CN115128201B - Gas chromatography-mass spectrometry fingerprint identification method for paulownia tomentosa leaves - Google Patents
Gas chromatography-mass spectrometry fingerprint identification method for paulownia tomentosa leaves Download PDFInfo
- Publication number
- CN115128201B CN115128201B CN202210884781.XA CN202210884781A CN115128201B CN 115128201 B CN115128201 B CN 115128201B CN 202210884781 A CN202210884781 A CN 202210884781A CN 115128201 B CN115128201 B CN 115128201B
- Authority
- CN
- China
- Prior art keywords
- paulownia
- temperature
- leaves
- peak
- fingerprint
- 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.)
- Active
Links
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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- 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
-
- 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Library & Information Science (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention discloses a gas chromatography-mass spectrometry fingerprint identification method of paulownia tomentosa leaves, which comprises the steps of (1) preparing a sample solution; (2) collection of GC/MS fingerprint patterns: respectively carrying out gas chromatography-mass spectrometry detection on the paulownia fortunei leaf sample solution and the sample solution to be identified, and recording fingerprint patterns of the paulownia fortunei leaf sample solution and the sample solution to be identified; (3) comparison of maps: the gas chromatography-mass spectrometry characteristic fingerprint of the paulownia fortunei leaf sample is shown in figure 1, 12 common peaks should be shown in the GC/MS spectrum of the sample to be identified, and correspond to the 12 common peaks in the GC/MS spectrum of the paulownia fortunei leaf, wherein the peak 5 is consistent with the retention time in the GC/MS spectrum of the paulownia fortunei leaf. The GC-MS fingerprint detection method for the fat-soluble components in the paulownia fortunei leaves provided by the invention has the advantages of high sensitivity and precision, good stability and repeatability, and can more objectively, comprehensively and accurately control the quality of the fat-soluble components in the paulownia fortunei leaves, has important significance for ensuring clinical efficacy, and fills a technical gap in the technical field of the identification of the paulownia fortunei leaves.
Description
Technical Field
The invention belongs to the technical field of quality control of traditional Chinese medicinal materials, and particularly relates to a gas chromatography-mass spectrometry fingerprint identification method of paulownia fortunei leaves.
Background
Chinese paulownia flowerPaulownia fortunei(seem.) hemsl.) is a falling tree of the genus paulownia of the family figwort, and is distributed in more than 10 provinces and cities in China. The paulownia fortunei serving as a high-quality fast-growing wood is widely applied to industrial and agricultural production, and is a common traditional Chinese medicine, wherein flowers, leaves, barks, roots and fruits of the paulownia fortunei can be used as medicines. The outline of the materia medica makes detailed records on pharmacological actions of all parts of the paulownia fortunei, and modern medical researches show that the paulownia fortunei has antibacterial, anti-inflammatory, anti-tumor and even insecticidal effects. The white paulownia has luxuriant branches and leaves, and the number of leaves is huge, thus being a rich resource. The paulownia leaves are used as feed, which not only can promote the growth of animals, but also can improve the disease resistance of animals. The medical value can be reflected in treating carbuncles, furuncles, wound bleeding and the like, and the paulownia fortunei leaves have important economic and social values.
Because of the high similarity of the leaves of the paulownia plants, particularly the traditional Chinese medicines are difficult to distinguish by naked eyes after being processed, sliced and powdered, no effective method is available at present for identifying the white paulownia leaves after being processed, sliced and powdered.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides a gas chromatography-mass spectrometry fingerprint identification method of paulownia fortunei leaves through a large number of experiments and researches.
The HPLC fingerprint identification method of the paulownia tomentosa leaves of the invention comprises the following steps:
(1) Preparation of sample solution: respectively weighing appropriate amount of paulownia tomentosa leaves and samples to be identified, crushing, sieving with a third sieve, precisely weighing 0.5 g, respectively placing in conical flasks with plugs, precisely adding petroleum ether 15 mL, sealing, performing ultrasonic treatment (power 500W, frequency 40 kHz) for 30min at 40 ℃, evaporating under reduced pressure to dryness, dissolving with 10 mL chromatographic n-hexane, adding appropriate amount of anhydrous sodium sulfate, drying, taking supernatant, and filtering with a 0.22 μm microporous filter membrane to obtain the paulownia tomentosa leaf sample solution and the samples to be identified.
(2) Collection of GC/MS spectra: GC/MS detection is carried out on the paulownia fortunei leaf sample solution and the sample solution to be identified respectively, and the fingerprint is recorded, and the chromatographic conditions are as follows: GC conditions: SH-Rtx-5MS gas chromatography column (30.0. 30.0 m X0.25 mm,0.25 μm); carrier gas high purity helium (purity > 99.99%); carrier gas volume flow rate 0.94 mL/min; the temperature of the sample inlet is 250 ℃; the initial temperature is 100 ℃, the temperature is kept constant for 2 min, the temperature is increased to 200 ℃ at the speed of 10 ℃/min, the temperature is kept constant for 5 min, the temperature is increased to 280 ℃ at the speed of 5 ℃/min, and the temperature is kept constant for 20 min; sample injection amount is 1.0 mu L; the split ratio is 1:10; mass spectrometry conditions: the ion source is an EI source; the ion source temperature is 230 ℃; the interface temperature is 250 ℃; ionization energy is 70 eV; delaying the solvent for 3 min; the scanning quality range m/z is 40-550.
(3) And (3) comparing the patterns: the gas chromatography-mass spectrometry characteristic fingerprint of the white paulownia leaf sample is shown in figure 1, 12 common peaks should be presented in the GC/MS spectrum of the sample to be identified, and correspond to 12 common peaks in the GC/MS spectrum of the white paulownia leaf, wherein the peak 5 is consistent with the retention time in the GC/MS spectrum of the white paulownia leaf, the 12 common peaks are respectively 2 (4H) -benzofurone, 5,6,7 a-tetrahydroo-4, 7a-trimethyl- (common peak 1), phytol (common peak 2), seventeen acetate (common peak 3), 2-methyldioctadecyl (common peak 4), squalene (common peak 5), eicosane (common peak 6), triacontane (common peak 7), triacontane (common peak 8), triacontane (common peak 9), tocopherols (common peak 10), tetracontane (common peak 11), beta-sitosterol (common peak 12), and the retention time is 62, 62-62, 7-tetracontane (common peak 3, 35-methyl-35-Z (common peak 3); the sample to be identified that satisfies these characteristics is identified as paulownia fortunei leaves, otherwise cannot be identified as paulownia fortunei leaves.
The invention is based on the GC/MS chromatographic conditions and fingerprint spectrograms of the paulownia tomentosa leaf extract, consumes a great deal of energy and creative labor, and has the following processes:
investigation of chromatographic conditions: we examined the chromatographic conditions from four aspects: flow dividing ratio investigation, temperature programming investigation, pre-column pressure and flow velocity investigation and sample inlet temperature investigation.
And (3) investigation of split ratio: the influence of split ratios of 1:10, 1:20 and 1:50 on GC-MS chromatograms of the petroleum ether sites of the paulownia fortunei leaves is examined respectively. Experimental study shows that the obtained map is ideal under the condition of the split ratio of 1:10. The split ratio was thus chosen to be 1:10.
Temperature programming investigation: the following three temperature programming methods were examined. When the method 3 is used, 17 peaks are detected together and the separation of the respective chromatographic peaks is optimal, so that the temperature-raising program is established as the GC temperature-raising program.
Method 1: the initial temperature is 60 ℃, the constant temperature is 5 min, the temperature is raised to 160 ℃ at the speed of 8 ℃/min, the constant temperature is 5 min, the temperature is raised to 280 ℃ at the speed of 12 ℃/min, and the constant temperature is 20 min.
Method 2: the initial temperature is 120 ℃, the temperature is kept constant for 3 min, the temperature is increased to 220 ℃ at the speed of 12 ℃/min, the temperature is kept constant for 5 min, the temperature is increased to 280 ℃ at the speed of 5 ℃/min, and the temperature is kept constant for 10 min.
Method 3: the initial temperature is 100 ℃, the temperature is kept constant for 2 min, the temperature is increased to 200 ℃ at the speed of 10 ℃/min, the temperature is kept constant for 5 min, the temperature is increased to 280 ℃ at the speed of 5 ℃/min, and the temperature is kept constant for 20 min.
Pre-column pressure and flow rate investigation: GC-MS analysis was examined under three different pre-column pressures of 60 KPa, 70 KPa, 80 KPa, respectively. The comparison shows that when the flow rate corresponding to the pre-column pressure of 70 KPa is 0.94 mL/min, the separation effect of the chromatographic peaks of the main components is better, so the experiment determines that the pre-column pressure is 70 KPa and the flow rate is 0.94 mL/min.
Temperature investigation of sample inlet: the effect of the sample inlet temperature of 250 ℃, 270 ℃ and 290 ℃ on the GC-MS chromatogram of the petroleum ether part of the paulownia fortunei leaves is examined respectively. Under the condition that other chromatographic conditions are consistent, the relative contents of 7 main components in the petroleum ether part of the paulownia fortunei leaf are smaller in RSD value at the temperature of the three sample inlets, which indicates that the relative contents of the main components are almost unchanged, and meanwhile, the separation effect of each main chromatographic peak is good. On the premise of meeting the vaporization requirement, the temperature should be selected as low as possible, and the temperature of the sample inlet is determined to be 250 ℃.
Thus, the chromatographic conditions were determined as: GC conditions: SH-Rtx-5MS gas chromatography column (30.0. 30.0 m X0.25 mm,0.25 μm); carrier gas high purity helium (purity > 99.99%); carrier gas volume flow rate 0.94 mL/min; the temperature of the sample inlet is 250 ℃; the initial temperature is 100 ℃, the temperature is kept constant for 2 min, the temperature is increased to 200 ℃ at the speed of 10 ℃/min, the temperature is kept constant for 5 min, the temperature is increased to 280 ℃ at the speed of 5 ℃/min, and the temperature is kept constant for 20 min; sample injection amount is 1.0 mu L; the split ratio is 1:10. mass spectrometry conditions: the ion source is an EI source; the ion source temperature is 230 ℃; the interface temperature is 250 ℃; ionization energy is 70 eV; delaying the solvent for 3 min; the scanning quality range m/z is 40-550.
Investigation of sample extraction method: the extraction mode examines two methods of ultrasonic extraction and Soxhlet extraction, and combines peak area results of 7 main components in petroleum ether extract, wherein the peak area of each main component of the ultrasonic extraction method is slightly larger than that of Soxhlet extraction in the two extraction methods, so the method is selected.
Investigation of feed liquid ratio: taking proper amount of paulownia fortunei leaves, crushing, sieving with a third sieve, precisely weighing three parts of 0.5 g fine powder, placing into a conical flask with a plug, respectively adding petroleum ether 5 mL, 10 mL and 15 mL, performing ultrasonic treatment (power 500W and frequency 40 kHz) at 40 ℃ for 30min, evaporating under reduced pressure, dissolving with 10 mL chromatographic n-hexane, adding proper amount of anhydrous sodium sulfate for drying, taking supernatant, and filtering with a microporous filter membrane of 0.22 mu m to obtain the paulownia fortunei. And combining peak area results of 7 main components in the petroleum ether extract, wherein the peak area of each common peak in the feed liquid ratio of 1:30 in the three feed liquid ratios is slightly larger than that of the other two feed liquid ratios, and finally selecting 1:30 as the extraction feed liquid ratio.
Investigation of extraction time: taking proper amount of paulownia fortunei leaves, crushing, sieving with a third sieve, precisely weighing three parts of 0.5 g fine powder, placing into a conical flask with a plug, adding petroleum ether 15 mL, respectively carrying out ultrasonic treatment (power 500W and frequency 40 kHz) for 30min, 40 min and 60 min at 40 ℃, evaporating under reduced pressure to dryness, dissolving with 10 mL chromatographic n-hexane, adding proper amount of anhydrous sodium sulfate for drying, taking supernatant, and filtering with a 0.22 mu m microporous membrane to obtain the paulownia fortunei. Combining peak area results of 7 main components in petroleum ether extract, wherein the peak area of each common peak is slightly larger than the other two extraction times in 30min in different extraction times, and finally 30min is selected as the extraction time.
Investigation of extraction temperature: taking proper amount of paulownia fortunei leaves, crushing, sieving with a third sieve, precisely weighing three parts of 0.5 g fine powder, placing into a conical flask with a plug, adding petroleum ether 15 mL, respectively carrying out ultrasonic treatment (power 500W and frequency 40 kHz) at 30 ℃ and 40 ℃ for 30min, evaporating under reduced pressure, dissolving with 10 mL chromatographic n-hexane, adding proper amount of anhydrous sodium sulfate for drying, taking supernatant, and filtering with a 0.22 mu m microporous filter membrane to obtain the paulownia fortunei. Combining peak area results of 7 main components in petroleum ether extract, wherein the peak area of each common peak in the three extraction temperatures is slightly larger than that of other two extraction temperatures, and finally 40 ℃ is selected as the extraction temperature.
Determination of the reference: one peak must be selected from the chromatographic fingerprint as a reference peak, and the corresponding substance is called a reference substance. The squalene (No. 5 peak) in the petroleum ether part of the paulownia fortunei is the peak with the largest peak area and the most stable, and can be well separated from the adjacent chromatographic peak, so the squalene can be used as a reference object of the fingerprint detection standard.
The research on the combined characteristic fingerprint of the paulownia fortunei She Qizhi is as follows:
precision experiment: taking a paulownia fortunei leaf sample, continuously sampling for 6 times according to the method of the invention, sampling 1 mu L each time, and recording the retention time and the peak area of 12 common chromatographic peaks. The relative retention time and relative peak area of each peak in the spectrum were calculated using squalene as a reference peak, and the values between them were calculated. The results show that the relative retention time (RSD) of each common peak is less than 0.72%, and the relative peak area (RSD) is less than 1.09%, which indicates that the precision of the instrument is good.
Repeatability experiments: 6 parts of paulownia fortunei leaf samples are taken, test solutions are prepared according to the method of the invention, and are respectively injected, 1 mu L of sample is injected each time, and the retention time and the peak area of 12 common chromatographic peaks are recorded. The relative retention time and relative peak area of each peak in the spectrum were calculated using squalene as a reference peak, and the values between them were calculated. The results showed that each of the common peaks had a relative retention time RSD of less than 0.87% and a relative peak area RSD of less than 2.49%, indicating good instrument reproducibility.
Stability experiment: taking a paulownia fortunei leaf sample, preparing the same test solution according to the method of the invention, injecting 1 mu L of sample each time when the test solution is placed in 0, 2, 4, 8, 12 and 24 h, and measuring according to the chromatographic conditions determined under 3.2.5 items. The retention time and peak area of 12 of the common chromatographic peaks were recorded. The relative retention time and relative peak area of each peak in the spectrum were calculated using squalene as a reference peak, and the values between them were calculated. The results showed that each of the common peaks had a relative retention time RSD of less than 1.66% and a relative peak area RSD of less than 3.81%, indicating good stability of sample 24 h.
Under the condition of the invention, GC/MS analysis is carried out on 19 batches of paulownia fortunei leaves, and the fingerprint spectrum of each batch of samples is recorded. And analyzing the fingerprint of 19 batches of paulownia fortunei leaves by adopting a traditional Chinese medicine chromatographic fingerprint similarity evaluation system (2012A edition), screening out chromatographic peaks with good repeatability as common peaks, and respectively calculating the relative peak areas of each common peak and reference peak in each sample fingerprint, wherein each total ion flow chart is shown in figure 2. The sample characteristic spectrum should show 12 common peaks and correspond to 12 common peaks in the chromatographic peaks of the reference substance of the control medicinal material, wherein peak 5 should be consistent with the retention time of the reference substance peaks, wherein the 12 common peaks are respectively 2 (4H) -Benzofuranone, 5,6,7 a-tetrahydroo-4, 7a-trimethyl- (common peak 1), phytol (common peak 2), heptadecyl acetate (common peak 3), 2-methyloctacosane (common peak 4), squalene (common peak 5), nonacosane (common peak 6), triacontane (common peak 7), triacontane (common peak 8), nonatriacontane (common peak 9), tocopherol (common peak 10), tetradecane (common peak 11), beta-sitosterol (common peak 12), and the retention time is 7.273 min (1, 7-Octadien-3-ol, 3, 7-dimethyl-), 26.404-3, 858-9, Z (common peak 35), and methyl-tetramethyide (common peak 35, 35).
GC/MS fingerprint similarity evaluation of paulownia fortunei leaves: and (3) performing similarity evaluation on the collected 19 batches of paulownia fortunei leaves by adopting a traditional Chinese medicine chromatographic fingerprint similarity evaluation system (2012A edition), and calculating the similarity between each batch of fingerprint and a reference fingerprint (R). The results show that the similarity between the finger print of each batch of paulownia fortunei leaves and the reference finger print (R) is between 0.973 and 1.
Compared with the prior art, the invention has the beneficial effects that:
the method can control the quality of the paulownia fortunei leaves stably, precisely and reproducibly and effectively identify the paulownia fortunei leaves for the first time by utilizing the GC/MS fingerprint spectrum, and fills a technical blank in the field.
The invention discloses a detection method of GC-MS finger print of paulownia fortunei leaves for the first time, wherein the retention time of a reference peak is 1, and the relative retention time of a common peak is calculated to obtain the GC-MS finger print or the GC-MS feature map of the paulownia fortunei leaves. The method establishes 12 characteristic peaks in the paulownia tomentosa leaves, selects squalene as a reference peak, determines the relative retention time of each characteristic peak, and determines four characteristic peaks of the paulownia tomentosa leaves as 7.273 min (1, 7-Octadien-3-ol, 3, 7-dimethyl-), 26.404 min (9, 12-Octadecadienoyl chloride, (Z, Z) -), 32.363 min (photonic acid, bis (7-methyl) ester) and 39.362 min (campestol) respectively. The GC-MS fingerprint detection method for the fat-soluble components in the paulownia fortunei leaves provided by the invention has the advantages of high sensitivity and precision, good stability and repeatability, and can more objectively, comprehensively and accurately control the quality of the fat-soluble components in the paulownia fortunei leaves, thereby having great significance for ensuring clinical efficacy.
Drawings
Fig. 1 is a gas chromatography-mass spectrometry fingerprint of paulownia tomentosa leaves.
Figure 2 is a gas chromatography-mass spectrometry fingerprint of 19 sets of paulownia fortunei leaves petroleum ether parts.
Fig. 3 is a comparative gas chromatography-mass spectrometry graph of four paulownia leaves of paulownia fortunei, paulownia orchioides, paulownia tomentosa and paulownia taiwan.
Detailed Description
The invention will now be further described in detail with reference to specific examples.
GC-MS fingerprint identification of paulownia fortunei, paulownia orchid, paulownia tomentosa and paulownia taiwan:
(1) Preparation of sample solution: respectively weighing appropriate amount of paulownia tomentosa leaves and samples to be identified, crushing, sieving with a third sieve, precisely weighing 0.5 g, respectively placing in conical flasks with plugs, precisely adding petroleum ether 15 mL, sealing, performing ultrasonic treatment (power 500W, frequency 40 kHz) for 30min at 40 ℃, evaporating under reduced pressure to dryness, dissolving with 10 mL chromatographic n-hexane, adding appropriate amount of anhydrous sodium sulfate, drying, taking supernatant, and filtering with a 0.22 μm microporous filter membrane to obtain the paulownia tomentosa leaf sample solution and the samples to be identified.
(2) Collection of HPLC profile: GC/MS detection is carried out on the paulownia fortunei leaf sample solution and the sample solution to be identified respectively, and the fingerprint is recorded, and the chromatographic conditions are as follows: GC conditions: SH-Rtx-5MS gas chromatography column (30.0. 30.0 m X0.25 mm,0.25 μm); carrier gas high purity helium (purity > 99.99%); carrier gas volume flow rate 0.94 mL/min; the temperature of the sample inlet is 250 ℃; the initial temperature is 100 ℃, the temperature is kept constant for 2 min, the temperature is increased to 200 ℃ at the speed of 10 ℃/min, the temperature is kept constant for 5 min, the temperature is increased to 280 ℃ at the speed of 5 ℃/min, and the temperature is kept constant for 20 min; sample injection amount is 1.0 mu L; the split ratio is 1:10; mass spectrometry conditions: the ion source is an EI source; the ion source temperature is 230 ℃; the interface temperature is 250 ℃; ionization energy is 70 eV; delaying the solvent for 3 min; the scanning quality range m/z is 40-550.
(3) And (3) comparing the patterns: the gas chromatography-mass spectrometry characteristic fingerprint of the white paulownia leaf sample is shown in figure 1, 12 common peaks should be presented in the GC/MS spectrum of the sample to be identified, and correspond to 12 common peaks in the GC/MS spectrum of the white paulownia leaf, wherein the peak 5 is consistent with the retention time in the GC/MS spectrum of the white paulownia leaf, the 12 common peaks are respectively 2 (4H) -benzofurone, 5,6,7 a-tetrahydroo-4, 7a-trimethyl- (common peak 1), phytol (common peak 2), seventeen acetate (common peak 3), 2-methyldioctadecyl (common peak 4), squalene (common peak 5), eicosane (common peak 6), triacontane (common peak 7), triacontane (common peak 8), triacontane (common peak 9), tocopherols (common peak 10), tetracontane (common peak 11), beta-sitosterol (common peak 12), and the retention time is 62, 62-62, 7-tetracontane (common peak 3, 35-methyl-35-Z (common peak 3); the sample to be identified that satisfies these characteristics is identified as paulownia fortunei leaves, otherwise cannot be identified as paulownia fortunei leaves.
Under the condition of the invention, GC/MS comparison chromatograms of the paulownia fortunei, the paulownia orchioides, the paulownia tomentosa and the paulownia taiwan are shown in figure 3 (from bottom to top, the paulownia fortunei, the paulownia tomentosa and the paulownia taiwan respectively). From comparison of four paulownia contrasts, the paulownia fortunei has four characteristic peaks compared with other three paulownia fortunei, and the retention time is 7.273 min of 1,7-Octadien-3-ol, 3,7-dimethyl-;26.404 min 9,12-Octadecadienoyl chloride, (Z, Z) -;32.363 The Phthalic acid of min, bis (7-methyl) ester;39.362 Campesterol for min. GC/MS fingerprints of the paulownia orchid, the paulownia tomentosa and the paulownia taiwan do not accord with the identification conditions of the GC/MS characteristic fingerprints of the paulownia fortunei leaves, and the paulownia fortunei leaves are distinguished.
The foregoing is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Many possible variations and modifications of the disclosed technology can be made by anyone skilled in the art, or equivalent embodiments with equivalent variations can be made, without departing from the scope of the invention. Therefore, any simple modification, equivalent variation and modification of the above embodiments according to the technical substance of the present invention shall fall within the scope of the technical solution of the present invention.
Claims (1)
1. A gas chromatography-mass spectrometry fingerprint identification method of paulownia fortunei leaves is characterized by comprising the following steps:
(1) Preparation of sample solution: respectively weighing appropriate amount of paulownia tomentosa leaves and a sample to be identified, crushing, sieving with a third sieve, precisely weighing 0.5 g, respectively placing in conical flasks with plugs, precisely adding petroleum ether 15 mL, sealing, performing ultrasonic treatment at a power of 500W and a frequency of 40 kHz and a temperature of 40 ℃ for 30min, evaporating under reduced pressure, dissolving with 10 mL chromatographic n-hexane, adding appropriate amount of anhydrous sodium sulfate, drying, taking supernatant, and filtering with a microporous membrane of 0.22 mu m to obtain an paulownia tomentosa leaf sample solution and a sample solution to be identified;
(2) Collection of GC/MS spectra: GC/MS detection is carried out on the paulownia fortunei leaf sample solution and the sample solution to be identified respectively, and the fingerprint is recorded, wherein the GC chromatographic conditions are as follows: SH-Rtx-5MS gas chromatographic column with parameters of 30.0. 30.0 m ×0.25. 0.25 mm and 0.25 μm; the carrier gas is high-purity helium with the purity more than 99.99%; carrier gas volume flow rate 0.94 mL/min; the temperature of the sample inlet is 250 ℃; the initial temperature is 100 ℃, the temperature is kept constant for 2 min, the temperature is increased to 200 ℃ at the speed of 10 ℃/min, the temperature is kept constant for 5 min, the temperature is increased to 280 ℃ at the speed of 5 ℃/min, and the temperature is kept constant for 20 min; sample injection amount is 1.0 mu L; the split ratio is 1:10; mass spectrometry conditions: the ion source is an EI source; the ion source temperature is 230 ℃; the interface temperature is 250 ℃; ionization energy is 70 eV; delaying the solvent for 3 min; scanning the mass range m/z to 40-550;
(3) And (3) comparing the patterns: comparing the GC/MS fingerprint of the white paulownia leaf sample solution with the GC/MS fingerprint of the sample solution to be identified, wherein 12 common peaks are required to be displayed in the GC/MS fingerprint of the sample solution to be identified and correspond to 12 common peaks in the GC/MS fingerprint of the white paulownia leaf sample solution, wherein the peak 5 is consistent with the retention time in the GC/MS fingerprint of the white paulownia leaf, the common peak 1 is 2 (4H) -Benzofuranone, 5,6,7 a-tetrahydrol-, the common peak 2 is phytol, the common peak 3 is heptadecyl acetate, the common peak 4 is 2-methyloctacosane, the common peak 5 is squalene, the common peak 6 is nonadecane, the common peak 7 is triacontane, the common peak 8 is triacontane, the common peak 9 is triacontane, the common peak 10 is tocopherols, the common peak 11 is decadecane, the common peak 12 is beta-sterols, and the retention times are 1, 6,7 a-tetrasterol, 35-35Z (methyl-35); the sample to be identified that satisfies these characteristics is identified as paulownia fortunei leaves, otherwise cannot be identified as paulownia fortunei leaves.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210884781.XA CN115128201B (en) | 2022-07-26 | 2022-07-26 | Gas chromatography-mass spectrometry fingerprint identification method for paulownia tomentosa leaves |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210884781.XA CN115128201B (en) | 2022-07-26 | 2022-07-26 | Gas chromatography-mass spectrometry fingerprint identification method for paulownia tomentosa leaves |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115128201A CN115128201A (en) | 2022-09-30 |
| CN115128201B true CN115128201B (en) | 2023-08-25 |
Family
ID=83386007
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210884781.XA Active CN115128201B (en) | 2022-07-26 | 2022-07-26 | Gas chromatography-mass spectrometry fingerprint identification method for paulownia tomentosa leaves |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115128201B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1963496A (en) * | 2006-11-21 | 2007-05-16 | 浙江林学院 | Method for setting up finger-print of pain-eliminate paste and standard finger-print thereof |
| CN112710765A (en) * | 2019-10-24 | 2021-04-27 | 泰州医药城国科化物生物医药科技有限公司 | Fingerprint detection method of gardenia medicinal material and application thereof |
| WO2021253160A1 (en) * | 2020-06-15 | 2021-12-23 | 陕西步长制药有限公司 | Fingerprint detection method for pharmaceutical preparation |
-
2022
- 2022-07-26 CN CN202210884781.XA patent/CN115128201B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1963496A (en) * | 2006-11-21 | 2007-05-16 | 浙江林学院 | Method for setting up finger-print of pain-eliminate paste and standard finger-print thereof |
| CN112710765A (en) * | 2019-10-24 | 2021-04-27 | 泰州医药城国科化物生物医药科技有限公司 | Fingerprint detection method of gardenia medicinal material and application thereof |
| WO2021253160A1 (en) * | 2020-06-15 | 2021-12-23 | 陕西步长制药有限公司 | Fingerprint detection method for pharmaceutical preparation |
Non-Patent Citations (1)
| Title |
|---|
| 丹七软胶囊脂溶性成分的GC-MS指纹图谱研究;庞颖;刘汉伟;高峰;杨健;;中国药房(第06期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115128201A (en) | 2022-09-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102091118B (en) | Quality detection method for rhodiola crenulata | |
| CN110702813B (en) | Miao medicine caulis et folium periplocae HPLC fingerprint spectrum research and multi-component content determination method | |
| CN109060985A (en) | A kind of Russia's color berry extract and its quality determining method | |
| CN110286169B (en) | Method for simultaneously extracting and respectively purifying 5 chemical components from processed ramulus mori and application thereof | |
| CN110297060B (en) | Fingerprint detection method and fingerprint thereof for ixeris sonchifolia medicinal materials | |
| CN108254470A (en) | In glutinous rehmannia while carbohydrate content measure and its fingerprint map construction method | |
| CN115128201B (en) | Gas chromatography-mass spectrometry fingerprint identification method for paulownia tomentosa leaves | |
| CN108267535B (en) | Green tangerine peel medicinal material fingerprint detection method and fingerprint thereof | |
| CN115078608B (en) | Method for identifying UPLC characteristic patterns of herba abri and herba abri | |
| CN102628838B (en) | Fujian herb cusia HPLC fingerprint construction method | |
| CN112946132B (en) | Chinese rose medicinal material fingerprint spectrum, construction method thereof and Chinese rose medicinal material quality detection method | |
| CN110441443B (en) | UPLC characteristic spectrum construction method and identification method of pyrrosia peduncularis, pyrrosia lingua, pyrrosia cottonii and pyrrosia huabeiensis | |
| CN109856254B (en) | Method for establishing ethyl acetate part HPLC fingerprint spectrum of longan leaf | |
| CN115060841B (en) | HPLC fingerprint identification method for paulownia tomentosa leaves | |
| CN113759029A (en) | Method for rapidly distinguishing mint water extract from spearmint water extract | |
| CN108982731B (en) | Method for constructing HPLC characteristic spectrum of sarcandra glabra buccal tablet and HPLC characteristic spectrum thereof | |
| CN112147265A (en) | Honeysuckle anti-inflammatory quality marker screening and quality identification method and application | |
| CN111721880A (en) | Method for establishing fingerprint of sarcandra glabra by using double-column tandem HPLC-MS | |
| CN111380989A (en) | Single-side needle quality control method based on ultra-high performance liquid chromatography | |
| CN115128200B (en) | HPLC quality detection method for paulownia tomentosa leaves | |
| CN114965776B (en) | Method for establishing characteristic spectrum of pediatric Huanglong granule and standard characteristic spectrum and application thereof | |
| CN110057950A (en) | A kind of construction method of the Pudilan sulfathiazole HPLC finger-print based on Detection wavelength switching | |
| CN111766309B (en) | Detection method of Anshenbao particle characteristic spectrum | |
| CN107389845B (en) | A kind of method for building up and its standard finger-print of the efficient liquid-phase chromatograph finger print atlas of pumpkin seeds | |
| CN116008456A (en) | Fingerprint detection method of wushen decoction |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |