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CN-120121754-B - Construction method of HPLC (high Performance liquid chromatography) characteristic spectrum of rhizoma Menispermi medicinal decoction pieces, standard decoction and formula granules

CN120121754BCN 120121754 BCN120121754 BCN 120121754BCN-120121754-B

Abstract

The invention provides a construction method of HPLC characteristic maps of rhizoma Menispermi medicinal decoction pieces, standard decoction pieces and formula particles, which comprises the steps of A) dissolving raw materials of a test product in a solvent, extracting to obtain a liquid to be tested, B) measuring the liquid to be tested by adopting a high performance liquid chromatography to obtain the HPLC characteristic maps of the rhizoma Menispermi medicinal decoction pieces, the standard decoction pieces and the formula particles, wherein the HPLC characteristic maps of the rhizoma Menispermi medicinal decoction pieces, the standard decoction pieces and the formula particles are obtained by the high performance liquid chromatography, chromatographic conditions are that a chromatographic column is a C18 column, a mobile phase A is acetonitrile solution, a mobile phase B is 0.05% trifluoroacetic acid aqueous solution, and gradient elution is carried out, the specific gradient elution is 0-15 min, a phase A is 5-11%, B phase is 95-89%, 15-40 min, a phase A is 11-18%, B phase is 89-60 min, a phase A is 18-21%, and B phase A is 82-79%. The invention adopts high performance liquid chromatography, selects acetonitrile-0.05% trifluoroacetic acid solution as mobile phase for gradient elution, takes magnaline, isocorydine, dauricine Ge Sulin base and dauricine as reference substances, establishes HPLC characteristic spectrum methods of rhizoma Menispermi medicinal decoction pieces, standard decoction pieces and formula particles, and provides more scientific technical means for controlling the medicinal quality of rhizoma Menispermi medicinal decoction pieces, standard decoction pieces and formula particles.

Inventors

  • ZHOU HOUCHENG
  • YANG LIAN
  • HUANG YU
  • ZHOU JINGWEI
  • OU YIJING
  • YU RONGPING
  • YANG LU
  • Aduroline
  • ZHAO HAITING
  • FEI WENBO
  • ZHONG LEI

Assignees

  • 四川新绿色药业科技发展有限公司

Dates

Publication Date
20260505
Application Date
20250427

Claims (10)

  1. 1. A construction method of HPLC characteristic spectrum of rhizoma Menispermi decoction pieces, standard decoction and formula granule comprises the following steps: A) Dissolving the raw materials of the test sample by using a solvent, and extracting to obtain a solution to be tested, wherein the solvent is methanol; B) Measuring the liquid to be measured by adopting a high performance liquid chromatography to obtain HPLC characteristic maps of rhizoma Menispermi medicinal decoction pieces, standard decoction pieces and formula particles; The high performance liquid chromatography has chromatographic conditions that a chromatographic column is a C18 column, a mobile phase A is acetonitrile solution, a mobile phase B is 0.05% trifluoroacetic acid aqueous solution, and gradient elution is carried out; the gradient elution specifically comprises the following steps: 0-15 min, 5-11% of phase A and 95-89% of phase B; 15-40 min, 11-18% of phase A and 89-82% of phase B; 40-60 min, 18-21% of phase A and 82-79% of phase B.
  2. 2. The method of claim 1, further comprising preparing a reference solution by dissolving magnolol, isocorydine, dauricine Ge Sulin, dauricine, and dauricine, respectively, with methanol to obtain a reference solution; and determining the reference substance solution by adopting a high performance liquid chromatography to obtain a chromatogram of the reference substance, and carrying out qualitative determination on the components of rhizoma Menispermi medicinal decoction pieces, standard decoction pieces and formula particles according to the chromatogram of the reference substance.
  3. 3. The method according to claim 2, wherein the concentration of the reference solution is specifically 100 μg/mL of magnaline, 100 μg/mL of isocorydine, 20 μg/mL of bats Ge Sulin base, and 20 μg/mL of bats puerarin.
  4. 4. The method according to claim 1, wherein the chromatographic column is C 18 x 4.6mm 5 μm, the column temperature is 20 to 30 ℃, and the theoretical plate number is not less than 5000 as calculated as magnolicine.
  5. 5. The method of claim 4, wherein the mobile phase flow rate is 0.6mL/min and the sample injection amount is 5-10. Mu.L.
  6. 6. The method of claim 4, wherein the detection wavelength is 250nm.
  7. 7. The method of claim 4, wherein the extraction in step A) is ultrasonic extraction, the power of the ultrasonic wave is 600W, the frequency is 40kHz, and the ultrasonic time is 30min.
  8. 8. The method according to claim 1, wherein the ratio of the mass g of the sample material to the volume mL of the solvent is 0.1-0.2:25; The raw materials of the test product are one or more of rhizoma Menispermi medicinal materials, rhizoma Menispermi decoction pieces, rhizoma Menispermi standard decoction or rhizoma Menispermi prescription granule.
  9. 9. The method of claim 1, wherein the similarity of HPLC characteristic patterns of rhizoma Menispermi decoction pieces, standard decoction pieces and formula particles is evaluated by a traditional Chinese medicine chromatographic fingerprint similarity evaluation system to obtain an HPLC standard characteristic pattern composed of 7 characteristic peaks, wherein peak 3 magnolol, peak 5 isocorydine, peak 6 bat Ge Sulin alkali, peak 7 dauricine; In the characteristic maps of the rhizoma Menispermi medicinal decoction pieces, the standard decoction pieces and the formula particles, magnaline is used as a reference peak S, and the relative retention time of each characteristic peak and S peak is calculated, wherein the relative retention time is within +/-10% of a specified value, and the specified values are respectively 0.35 (peak 1), 0.60 (peak 2) and 1.12 (peak 4).
  10. 10. A method for identifying characteristic patterns of rhizoma Menispermi decoction pieces, standard decoction pieces and formula particles, which is characterized in that the method is adopted for detection and analysis of detection results.

Description

Construction method of HPLC (high Performance liquid chromatography) characteristic spectrum of rhizoma Menispermi medicinal decoction pieces, standard decoction and formula granules Technical Field The invention relates to the technical field of analysis and detection, in particular to a construction method of HPLC characteristic maps of rhizoma Menispermi medicinal decoction pieces, standard decoction pieces and formula particles. Background Rhizoma Menispermi is decoction piece obtained by removing impurities, cleaning, moistening, slicing, and drying. The rhizoma Menispermi standard decoction is lyophilized powder prepared from the medicinal materials by fixed preparation process, and rhizoma Menispermi granule is prepared from rhizoma Menispermi medicinal materials by processing and processing according to main quality index of standard decoction. In order to ensure the uniformity and stability of the quality of rhizoma Menispermi medicinal materials, standard decoction and prescription granules. Therefore, it is necessary to establish a new characteristic spectrum method to control the quality and to identify the rhizoma Menispermi and its preparation obviously. Disclosure of Invention In view of the above, the technical problem to be solved by the invention is to provide a construction method of HPLC characteristic maps of rhizoma Menispermi medicinal decoction pieces, standard decoction and formula particles. The invention provides a construction method of HPLC characteristic maps of rhizoma Menispermi medicinal decoction pieces, standard decoction and prescription granule, which comprises the following steps: A) Dissolving the raw materials of the test sample by using a solvent, and extracting to obtain a solution to be tested; B) Measuring the liquid to be measured by adopting a high performance liquid chromatography to obtain HPLC characteristic maps of rhizoma Menispermi medicinal decoction pieces, standard decoction pieces and formula particles; the high performance liquid chromatography has chromatographic conditions that a chromatographic column is a C18 column, a mobile phase A is acetonitrile solution, a mobile phase B is 0.05% trifluoroacetic acid aqueous solution, and gradient elution is carried out; the gradient elution specifically comprises the following steps: 0-15 min, 5-11% of phase A and 95-89% of phase B; 15-40 min, 11-18% of phase A and 89-82% of phase B; 40-60 min, 18-21% of phase A and 82-79% of phase B. The raw materials of the test sample are one or more of rhizoma Menispermi medicinal materials, rhizoma Menispermi medicinal decoction pieces, rhizoma Menispermi medicinal standard decoction or rhizoma Menispermi medicinal formula granules. The method for constructing HPLC characteristic maps of asiatic moonseed rhizome medicinal materials, decoction pieces, standard decoction and prescription granule comprises the steps of firstly dissolving the raw materials of a test sample by using a solvent, and extracting to obtain a liquid to be tested. Specifically, the preparation method comprises dissolving the raw materials of the sample in solvent, extracting, cooling, shaking, and filtering. The extraction is heating reflux extraction or ultrasonic extraction, preferably ultrasonic extraction. The power of the ultrasonic wave is 600W, the frequency is 40kHz, and the ultrasonic wave time is 30min. The ratio of the mass g of the raw materials of the sample to the volume mL of the solvent is 0.1-0.2:25. In some embodiments, the ratio of the mass g of the sample material to the volume mL of the solvent according to the invention is 0.2:25. Specifically, the preparation of the test solution is to take rhizoma Menispermi medicinal materials, decoction pieces, standard decoction and formula particles, precisely weigh, put into a conical flask with a plug, precisely add methanol, seal, weigh, ultrasonically treat for 30 minutes, take out, cool, shake uniformly, filter, and take the subsequent filtrate. The quality of the raw materials can be controlled by the method. The invention also comprises the steps of preparing reference substance solutions, namely respectively taking magnolol, isocorydine, dauricine Ge Sulin alkali and dauricine, and dissolving the magnolol with methanol to obtain the reference substance solutions; and determining the reference substance solution by adopting a high performance liquid chromatography to obtain a chromatogram of the reference substance, and carrying out qualitative determination on the components of rhizoma Menispermi medicinal decoction pieces, standard decoction pieces and formula particles according to the chromatogram of the reference substance. The concentration of the reference solution is preferably specifically 100 mug/mL of magnaline, 100 mug/mL of isocorydine, 20 mug/mL of dauricine Ge Sulin alkali and 20 mug/mL of dauricine. And (3) measuring the liquid to be measured by adopting a high performance liquid chromatography to obtain HPLC chara