CN108519446A - A method of measuring Salanesol in tobacco - Google Patents
A method of measuring Salanesol in tobacco Download PDFInfo
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- CN108519446A CN108519446A CN201810146540.9A CN201810146540A CN108519446A CN 108519446 A CN108519446 A CN 108519446A CN 201810146540 A CN201810146540 A CN 201810146540A CN 108519446 A CN108519446 A CN 108519446A
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- 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
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- 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
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- 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
- G01N2030/062—Preparation extracting sample from raw material
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Abstract
The invention discloses a kind of methods of Salanesol in measurement tobacco, use following device：Sample extraction bottle, including：Outer tube (1)；Inner sleeve (2)；Catheter (4；Sieve plate (3) is located in the lower ending opening of the inner sleeve (2), for one or more；This approach includes the following steps：1. tobacco ultrasonic extraction；2. filtering and transfer；3. making chromatogram；4. the peak area with standard chromatogram, you can obtain the content of Salanesol in tobacco.The device that the present invention uses have the advantages that processing it is simple, conveniently, effect it is good.The assay method of the present invention has the advantages that quantitatively accurate, reproducible, detection limit is low etc., is suitable for the accurate test of batch samples.Effective technical support is provided for the safety supervision during Tobacco.
Description
Technical field
The invention belongs to technical field of analytical chemistry, and in particular to a method of measuring Salanesol in tobacco.
Background technology
Salanesol is one of main terpenoid ingredient in tobacco, is polyisoprene alcohols compound, is had anti-
A variety of physiological activity such as bacterium, anti-inflammatory, hemostasis.Meanwhile Salanesol or a kind of important medicine intermediate, it is in ubiquinone class drug
The irreplaceable ingredient of mesosome, and the side chain of synthesis vitamin K and Co-Q10 and synthesis anti-ulcer medicament, anticancer drug
Irreplaceable natural material has very high medical value.In tobacco, Salanesol is benzo in cigarette smoke [a] pyrene shape
At important as precursors compound, its content directly affects the burst size of benzo in cigarette smoke [a] pyrene, and then to cigarette cigarette
The safety of gas has a direct impact.Therefore, have to tobacco quality control to the Accurate Determining of Salanesol in tobacco very heavy
The meaning wanted.
According to the literature, it is high performance liquid chromatography to measure the most common method of Salanesol, including normal-phase chromatography and anti-
Two kinds of clastotypes of phase chromatography, detection method mainly have differential pulse polarograpll, ultraviolet detection and Evaporative light scattering detector etc..Mesh
Before, standard GB/T/T 31758-2015 are also using the eggplant Buddhist nun in Tobacco Leaves by High Performance Liquid Chromatography and leaf tobacco extract
Alcohol.But Traditional liquid phase chromatography can have many problems when analyzing Salanesol, such as：Chromatography overlong time, before sample
Processing need to through sodium hydroxide methanol solution saponification process, n-hexane extraction, water remove impurity, step is various, operation
Complexity trouble, and accuracy is poor.Therefore, current to be badly in need of exploitation one kind simply, quickly, accurate, reliable high throughput analysis side
Method is used to measure the Salanesol in tobacco.
Invention content
In view of the above-mentioned deficiencies in the prior art, it is an object of the present invention to provide a kind of method measuring Salanesol in tobacco, it is
Tobacco quality control provides technical support.
It is used cooperatively, tobacco sample can be carried out quick the present invention also aims to provide a kind of and above method
Saponification, extraction, dry filter and the device for directly collecting filtrate, to simplify sample pre-treatments step, the pre-treatment entirely detected
Process need not shift sample, improve analysis efficiency, can provide effective guarantee for the Accurate Determining of Salanesol in tobacco.
The purpose of the present invention is what is be achieved by the following technical programs.
A method of measuring Salanesol in tobacco, which is characterized in that use following device：
Sample extraction bottle, including：Outer tube 1, sealed bottom and to be flat, oral area has a sealing ring 11；Inner sleeve
2, outer diameter is adapted with the internal diameter of the outer tube 1, length be more than the outer tube 1 length；Catheter 4, is located at
In the inner sleeve 2, lower end has an extension mouth and the lower end inner wall of the inner sleeve 2 to be sealedly and fixedly connected, and described interior
2 upper port of casing is fixedly connected to form compression face 21, and upper part is stretched out outside the inner sleeve 2, and back bending is downward；Sieve plate 3, position
In in the lower ending opening of the inner sleeve 2, for one or more；
This approach includes the following steps：
1. crushing the tobacco sample of freeze-drying, 40 mesh sieve is crossed, 0.1~0.5g is weighed, pours into outer tube, the soap of 10mL is added
Change reagent, 30~50min of ultrasonic saponification in 50~70 DEG C of constant-temperature ultrasonic water-baths；
2. after saponification is complete, then it is separately added into 20mL n-hexanes and 5mL ultra-pure waters into the outer tube of step 1.,
5~15min of oscillation extraction stands 5~15min after solution layering, is inserted into inner sleeve and firmly pushes, make the n-hexane on upper layer
Extract liquor is entered into catheter in sample collection bottle by filtering sieve plate, is cooled to room temperature to obtain sample filtrate to be analyzed；
3. being analyzed 2. sample filtrate that step obtains using liquid chromatogram, quantitatively surveyed with evaporative light scattering detector
Determine Salanesol, makes the chromatogram of Salanesol in tobacco；
4. the peak of the peak area and standard solution chromatogram of Salanesol in the sample in 3. chromatogram that comparison step obtains
Area, you can obtain the Salanesol content in tobacco.
Preferably, the step potassium hydroxide solution that 1. saponification reagent is 0.6mol/L, solvent is ethyl alcohol.
Preferably, step discards the 2~3mL filtrates initially flowed out 2. in extract liquor filter process, collects 1~1.5mL filters
Liquid is as sample filtrate.
Preferably, 3. the liquid phase chromatogram condition is step：Chromatographic column：Waters Acquity UPLCTM BEH C18
(2.1 × 50mm, 1.7 μm) analytical column；Mobile phase：Volume ratio is 50:50 methanol:Acetonitrile；Type of elution：Isocratic elution；Stream
Speed：0.8mL/min；Sample size：5μL；Column temperature：30℃；Detector：Evaporative light scattering detector；Drift tube temperature：80℃；It carries
Gas：Nitrogen；Flow velocity：2.0mL/min.
Preferably, the sieve plate 3 is two, and aperture is 0.45 μm, and anhydrous sodium sulfate is filled between two sieve plates.
The internal diameter of the inner sleeve is preferably respectively 1~3cm, and length is 4~8cm.
The method of the present invention integrates extraction, filtering, transfer using sample extraction bottle, and sample extracts in outer tube 1
After；Inner sleeve 2 is firmly pushed at compression face 21, extract liquor enters after being filtered by sieve plate 3 in catheter 4, and from
The downward mouth of back bending of catheter 4 exits into sample bottle, obtains sample filtrate and carries out chromatography.The extraction of this process sample
It takes, filter, shifting and be integrated, avoiding experimental error caused by operation.
1 oral area of outer tube dress fixes inner sleeve 2 there are one sealing ring 11 when for extracting, with prevent extract liquor from splashing out with
And it is buffered when sample filtering.
In the present invention, 4 diameter of catheter is preferably 1-2mm.
Beneficial effects of the present invention are：
(1) device that uses of the present invention, need not shift in the pretreatment process of sample sample can directly extract, mistake
It filters and collects to obtain sample to be analysed, effectively prevent operating caused experimental error.Salanesol in tobacco of the present invention exists
It can be detached completely in 4.0min, compared with the analysis method that current GB/T 31758-2015 are used, analysis time contracting
Short by 50%, the method compared to report can be improved 5 times or more, be suitable for the accurate test of batch samples；With processing letter
It is single, conveniently, the good advantage of effect.
(2) assay method sample extraction effect and precision of the invention improve a lot than conventional method, analysis knot
Fruit is consistent with the result that standard method measures.Have the advantages that quantitatively accurate, reproducible, detection limit is low etc., is suitable for large quantities of
Measure the accurate test of sample.The assay method sample analysis efficiency improves a lot than conventional method, can be Salanesol in tobacco
Detection accurately and reliably method is provided, control of the effective guarantee to tobacco quality.
Description of the drawings
Fig. 1 is the outer tube schematic diagram of the present invention；
Fig. 2 is the inner sleeve schematic diagram of the present invention；
Inner sleeve is inserted into housing spout part schematic diagram when Fig. 3 is the extraction mixing of the present invention；
Inner sleeve is inserted into outer tube schematic internal view when Fig. 4 is the extract liquor filtering of the present invention, transfer；
Fig. 5 is that the sample extraction bottle of the present invention is placed on sample extraction frame and sample collection bottle is placed on sample receipts
Collect the schematic diagram of sample transfer step when on frame；
Fig. 6 is Salanesol standard solution chromatogram；
Fig. 7 is the chromatogram of Salanesol in 1 sample solution of embodiment.
Description of the drawings：1- outer tubes；2- inner sleeves；21- compression faces；3- sieve plates；4- catheters；5- sample extraction framves；51-
Sample extraction bottle putting hole；6- sample collection framves；61- sample bottle putting holes；62- waste collection bottle putting holes；610- samples are received
Collect bottle；620- waste collection bottles；11- sealing rings；12- samples.
Specific implementation mode
With reference to embodiment, the present invention is described in further detail.When to understand, embodiment is not to this hair
The restriction of bright protection domain.
Embodiment 1
Tobacco sample 1# is sampled, the method for measuring wherein Salanesol content includes the following steps：
1. sample pre-treatments：Tobacco sample 1# is sampled, after freeze-drying crushes, accurately weighs 0.1g tobacco samples
(being accurate to 0.1mg) after the potassium hydroxide-ethanol solution of 10mL 0.6mol/L is added, then, utilizes sealing ring in outer tube
Inner sleeve is stuck in the oral area of outer tube, the ultrasonic saponification 30min in 50 DEG C of constant-temperature ultrasonic water-baths.Wait for that saponification is complete
Afterwards, then it is separately added into 20mL n-hexanes and 5mL ultra-pure waters, oscillation extraction 5min stands 5min after solution layering.Firmly push
Inner sleeve, at this time upper layer n-hexane extract just by filter sieve plate enter in catheter, that is loaded between two layers of sieve plate is anhydrous
Sodium sulphate can absorb the moisture content in n-hexane extract, and from catheter thin bend pipe outflow, reach filtering and
The effect of dry n-hexane extract.The downward mouth of back bending from catheter collects extract liquor, and the 2mL initially flowed out, which is discarded, (to be collected
Into waste collection bottle 620), it is impossible to be used in then analysis directly collects 1.0mL with chromatography column feed materials bottle (sample collection bottle 610)
Sample filtrate, after being cooled to room temperature, for liquid-phase chromatographic analysis.
2. liquid-phase chromatographic analysis：Chromatographic column：Waters Acquity UPLCTMBEH C18 (2.1 × 50mm, 1.7 μm) points
Analyse column；Mobile phase：Methanol-acetonitrile (50:50, volume ratio)；Type of elution：Isocratic elution；Flow velocity：0.8mL/min；Sample size：5
μL；Column temperature：30℃；Detector：Evaporative light scattering detector；Drift tube temperature：80℃；Carrier gas：Nitrogen；Flow velocity：2.0mL/
min。
By detection, the content of Salanesol is 5.27mg/g in 1# tobacco samples.
Embodiment 2
The method for measuring Salanesol content in tobacco sample 2#, includes the following steps：
1. sample pre-treatments：Tobacco sample 2# is sampled, after freeze-drying crushes, accurately weighs 0.5g tobacco samples
(being accurate to 0.1mg) after the potassium hydroxide-ethanol solution of the 0.6mol/L of 10mL is added, then, utilizes sealing in outer tube
Inner sleeve is stuck in the oral area of outer tube by circle, the ultrasonic saponification 50min in 70 DEG C of constant-temperature ultrasonic water-baths.Wait for that saponification is complete
Quan Hou, then it is separately added into 20mL n-hexanes and 5mL ultra-pure waters, oscillation extraction 5min stands 5min after solution layering.Under firmly
Inner sleeve is pressed, upper layer n-hexane extract is just entered by filtering sieve plate in catheter at this time, the nothing loaded between two layers of sieve plate
Aqueous sodium persulfate can absorb the moisture content in n-hexane extract, and n-hexane extract is from the thin bend pipe stream in catheter
Go out, achievees the effect that filtering and dry n-hexane extract.The downward mouth of back bending from catheter collects extract liquor, initially flows out
2mL is discarded and (is collected into waste collection bottle 620), it is impossible to be used in then analysis uses chromatography column feed materials bottle (sample collection bottle 610) straight
The sample filtrate for receiving collection 1.0mL, after being cooled to room temperature, for liquid-phase chromatographic analysis.
2. liquid-phase chromatographic analysis：Chromatographic column：Waters Acquity UPLCTMBEH C18 (2.1 × 50mm, 1.7 μm) points
Analyse column；Mobile phase：Methanol-acetonitrile (50:50, volume ratio)；Type of elution：Isocratic elution；Flow velocity：0.8mL/min；Sample size：5
μL；Column temperature：30℃；Detector：Evaporative light scattering detector；Drift tube temperature：80℃；Carrier gas：Nitrogen；Flow velocity：2.0mL/
min。
By detection, the content of Salanesol is 8.74mg/g in 2# tobacco samples.
Embodiment 3
The method for measuring Salanesol content in tobacco sample 3#, includes the following steps：
1. sample pre-treatments：Tobacco sample 3# is sampled, after freeze-drying crushes, accurately weighs 0.25g tobacco samples
(being accurate to 0.1mg) after the potassium hydroxide-ethanol solution of the 0.6mol/L of 10mL is added, then, utilizes sealing in outer tube
Inner sleeve is stuck in the oral area of outer tube by circle, the ultrasonic saponification 35min in 60 DEG C of constant-temperature ultrasonic water-baths.Wait for that saponification is complete
Quan Hou, then it is separately added into 20mL n-hexanes and 5mL ultra-pure waters, oscillation extraction 10min stands 10min after solution layering.Firmly
Inner sleeve is pushed, upper layer n-hexane extract is just entered by filtering sieve plate in catheter at this time, is loaded between two layers of sieve plate
Anhydrous sodium sulfate can absorb the moisture content in n-hexane extract, and n-hexane extract is from the thin bend pipe stream in catheter
Go out, achievees the effect that filtering and dry n-hexane extract.The downward mouth of back bending from catheter collects extract liquor, initially flows out
2mL is discarded and (is collected into waste collection bottle 620), it is impossible to be used in then analysis uses chromatography column feed materials bottle (sample collection bottle 610) straight
The sample filtrate for receiving collection 1.0mL, after being cooled to room temperature, for liquid-phase chromatographic analysis.
2. liquid-phase chromatographic analysis：Chromatographic column：Waters Acquity UPLCTMBEH C18 (2.1 × 50mm, 1.7 μm) points
Analyse column；Mobile phase：Methanol-acetonitrile (50:50, volume ratio)；Type of elution：Isocratic elution；Flow velocity：0.8mL/min；Sample size：5
μL；Column temperature：30℃；Detector：Evaporative light scattering detector；Drift tube temperature：80℃；Carrier gas：Nitrogen；Flow velocity：2.0mL/
min。
By detection, the content of Salanesol is 3.90mg/g in 3# tobacco samples.
Claims (6)
1. a kind of method measuring Salanesol in tobacco, which is characterized in that use following device：
Sample extraction bottle, including：Outer tube (1), sealed bottom and to be flat, oral area has a sealing ring (11)；Inner sleeve
(2), outer diameter is adapted with the internal diameter of the outer tube (1), length be more than the outer tube (1) length；Catheter
(4), it is located in the inner sleeve (2), lower end has the lower end inner wall of an extension mouth and the inner sleeve (2) to seal fixed connect
It connecing, compression face (21) is fixedly connected to form with the inner sleeve (2) upper port, upper part stretches out inner sleeve (2) outside,
And back bending is downward；Sieve plate (3) is located in the lower ending opening of the inner sleeve (2), for one or more；
This approach includes the following steps：
1. weighing 0.1~0.5g of tobacco powder, pour into outer tube, the saponification reagent of 10mL is added, it is super in 50~70 DEG C of constant temperature
30~50min of ultrasonic saponification in sound wave water-bath；
2. 20mL n-hexanes and 5mL water are separately added into step outer tube 1. again, 5~15min of oscillation extraction, stand 5~
15min is inserted into inner sleeve and firmly pushes after solution layering, and the n-hexane extract on upper layer is made to pass through filtering sieve plate into drain
Pipe enters in sample collection bottle, is cooled to room temperature to obtain sample filtrate to be analyzed；
3. being analyzed 2. sample filtrate that step obtains using liquid chromatogram, eggplant is quantitative determined with evaporative light scattering detector
Buddhist nun's alcohol makes the chromatogram of Salanesol in tobacco；
4. the peak area of the peak area and standard solution chromatogram of Salanesol in the sample in 3. chromatogram that comparison step obtains,
It can be obtained the Salanesol content in tobacco.
2. according to the method described in claim 1, it is characterized in that, the step hydrogen-oxygen that 1. saponification reagent is 0.6mol/L
Change potassium solution, solvent is ethyl alcohol.
3. according to the method described in claim 1, it is characterized in that, step 2. in extract liquor filter process, discards initial outflow
2~3mL filtrates, collect 1~1.5mL filtrates as sample filtrate.
4. according to the method described in claim 1, it is characterized in that, step 3. the liquid phase chromatogram condition is：Chromatographic column：
Waters Acquity UPLCTMBEH C18 (2.1 × 50mm, 1.7 μm) analytical column；Mobile phase：Volume ratio is 50:50 first
Alcohol:Acetonitrile；Type of elution：Isocratic elution；Flow velocity：0.8mL/min；Sample size：5μL；Column temperature：30℃；Detector：Evaporative light dissipates
Penetrate detector；Drift tube temperature：80℃；Carrier gas：Nitrogen；Flow velocity：2.0mL/min.
5. according to the method described in claim 1, it is characterized in that, the sieve plate (3) is two, nothing is filled between two sieve plates
Aqueous sodium persulfate.
6. according to the method described in claim 5, it is characterized in that, the aperture of the sieve plate (3) is 0.45 μm, by polytetrafluoro
Vinyl material is made.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201810146540.9A CN108519446A (en) | 2018-02-12 | 2018-02-12 | A method of measuring Salanesol in tobacco |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810146540.9A CN108519446A (en) | 2018-02-12 | 2018-02-12 | A method of measuring Salanesol in tobacco |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109738553A (en) * | 2019-03-08 | 2019-05-10 | 云南中烟工业有限责任公司 | A kind of pre-treating method for the measurement simultaneously of free, reference state sterol in tobacco |
| CN112147256A (en) * | 2020-09-25 | 2020-12-29 | 上海普康药业有限公司 | Coenzyme Q determination based on LC-MS10Method for preparing solanesol chloride in capsule |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103399102A (en) * | 2013-07-08 | 2013-11-20 | 中国农业科学院烟草研究所 | Method for determining total solanesol in tobaccos and tobacco products |
| CN105784455A (en) * | 2016-05-16 | 2016-07-20 | 张起香 | Extraction structure and liquid pretreatment device |
-
2018
- 2018-02-12 CN CN201810146540.9A patent/CN108519446A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103399102A (en) * | 2013-07-08 | 2013-11-20 | 中国农业科学院烟草研究所 | Method for determining total solanesol in tobaccos and tobacco products |
| CN105784455A (en) * | 2016-05-16 | 2016-07-20 | 张起香 | Extraction structure and liquid pretreatment device |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109738553A (en) * | 2019-03-08 | 2019-05-10 | 云南中烟工业有限责任公司 | A kind of pre-treating method for the measurement simultaneously of free, reference state sterol in tobacco |
| CN112147256A (en) * | 2020-09-25 | 2020-12-29 | 上海普康药业有限公司 | Coenzyme Q determination based on LC-MS10Method for preparing solanesol chloride in capsule |
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