CN109725098B - Method for measuring rhynchophylline transfer rate and rhynchophylline retention rate of filter tip of smoke - Google Patents
Method for measuring rhynchophylline transfer rate and rhynchophylline retention rate of filter tip of smoke Download PDFInfo
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- CN109725098B CN109725098B CN201910165684.3A CN201910165684A CN109725098B CN 109725098 B CN109725098 B CN 109725098B CN 201910165684 A CN201910165684 A CN 201910165684A CN 109725098 B CN109725098 B CN 109725098B
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Abstract
The invention discloses a method for measuring rhynchophylline transfer rate and filter tip rhynchophylline retention rate, which comprises the steps of carrying out balance treatment on a cigarette containing rhynchophylline feed liquid, then respectively extracting tobacco shreds, filter discs and a cigarette holder to obtain extract, measuring rhynchophylline in cigarette smoke by adopting a gas chromatography-mass spectrometry ion selection mode, improving the sensitivity and accuracy of detection, calculating the transfer rate and the retention rate of rhynchophylline in the cigarette smoke by using the area ratio of the rhynchophylline in the filter discs and the filter tips to the area ratio of the rhynchophylline in the cigarette tobacco shreds, providing an important reference basis for the efficiency research of novel natural cigarette perfume in the cigarette, and having important significance for the research and development of functional or medicinal cigarettes. Meanwhile, a reference method is provided for quantitative evaluation of other functional cigarettes.
Description
Technical Field
The invention belongs to the technical field of chemical analysis and detection, and particularly relates to a method for measuring a rhynchophylline transfer rate of smoke and a rhynchophylline retention rate of a filter tip.
Background
The rhynchophylline is alkaloid extracted from hooked branches of uncaria rhynchophylla of Rubiaceae, is one of main components of effective components of plants in uncaria of traditional Chinese medicine, has definite biological activity on human cardiovascular system, central nervous system, blood system and the like, and is a common medicine source of a prescription for treating hypertension. In recent years, with the gradually known nutritive value and medicinal function of uncaria rhynchophylla, the uncaria rhynchophylla is applied to traditional Chinese medicines, but the uncaria rhynchophylla is relatively little in the research aspect of cigarettes, and people like zhangyin in 2011 have studied the analysis of components of uncaria rhynchophylla ultrasonic extract and the application of uncaria rhynchophylla ultrasonic extract in cigarettes, while the research on the uncaria rhynchophylla alkali transfer rate in smoke is reported.
Disclosure of Invention
The invention aims to provide a method for measuring the rhynchophylline transfer rate of smoke and the rhynchophylline retention rate of a filter tip, so as to overcome the defects of the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for measuring the rhynchophylline transfer rate and the filter tip rhynchophylline retention rate of smoke comprises the following steps:
step 1), balancing the cigarettes added with the rhynchophylline feed liquid for at least 48 hours at 21-23 ℃ and RH in an environment of 58-62%, and then equally dividing the cigarettes into two groups;
step 2), stripping cut tobacco in cigarettes from the first group of cigarettes subjected to balance treatment, removing filtrate after extracting alkane compounds from the stripped cut tobacco by using petroleum ether, adding an ammonia water solution into the cut tobacco subjected to alkane compound extraction to completely infiltrate the cut tobacco, adding a dichloromethane solution to shake and extract the cut tobacco for not less than 1 hour to obtain an extract, and performing GC/MS analysis on the extract to obtain the area of rhynchophylline in the cut tobacco of the cigarettes;
step 3), smoking the second group of cigarettes after the balance treatment, filtering the smoking smoke by using a smoke trapping filter disc, then respectively performing oscillation extraction on the cigarette filter tip and the smoke trapping filter disc after smoke filtration in methanol for at least 1 hour to obtain extract liquor, and then respectively performing GC/MS analysis on the extract liquor to respectively obtain the area of rhynchophylline in the filter tip and the area of rhynchophylline in the filter disc;
and 4) adopting an area calculation method for the rhynchophylline transfer rate in the smoke, namely the rhynchophylline transfer rate in the cigarette smoke is equal to the area of rhynchophylline in the filter disc divided by the area of rhynchophylline in the cigarette tobacco shreds, and the rhynchophylline transfer rate in the cigarette filter is equal to the area of rhynchophylline in the filter tip divided by the area of rhynchophylline in the cigarette tobacco shreds.
Furthermore, the concentration of the ammonia water solution is 5-10%.
Further, the specific process of GC/MS analysis is as follows: after a sample to be detected is injected by an automatic injector of a 7890B gas chromatograph, the sample is instantly gasified at the temperature of a sample injection port of 280 ℃, then the gasified gas is carried into the gas chromatograph-mass spectrometer by carrier gas, and separation and detection are carried out by utilizing a capillary chromatographic column in the gas chromatograph-mass spectrometer, so that a rhynchophylline component content result can be obtained.
Further, the gas chromatography-mass spectrometer (GC/MS) employs 7890B-5977A gas chromatography-mass spectrometer.
Further, the capillary chromatographic column adopts HP-5MS capillary column.
Further, the carrier gas in the GC/MS analysis is He, and the carrier gas flow rate is as follows: 1.2 mL/min.
Further, the initial temperature of the gas chromatograph column oven is increased from 80 ℃ to 290 ℃ at a heating rate of 5 ℃/min, and is kept at 290 ℃ for 30 min.
Further, the analysis conditions by GC/MS are as follows: transmission line temperature: 280 ℃; an ionization mode: an EI source; ionization energy: 70 eV; ion source temperature: 230 ℃; quadrupole temperature: 150 ℃; solvent retardation: 3.5 min; scanning range: 35-500 amu; the detection mode is as follows: qualitative in full scan mode and quantitative in SIM mode; characteristic ion (m/z) of rhynchophylline: 384.
compared with the prior art, the invention has the following beneficial technical effects:
the method comprises the steps of carrying out balance treatment on a cigarette containing rhynchophylline feed liquid, extracting tobacco shreds, a filter disc and a cigarette holder respectively to obtain extract liquid, adding a certain amount of dilute alkali solution to enable rhynchophylline to be dissociated, wherein the free alkaloid is easier to extract than combined alkaloid, the extraction effect is improved, then determining the rhynchophylline area in the extract liquid by adopting a gas chromatography-mass spectrometry selected ion mode, the detection sensitivity and accuracy are improved, the transfer rate and the interception rate of the rhynchophylline in the cigarette smoke are calculated according to the area ratio of the rhynchophylline area in the tobacco shreds of the cigarette occupied by the rhynchophylline area in the filter disc and the filter holder, the algorithm is simple, the accuracy is high, an important reference basis is provided for the efficacy research of novel natural cigarette spice in the cigarette, and the method is significant for the research and development of functional or medicinal cigarettes, the rhynchophylline is added into the tobacco shreds, and the cigarette is transferred into cigarette smoke and trapped in a cigarette filter after being combusted, so that the effect of the rhynchophylline in sensory evaluation of cigarettes is specifically and quantitatively evaluated, and a reference method is provided for quantitative evaluation of other functional cigarettes.
Drawings
FIG. 1 is a SIM diagram of the ion scanning of rhynchophylline in cut tobacco of cigarette.
Figure 2 is a SIM of rhynchophylline in a cigarette smoke capture filter and cigarette filter.
FIG. 3 is a SIM diagram of rhynchophylline in cigarettes of different quantities according to an embodiment.
Detailed Description
The invention is described in further detail below with reference to the accompanying drawings:
the invention aims to provide a method for measuring the rhynchophylline transfer rate and the rhynchophylline rejection rate of a filter tip, which adopts gas chromatography-mass spectrometry and selects an ion scanning detection method to improve the detection sensitivity and accuracy of rhynchophylline, so that the calculation of the transfer rate and rejection rate of rhynchophylline is simpler and more convenient, and the rhynchophylline is quantitatively evaluated in cigarette smoke.
A method for measuring the rhynchophylline transfer rate and the filter tip rhynchophylline retention rate of smoke comprises the following steps:
step 1), balancing the cigarettes added with the rhynchophylline feed liquid for at least 48 hours at 21-23 ℃ and RH in an environment of 58-62%, and then equally dividing the cigarettes into two groups to keep the moisture of the cigarettes consistent;
step 2), stripping cut tobacco in cigarettes from the first group of cigarettes after the balance treatment, extracting alkane compounds from the stripped cut tobacco by using petroleum ether, removing filtrate, adding an ammonia water solution to completely soak the cut tobacco, extracting the cut tobacco by using a dichloromethane solution in a shaking way for not less than 1 hour, performing GC/MS analysis on the extract, and analyzing the amount of rhynchophylline in the cut tobacco of the cigarettes to obtain the cigarette;
adopting ammonia water solution with the concentration of 5-10%;
step 3), smoking the second group of cigarettes after the balance treatment, filtering the smoking smoke by using a smoke trapping filter disc, then respectively shaking and extracting the smoke trapping filter disc and the cigarette filter tip after the smoke is filtered in methanol for at least 1 hour, then respectively carrying out GC/MS analysis on the extract liquor to respectively obtain the area of rhynchophylline in the filter disc and the area of rhynchophylline in the filter tip, and detecting the transfer rate of rhynchophylline in the total particulate matters of the cigarette smoke;
and 4) adopting an area calculation method for the rhynchophylline transfer rate in the smoke, namely the rhynchophylline transfer rate in the cigarette smoke is equal to the area of rhynchophylline in the filter disc divided by the area of rhynchophylline in the cigarette tobacco shreds, and the rhynchophylline transfer rate in the cigarette filter is equal to the area of rhynchophylline in the filter tip divided by the area of rhynchophylline in the cigarette tobacco shreds.
The specific process of GC/MS analysis is as follows: after a sample to be detected is injected by an automatic injector of a 7890B gas chromatograph, the sample is instantly gasified at the temperature of a sample injection port of 280 ℃, then the gasified gas is carried into the gas chromatograph-mass spectrometer by carrier gas, and separation and detection are carried out by utilizing a capillary chromatographic column in the gas chromatograph-mass spectrometer, so that a rhynchophylline component content result can be obtained.
The gas chromatography-mass spectrometer (GC/MS) adopts a 7890B-5977A gas chromatography-mass spectrometer;
the capillary chromatographic column adopts HP-5MS capillary column;
analysis conditions were performed by GC/MS: a chromatographic column: HP-5MS capillary column (30 m.times.0.25 mm.times.0.25 μm); the carrier gas is He; carrier gas flow: 1.2 mL/min; sample inlet temperature: 280 ℃; the split ratio is as follows: 10: 1; sample introduction amount: 1 mu L of the solution; temperature programming: the initial temperature is increased from 80 to 290 ℃ at the heating rate of 5 ℃/min, and is kept at 290 ℃ for 30 min; transmission line temperature: 280 ℃; an ionization mode: an EI source; ionization energy: 70 eV; ion source temperature: 230 ℃; quadrupole temperature: 150 ℃; solvent retardation: 3.5 min; scanning range: 35-500 amu; the detection mode is as follows: qualitative in full scan mode and quantitative in SIM mode; characteristic ion (m/z) of rhynchophylline: 384.
two organic solvents of methanol and dichloromethane are selected in experiments to respectively extract rhynchophylline in the cigarette tobacco shreds, the smoke trapping filter disc and the cigarette filter tip, and experimental results show that when the filter tip is extracted by the dichloromethane, the filter tip is dissolved and is not suitable for extracting the rhynchophylline in the filter tip, so that the cigarette tobacco shreds are finally selected to be extracted by the dichloromethane, and the filter disc and the filter tip are extracted by the methanol.
The experiment compares the ultrasonic extraction mode and the oscillation extraction mode, and the result shows that the water temperature can be increased along with the extraction time in the ultrasonic extraction process to influence the analysis result, so the extraction mode is selected as the oscillation extraction mode to ensure the consistency of the test conditions.
In order to enable rhynchophylline to be better dissociated, in the extraction process of rhynchophylline in cigarette tobacco shreds, a test is compared with a test without adding ammonia water, and the result shows that rhynchophylline is not detected in a cigarette tobacco shred extract without adding ammonia water, and rhynchophylline is detected in a cigarette tobacco shred extract with a certain amount of 5% ammonia water, so that a proper amount of 5% ammonia water is selectively added in the treatment of the cigarette tobacco shreds, rhynchophylline belongs to alkaloid, a certain amount of dilute alkali solution is added, rhynchophylline can be dissociated, the dissociated alkaloid is easier to extract than combined alkaloid, and the extraction effect is improved;
in the specific implementation process: shaking and extracting the sucked cigarette smoke trapping filter disc for 1 hour by 50mL of methanol, and concentrating the extract liquid to 0.5mL for GC/MS analysis; adding 50mL of methanol into the sucked cigarette filter, shaking and extracting for 1 hour, concentrating the extract liquid to 0.5mL, and then carrying out GC/MS analysis;
experiments investigate the content determination conditions of rhynchophylline in the cut tobacco of 5, 10 and 20 cigarettes, the smoke trapping filter sheet and the cigarette filter, and test results show that the rhynchophylline extraction effect of 20 cigarettes is good, as shown in fig. 1 to 3.
Table 1 comparison of extraction effects of rhynchophylline in different quantities of cigarette cigarettes, the area refers to the area of 384 peaks of rhynchophylline characteristic ions:
calculation of Uncariaine transfer Rate
The test adopts an area calculation method for the rhynchophylline transfer rate, namely the rhynchophylline transfer rate in cigarette smoke is equal to the area of rhynchophylline in the filter disc divided by the area of rhynchophylline in cigarette tobacco shreds, and the rhynchophylline transfer rate in the cigarette filter is equal to the area of rhynchophylline in the filter tip divided by the area of rhynchophylline in the cigarette tobacco shreds.
The transfer rate of rhynchophylline in smoke is equal to the area of rhynchophylline in filter sheet/area of rhynchophylline in cigarette tobacco shred
The entrapment rate of the rhynchophylline in the filter tip is equal to the area of the rhynchophylline in the filter tip/the area of the rhynchophylline in the cigarette cut tobacco
Determination results of rhynchophylline transfer rate and interception rate in cigarette smoke
TABLE 2 transfer and retention of rhynchophylline in cigarette smoke
| Median area of tobacco | Area in filter disc | Area in filter tip | Transfer Rate (%) | Retention (%) |
| 187941 | 11507 | 10200 | 6.12 | 5.43 |
The method is characterized in that the transfer rate and the retention rate of the rhynchophylline in the cigarette smoke are calculated by measuring the rhynchophylline in the cigarette tobacco shreds, the rhynchophylline in the smoke trapping filter disc and the rhynchophylline in the cigarette filter tip by using an area calculation method, has certain innovativeness, provides an important reference basis for the efficacy research of novel natural cigarette flavors in cigarettes, and has important significance for the research and development of functional or medicinal cigarettes.
Claims (4)
1. A method for measuring the rhynchophylline transfer rate and the filter tip rhynchophylline rejection rate of smoke is characterized by comprising the following steps:
step 1), balancing the cigarettes added with the rhynchophylline feed liquid for at least 48 hours at 21-23 ℃ and RH in an environment of 58-62%, and then equally dividing the cigarettes into two groups;
step 2), stripping cut tobacco in cigarettes from the first group of cigarettes subjected to balance treatment, removing filtrate after extracting alkane compounds from the stripped cut tobacco by using petroleum ether, adding an ammonia water solution into the cut tobacco subjected to alkane compound extraction to completely infiltrate the cut tobacco, adding a dichloromethane solution to shake and extract the cut tobacco for not less than 1 hour to obtain an extract, and performing GC/MS analysis on the extract to obtain the area of rhynchophylline in the cut tobacco of the cigarettes; adopting ammonia water solution with the concentration of 5-10%;
step 3), smoking the second group of cigarettes after the balance treatment, filtering the smoking smoke by using a smoke trapping filter disc, then respectively performing oscillation extraction on the cigarette filter tip and the smoke trapping filter disc after smoke filtration in methanol for at least 1 hour to obtain extract liquor, and then respectively performing GC/MS analysis on the extract liquor to respectively obtain the area of rhynchophylline in the filter tip and the area of rhynchophylline in the filter disc; the specific process of GC/MS analysis is as follows: after a sample to be detected is injected by an automatic injector of a 7890B gas chromatograph, instantly gasifying the sample at the temperature of a 280 ℃ injection port, then introducing gasified gas into the gas chromatograph-mass spectrometer through carrier gas, and performing separation detection by using a capillary chromatographic column in the gas chromatograph-mass spectrometer to obtain a rhynchophylline component content result; the capillary chromatographic column adopts HP-5MS capillary column; the initial temperature of the column oven of the gas chromatograph is increased to 290 ℃ from 80 ℃ at the heating rate of 5 ℃/min, and is kept at 290 ℃ for 30 min;
and 4) adopting an area calculation method for the rhynchophylline transfer rate in the smoke, namely the rhynchophylline transfer rate in the cigarette smoke is equal to the area of rhynchophylline in the filter disc divided by the area of rhynchophylline in the cigarette tobacco shreds, and the rhynchophylline transfer rate in the cigarette filter is equal to the area of rhynchophylline in the filter tip divided by the area of rhynchophylline in the cigarette tobacco shreds.
2. The method for measuring the rhynchophylline transfer rate and the rhynchophylline rejection rate of the filter tip according to claim 1, wherein a 7890B-5977A gas chromatograph-mass spectrometer is adopted by the gas chromatograph-mass spectrometer.
3. The method for measuring the rhynchophylline transfer rate and the rhynchophylline retention rate of the filter tip according to claim 1, wherein a carrier gas in GC/MS analysis is He, and the carrier gas flow rate is as follows: 1.2 mL/min.
4. The method for measuring the rhynchophylline transfer rate and the rhynchophylline rejection rate of the filter tip according to claim 1, wherein the GC/MS analysis conditions are as follows: transmission line temperature: 280 ℃; an ionization mode: an EI source; ionization energy: 70 eV; ion source temperature: 230 ℃; quadrupole temperature: 150 ℃; solvent retardation: 3.5 min; scanning range: 35-500 amu; characteristic ion m/z of rhynchophylline: 384.
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