CN111983047A - Fingerprint detection method for volatile oil intermediate of perilla yellow cough-relieving capsule and application of fingerprint detection method - Google Patents
Fingerprint detection method for volatile oil intermediate of perilla yellow cough-relieving capsule and application of fingerprint detection method Download PDFInfo
- Publication number
- CN111983047A CN111983047A CN202010644034.XA CN202010644034A CN111983047A CN 111983047 A CN111983047 A CN 111983047A CN 202010644034 A CN202010644034 A CN 202010644034A CN 111983047 A CN111983047 A CN 111983047A
- Authority
- CN
- China
- Prior art keywords
- temperature
- percent
- detection method
- volatile oil
- pinene
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
Landscapes
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- General Physics & Mathematics (AREA)
- Engineering & Computer Science (AREA)
- Library & Information Science (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a detection method of a suhuang cough relieving capsule volatile oil intermediate fingerprint spectrum and application thereof. The detection method comprises the following steps: (1) preparing a test solution; (2) determining GC-MS chromatographic conditions; (3) determining a GC-MS spectrum; (4) and establishing a standard fingerprint spectrum to obtain the suhuang cough relieving capsule volatile oil intermediate fingerprint spectrum. The detection method can be used for quality control of volatile oil intermediate of SUHUANGZHIKE Capsule.
Description
Technical Field
The invention relates to but is not limited to a pharmaceutical analysis technology, in particular to a method for detecting a fingerprint of a threo Huang cough relieving capsule volatile oil intermediate and application thereof.
Background
The prescription of the cough-relieving capsule of perilla yellow comprises honey-fried ephedra, perilla leaf, earthworm, fried loquat leaf, perilla seed, cicada slough, radix peucedani, burdock and schisandra fruit. Wherein, the ephedra herb, the earthworm, the cicada slough, the perilla leaf and the whiteflower hogfennel root are used for dispelling wind and dispersing wind and are used for relieving cough, relieving sore throat and relieving itching; the perilla fruit and the Chinese magnoliavine fruit relieve spasm and spasm; the burdock and the loquat leaf can moisten lung and clear heat. The medicines are used together, the medicine has the effects of dispersing lung qi with pungent and warm natured drugs, dispelling wind and relieving spasm, dredging orifices and lowering qi, eliminating phlegm and relieving asthma, dispelling phlegm and eliminating spasm, and lung qi can be dispersed and lowered, and qi movement is smooth, so that the aims of integrally adjusting wind cough and relieving cough and treating both principal and secondary aspect of disease are fulfilled, and the medicine is used for treating cough variant asthma and cough after cold. Clinical tests show that the medicine has good effect and less adverse reaction compared with the current common western medicines.
Chinese patent application CN104007222A discloses a detection method of Suhuang cough-relieving capsules, which is used for quality control of the Suhuang cough-relieving capsules; the method adopts thin-layer chromatography to perform thin-layer qualitative detection on perilla leaf volatile oil in the perilla yellow cough-relieving capsule.
Although the prior art discloses a quality detection method of a perilla yellow cough-relieving capsule, no detection method of a perilla yellow cough-relieving capsule volatile oil intermediate fingerprint spectrum exists or the existing detection method of the perilla yellow cough-relieving capsule volatile oil cannot reflect the change of the quality of the perilla yellow cough-relieving capsule, and meanwhile, the relationship among effective components is not established yet.
Disclosure of Invention
The following is a summary of the subject matter described in detail herein. This summary is not intended to limit the scope of the claims.
The application provides a detection method of a suhuang cough relieving capsule volatile oil intermediate fingerprint spectrum and application thereof, a marker for controlling the quality of the suhuang cough relieving capsule volatile oil intermediate is established, and meanwhile, the quality of a product and the volatile oil intermediate can be better controlled.
The application provides a detection method of a suhuang cough relieving capsule volatile oil intermediate fingerprint spectrum, which comprises the following steps:
(1) preparation of a test solution:
precisely absorbing about 10 mu L of the volatile oil intermediate of the suhuang cough-relieving capsule, placing the volatile oil intermediate into a 2.0mL sample bottle, adding 990 mu L of n-hexane, and swirling for 30s to obtain a test solution;
(2) establishment of GC-MS analysis conditions
Capillary column: 5% -phenyl-methyl polysiloxane, HP-5MS capillary column with specification of 0.25 μm × 30m,0.25 mm;
carrier gas: high-purity He;
column flow rate: 1 mL;
sample inlet temperature: 230-270 ℃;
the split ratio is as follows: (15-25) 1;
interface temperature: 200 ℃;
pressing the column in front: 7psi-11 psi;
solvent retardation: 2.5 min;
mass spectrum conditions: EI electron source, the ion source temperature is 250 ℃, and the scanning range is 0-500 m/z;
sample introduction volume: 0.6-1.2 μ L;
temperature rising procedure: the initial temperature is 80 ℃, and the temperature is kept for 5 min; raising the temperature to 92 ℃ at the speed of 2 ℃/min, and keeping the temperature for 2 min; heating to 120 deg.C at 2 deg.C/min, and maintaining for 15 min; heating to 200 deg.C at 5 deg.C/min, and maintaining for 7 min;
(3) determination of GC-MS spectra
Sampling the sample solution according to the GC-MS analysis condition to obtain a GC-MS spectrum;
(4) establishment of standard fingerprint
Identifying common characteristic peaks in the above GC-MS profile, preferably 12 common characteristic peaks are established, including common characteristic peaks of the following components:
alpha-pinene, 5-methyl-2-furaldehyde, beta-pinene, para-cymene, limonene, linalool, perillanone, alpha-terpineol, (-) -trans-caryophyllene, gamma-curcumene, (+) -thujene and caryophyllene oxide.
In embodiments of the present application, the relative retention times of α -pinene, 5-methyl-2-furaldehyde, β -pinene, p-cymene, limonene, linalool, perillyl ketone, α -terpineol, (-) -trans-caryophyllene, γ -curcumene, (+) -thujene, and caryophyllene oxide in the GC-MS spectrum relative to perillyl ketone are respectively:
0.23 percent of alpha-pinene, 0.29 percent of 5-methyl-2-furaldehyde, 0.35 percent of beta-pinene, 0.37 percent of para-cymene, 0.38 percent of limonene, 0.54 percent of linalool, 1.00 percent of perillanone, 1.16-1.17 percent of alpha-terpineol, 1.53-1.54 percent of (-) -trans-caryophyllene, 1.64 percent of gamma-curcumene, (+) -thujene and 2.22-2.23 percent of caryophyllene oxide.
In a preferred embodiment of the application, the application provides a method for detecting the fingerprint of the volatile oil intermediate of the suhuang cough relieving capsule, wherein the GC-MS analysis conditions are as follows:
capillary column: 5% -phenyl-methyl polysiloxane, HP-5MS capillary column with specification of 0.25 μm × 30m,0.25 mm;
carrier gas: high-purity He;
column flow rate: 1 mL;
sample inlet temperature: 250 ℃;
the split ratio is as follows: 20: 1;
interface temperature: 200 ℃;
pressing the column in front: 10 psi;
solvent retardation: 2.5 min;
mass spectrum conditions: EI electron source, the ion source temperature is 250 ℃, and the scanning range is 0-500 m/z;
sample introduction volume: 1 mu L of the solution;
temperature rising procedure: the initial temperature is 80 ℃, and the temperature is kept for 5 min; raising the temperature to 92 ℃ at the speed of 2 ℃/min, and keeping the temperature for 2 min; heating to 120 deg.C at 2 deg.C/min, and maintaining for 15 min; raising the temperature to 200 ℃ at a speed of 5 ℃/min, and keeping the temperature for 7 min.
In some embodiments, the present application provides a method for detecting a fingerprint of a suhuang cough relieving capsule volatile oil intermediate, wherein the method comprises the following steps:
(1) preparation of a test solution:
sucking 10 μ L of volatile oil intermediate of SUHUANGZHIKE Capsule, placing in 2.0mL sample bottle, adding 990 μ L n-hexane, and vortexing for 30s to obtain test solution;
(2) establishment of GC-MS analysis conditions
Capillary column: 5% -phenyl-methyl polysiloxane, HP-5MS capillary column with specification of 0.25 μm × 30m,0.25 mm;
carrier gas: high-purity He;
column flow rate: 1 mL;
sample inlet temperature: 250 ℃;
the split ratio is as follows: 20: 1;
interface temperature: 200 ℃;
pressing the column in front: 10 psi;
solvent retardation: 2.5 min;
mass spectrum conditions: EI electron source, the ion source temperature is 250 ℃, and the scanning range is 0-500 m/z;
sample introduction volume: 1 mu L of the solution;
temperature rising procedure: the initial temperature is 80 ℃, and the temperature is kept for 5 min; raising the temperature to 92 ℃ at the speed of 2 ℃/min, and keeping the temperature for 2 min; heating to 120 deg.C at 2 deg.C/min, and maintaining for 15 min; heating to 200 deg.C at 5 deg.C/min, and maintaining for 7 min;
(3) determination of GC-MS spectra
Sampling the sample solution according to the GC-MS analysis condition to obtain a GC-MS spectrum;
(4) establishment of standard fingerprint
Determining 12 common characteristic peaks in the GC-MS spectrum, wherein the common characteristic peaks comprise common characteristic peaks of the following components:
alpha-pinene, 5-methyl-2-furfural, beta-pinene, p-cymene, limonene, linalool, perillanone, alpha-terpineol, (-) -trans-caryophyllene, gamma-curcumene, (+) -thujene and caryophyllene oxide; preferably, the relative retention times in the GC-MS spectrum relative to perillaketone are respectively:
0.23 percent of alpha-pinene, 0.29 percent of 5-methyl-2-furaldehyde, 0.35 percent of beta-pinene, 0.37 percent of para-cymene, 0.38 percent of limonene, 0.54 percent of linalool, 1.00 percent of perillanone, 1.16-1.17 percent of alpha-terpineol, 1.53-1.54 percent of (-) -trans-caryophyllene, 1.64 percent of gamma-curcumene, (+) -thujene and 2.22-2.23 percent of caryophyllene oxide.
In the embodiment of the application, after the standard fingerprint is determined, the evaluation is carried out by adopting the Chinese national pharmacopoeia committee 'Chinese medicine fingerprint similarity evaluation system software 2004 version A'.
On the other hand, the application provides the application of the fingerprint obtained by the detection method in the quality control of the Suhuang cough relieving capsule in the production process.
The established fingerprint spectrum measuring method can systematically monitor the overall quality of the suhuang cough-relieving capsule intermediate, and meanwhile, the common chromatographic peak of the volatile oil identified in the established fingerprint spectrum can be used for controlling and producing stable suhuang cough-relieving capsules and simultaneously ensuring the stability of the suhuang drug effect.
Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The objectives and other advantages of the invention will be realized and attained by the structure particularly pointed out in the written description and claims hereof as well as the appended drawings.
Drawings
The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the example serve to explain the principles of the invention and not to limit the invention.
FIG. 1 is a comparative chromatogram of the volatile oil intermediate of Suhuang cough-relieving capsule of example 1 with different temperature-raising procedures;
FIG. 2 is a comparative chromatogram of the volatile oil intermediate of the Suhuang cough relieving capsule from different chromatographic columns of example 1;
FIG. 3 is a comparative chromatogram of the volatile oil intermediate of Suhuang cough relieving capsule of example 1 at different split ratios;
FIG. 4 is a comparative chromatogram of the volatile oil intermediate of the Suhuang Zhike capsule in different sample volumes of example 1;
FIG. 5 is the fingerprint of the volatile oil intermediate of 13 Suhuang Zhike capsules of example 1;
FIG. 6 is a schematic representation of the essential oil intermediate of 13 Suhuang Zhike capsules of example 1;
FIG. 7 is the fingerprint of the volatile oil intermediate of 29 batches of Suhuang Zhike capsules of example 1;
FIG. 8 is a schematic representation of the essential oil intermediate of 29 Suhuang Zhike capsules of example 1.
Detailed Description
Hereinafter, embodiments of the present invention will be described in detail in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be noted that the embodiments and features of the embodiments in the present application may be arbitrarily combined with each other without conflict.
The instrument comprises the following steps: agilent GC-MS (containing 7890B gas chromatograph, 5977 mass spectrometer detector and 7693 autosampler) equipped with MassHunter 08B.00 workstation and Nist2017 mass spectrometry database; HP-5MS quartz capillary column (30m × 0.25mm, 0.25 mm); ZNHW-II type electric jackets (Shanghai Yuzhong instruments & Equipment Co., Ltd.); METTLER TOLEDO XS105DU ten-thousandth electronic balance (METTLER-TOLEDO group); Milli-Q water purifiers (Bedford, MA, USA); volatile oil extractor.
Material reagent: the Suhuang cough relieving capsule intermediate is provided by Beijing Haiyan pharmaceutical Co., Ltd of Yangziang pharmaceutical industry; chromatographically pure hexane (Shanghai Kangshenghua Biochemical high purity solvent Co., Ltd.); C7-C40 normal paraffin (USA 02si company). (-) -trans-caryophyllene is greater than or equal to 98% (Sigma), perillanone is greater than or equal to 99.0% (Santa Cruz Biotechnology), (+) -thujene is greater than or equal to 99% (Sigma), alpha-pyrolene (Sigma).
Example 1
Preparing a volatile oil intermediate of the perilla yellow cough relieving capsule: refer to the Chinese patent CN 200310100103.7.
Preparing a test solution of a volatile oil intermediate of a perilla yellow cough relieving capsule: precisely absorbing about 10 mu L of the volatile oil intermediate of the suhuang cough-relieving capsule, placing the volatile oil intermediate into a 2.0mL sample bottle, adding 990 mu L of n-hexane, and vortexing for 30s to obtain a test solution.
Comparison of GC-MS detection conditions
Quartz capillary column HP-5MS (0.25 μm × 30m,0.25mm, 5% -phenyl-methyl polysiloxane); the carrier gas is high-purity He, the flow rate of the column is 1mL, the injection port temperature is 250 ℃, the split ratio is 20:1, the interface temperature is 200 ℃, and the solvent delay is 2.5 min. Mass spectrum conditions: EI electron source, ion source temperature 250 deg.C, scanning range 0-500m/z, database NIST 2017. The injection volume was 1. mu.L.
Adopting a Suhuang cough relieving capsule volatile oil intermediate sample (18022221), and comparing 4 temperature raising programs, wherein the comparison is as follows: (1) maintaining the initial temperature at 120 deg.C for 5 min; reducing the temperature to 80 ℃ at a speed of 5 ℃/min, and keeping the temperature for 1 min; then raising the temperature to 120 ℃ at the speed of 1 ℃/min, and keeping the temperature for 10 min; raising the temperature to 200 ℃ at a speed of 5 ℃/min, and keeping the temperature for 5 min. (2) The initial temperature is 70 ℃, and the temperature is kept for 10 min; heating to 120 deg.C at 2 deg.C/min, and maintaining for 20 min; then raising the temperature to 200 ℃ at a speed of 5 ℃/min, and keeping the temperature for 8 min. (3) The initial temperature is 110 ℃, the temperature is kept for 4min, the temperature is reduced to 80 ℃ at the speed of 5 ℃/min, and the temperature is kept for 1 min; then raising the temperature to 120 ℃ at the speed of 2 ℃/min, and keeping the temperature for 20 min; raising the temperature to 200 ℃ at a speed of 5 ℃/min, and keeping the temperature for 8 min; then raising the temperature to 280 ℃ at a speed of 10 ℃/min, and keeping the temperature for 2 min. (4) The initial temperature is 80 ℃, and the temperature is kept for 5 min; raising the temperature to 92 ℃ at the speed of 2 ℃/min, and keeping the temperature for 2 min; heating to 120 deg.C at 2 deg.C/min, and maintaining for 15 min; raising the temperature to 200 ℃ at a speed of 5 ℃/min, and keeping the temperature for 7 min. The results are shown in FIG. 1.
The comparison result shows that when the temperature raising program is compared with the investigated temperature raising program, the chromatographic peak separation of the volatile oil intermediate sample of the suhuang cough relieving capsule is better when the temperature raising program is (4).
1.2. Comparison of chromatography columns
Samples of the same batch of the volatile oil intermediate of the suhuang cough-relieving capsule were taken and injected into a volume of 1 μ L using (1) an HP-5MS capillary column (0.25 μm × 30m,0.25mm, 5% -phenyl-methyl polysiloxane) and (2) DB-5MS (0.25 μm × 30m × 0.25mm, 5% -phenyl-methyl polysiloxane), respectively. The results are shown in FIG. 2.
The results show that the separation effect of each chromatographic peak of the volatile oil intermediate sample is better when the type of the chromatographic column is an HP-5MS capillary column (0.25 mu m multiplied by 30m multiplied by 0.25mm, 5% -phenyl-methyl polysiloxane).
1.3. Comparison of the shunt ratio
Taking the volatile oil intermediate sample of the same batch of Suhuang cough relieving capsules, and injecting 1 mu L of sample by using an HP-5MS capillary column (0.25 mu m multiplied by 30m multiplied by 0.25mm, 5% -phenyl-methyl polysiloxane) under the conditions of the split ratio of 10: 1, 20:1, 30: 1 and 40: 1 respectively. The results are shown in FIG. 3.
By comparing different split ratios, the result shows that the effect of the split ratio of 20:1 and 30: 1 is superior to that of the split ratio of 10: 1 and 40: 1; however, at a split ratio of 20:1, the number of peaks detected by the mass spectrometer (167) is significantly greater than the number of peaks detected at a split ratio of 30: 1 (113).
1.4. Comparison of sample volumes
The sample introduction volume is related to the number of chromatographic peaks detected by the mass spectrum and the separation condition among the chromatographic peaks, so that the sample separation condition is investigated in experiments at different sample introduction volumes. Taking the same batch of Suhuang cough relieving capsule volatile oil intermediate samples, and inspecting the influence of injection volumes of 0.5 muL, 1 muL, 1.5 muL and 2 muL on the separation condition of each chromatographic peak in a spectrogram by using an HP-5MS capillary column (0.25μm multiplied by 30m,0.25mm, 5% -phenyl-methyl polysiloxane) under the condition that the split flow ratio is 20: 1. The results are shown in FIG. 4.
Experimental results show that when the sample injection amount is 0.5 muL, chromatographic peaks are well separated, but the number of peaks is small, the condition of sample components cannot be fully reflected, and when the sample injection volumes are respectively 1.5 muL and 2 muL, chromatographic peaks in a total ion flow graph are not well separated due to overlarge sample injection volumes. Therefore, the injection volume was selected to be 1. mu.L.
In addition, the influence of the sample inlet temperature such as 230 ℃, 240 ℃, 250 ℃, 260 ℃ and 270 ℃ on the separation condition of each spectrum peak in the spectrogram is also considered; effect of column front pressures, e.g., 7psi, 8psi, 9psi, 10psi, and 11psi, on the separation of the spectral peaks in the spectra.
By comprehensive comparison, the temperature is maintained for 5min under the condition that the initial temperature is 80 ℃ by using a temperature rising program; raising the temperature to 92 ℃ at the speed of 2 ℃/min, and keeping the temperature for 2 min; heating to 120 deg.C at 2 deg.C/min, and maintaining for 15 min; heating to 200 deg.C at 5 deg.C/min, and maintaining for 7 min; the chromatographic column is HP-5MS capillary column (0.25 μm × 30m,0.25mm, 5% -phenyl-methyl polysiloxane); the split ratio is 20: 1; the temperature of a sample inlet is 250 ℃; the sample injection volume is 1 mu L; the separation condition of the volatile oil intermediate sample of the Suhuang cough relieving capsule is better when the pre-column pressure is 10 psi.
2. Methodology validation
A sample of the Suhuang cough-relieving capsule intermediate is prepared into a test solution for GC-MS analysis according to the sample solution preparation method under 'example 1'.
2.1 precision test
And 6 groups of tests are carried out, the relative content of each chromatographic peak is calculated according to the relative content of the peak area of each chromatographic peak relative to the perillaketone in the spectrogram, and the relative retention time and the relative peak area RSD value of 12 chromatographic peaks with larger response and relatively stable content are all less than 5 percent. The results are shown in Table 2-1 relative to the retention time RSD values; the relative peak area RSD values are shown in Table 2-2.
Table 2-1 precision experiments relative retention time results (n ═ 6)
Table 2-2 results of relative peak area for precision experiments (n ═ 6)
2.2 stability test
6 groups of tests are carried out, and the stability of the sample within 0-24 h is inspected. And calculating the relative content of each chromatographic peak according to the relative content of each chromatographic peak to the area of the perillanone peak in the spectrogram, and calculating the relative retention time and the relative peak area RSD (maximum likelihood of false alarm) value of 12 chromatographic peaks with larger response and relatively stable content which are all less than 5 percent. The results are shown in tables 2-3 relative to the retention time RSD values; the relative peak area RSD values are shown in tables 2-4.
Table 2-3 stability experiments relative retention time results (n ═ 6)
Tables 2-4 results of relative peak area for stability experiments (n ═ 6)
2.3 repeatability test
6 sets of experiments were performed to investigate experimental reproducibility. And calculating the relative content of each chromatographic peak according to the relative content of each chromatographic peak to the area of the perillanone peak in the spectrogram, and calculating the relative retention time and the relative peak area RSD (maximum likelihood of false alarm) value of 12 chromatographic peaks with larger response and relatively stable content which are all less than 5 percent. The results are shown in tables 2-5 relative to the retention time RSD values; the relative peak area RSD values are shown in tables 2-6.
Table 2-5 relative retention time results of repeated experiments (n ═ 6)
Table 2-6 relative peak area results of repeated experiments (n ═ 6)
3 fingerprint spectra of volatile oil intermediate of Suhuang cough relieving capsule in different batches
3.1 establishment of fingerprint of volatile oil intermediate of Suhuang cough relieving Capsule
Taking 13 batches of volatile oil intermediate samples of the suhuang cough-relieving capsules in different batches, preparing a test solution according to the method in the 'example 1', carrying out GC-MS analysis by applying the chromatographic conditions to obtain a GC-MS fingerprint, and analyzing the result. And (3) calibrating by taking 12 chromatographic peaks with the peak area content of the perillaketone being more than 0.01% in a spectrogram as a common peak, and analyzing GC-MS chromatograms of volatile oil intermediates of the Suhuang cough relieving capsules in 13 different production batches. As a result, the common characteristic peaks identified were: 0.23 percent of alpha-pinene, 0.29 percent of 5-methyl-2-furaldehyde, 0.35 percent of beta-pinene, 0.37 percent of para-cymene, 0.38 percent of limonene, 0.54 percent of linalool, 1.00 percent of perillanone, 1.16-1.17 percent of alpha-terpineol, 1.53-1.54 percent of (-) -trans-caryophyllene, 1.64 percent of gamma-curcumene, (+) -thujene and 2.22-2.23 percent of caryophyllene oxide. GC-MS data of 13 batches of samples are processed and then led into a traditional Chinese medicine chromatography fingerprint computer similarity evaluation system, and 13 batches of volatile oil intermediate total ion flow diagram superposition graphs after multi-point correction are shown in an attached figure 5.
From the results of GC-MS of 13 batches of suhuang cough relieving capsule volatile oil intermediates in table 3-1, the volatile oil intermediates mainly comprise α -pinene (0.0560-0.1004), 5-methyl-2-furaldehyde (0.0185-0.0360), β -pinene (0.0063-0.0117), p-cymene (0.0040-0.0083), limonene (0.0039-0.0111), linalool (0.0119-0.0234), perilla ketone (1.0000), α -terpineol (0.0815-0.1810), (-) -trans-caryophyllene (0.0434-0.0637), γ -curcumene (0.0053-0.0086), (+) -thujopside-arborvitae (0.0109-0.0320) and caryophyllene oxide (0.0103-0.0).
3.2 fingerprint similarity evaluation
The sample measurement result is calculated by adopting a traditional Chinese medicine fingerprint similarity evaluation system 2004A edition to obtain a fingerprint pattern chart and a similarity analysis result of the volatile oil intermediate of the suhuang cough relieving capsule. Wherein the common mode diagram is shown in figure 6, and the similarity analysis results of 13 volatile oil intermediates of different batches are shown in table 3.2.
TABLE 3.2 Suhuang antitussive capsules 13 batches of volatile oil intermediate samples similarity
The result shows that the similarity of 13 batches of the volatile oil intermediate of the suhuang cough relieving capsule is between 0.993 and 0.999, the result meets the requirement of the national food and drug administration on the similarity of the traditional Chinese medicine fingerprint, and the quality of the volatile oil intermediate of the suhuang cough relieving capsule is stable.
3.3 Perilla yellow cough relieving capsule volatile oil intermediate GC-MS fingerprint verification experiment
The practicability of the established method is considered, 30 batches of different batches of Suhuang cough relieving capsule volatile oil intermediate samples are selected, the selected samples are determined according to the sample solution preparation method and the GC-MS condition optimization result under the item of 'example 1', and the determination results are analyzed. Results the overlay of the fingerprint of the volatile oil intermediate of 29 threo Huang Zhike capsules is shown in figure 7, except that the second batch (batch No. 18042923) cannot be introduced into the similarity analysis software during data processing.
The sample measurement result is calculated by adopting a traditional Chinese medicine fingerprint similarity evaluation system 2004A edition to obtain a fingerprint pattern chart and a similarity analysis result of the volatile oil intermediate of the suhuang cough relieving capsule. Wherein the common mode diagram is shown in figure 8, and the similarity analysis results of different batches of volatile oil intermediates are shown in table 3.3.
TABLE 3.3 Suhuang antitussive Capsule 29 batches of volatile oil intermediate samples similarity
The result shows that the similarity of the 29 batches of the volatile oil intermediate of the suhuang cough-relieving capsule is between 0.992 and 1, the result meets the requirement that the similarity of the fingerprint spectrum of the traditional Chinese medicine is more than 0.8 by the national food and drug administration, and the quality of the volatile oil intermediate of the suhuang cough-relieving capsule is further proved to be stable.
The method establishes the suhuang cough relieving capsule volatile oil intermediate fingerprint spectrum on the basis of comparing chromatographic conditions such as chromatographic columns, programmed heating conditions and the like, verifies the established method, establishes 42 batches of suhuang cough relieving capsule volatile oil intermediate fingerprint spectra, and performs similarity analysis and the like on the determination result. The results show that: and taking the perillaketone peak area as a reference peak, calculating the relative peak area of each chromatographic peak, and finally determining 12 chromatographic peaks containing perillaketone and having relatively large content as a common peak.
Meanwhile, the fingerprint chromatogram method for the volatile oil intermediate of the suhuang cough relieving capsule is determined as follows: the temperature raising procedure is that the initial temperature is 80 ℃ and the temperature is kept for 5 min; raising the temperature to 92 ℃ at the speed of 2 ℃/min, and keeping the temperature for 2 min; heating to 120 deg.C at 2 deg.C/min, and maintaining for 15 min; heating to 200 deg.C at 5 deg.C/min, and maintaining for 7 min; the chromatographic column is HP-5MS capillary column (0.25 μm × 30m,0.25mm, 5% -phenyl-methyl polysiloxane); the split ratio is 20: 1; the temperature of a sample inlet is 250 ℃; the sample injection volume is 1 mu L; the column front pressure was 10 psi.
Furthermore, the methodological investigation result shows that the retention time of 12 selected chromatographic peaks and the RSD value of the relative peak area are all less than 5 percent. The similarity analysis result shows that the similarity of 42 batches of the suhuang cough relieving capsule volatile oil intermediate samples is between 0.992 and 1. From 12 common peaks in 42 batches of volatile oil intermediate samples, it can be seen that the relative content of alpha-terpineol and caryophyllene oxide is increased in 2015 compared with 2018. The method can be used for distinguishing volatile oil intermediate samples with large production time distance.
Although the embodiments disclosed in the present application are described above, the descriptions are only for the convenience of understanding the present application, and are not intended to limit the present application. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the disclosure as defined by the appended claims.
Claims (7)
1. A detection method of a suhuang cough relieving capsule volatile oil intermediate fingerprint spectrum comprises the following steps:
(1) preparation of test solution
Sucking 10 μ L of volatile oil intermediate of SUHUANGZHIKE Capsule, placing in 2.0mL sample bottle, adding 990 μ L n-hexane, and vortexing for 30s to obtain test solution;
(2) establishment of GC-MS analysis conditions
Capillary column: 5% -phenyl-methyl polysiloxane, HP-5MS capillary column with specification of 0.25 μm × 30m,0.25 mm;
carrier gas: high-purity He;
column flow rate: 1 mL;
sample inlet temperature: 230-270 ℃;
the split ratio is as follows: (15-25) 1;
interface temperature: 200 ℃;
pressing the column in front: 7psi-11 psi;
solvent retardation: 2.5 min;
mass spectrum conditions: EI electron source, the ion source temperature is 250 ℃, and the scanning range is 0-500 m/z;
sample introduction volume: 0.6-1.2 μ L;
temperature rising procedure: the initial temperature is 80 ℃, and the temperature is kept for 5 min; raising the temperature to 92 ℃ at the speed of 2 ℃/min, and keeping the temperature for 2 min; heating to 120 deg.C at 2 deg.C/min, and maintaining for 15 min; heating to 200 deg.C at 5 deg.C/min, and maintaining for 7 min;
(3) determination of GC-MS spectra
Sampling the sample solution according to the GC-MS analysis condition to obtain a GC-MS spectrum;
(4) establishment of standard fingerprint
Common characteristic peaks were determined in the above GC-MS spectra.
2. The detection method according to claim 1, wherein the injection port temperature is 250 ℃;
the split ratio is 20: 1;
the column front pressure was 10 psi; and
the injection volume was 1. mu.L.
3. The detection method according to claim 1, wherein the determining of the common characteristic peaks establishes 12 common characteristic peaks comprising common characteristic peaks of:
alpha-pinene, 5-methyl-2-furaldehyde, beta-pinene, para-cymene, limonene, linalool, perillanone, alpha-terpineol, (-) -trans-caryophyllene, gamma-curcumene, (+) -thujene and caryophyllene oxide.
4. The detection method according to claim 3, wherein the relative retention times of the α -pinene, 5-methyl-2-furfural, β -pinene, p-cymene, limonene, linalool, perillanone, α -terpineol, (-) -trans-caryophyllene, γ -curcumene, (+) -thujene and caryophyllene oxide in the GC-MS spectrum with respect to perillanone are respectively:
0.23 percent of alpha-pinene, 0.29 percent of 5-methyl-2-furaldehyde, 0.35 percent of beta-pinene, 0.37 percent of para-cymene, 0.38 percent of limonene, 0.54 percent of linalool, 1.00 percent of perillanone, 1.16-1.17 percent of alpha-terpineol, 1.53-1.54 percent of (-) -trans-caryophyllene, 1.64 percent of gamma-curcumene, (+) -thujene and 2.22-2.23 percent of caryophyllene oxide.
5. The detection method according to claim 1, wherein the GC-MS analysis conditions are:
capillary column: 5% -phenyl-methyl polysiloxane, HP-5MS capillary column with specification of 0.25 μm × 30m,0.25 mm;
carrier gas: high-purity He;
column flow rate: 1 mL;
sample inlet temperature: 250 ℃;
the split ratio is as follows: 20: 1;
interface temperature: 200 ℃;
pressing the column in front: 10 psi;
solvent retardation: 2.5 min;
mass spectrum conditions: EI electron source, the ion source temperature is 250 ℃, and the scanning range is 0-500 m/z;
sample introduction volume: 1 mu L of the solution;
temperature rising procedure: the initial temperature is 80 ℃, and the temperature is kept for 5 min; raising the temperature to 92 ℃ at the speed of 2 ℃/min, and keeping the temperature for 2 min; heating to 120 deg.C at 2 deg.C/min, and maintaining for 15 min; raising the temperature to 200 ℃ at a speed of 5 ℃/min, and keeping the temperature for 7 min.
6. The detection method according to any one of claims 1 to 5, wherein the detection method comprises the steps of:
(1) preparation of a test solution:
sucking 10 μ L of volatile oil intermediate of SUHUANGZHIKE Capsule, placing in 2.0mL sample bottle, adding 990 μ L n-hexane, and vortexing for 30s to obtain test solution;
(2) establishment of GC-MS analysis conditions
Capillary column: 5% -phenyl-methyl polysiloxane, HP-5MS capillary column with specification of 0.25 μm × 30m,0.25 mm;
carrier gas: high-purity He;
column flow rate: 1 mL;
sample inlet temperature: 250 ℃;
the split ratio is as follows: 20: 1;
interface temperature: 200 ℃;
pressing the column in front: 10 psi;
solvent retardation: 2.5 min;
mass spectrum conditions: EI electron source, the ion source temperature is 250 ℃, and the scanning range is 0-500 m/z;
sample introduction volume: 1 mu L of the solution;
temperature rising procedure: the initial temperature is 80 ℃, and the temperature is kept for 5 min; raising the temperature to 92 ℃ at the speed of 2 ℃/min, and keeping the temperature for 2 min; heating to 120 deg.C at 2 deg.C/min, and maintaining for 15 min; heating to 200 deg.C at 5 deg.C/min, and maintaining for 7 min;
(3) determination of GC-MS spectra
Sampling the sample solution according to the GC-MS analysis condition to obtain a GC-MS spectrum; (4) establishment of standard fingerprint
Determining 12 common characteristic peaks in the GC-MS spectrum, wherein the common characteristic peaks comprise common characteristic peaks of the following components: alpha-pinene, 5-methyl-2-furfural, beta-pinene, p-cymene, limonene, linalool, perillanone, alpha-terpineol, (-) -trans-caryophyllene, gamma-curcumene, (+) -thujene and caryophyllene oxide; preferably, the relative retention times in the GC-MS spectrum relative to perillaketone are respectively:
0.23 percent of alpha-pinene, 0.29 percent of 5-methyl-2-furaldehyde, 0.35 percent of beta-pinene, 0.37 percent of para-cymene, 0.38 percent of limonene, 0.54 percent of linalool, 1.00 percent of perillanone, 1.16-1.17 percent of alpha-terpineol, 1.53-1.54 percent of (-) -trans-caryophyllene, 1.64 percent of gamma-curcumene, (+) -thujene and 2.22-2.23 percent of caryophyllene oxide.
7. The application of the fingerprint obtained by the detection method of any one of claims 1-6 in quality control of the suhuang cough-relieving capsule intermediate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010644034.XA CN111983047A (en) | 2020-07-03 | 2020-07-03 | Fingerprint detection method for volatile oil intermediate of perilla yellow cough-relieving capsule and application of fingerprint detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010644034.XA CN111983047A (en) | 2020-07-03 | 2020-07-03 | Fingerprint detection method for volatile oil intermediate of perilla yellow cough-relieving capsule and application of fingerprint detection method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111983047A true CN111983047A (en) | 2020-11-24 |
Family
ID=73438490
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010644034.XA Pending CN111983047A (en) | 2020-07-03 | 2020-07-03 | Fingerprint detection method for volatile oil intermediate of perilla yellow cough-relieving capsule and application of fingerprint detection method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111983047A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104007222A (en) * | 2014-06-03 | 2014-08-27 | 扬子江药业集团北京海燕药业有限公司 | Method for detecting ephedra-perilla leaf cough relieving capsule |
| US20190145942A1 (en) * | 2016-05-30 | 2019-05-16 | Givaudan Sa | Improvements in or Relating to Organic Compounds |
| CN110687219A (en) * | 2019-08-23 | 2020-01-14 | 扬子江药业集团北京海燕药业有限公司 | Detection method of suhuang cough-relieving capsule fingerprint and application thereof |
-
2020
- 2020-07-03 CN CN202010644034.XA patent/CN111983047A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104007222A (en) * | 2014-06-03 | 2014-08-27 | 扬子江药业集团北京海燕药业有限公司 | Method for detecting ephedra-perilla leaf cough relieving capsule |
| US20190145942A1 (en) * | 2016-05-30 | 2019-05-16 | Givaudan Sa | Improvements in or Relating to Organic Compounds |
| CN110687219A (en) * | 2019-08-23 | 2020-01-14 | 扬子江药业集团北京海燕药业有限公司 | Detection method of suhuang cough-relieving capsule fingerprint and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| 任淑清等: "紫苏梗挥发油的GC-MS定性分析", 《中国药房》 * |
| 张桂芝等: "羌活饮片挥发油化学成分GC-MS数字化指纹图谱研究", 《中成药》 * |
| 李舒丰等: "苏黄止咳胶囊及其挥发油中间体中挥发性成分的分析", 《中成药》 * |
| 蔡伟等: "3种紫苏属植物挥发油化学成分的GC-MS分析", 《云南中医中药杂志》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112730696A (en) | Method for detecting 5 cannabinol compounds in cannabis sativa oil by using HPLC (high performance liquid chromatography) method | |
| CN108896681B (en) | Nerve-soothing brain-tonifying liquid multi-index quantitative fingerprint establishment method and application thereof | |
| CN111983092A (en) | Method for detecting fructus amomi medicinal material fingerprint | |
| CN113049720A (en) | Method for detecting residual quantity of ethanol in compound fresh bamboo juice | |
| CN111089916B (en) | Method for detecting content of paeoniflorin, liquiritin and ammonium glycyrrhizinate in radix bupleuri and radix paeoniae alba oral liquid | |
| CN116429943A (en) | Quality analysis method of traditional Chinese medicine radix scutellariae based on dual-wavelength equal-baseline differential fusion map | |
| CN114778728A (en) | Determination method for curcumin compound content | |
| CN111983047A (en) | Fingerprint detection method for volatile oil intermediate of perilla yellow cough-relieving capsule and application of fingerprint detection method | |
| CN108918696B (en) | Establishment method and quality control method of spleen-tonifying and lung-benefiting prescription fingerprint | |
| AU2021106279A4 (en) | Method for establishing hplc-elsd fingerprints of shenlingbaizhu pills and standard fingerprints thereof | |
| CN113671099B (en) | Detection method of ziye Dan capsules | |
| CN114563497A (en) | Quantitative fingerprint detection method for bupleurum tenue capsule components | |
| CN114965802B (en) | Quality control method of climacteric syndrome relieving tablet | |
| CN113866318B (en) | Detection method of (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride | |
| CN114646690B (en) | Detection method for chemical components in immature bitter orange, longstamen onion bulb and cassia twig decoction and fingerprint establishment method | |
| CN113917009A (en) | Construction method and application of bupleurum chinense non-saponin component HPLC fingerprint | |
| CN108152384B (en) | Analysis method for distinguishing pericarpium citri reticulatae and pericarpium citri reticulatae by combining fingerprint spectrum with chemometrics | |
| CN109187780B (en) | Detection method of compound motherwort granules | |
| CN109239249B (en) | Method for measuring gas-phase fingerprint of Xingnaojing injection and standard gas-phase fingerprint thereof | |
| CN109856254B (en) | Method for establishing ethyl acetate part HPLC fingerprint spectrum of longan leaf | |
| CN114720568A (en) | Method for establishing fingerprint spectrum of radix pseudostellariae medicinal material and application thereof | |
| CN114563498B (en) | Quantitative fingerprint detection method for gingerol components in ginger and ginger processed pinellia percolate | |
| Tan et al. | A practical method for rapid screening and quantitative analysis of 130 pesticide residues in herbal medicines based on the Kovats retention index principle: an exemplary study using Panacis quinquefolii radix | |
| Chen et al. | Effects of processing method changes in main volatile compounds of Qixue Shuangbu Prescription by needle trap device coupled with gas chromatography-triple quadrupole mass spectrometry | |
| Humston-Fulmer et al. | Improving cannabis differentiation by expanding coverage of the chemical profile with GCxGC-TOFMS |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |