CN-122017072-A - Liquid chromatography characteristic spectrum method for identifying authenticity of Shuanghuanglian oral liquid for livestock

CN122017072ACN 122017072 ACN122017072 ACN 122017072ACN-122017072-A

Abstract

The invention discloses a liquid chromatography characteristic spectrum method for identifying the authenticity of veterinary Shuanghuanglian oral liquid, which relates to the technical field of high performance liquid chromatography, and establishes characteristic chromatographic peaks for simultaneously distinguishing weeping forsythiae capsule from weeping forsythia leaves, honeysuckle and honeysuckle administration, thereby fundamentally solving the technical problem of the current market, definitely identifying illegal production behaviors, providing a direct and reliable technical means for hitting the potential rules of the veterinary drug industry, and greatly improving the fineness and scientificity of quality evaluation by providing a special characteristic spectrum and definitely tracing the attribution and the sources of 10 main chromatographic peaks in the spectrum compared with the prior standard method.

Inventors

CHEN JING
YANG YANCHAO
ZHANG NA
GUO LI
YANG HEYUN
LI HUIPING
ZHANG QIUNA
LIU RUONAN
LIN CUI
ZHANG PEIXUN

Assignees

保定冀中药业有限公司
保定阳光本草药业有限公司

Dates

Publication Date: 20260512
Application Date: 20260225

Claims (6)

1. The liquid chromatography characteristic spectrum method for identifying the authenticity of the Shuanghuanglian oral liquid for animals is characterized by comprising the following steps: s1, setting chromatographic conditions, performing gradient elution by taking octadecylsilane chemically bonded silica as a stationary phase and methanol and 0.2% phosphoric acid solution as a mobile phase, and performing analysis under a specified detection wavelength; s2, preparing new chlorogenic acid, cryptochlorogenic acid, forsythoside E, forsythoside I, forsythoside B, forsythoside H, forsythoside A, forsythoside and baicalin reference substance solution; S3, precisely measuring Shuanghuanglian oral liquid to prepare a sample solution; S4, obtaining a chromatogram of the solution of the sample, and calculating the relative retention time of a peak 2, a peak 6 and a peak S in the chromatogram, wherein if the peak 2 is not detected in the chromatogram, the fructus forsythiae is not used for administration according to the requirement in the prescription feeding, and if the peak 6 is not detected in the graph, the flos lonicerae is not used for administration according to the requirement in the prescription feeding.
2. The liquid chromatography characteristic spectrum method for identifying the true and false of the veterinary Shuanghuanglian oral liquid according to claim 1, wherein the step S1 specifically comprises the following steps: S101, setting chromatographic conditions, wherein octadecylsilane chemically bonded silica is used as a filler, methanol is used as a mobile phase A, and 0.2% phosphoric acid is used as a mobile phase B; S102, performing gradient elution according to preset rules, wherein the flow rate is 0.8ml per minute, the column temperature is 30 ℃, the detection wavelength is 235nm, and the theoretical plate number is not less than 6000 according to chlorogenic acid peaks.
3. The liquid chromatography characteristic spectrum method for identifying the true and false of the veterinary Shuanghuanglian oral liquid according to claim 1, wherein the step S2 specifically comprises the following steps: s201, taking new chlorogenic acid, cryptochlorogenic acid, forsythoside E, forsythoside I, forsythoside B, forsythoside H, forsythoside A, forsythoside and baicalin reference substances; s202, adding methanol to prepare solutions containing 40ug of chlorogenic acid, 40ug of cryptochlorogenic acid, 200ug of forsythoside E, 40ug of forsythoside I, 40ug of forsythoside B, 40ug of forsythoside H, 40ug of forsythoside A, 60ug of forsythoside and 100ug of baicalin per 1ml of the solution as reference solutions.
4. The liquid chromatography characteristic spectrum method for identifying the true and false of the veterinary Shuanghuanglian oral liquid according to claim 1, wherein the step S3 specifically comprises the following steps: S301, measuring 1ml to 50ml of Shuanghuanglian oral liquid in a volumetric flask, adding 50% methanol, and treating the obtained solution by using ultrasonic waves for 20min; S302, after cooling, adding 50% methanol into the volumetric flask until the liquid level reaches a specified scale mark, uniformly shaking the solution, and then filtering, and taking the subsequent filtrate, thereby obtaining the sample solution.
5. The liquid chromatography characteristic spectrum method for identifying the true and false of the veterinary Shuanghuanglian oral liquid according to claim 1, wherein the step S4 specifically comprises the following steps: s401, obtaining a chromatogram of a solution of a sample, wherein the solution of the chromatogram shows 12 characteristic peaks, and 10 characteristic peaks respectively correspond to retention time of reference peaks of corresponding reference solutions; S402, the peak corresponding to the chlorogenic acid reference peak is an S peak, and the relative retention time of the peak 2, the peak 6 and the S peak in the chromatogram is calculated, wherein the relative retention time is within the constraint range of a specified value.
6. The liquid chromatography characteristic spectrum method for identifying true and false of veterinary Shuanghuanglian oral liquid according to claim 5, wherein the step S4 specifically further comprises the following steps: S403, in the chromatogram, peak 2 is a characteristic peak which is derived from fructus forsythiae, can be transferred to the Shuanghuanglian oral liquid and is different from a characteristic peak of fructus forsythiae leaves, and peak 6 is a characteristic peak which is derived from honeysuckle, can be transferred to the Shuanghuanglian oral liquid and is different from honeysuckle; S404, if peak 2 is not detected in the chromatogram of the sample solution, fructus forsythiae is not used for administration according to the requirement in the prescription, and if peak 6 is not detected in the chromatogram of the sample solution, flos Lonicerae is not used for administration according to the requirement in the prescription.

Description

Liquid chromatography characteristic spectrum method for identifying authenticity of Shuanghuanglian oral liquid for livestock Technical Field The invention relates to the technical field of high performance liquid chromatography, in particular to a liquid chromatography characteristic spectrum method for identifying the authenticity of Shuanghuanglian oral liquid for livestock. Background The Shuanghuanglian oral liquid is used as a classical Chinese veterinary medicine preparation consisting of three medicinal materials of honeysuckle, baicalin and weeping forsythiae capsule, has obvious effects of relieving exterior syndrome by pungent and cool, clearing heat and detoxicating, is widely used for treating common cold and fever of chickens, is deeply trusted by farmers, and has the curative effect dependent on the synergistic effect of active ingredients such as chlorogenic acid, baicalin, forsythin and the like in the medicinal materials; However, as market demands expand to cause severe fluctuation of prices of raw materials such as fructus forsythiae and the like, partial enterprises are in order to reduce cost and obtain violence, pseudo-inferior products are produced by adopting a mode of replacing honeysuckle with lonicera japonica, replacing fructus forsythiae fruits with fructus forsythiae leaves and even directly adding chemical monomer components such as chlorogenic acid, baicalin, forsythin and the like, the behaviors not only seriously affect the quality and clinical safety of the products, but also seriously disturb market order, damage interests of normative production enterprises and farmers and restrict healthy development of the veterinary drug industry; therefore, in order to effectively suppress the phenomena of illegal feeding and adulteration and maintain a fair competitive market environment, a rapid and accurate method for identifying the authenticity of the Shuanghuanglian oral liquid is needed to be established. Disclosure of Invention The invention aims to provide a liquid chromatography characteristic spectrum method for identifying the authenticity of Shuanghuanglian oral liquid for animals, which solves the problems in the background technology. The invention provides a liquid chromatography characteristic spectrum method for identifying the authenticity of Shuanghuanglian oral liquid for livestock, which comprises the following steps: s1, setting chromatographic conditions, performing gradient elution by taking octadecylsilane chemically bonded silica as a stationary phase and methanol and 0.2% phosphoric acid solution as a mobile phase, and performing analysis under a specified detection wavelength; s2, preparing new chlorogenic acid, cryptochlorogenic acid, forsythoside E, forsythoside I, forsythoside B, forsythoside H, forsythoside A, forsythoside and baicalin reference substance solution; S3, precisely measuring Shuanghuanglian oral liquid to prepare a sample solution; S4, obtaining a chromatogram of the solution of the sample, and calculating the relative retention time of a peak 2, a peak 6 and a peak S in the chromatogram, wherein if the peak 2 is not detected in the chromatogram, the fructus forsythiae is not used for administration according to the requirement in the prescription feeding, and if the peak 6 is not detected in the graph, the flos lonicerae is not used for administration according to the requirement in the prescription feeding. Preferably, the step S1 specifically includes the following steps: S101, setting chromatographic conditions, wherein octadecylsilane chemically bonded silica is used as a filler, methanol is used as a mobile phase A, and 0.2% phosphoric acid is used as a mobile phase B; S102, performing gradient elution according to preset rules, wherein the flow rate is 0.8ml per minute, the column temperature is 30 ℃, the detection wavelength is 235nm, and the theoretical plate number is not less than 6000 according to chlorogenic acid peaks. Preferably, the step S2 specifically includes the following steps: s201, taking new chlorogenic acid, cryptochlorogenic acid, forsythoside E, forsythoside I, forsythoside B, forsythoside H, forsythoside A, forsythoside and baicalin reference substances; s202, adding methanol to prepare solutions containing 40ug of chlorogenic acid, 40ug of cryptochlorogenic acid, 200ug of forsythoside E, 40ug of forsythoside I, 40ug of forsythoside B, 40ug of forsythoside H, 40ug of forsythoside A, 60ug of forsythoside and 100ug of baicalin per 1ml of the solution as reference solutions. Preferably, the step S3 specifically includes the following steps: S301, measuring 1ml to 50ml of Shuanghuanglian oral liquid in a volumetric flask, adding 50% methanol, and treating the obtained solution by using ultrasonic waves for 20min; S302, after cooling, adding 50% methanol into the volumetric flask until the liquid level reaches a specified scale mark, uniformly shaking the solution, and then filtering, and taking the subsequent fi