3), 1.117 (peak 4), 2.053 (peak 5), 2.150 (peak 6), 2.348 (peak 7).

The chromatogram of the test sample of the constructed pyrrosia petiolata formula particle shows 7 characteristic peaks, and the peak corresponding to the chlorogenic acid reference substance is an S peak. The relative retention time of each characteristic peak to the S peak is calculated and should be within ± 10% of the specified value. The specified values are: 0.62 (peak 1), 1.06 (peak 3), 1.10 (peak 4), 2.10 (peak 5), 2.19 (peak 6), 2.40 (peak 7).

Example 3

The embodiment relates to identification of pyrrosia petiolata medicinal material, decoction pieces, standard decoction and formula granules of pyrrosia petiolata medicinal material. The method comprises the following specific steps:

a. taking pyrrosia peduncularis medicinal material to be identified and decoction pieces, standard decoction and formula granules thereof, and respectively preparing sample solutions of the pyrrosia peduncularis medicinal material and the decoction pieces, the standard decoction and the formula granules thereof according to the preparation method of the test sample in the step b of the embodiment 2;

b. respectively carrying out high performance liquid chromatography detection on the sample solution obtained in the step a according to the detection conditions in the step c in the embodiment 2 to obtain corresponding characteristic maps to be identified;

c. and c, comparing the characteristic spectrum to be identified obtained in the step b with the corresponding characteristic spectrum constructed in the step d in the embodiment 2, and identifying the characteristic spectrum to be identified as qualified if the characteristic spectrum to be identified is consistent with the corresponding characteristic spectrum constructed in the step d.

Example 4

Investigation of wavelength

On the basis of the experimental conditions, a diode array detector is utilized to respectively carry out full-wave-band scanning on a neochlorogenic acid reference solution, a chlorogenic acid reference solution, a cryptochlorogenic acid reference solution, a caffeic acid reference solution, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C reference solution and a test solution, and chromatograms of the test solution at wavelengths of 230nm, 254nm, 270nm, 290nm, 310nm, 330nm and 360nm are extracted. As a result, the amount of information on the chromatographic peak was large at a detection wavelength of 330nm, and the detection wavelength was determined to be 330 nm. As shown in fig. 1, it is a wavelength diagram of pyrrosia petiolata standard decoction for reference.

Example 5

Investigation of column temperature

Under the chromatographic conditions set forth above, the column temperature was examined at 25 ℃, 30 ℃ and 35 ℃. See fig. 2 and table 1.

TABLE 1

The results show that the chromatogram peaks are symmetrical when the column temperature is 25, 30 and 35 ℃, and the separation degree meets the requirements. Therefore, the column temperature is selected to be 30 ℃ for subsequent investigation.

Example 6

Investigation of flow Rate

The flow rates were 0.8ml/min, 1.0ml/min, and 1.2ml/min under the chromatographic conditions set forth above. See fig. 3 and table 2.

TABLE 2

The result shows that when the flow rates are respectively 0.8ml/min and 1.0ml/min, the relative retention time chromatogram of each characteristic peak has symmetrical peak shapes, and the separation degree meets the requirement. A flow rate of 1.0ml/min was chosen for subsequent investigation.

Example 7

Examination of extraction methods

Taking about 0.3g of the product (batch number SWBT 181216), precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of 50% methanol respectively, sealing, weighing, respectively examining the reflux and ultrasound of the extraction mode, extracting for 30min, cooling, weighing again, supplementing the lost weight with 50% methanol, shaking up, filtering, and taking the subsequent filtrate. See fig. 4.

The results show that the ultrasonic extraction of the test sample has better effect than the reflux extraction. Therefore, the test sample extraction method is determined as ultrasonic extraction.

Example 8

Examination of extraction solvent

Weighing about 0.3g of the product (batch No. SWBT 181216), precisely weighing, placing in a conical flask with a plug, respectively adding sample extraction solvents of methanol, 80% methanol, 50% methanol, water, ethanol, and 50% ethanol each 20ml, sealing, weighing, ultrasonically extracting for 30min, cooling, weighing, supplementing lost weight with corresponding extraction solvent, shaking, filtering, and collecting filtrate. See fig. 5.

The result shows that when the extraction solvent is 50% methanol, the chromatographic peak information amount is large and the separation degree is good. Therefore, the extraction solvent of the test sample was determined to be 50% methanol.

Example 9

Investigation of extraction time

Taking about 0.3g of the product (batch number SWBT 181216), precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of 50% methanol respectively, sealing the plug, weighing, ultrasonically extracting, inspecting the extraction time of the sample at 20 minutes, 30 minutes and 40 minutes respectively, cooling, weighing again, supplementing the lost weight with 50% methanol, shaking uniformly, filtering, and taking the subsequent filtrate to obtain the final product. See fig. 6.

The result shows that the extraction time is only 20 minutes, but the extraction time of the test sample is determined to be 30 minutes by combining the conventional traditional Chinese medicine extraction process in order to ensure that the solution can be fully extracted.

Example 10

Material ratio investigation

Respectively taking about 0.2g, 0.3g and 0.4g of the product (batch number SWBT 181216), precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of 50% methanol, sealing the plug, weighing, ultrasonically extracting for 30 minutes, cooling, weighing again, supplementing the lost weight with 50% methanol, shaking up, filtering, and taking the subsequent filtrate to obtain the final product. See fig. 7.

As a result, it was found that the ratio of the chromatographic peak heights was suitable when the sample amount was 0.3 g. Therefore, the sample amount of the sample is determined to be 0.3 g.

In conclusion, the preparation method of the test solution with the pyrrosia petiolata standard decoction characteristic spectrum is determined as follows: taking about 0.3g of the product, accurately weighing, placing in a conical flask with a plug, accurately adding 20ml of 50% methanol, sealing the plug, weighing, ultrasonically extracting for 30 minutes, cooling, weighing again, supplementing the lost weight with 50% methanol, shaking uniformly, filtering, and taking the subsequent filtrate.

Example 11

Identification of chromatographic peaks

Preparation of a test solution: according to the experimental conditions formulated above, a pyrrosia petiolata standard decoction test sample solution is prepared.

Preparation of reference solutions: respectively adding 50% methanol into neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C to obtain solutions each containing 40 μ g per 1 ml.

Preparation of negative control solution: preparing negative control solution of pyrrosia petiolata standard decoction according to the experimental conditions.

And identifying characteristic spectrum peaks of pyrrosia petiolata standard decoction. See fig. 8.

The results show that peak 1 is neochlorogenic acid, peak 2 is chlorogenic acid, peak 3 is cryptochlorogenic acid, peak 4 is caffeic acid, peak 5 is isochlorogenic acid B, peak 6 is isochlorogenic acid a, peak 7 is isochlorogenic acid C, and in the following methodological investigation, 7 peaks in the sample spectrum of fig. 8 were investigated.

Example 12

Examination of precision

Sample solution of standard Pogostemon petiolatus decoction (batch No. SWBT 181216) is continuously injected for 6 times, 10 μ l each time, according to a formulated experimental method, and retention time and peak area are calculated, as shown in tables 3 and 4.

Table 3: precision investigation-retention time

Table 4: precision survey-Peak area

The result shows that the retention time of each characteristic peak and the RSD value of the peak area meet the requirements, and the instrument has good precision.

Example 13

Examination of repeatability

6 parts of pyrrosia lingua standard decoction freeze-dried powder (batch number SWBT 181216) are precisely weighed, and prepared and measured according to a formulated experimental method. See tables 5 and 6.

Table 5: repeatability test-relative retention time

Table 6: repeatability test-relative peak area

The result shows that the relative retention time of each characteristic peak and the RSD value of the relative peak area meet the requirements, and the repeatability of the method is good.

Example 14

Intermediate precision investigation

Investigation of different persons and time

On the basis of the experimental conditions formulated above, two portions of pyrrosia lingua standard soup freeze-dried powder (batch number SWBT 181216) are precisely weighed under the conditions of different personnel (A, B) and different time (T1, T2) respectively, and the samples are prepared and measured. The results are shown in tables 7 and 8.

Table 7: personnel and time review-relative retention time

Table 8: personnel and time review-relative peak area

The result shows that different people can determine the same sample at different time, and the method has better stability.

Example 15

Investigation of different instruments

On the basis of the experimental conditions formulated above, two parts of pyrrosia petiolata standard decoction lyophilized powder (batch number SWBT 181216) are precisely weighed respectively to prepare test solution, and the test solution is measured on Shimadzu LC-20AD type high performance liquid chromatograph, Agilent 1260 type high performance liquid chromatograph, and Waters 2695-. See FIG. 9, tables 9-10.

Table 9: instrument durability examination-relative retention time

Table 10: instrument durability examination-relative peak area

The result shows that the RSD of each characteristic peak relative retention time is 0.51-4.05% under different instrument conditions; the RSD of the relative peak area of each characteristic peak is 0.26-11.12%, so that the durability of 3 brands of instruments is good.

Example 16

Chromatographic column durability investigation

Based on the experimental conditions set forth above, the columns were analyzed to be Phenomenex Luna 5um C18(2) 100A 4.6X 250mm, InertSustain C185. mu.m 4.6X 250mm, Inertsil ODS-33. mu.m 4.6X 250mm, respectively. The results are shown in FIG. 10, tables 11-12.

Table 11: chromatographic column durability study-relative retention time

Table 12: column durability test-relative peak area

The results show that the RSD of the characteristic peak relative retention time is 2.68-7.01%, and the RSD of the characteristic peak relative peak area is 2.23-29.36% when the sample is detected by using the 3 chromatographic columns.

Example 17

Stability survey

Based on the experimental conditions, the same test solution is taken and respectively measured at 0h, 4h, 8h, 12h, 18h and 24 h. See tables 13 and 14.

Table 13: stability survey-retention time

Table 14: stability survey-Peak area

The result shows that the RSD of the retention time of the corresponding characteristic peak is 0.08-0.18%, and the sample solution is stable within 24 hours.

In summary, the RSD of each characteristic peak relative retention time meets the requirements in the above studies, and the method is good.

Example 18

Standard decoction characteristic map verification

According to the preparation method of the test solution of the standard decoction in example 1, 20 batches of the product were subjected to characteristic spectrum measurement, and the relative retention time and the relative peak area were calculated, and the verification results are shown in tables 15 to 16.

Table 15: retention time of 20 batches of pyrrosia petiolata standard decoction

Table 16: relative peak area of 20 batches of pyrrosia petiolata standard decoction

Serial number	Batch number	Peak 1	Peak 2 (S)	Peak 3	Peak 4	Peak 5	Peak 6	Peak 7
									1	SWBT181210	0.677	1.000	0.65	0.065	0.106	0.071	0.195
2	SWBT181211	0.705	1.000	0.666	0.083	0.117	0.076	0.218
									3	SWBT181212	0.75	1.000	0.7	0.078	0.119	0.078	0.203
4	SWBT181213	0.789	1.000	0.728	0.072	0.111	0.076	0.187
									5	SWBT181214	0.768	1.000	0.713	0.074	0.121	0.081	0.209
6	SWBT181215	0.698	1.000	0.658	0.067	0.106	0.071	0.196
									7	SWBT181216	0.702	1.000	0.67	0.061	0.107	0.077	0.174
8	SWBT181217	0.713	1.000	0.676	0.066	0.104	0.071	0.183
									9	SWBT181218	0.744	1.000	0.704	0.073	0.112	0.061	0.172
10	SWBT181219	0.763	1.000	0.705	0.068	0.113	0.078	0.197
									11	SWBT190301	0.817	1.000	0.758	0.132	0.129	0.101	0.17
12	SWBT190302	0.86	1.000	0.787	0.083	0.151	0.119	0.186
									13	SWBT190303	0.827	1.000	0.756	0.08	0.119	0.096	0.159
14	SWBT190304	0.903	1.000	0.805	0.073	0.122	0.095	0.153
									15	SWBT190305	0.837	1.000	0.772	0.08	0.126	0.099	0.16
16	SWBT190306	0.897	1.000	0.798	0.072	0.13	0.103	0.166
									17	SWBT190307	0.907	1.000	0.807	0.081	0.117	0.101	0.165
18	SWBT190308	0.829	1.000	0.764	0.084	0.121	0.099	0.164
									19	SWBT190309	0.836	1.000	0.768	0.069	0.14	0.108	0.169
20	SWBT190310	0.898	1.000	0.81	0.116	0.149	0.113	0.176
										RSD%	9.5	0.00	7.38	21.56	11.11	18.56	10.08

According to the principle that the relative retention time is stable, samples of each batch can be detected, and the peaks are relatively high, 7 peaks with good repeatability are selected as characteristic peaks. The results show that when the peak 2 is taken as the S peak, the relative retention time RSD of the characteristic peak of the pyrrosia petiolata standard decoction of 20 batches is less than 2.0 percent. Finally, the following steps are provided: the chromatogram of the test sample should show 7 characteristic peaks corresponding to the retention time of 7 characteristic peaks in the chromatogram peaks of the reference substance of the reference medicinal material, wherein the peak corresponding to the chlorogenic acid reference substance is the S peak, and the relative retention time of the S peak of each characteristic peak is calculated and should be within + -10% of the specified value. The specified values are: 0.633 (peak 1), 1.066 (peak 3), 1.117 (peak 4), 2.053 (peak 5), 2.150 (peak 6), 2.348 (peak 7).

A traditional Chinese medicine chromatogram fingerprint similarity evaluation system (2012 edition) is adopted to synthesize 20 batches of pyrrosia petiolata standard decoction, a contrast characteristic spectrum of the pyrrosia petiolata standard decoction characteristic spectrum is established, and the quality of the pyrrosia petiolata standard decoction can be controlled integrally and accurately. See fig. 11.

Example 19

Formula particle characteristic spectrum verification

According to the preparation method of the test solution of the pyrrosia petiolata formula particle in the example 1, the characteristic spectrum of 3 batches of samples of the product is measured, and the relative retention time and the relative peak area are calculated and shown in tables 17-18.

Table 17: relative retention time of 3 batches of pyrrosia pedunculata formula granules

Table 18: relative peak area of 3 batches of pyrrosia petiolata formula granules

According to the principle that the relative retention time is stable, samples of each batch can be detected, and the peaks are relatively high, 7 peaks with good repeatability are selected as characteristic peaks. When peak 2 is taken as the S peak, the 7 characteristic peaks relative retention time RSD of 3 batches of pyrrosia petiolata formulation granules are all less than 1.0%. Finally, the following steps are provided: the chromatogram of the test sample should show 7 characteristic peaks, and the relative retention time of the S peak should be within + -10% of the specified value. The specified values are: 0.62 (peak 1), 1.06 (peak 3), 1.10 (peak 4), 2.10 (peak 5), 2.19 (peak 6), 2.40 (peak 7).

Synthesizing 3 batches of pyrrosia peduncularis formula granules by adopting a traditional Chinese medicine chromatography fingerprint similarity evaluation system (2012 edition), and establishing a control spectrum of characteristic spectrum of pyrrosia peduncularis formula granules. See fig. 12, where peak 1: neochlorogenic acid, peak 2 (S): chlorogenic acid, peak 3: cryptochlorogenic acid, peak 4: caffeic acid, peak 5: isochlorogenic acid B, peak 6: isochlorogenic acid a, peak 7: isochlorogenic acid C.

Example 20

Verification of medicinal material characteristic map

According to the preparation method of the test solution of pyrrosia petiolata medicinal material in the example 1, 20 batches of samples are measured, and the relative retention time and the ratio of relative peak areas are calculated. The results are shown in tables 19 and 20.

Table 19: characteristic spectrum relative retention time ratio of 20 batches of pyrrosia pedunculosa medicinal materials

Table 20: relative peak area of characteristic spectrum of 20 batches of pyrrosia pedunculosa medicinal materials

According to the principle that the relative retention time is stable, samples of each batch can be detected, and the peaks are relatively high, 7 peaks with good repeatability are selected as characteristic peaks. When the peak 2 is taken as the S peak, the relative retention time RSD of 7 characteristic peaks of 20 batches of pyrrosia petiolata medicinal materials is less than 2.0 percent. Finally, the following steps are provided: the chromatogram of the test sample should show 7 characteristic peaks, the peak corresponding to the chlorogenic acid reference is S peak, and the relative retention time of each characteristic peak and S peak is calculated and should be within + -10% of the specified value. The specified values are: 0.633 (peak 1), 1.067 (peak 3), 1.117 (peak 4), 2.058 (peak 5), 2.156 (peak 6), 2.354 (peak 7).

Synthesizing the samples by adopting a traditional Chinese medicine chromatography fingerprint similarity evaluation system (2012 edition), and establishing a control characteristic spectrum of the pyrrosia pedunculosa medicinal material. See fig. 13, where peak 1: neochlorogenic acid, peak 2 (S): chlorogenic acid, peak 3: cryptochlorogenic acid, peak 4: caffeic acid, peak 5: isochlorogenic acid B, peak 6: isochlorogenic acid a, peak 7: isochlorogenic acid C.

Example 21

Decoction piece characteristic spectrum verification

The 20 batches were tested as above and the relative retention time, relative peak area ratios were calculated. The results are shown in tables 21 and 22.

Table 21: characteristic spectrum relative retention time ratio of 20 batches of pyrrosia pedunculosa decoction pieces

Table 22: relative peak area of characteristic spectrum of 20 batches of pyrrosia lingua decoction pieces

According to the principle that the relative retention time is stable, samples of each batch can be detected, and the peaks are relatively high, 7 peaks with good repeatability are selected as characteristic peaks. When the peak 2 is taken as the S peak, the RSD of the characteristic peak relative to the retention time of 20 batches of pyrrosia petiolata decoction pieces is less than 1.0 percent. Finally, the following steps are provided: the chromatogram of the test sample should show 7 characteristic peaks, the peak corresponding to the chlorogenic acid reference is S peak, and the relative retention time of each characteristic peak and S peak is calculated and should be within + -10% of the specified value. The specified values are: 0.633 peak 1), 1.066 (peak 3), 1.117 (peak 4), 2.057 (peak 5), 2.154 (peak 6), 2.352 (peak 7).

Synthesizing 20 batches of pyrrosia pedunculosa decoction piece characteristic spectrums by adopting a traditional Chinese medicine chromatography fingerprint similarity evaluation system (2012 edition), and establishing a control spectrum of the pyrrosia pedunculosa decoction piece characteristic spectrums. See fig. 14, where peak 1: neochlorogenic acid, peak 2 (S): chlorogenic acid, peak 3: cryptochlorogenic acid, peak 4: caffeic acid, peak 5: isochlorogenic acid B, peak 6: isochlorogenic acid a, peak 7: isochlorogenic acid C.

Example 22

A traditional Chinese medicine chromatogram fingerprint similarity evaluation system (2012 edition) is adopted to respectively synthesize 20 batches of pyrrosia pedunculosa medicinal material characteristic spectrums into a medicinal material contrast spectrum, referring to fig. 15, 20 batches of pyrrosia pedunculosa decoction piece characteristic spectrums into a decoction piece contrast spectrum, 20 batches of pyrrosia pedunculosa standard decoction characteristic spectrums into a standard decoction piece contrast spectrum, and 3 batches of pyrrosia pedunculosa formula granule characteristic spectrums into a formula granule contrast spectrum. The pyrrosia peduncularis medicinal materials, decoction pieces, standard decoction and formula granules are compared with the reference map, and the result is shown in figure 15, wherein the peak 1: neochlorogenic acid, peak 2 (S): chlorogenic acid, peak 3: cryptochlorogenic acid, peak 4: caffeic acid, peak 5: isochlorogenic acid B, peak 6: isochlorogenic acid a, peak 7: isochlorogenic acid C.

The results show that 7 characteristic peaks can be detected in pyrrosia pedunculata medicinal materials, decoction pieces, standard decoction and formula granules, the material bases of the characteristic peaks are consistent, the method can accurately and efficiently detect the characteristic components in the pyrrosia pedunculata medicinal materials, the decoction pieces, the standard decoction and the formula granules, and the aim of integrally controlling the quality of the pyrrosia pedunculata medicinal materials, the decoction pieces, the standard decoction and the formula granules is fulfilled.

In conclusion, the detection method is suitable for the high performance liquid characteristic spectrum of pyrrosia peduncularis medicinal materials, decoction pieces, standard decoction and formula granules, can integrally control the characteristic components in the pyrrosia peduncularis medicinal materials, the decoction pieces, the standard decoction and the formula granules, ensures the integral stability of the quality of the pyrrosia peduncularis medicinal materials, the decoction pieces, the standard decoction and the formula granules, and has the advantages of simple operation, high precision, good stability, good repeatability and high accuracy.