CN109580841B - Detection method for detecting essential oil content in rose product by solid-phase microextraction - Google Patents
Detection method for detecting essential oil content in rose product by solid-phase microextraction Download PDFInfo
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Abstract
The invention relates to the technical field of rose essential oil detection, in particular to a detection method for detecting the content of essential oil in a rose product by solid-phase microextraction, which can quickly and conveniently detect the content of essential oil in rose. The technical key points comprise that S1: preparing an internal standard solution; s2: pretreating a sample, extracting the sample with water to obtain water extract, extracting the water extract, and adding an internal standard solution into the water extract in the extraction process; s3: carrying out chromatography-mass spectrometry on the pretreated sample; s4: according to the result of the chromatographic-mass spectrometric analysis and in combination with the concentration of the internal standard solution, calculating the concentrations of citronellol, beta-phenethyl alcohol, geraniol and eugenol in the rose water extract to be M, mg/L; s5: calculating the concentration N, mg/L, N-M/0.92 of essential oil substances in the rose water extract according to the calculated M value; calculating the concentration of essential oil substance in rose, R, mg/g, R-M x (200/25) x 10‑3÷0.92。
Description
Technical Field
The invention belongs to the technical field of rose essential oil detection, and particularly relates to a detection method for detecting the content of essential oil in a rose product by solid-phase microextraction.
Background
Roses (Rosa rugosa) are plants of Rosaceae and Rosa, have beautiful and fresh color and rich fragrance, have multiple purposes of eating, medical treatment, health care, beauty treatment, appreciation and the like, are one of ten major flowers in China and are also one of four major cut flowers in the world. The rose essential oil is the crown of fresh flower oil, contains various effective aromatic components, is used as a traditional beauty product, and has the effects of improving skin and lightening melanin; can also regulate endocrine, nourish uterus, and relieve dysmenorrhea. The rose essential oil has fragrant smell, and is an essential perfume for women as a traditional perfume. Researches show that the rose essential oil also has antibacterial and bacteriostatic functions, is relatively sensitive to escherichia coli, staphylococcus aureus and salmonella typhimurium, has an anxiolytic effect and has great development value.
Chinese patent publication No. CN103063632B discloses a method for detecting the content of rose essential oil, which comprises treating rose with liquid nitrogen, dimethyl sulfoxide, etc. to make essential oil flow out, and then treating the sample with fluorescent dye, etc. The detection method is complex to operate, and is difficult to implement when dangerous goods such as liquid nitrogen, dimethyl sulfoxide and the like are used.
Disclosure of Invention
The invention aims to provide a detection method for detecting the content of essential oil in a rose product by solid-phase microextraction, which can quickly and conveniently detect the content of the essential oil in rose.
The technical purpose of the invention is realized by the following technical scheme:
a detection method for detecting the content of essential oil in a rose product by solid-phase microextraction comprises the following steps:
s1: preparing an internal standard solution;
s2: pretreating a sample, extracting the sample with water to obtain water extract, extracting the water extract, and adding an internal standard solution into the water extract in the extraction process;
s3: carrying out chromatography-mass spectrometry on the pretreated rose water extract;
s4: according to the result of the chromatographic-mass spectrometric analysis and in combination with the concentration of the internal standard solution, calculating the concentrations of citronellol, beta-phenethyl alcohol, geraniol and eugenol in the rose water extract to be M, mg/L;
s5: calculating the concentration N, mg/L, N-M/0.92 of essential oil substances in the rose water extract according to the calculated M value; calculating the concentration of essential oil substance in rose, R, mg/g, R-M x (200/25) x 10-3÷0.92；
S2, the water extraction process comprises the steps of putting a sample into a water bath kettle at the temperature of 60-70 ℃, and extracting twice;
during the water extraction process, magnesium formate is added to the solution.
Further, adding S6 after S5, and calculating the relative concentration W,% per thousand of the total amount of essential oil in the rose water extract according to the calculated N value; w ÷ N ÷ 629180 × 1000.
Further, in the step S1, the internal standard solution is a 2-nonanol solution.
Further, in the step S2, the sample pretreatment process includes adding the sample into a headspace bottle, adding sodium chloride and an internal standard solution, sealing, and magnetically stirring; the extraction head was inserted, extracted under water bath conditions, and then analyzed.
Further, in the step S3, the chromatographic conditions are that the capillary column: WAXETR30m × 0.25mm × 0.50 μm; column temperature procedure: the initial temperature is 50 ℃, the temperature is kept constant for 2min, the temperature is increased to 80 ℃ at the speed of 3 ℃/min, the temperature is increased to 115 ℃ at the speed of 10 ℃/min, the temperature is increased to 240 ℃ at the speed of 3 ℃/min, and the temperature is kept for 5 min; carrier gas, high-purity helium gas: the purity is more than or equal to 99.999 percent; flow rate: 1mL/min, no shunt sample introduction; sample inlet temperature: 230 ℃ to 230 ℃.
Further, in step S3, the mass spectrometry conditions are ion source temperature: 230 ℃; transmission line temperature: 240 ℃; electron bombardment source: 70 eV; scanning range: 29 to 300 amu.
Further, in the step S2, allyl caproate is added to the aqueous extract when the extraction is performed.
Furthermore, the volume ratio of the allyl hexanoate to the rose water extract is 1: 20.
The invention has the beneficial effects that:
1. the method developed by the invention can calculate and deduce the content of the essential oil in the roses and the relative concentration of the essential oil by detecting the contents of the four substances in the roses, has simple method and convenient operation, and creates the precedent for detecting the content of the essential oil in the roses in the field.
2. In the invention, 2-nonanol is selected as an internal standard liquid, the molecular structure and the property of the 2-nonanol are similar to those of the component to be detected, and a peak appears near the component to be detected; meanwhile, the rose does not contain the substance 2-nonanol, the 2-nonanol can be completely separated from other components in the sample, and peaks can be generated on a chromatogram map; the 2-nonanol has good mutual dissolving effect with the sample, and can not generate irreversible chemical reaction with a rose beverage, which is a complex sample.
3. Under the condition of the chromatographic-mass spectrometry, the separation effect of each substance in the sample is good, and no obvious peak dragging or overlapping phenomenon exists.
4. The allyl hexanoate is a low-polarity substance, and the addition of the allyl hexanoate into the rose water extract during extraction can change the polarity of the whole extraction system, enlarge the extraction range and improve the extraction effect.
5. The magnesium formate can destroy the lignified wall layer of the cell wall of the vegetable oil cell, so that the volatile oil in the oil cell can flow out quickly, the water extraction time can be further reduced, and the production cost of an enterprise can be saved.
Detailed Description
The technical solution of the present invention will be clearly and completely described below with reference to the specific embodiments. It is to be understood that the described embodiments are merely a few embodiments of the invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments of the present invention without making any creative effort, shall fall within the protection scope of the present invention.
A method for detecting the content of essential oil in a rose product by solid-phase microextraction is disclosed in the following embodiments.
Example 1
S1: preparing an internal standard solution. Transferring 100 mu L to 100mL volumetric flask of 2-nonanol, and preparing 823mg/L standard stock solution by using absolute ethyl alcohol to fix the volume. Transferring the standard stock solution into a volumetric flask with the volume of 2.5mL to 25mL, using absolute ethyl alcohol for constant volume, preparing an internal standard solution with the volume of 82.3mg/L, and storing the internal standard solution in a low-temperature dark place for later use.
S2: sample pretreatment, weighing 25g of fresh rose, extracting twice with 200mL of water in a 70 ℃ water bath kettle for 1h each time, and combining the two water extracts.
2mL of the water extract is transferred into a 20mL headspace bottle, 3g of NaCl is added, 10 mu L of internal standard solution is added, a magnetic stirring rotor is placed, and the mixture is sealed; stirring at 750r/min for 1 hr, inserting extraction head, placing fiber head in the upper space 20mm away from the sample surface, extracting at 50 deg.C in water bath for 30min, taking out handle, and directly performing the subsequent detection steps.
S3: and (3) carrying out chromatography-mass spectrometry on the pretreated rose water extract. The chromatographic conditions are, capillary chromatographic column: WAXETR30m × 0.25mm × 0.50 μm; column temperature procedure: the initial temperature is 50 ℃, the temperature is kept constant for 2min, the temperature is increased to 80 ℃ at the speed of 3 ℃/min, the temperature is increased to 115 ℃ at the speed of 10 ℃/min, the temperature is increased to 240 ℃ at the speed of 3 ℃/min, and the temperature is kept for 5 min; carrier gas, high-purity helium gas: the purity is more than or equal to 99.999 percent; flow rate: 1mL/min, no shunt sample introduction; sample inlet temperature: 230 ℃ to 230 ℃. Mass spectrometry conditions were ion source temperature: 230 ℃; transmission line temperature: 240 ℃; electron bombardment source: 70 eV; scanning range: 29 to 300 amu.
S4: according to the peak areas of various substances detected by the chromatographic-mass spectrometric analysis, and the concentration of four substances, namely citronellol, beta-phenethyl alcohol, geraniol and eugenol in the rose water extract is calculated to be M, mg/L by combining the concentration of the internal standard solution.
S5: according to the calculated M value, the concentration N, mg/L, N-M/0.92 of essential oil substances in the rose water extract is calculated. Wherein, 0.92 is the ratio of four substances of citronellol, beta-phenethyl alcohol, geraniol and eugenol in the known 20 essential oil substances in the rose product which is detected and calculated. Calculating the concentration of essential oil substance in rose, R, mg/g, R-M x (200/25) x 10-3÷0.92。
S6: calculating the relative concentration W,% per thousand of the total amount of essential oil in the rose water extract according to the calculated N value; w ÷ N ÷ 629180 × 1000. Wherein 629180mg/L is the content of known 20 effective substances in 100% pure essential oil.
Example 2
S1: preparing an internal standard solution. Transferring 100 mu L to 100mL volumetric flask of 2-nonanol, and preparing 823mg/L standard stock solution by using absolute ethyl alcohol to fix the volume. Transferring the standard stock solution into a volumetric flask with the volume of 2.5mL to 25mL, using absolute ethyl alcohol for constant volume, preparing an internal standard solution with the volume of 82.3mg/L, and storing the internal standard solution in a low-temperature dark place for later use.
S2: sample pretreatment, weighing 25g of fresh rose, extracting twice with 200mL of water in a 60 ℃ water bath kettle for 1h each time, and combining the two water extracts.
2mL of the water extract is transferred into a 20mL headspace bottle, 3g of NaCl is added, 10 mu L of internal standard solution is added, a magnetic stirring rotor is placed, and the mixture is sealed; stirring at 750r/min for 1 hr, inserting extraction head, placing fiber head in the upper space 20mm away from the sample surface, extracting at 50 deg.C in water bath for 30min, taking out handle, and directly performing the subsequent detection steps.
S3: and (3) carrying out chromatography-mass spectrometry on the pretreated rose water extract. The chromatographic conditions are, capillary chromatographic column: WAXETR30m × 0.25mm × 0.50 μm; column temperature procedure: the initial temperature is 50 ℃, the temperature is kept constant for 2min, the temperature is increased to 80 ℃ at the speed of 3 ℃/min, the temperature is increased to 115 ℃ at the speed of 10 ℃/min, the temperature is increased to 240 ℃ at the speed of 3 ℃/min, and the temperature is kept for 5 min; carrier gas, high-purity helium gas: the purity is more than or equal to 99.999 percent; flow rate: 1mL/min, no shunt sample introduction; sample inlet temperature: 230 ℃ to 230 ℃. Mass spectrometry conditions were ion source temperature: 230 ℃; transmission line temperature: 240 ℃; electron bombardment source: 70 eV; scanning range: 29 to 300 amu.
S4: according to the peak areas of various substances detected by the chromatographic-mass spectrometric analysis, and the concentration of four substances, namely citronellol, beta-phenethyl alcohol, geraniol and eugenol in the rose water extract is calculated to be M, mg/L by combining the concentration of the internal standard solution.
S5: according to the calculated M value, the concentration N, mg/L, N-M/0.92 of essential oil substances in the rose water extract is calculated. Wherein, 0.92 is the ratio of four substances of citronellol, beta-phenethyl alcohol, geraniol and eugenol in the known 20 essential oil substances which are detected and calculated. Calculating the concentration of essential oil substance in rose, R, mg/g, R-M x (200/25) x 10-3÷0.92。
S6: calculating the relative concentration W,% per thousand of the total amount of essential oil in the rose water extract according to the calculated N value; w ÷ N ÷ 629180 × 1000. Wherein 629180mg/L is the content of known 20 effective substances in 100% pure essential oil.
Example 3
S1: preparing an internal standard solution. Transferring 100 mu L to 100mL volumetric flask of 2-nonanol, and preparing 823mg/L standard stock solution by using absolute ethyl alcohol to fix the volume. Transferring the standard stock solution into a volumetric flask with the volume of 2.5mL to 25mL, using absolute ethyl alcohol for constant volume, preparing an internal standard solution with the volume of 82.3mg/L, and storing the internal standard solution in a low-temperature dark place for later use.
S2: sample pretreatment, weighing 25g of fresh rose, extracting twice with 200mL of water in a 70 ℃ water bath kettle for 1h each time, and combining the two water extracts.
2mL of the water extract is transferred into a 20mL headspace bottle, 3g of NaCl is added, 10 mu L of internal standard liquid and allyl caproate liquid are added, wherein the volume ratio of the allyl caproate to the rose water extract is 1:20, a magnetic stirring rotor is placed, and the magnetic stirring rotor is sealed; stirring at 750r/min for 1 hr, inserting extraction head, placing fiber head in the upper space 20mm away from the sample surface, extracting at 50 deg.C in water bath for 30min, taking out handle, and directly performing the subsequent detection steps.
S3: and (3) carrying out chromatography-mass spectrometry on the pretreated rose water extract. The chromatographic conditions are, capillary chromatographic column: WAXETR30m × 0.25mm × 0.50 μm; column temperature procedure: the initial temperature is 50 ℃, the temperature is kept constant for 2min, the temperature is increased to 80 ℃ at the speed of 3 ℃/min, the temperature is increased to 115 ℃ at the speed of 10 ℃/min, the temperature is increased to 240 ℃ at the speed of 3 ℃/min, and the temperature is kept for 5 min; carrier gas, high-purity helium gas: the purity is more than or equal to 99.999 percent; flow rate: 1mL/min, no shunt sample introduction; sample inlet temperature: 230 ℃ to 230 ℃. Mass spectrometry conditions were ion source temperature: 230 ℃; transmission line temperature: 240 ℃; electron bombardment source: 70 eV; scanning range: 29 to 300 amu.
S4: according to the peak areas of various substances detected by the chromatographic-mass spectrometric analysis, and the concentration of four substances, namely citronellol, beta-phenethyl alcohol, geraniol and eugenol in the rose water extract is calculated to be M, mg/L by combining the concentration of the internal standard solution.
S5: according to the calculated M value, the concentration N, mg/L, N-M/0.92 of essential oil substances in the rose water extract is calculated. WhereinAnd 0.92 is the ratio of four substances of citronellol, beta-phenethyl alcohol, geraniol and eugenol in the 20 known essential oil substances which are detected and calculated. Calculating the concentration of essential oil substance in rose, R, mg/g, R-M x (200/25) x 10-3÷0.92。
S6: calculating the relative concentration W,% per thousand of the total amount of essential oil in the rose water extract according to the calculated N value; w ÷ N ÷ 629180 × 1000. Wherein 629180mg/L is the content of known 20 effective substances in 100% pure essential oil.
Example 4
S1: preparing an internal standard solution. Transferring 100 mu L to 100mL volumetric flask of 2-nonanol, and preparing 823mg/L standard stock solution by using absolute ethyl alcohol to fix the volume. Transferring the standard stock solution into a volumetric flask with the volume of 2.5mL to 25mL, using absolute ethyl alcohol for constant volume, preparing an internal standard solution with the volume of 82.3mg/L, and storing the internal standard solution in a low-temperature dark place for later use.
S2: sample pretreatment, weighing 25g of fresh rose, extracting twice with 200mL of water in a 70 ℃ water bath kettle for 1h each time, and combining the two water extracts. 2g of magnesium formate was dissolved in 200mL of water for extraction.
2mL of the water extract is transferred into a 20mL headspace bottle, 3g of NaCl is added, 10 mu L of internal standard solution is added, a magnetic stirring rotor is placed, and the mixture is sealed; stirring at 750r/min for 1 hr, inserting extraction head, placing fiber head in the upper space 20mm away from the sample surface, extracting at 50 deg.C in water bath for 30min, taking out handle, and directly performing the subsequent detection steps.
S3: and (3) carrying out chromatography-mass spectrometry on the pretreated rose water extract. The chromatographic conditions are, capillary chromatographic column: WAXETR30m × 0.25mm × 0.50 μm; column temperature procedure: the initial temperature is 50 ℃, the temperature is kept constant for 2min, the temperature is increased to 80 ℃ at the speed of 3 ℃/min, the temperature is increased to 115 ℃ at the speed of 10 ℃/min, the temperature is increased to 240 ℃ at the speed of 3 ℃/min, and the temperature is kept for 5 min; carrier gas, high-purity helium gas: the purity is more than or equal to 99.999 percent; flow rate: 1mL/min, no shunt sample introduction; sample inlet temperature: 230 ℃ to 230 ℃. Mass spectrometry conditions were ion source temperature: 230 ℃; transmission line temperature: 240 ℃; electron bombardment source: 70 eV; scanning range: 29 to 300 amu.
S4: according to the peak areas of various substances detected by the chromatographic-mass spectrometric analysis, and the concentration of four substances, namely citronellol, beta-phenethyl alcohol, geraniol and eugenol in the rose water extract is calculated to be M, mg/L by combining the concentration of the internal standard solution.
S5: according to the calculated M value, the concentration N, mg/L, N-M/0.92 of essential oil substances in the rose water extract is calculated. Wherein, 0.92 is the ratio of four substances of citronellol, beta-phenethyl alcohol, geraniol and eugenol in the known 20 essential oil substances which are detected and calculated. Calculating the concentration of essential oil substance in rose, R, mg/g, R-M x (200/25) x 10-3÷0.92。
S6: calculating the relative concentration W,% per thousand of the total amount of essential oil in the rose water extract according to the calculated N value; w ÷ N ÷ 629180 × 1000. Wherein 629180mg/L is the content of known 20 effective substances in 100% pure essential oil.
Example 5
S1: preparing an internal standard solution. Transferring 100 mu L to 100mL volumetric flask of 2-nonanol, and preparing 823mg/L standard stock solution by using absolute ethyl alcohol to fix the volume. Transferring the standard stock solution into a volumetric flask with the volume of 2.5mL to 25mL, using absolute ethyl alcohol for constant volume, preparing an internal standard solution with the volume of 82.3mg/L, and storing the internal standard solution in a low-temperature dark place for later use.
S2: pretreating the sample, weighing 25g of fresh rose, extracting twice with 200mL of water in a water bath kettle at 80 ℃ for 40min each time, and combining the two water extracts. 2g of magnesium formate was dissolved in 200mL of water for extraction.
2mL of the water extract is transferred into a 20mL headspace bottle, 3g of NaCl is added, 10 mu L of internal standard solution is added, a magnetic stirring rotor is placed, and the mixture is sealed; stirring at 750r/min for 1 hr, inserting extraction head, placing fiber head in the upper space 20mm away from the sample surface, extracting at 50 deg.C in water bath for 30min, taking out handle, and directly performing the subsequent detection steps.
S3: and (3) carrying out chromatography-mass spectrometry on the pretreated rose water extract. The chromatographic conditions are, capillary chromatographic column: WAXETR30m × 0.25mm × 0.50 μm; column temperature procedure: the initial temperature is 50 ℃, the temperature is kept constant for 2min, the temperature is increased to 80 ℃ at the speed of 3 ℃/min, the temperature is increased to 115 ℃ at the speed of 10 ℃/min, the temperature is increased to 240 ℃ at the speed of 3 ℃/min, and the temperature is kept for 5 min; carrier gas, high-purity helium gas: the purity is more than or equal to 99.999 percent; flow rate: 1mL/min, no shunt sample introduction; sample inlet temperature: 230 ℃ to 230 ℃. Mass spectrometry conditions were ion source temperature: 230 ℃; transmission line temperature: 240 ℃; electron bombardment source: 70 eV; scanning range: 29 to 300 amu.
S4: according to the peak areas of various substances detected by the chromatographic-mass spectrometric analysis, and the concentration of four substances, namely citronellol, beta-phenethyl alcohol, geraniol and eugenol in the rose water extract is calculated to be M, mg/L by combining the concentration of the internal standard solution.
S5: according to the calculated M value, the concentration N, mg/L, N-M/0.92 of essential oil substances in the rose water extract is calculated. Wherein, 0.92 is the ratio of four substances of citronellol, beta-phenethyl alcohol, geraniol and eugenol in the known 20 essential oil substances which are detected and calculated. Calculating the concentration of essential oil substance in rose, R, mg/g, R-M x (200/25) x 10-3÷0.92。
S6: calculating the relative concentration W,% per thousand of the total amount of essential oil in the rose water extract according to the calculated N value; w ÷ N ÷ 629180 × 1000. Wherein 629180mg/L is the content of known 20 effective substances in 100% pure essential oil.
Example 6
S1: preparing an internal standard solution. Transferring 100 mu L to 100mL volumetric flask of 2-nonanol, and preparing 823mg/L standard stock solution by using absolute ethyl alcohol to fix the volume. Transferring the standard stock solution into a volumetric flask with the volume of 2.5mL to 25mL, using absolute ethyl alcohol for constant volume, preparing an internal standard solution with the volume of 82.3mg/L, and storing the internal standard solution in a low-temperature dark place for later use.
S2: sample pretreatment, weighing 25g of fresh rose, extracting twice with 200mL of water in a water bath kettle at 80 ℃ for 1h each time, and combining the two water extracts. 2g of magnesium formate was dissolved in 200mL of water for extraction.
2mL of the water extract is transferred into a 20mL headspace bottle, 3g of NaCl is added, 10 mu L of internal standard liquid and allyl caproate liquid are added, wherein the volume ratio of the allyl caproate to the rose water extract is 1:20, a magnetic stirring rotor is placed, and the magnetic stirring rotor is sealed; stirring at 750r/min for 1 hr, inserting extraction head, placing fiber head in the upper space 20mm away from the sample surface, extracting at 50 deg.C in water bath for 30min, taking out handle, and directly performing the subsequent detection steps.
S3: and (3) carrying out chromatography-mass spectrometry on the pretreated rose water extract. The chromatographic conditions are, capillary chromatographic column: WAXETR30m × 0.25mm × 0.50 μm; column temperature procedure: the initial temperature is 50 ℃, the temperature is kept constant for 2min, the temperature is increased to 80 ℃ at the speed of 3 ℃/min, the temperature is increased to 115 ℃ at the speed of 10 ℃/min, the temperature is increased to 240 ℃ at the speed of 3 ℃/min, and the temperature is kept for 5 min; carrier gas, high-purity helium gas: the purity is more than or equal to 99.999 percent; flow rate: 1mL/min, no shunt sample introduction; sample inlet temperature: 230 ℃ to 230 ℃. Mass spectrometry conditions were ion source temperature: 230 ℃; transmission line temperature: 240 ℃; electron bombardment source: 70 eV; scanning range: 29 to 300 amu.
S4: according to the peak areas of various substances detected by the chromatographic-mass spectrometric analysis, and the concentration of four substances, namely citronellol, beta-phenethyl alcohol, geraniol and eugenol in the rose water extract is calculated to be M, mg/L by combining the concentration of the internal standard solution.
S5: according to the calculated M value, the concentration N, mg/L, N-M/0.92 of essential oil substances in the rose water extract is calculated. Wherein, 0.92 is the ratio of four substances of citronellol, beta-phenethyl alcohol, geraniol and eugenol in the known 20 essential oil substances which are detected and calculated. Calculating the concentration of essential oil substance in rose, R, mg/g, R-M x (200/25) x 10-3÷0.92。
S6: calculating the relative concentration W,% per thousand of the total amount of essential oil in the rose water extract according to the calculated N value; w ÷ N ÷ 629180 × 1000. Wherein 629180mg/L is the content of known 20 effective substances in 100% pure essential oil.
Example 7
S1: preparing an internal standard solution. Transferring 100 mu L to 100mL volumetric flask of 2-nonanol, and preparing 823mg/L standard stock solution by using absolute ethyl alcohol to fix the volume. Transferring the standard stock solution into a volumetric flask with the volume of 2.5mL to 25mL, using absolute ethyl alcohol for constant volume, preparing an internal standard solution with the volume of 82.3mg/L, and storing the internal standard solution in a low-temperature dark place for later use.
S2: pretreating the sample, weighing 25g of fresh rose, extracting with 200mL of water twice in a water bath kettle at 80 ℃ for 30min each time, and combining the two water extracts. 2g of magnesium formate was dissolved in 200mL of water for extraction.
2mL of the water extract is transferred into a 20mL headspace bottle, 3g of NaCl is added, 10 mu L of internal standard liquid and allyl caproate liquid are added, wherein the volume ratio of the allyl caproate to the rose water extract is 1:20, a magnetic stirring rotor is placed, and the magnetic stirring rotor is sealed; stirring at 750r/min for 1 hr, inserting extraction head, placing fiber head in the upper space 20mm away from the sample surface, extracting at 50 deg.C in water bath for 30min, taking out handle, and directly performing the subsequent detection steps.
S3: and (3) carrying out chromatography-mass spectrometry on the pretreated rose water extract. The chromatographic conditions are, capillary chromatographic column: WAXETR30m × 0.25mm × 0.50 μm; column temperature procedure: the initial temperature is 50 ℃, the temperature is kept constant for 2min, the temperature is increased to 80 ℃ at the speed of 3 ℃/min, the temperature is increased to 115 ℃ at the speed of 10 ℃/min, the temperature is increased to 240 ℃ at the speed of 3 ℃/min, and the temperature is kept for 5 min; carrier gas, high-purity helium gas: the purity is more than or equal to 99.999 percent; flow rate: 1mL/min, no shunt sample introduction; sample inlet temperature: 230 ℃ to 230 ℃. Mass spectrometry conditions were ion source temperature: 230 ℃; transmission line temperature: 240 ℃; electron bombardment source: 70 eV; scanning range: 29 to 300 amu.
S4: according to the peak areas of various substances detected by the chromatographic-mass spectrometric analysis, and the concentration of four substances, namely citronellol, beta-phenethyl alcohol, geraniol and eugenol in the rose water extract is calculated to be M, mg/L by combining the concentration of the internal standard solution.
S5: according to the calculated M value, the concentration N, mg/L, N-M/0.92 of essential oil substances in the rose water extract is calculated. Wherein, 0.92 is the ratio of four substances of citronellol, beta-phenethyl alcohol, geraniol and eugenol in the known 20 essential oil substances which are detected and calculated. Calculating the concentration of essential oil substance in rose, R, mg/g, R-M x (200/25) x 10-3÷0.92。
S6: calculating the relative concentration W,% per thousand of the total amount of essential oil in the rose water extract according to the calculated N value; w ÷ N ÷ 629180 × 1000. Wherein 629180mg/L is the content of known 20 effective substances in 100% pure essential oil.
Detection method
1. The method comprises the steps of respectively taking nine rose beverages, namely, a Huasu drinking water nectarine flavor plant beverage, a Jiu rose beverage, a Huasu drinking grapefruit flavor plant beverage and a Jiu rose juice beverage, which are produced by Ningshi International Rose industries, Ltd, and fresh rose juice produced by Yunnan Kunming and Baoshijiang, performing chromatographic mass spectrometry, wherein the results show that the peak areas of four substances, namely citronellol, beta-phenethyl alcohol, geraniol and eugenol, are large, calculating the proportion of the four substances in 20 known essential oil substances in each sample respectively, and finally calculating the average value to be 0.92.
2. After the pure essential oil is diluted by absolute ethyl alcohol in a gradient way, the sample injection detection is directly carried out, and 20 known essential oil substances are determined: alpha-pinene; beta-pinene; methyl geranate; citronellyl acetate; alpha-terpineol; citronellol; nerol; geraniol; nerolidol; heptadecane; linalool; beta-phenylethyl alcohol; methyl eugenol; twenty three of ten; rose oxide; linalool; beta-myrcene; d-limonene; heneicosane; nonadecane. The measured value content of known 20 effective substances in 100% pure essential oil is 629180mg/L according to peak area calculation.
3.M＝82.3×5×10-3And (B/A) mg/L, wherein B is the sum of the peak areas of four substances, namely citronellol, beta-phenethyl alcohol, geraniol and eugenol, and A is the peak area of an internal standard substance.
The result of the detection
Experimental results show that the detection and calculation method can conveniently and quickly detect the content and the concentration of the essential oil in the roses. In example 3, compared with example 1, the peak area values of all the substances shown by the chromatographic-mass spectrometric detection result are changed, but the concentration values M of the four substances, namely citronellol, beta-phenylethyl alcohol, geraniol and eugenol in the final sample are not obviously changed, the addition of allyl caproate can promote the extraction effect when the sample is pretreated, and the improvement of the extraction effect shows that the sample pretreatment method can be applied to the sample with lower concentration, so that the application range of the method disclosed by the invention is expanded. Compared with example 1, the peak area of the substance has no obvious change when magnesium formate is added in water extraction of example 4, but compared with example 4, the water extraction time is shorter and the peak area of the substance has no obvious change in example 5, which shows that the magnesium formate can accelerate the release of essential oil in roses.
Claims (2)
1. A detection method for detecting the content of essential oil in a rose product by solid-phase microextraction is characterized by comprising the following steps:
s1: preparing an internal standard solution, wherein the internal standard solution is a 2-nonanol solution;
s2: pre-treating a sample, namely adding the sample into a headspace bottle, adding sodium chloride and an internal standard solution, adding allyl caproate, wherein the volume ratio of the allyl caproate to the rose water extract is 1:20, sealing, and magnetically stirring; inserting an extraction head, extracting under a water bath condition, and then analyzing;
s3: carrying out chromatography-mass spectrometry on the pretreated rose water extract under the following chromatographic conditions of a capillary chromatographic column: WAXETR30m × 0.25mm × 0.50 μm; column temperature procedure: the initial temperature is 50 ℃, the temperature is kept constant for 2min, the temperature is increased to 80 ℃ at the speed of 3 ℃/min, the temperature is increased to 115 ℃ at the speed of 10 ℃/min, the temperature is increased to 240 ℃ at the speed of 3 ℃/min, and the temperature is kept for 5 min; carrier gas, high-purity helium gas: the purity is more than or equal to 99.999 percent; flow rate: 1mL/min, no shunt sample introduction; sample inlet temperature: 230 ℃, mass spectrometry conditions are ion source temperature: 230 ℃; transmission line temperature: 240 ℃; electron bombardment source: 70 eV; scanning range: 29-300 amu;
s4: according to the result of the chromatographic-mass spectrometric analysis and in combination with the concentration of the internal standard solution, calculating the concentrations of citronellol, beta-phenethyl alcohol, geraniol and eugenol in the rose water extract to be M, mg/L;
s5: calculating the concentration N, mg/L, N-M/0.92 of essential oil substances in the rose water extract according to the calculated M value; calculating the concentration of essential oil substance in rose, R, mg/g, R-M x (200/25) x 10-3÷0.92；
S2, the water extraction process comprises the steps of putting a sample into a water bath kettle at the temperature of 60-70 ℃, and extracting twice;
during the water extraction process, magnesium formate is added to the solution.
2. The method for detecting the content of essential oil in rose products by solid-phase microextraction according to claim 1, which is characterized in that: adding S6 behind S5, and calculating the relative concentration W,% per thousand of the total amount of essential oil in the rose water extract according to the calculated N value; w ÷ N ÷ 629180 × 1000.
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