CN104950051A - Method for detecting isothiazolinone compounds in water-based glue for cigarettes - Google Patents

Abstract

The invention relates to a method for detecting isothiazolinone compounds in water-based glue for cigarettes. The method comprises the following steps: (1) adding an extracting agent and an internal standard solution into a sample of the water-based glue for cigarettes, carrying out ultrasonic extraction and centrifuging to obtain liquid supernatant; (2) filtering the liquid supernatant, adding a derivatization reagent to carry out a derivatization reaction, and after the reaction is finished, cooling to room temperature; and (3) carrying out quantitative analysis on the isothiazolinone compounds in a reaction solution of the step (2) by using a GC-MS (Gas Chromatography-Mass Spectrometer). The method provided by the invention can be used for simultaneously detecting the five isothiazolinone compounds in the water-based glue for cigarettes, is rapid and accurate, and can provide technical supports to the quality safety of cigarette products.

Description

A kind of method detecting OIT compounds in smoke aqueous gel

Technical field

The present invention relates to a kind of method detecting OIT compounds (MI, CMI, BIT, OI, DCOI) in smoke aqueous gel, belong to isothiazolinone compound test technical field.

Background technology

Smoke aqueous gel is one of material required in production of cigarettes, as glue for connecting, overlapping glue, and packaging glue etc.Because this material water dissolution properties is stronger; producing and depositing very easily rotten because of bacterium in process change; thus cause product quality inferior; unnecessary loss is brought directly to enterprise; therefore smoke aqueous gel usually can add sterilization antiseptic to control microbial growth breeding or to kill bacterium, in case the corruption of organic matter in production and process.Isothiazolinone (Isothiazolones) compound is widely used in smoke aqueous gel because having good cost performance.

But research finds, as being exposed to high concentration OIT compounds for a long time, may there is healthy hidden danger, as methylisothiazolinone has certain cytotoxicity and neurotoxicity, can produce the situations such as allergic reaction with skin contact.Therefore, suck security in order to what improve cigarette product, need the strict content controlling OIT compounds in smoke aqueous gel.Based on this, the detection of OIT compounds in smoke aqueous gel is also become particularly necessary.

At present, the detection method of OIT compounds mainly contains liquid phase chromatography, Liquid Chromatography-Tandem Mass Spectrometry, gas chromatography-mass spectrography etc., is widely used in the fields such as cosmetics, environmental water sample, toy for children, paper food wrappage.But due to the singularity of smoke aqueous gel sample, directly adopt said method testing result error larger.And the analysis and research that also there is not yet for OIT compounds in smoke aqueous gel are at present reported.

Summary of the invention

The object of the invention is to propose a kind of special detection method for OIT compounds in smoke aqueous gel, by carrying out pre-service to smoke aqueous gel sample, particularly to the optimization of derivatization reagent, again in conjunction with gas chromatography-mass spectrometry instrument, can the 5 kinds of industrial conventional OIT compounds in smoke aqueous gel be detected.The method of the invention is quick, accurate, for cigarette product quality safety provides technical support.

For achieving the above object, the present invention adopts following scheme to realize:

Detect a method for OIT compounds in smoke aqueous gel, comprise the steps:

1) in smoke aqueous gel sample, extractant and inner mark solution is added, ultrasonic extraction, centrifugal, obtain supernatant;

2) by supernatant liquid filtering, add derivatization reagent and carry out derivative reaction, after reaction terminates, be cooled to room temperature;

3) GC-MS is utilized to step 2) OIT compounds carries out quantitative test in gained reactant liquor.

The present invention by extracting and derivatization treatment smoke aqueous gel sample, more fast, accurately detects at least five kinds of OIT compounds (MI, CMI, BIT, OI, DCOI) in water base adhesive in conjunction with GC-MS ion mode.

In detection method of the present invention, step 1) in, described extraction solvent is selected from the one in methyl alcohol, normal hexane, methylene chloride, methyl tert-butyl ether, ethyl acetate, preferred methylene chloride.Described extractant is 5-15mL:0.2g, preferred 10mL:0.2g with the volume mass ratio of smoke aqueous gel sample.

In detection method of the present invention, step 1) in, described inner mark solution is 5-chloro-2-methyl-3-isothiazolone-d3 (CMI-d3) (No. CAS is 1329611-34-8) solution, and concentration is 0.2mg/mL; Described inner mark solution is 50-100 μ L:0.2g, preferably 80 μ L:0.2g with the volume mass ratio of smoke aqueous gel sample.

In detection method of the present invention, step 1) in, the described ultrasonic extraction time is 5-45min, preferred 30min.

In detection method of the present invention, step 2) in, described filtration employing 0.45 μm of organic phase filter membrane.

In detection method of the present invention, step 2) in, described derivatization reagent is two (TMS) trifluoroacetamide (BSTFA) of N-methyl-N-(trimethyl silane) trifluoroacetamide (MSTFA) or N, O-; Described derivatization reagent is 10-60 μ L:0.2g, preferably 30 μ L:0.2g with the volume mass ratio of smoke aqueous gel sample.

In detection method of the present invention, step 2) in, described derivative reaction condition: temperature 40-80 DEG C, time 10-50min, preferable temperature 60 DEG C.

In detection method of the present invention, the design parameter condition of described GC-MS, comprise chromatographic column kind, the conventional parameter condition of detection OIT compounds that sample introduction pattern, injector temperature, heating schedule are grasped for those skilled in the art, the present invention repeats no more this.

In the present invention, selected chromatographic condition is as follows: chromatographic column: chromatographic column is selected from the one in nonpolar HP-5MS (30m × 0.25mm × 0.25 μm), the DB-624 (30m × 0.25mm × 1.4 μm) of middle polarity and the HP-INNOWAX (30m × 0.25mm × 0.25 μm) of strong polarity; Injector temperature: 250 DEG C; Carrier gas: high-purity helium, purity >=99.999%; Sample size: 2 μ L; Flow velocity: 1.2mL/min (constant current mode); Split ratio: 5:1; Heating schedule: initial temperature 80 DEG C keeps 2min, rises to 220 DEG C, then rise to 240 DEG C with the programming rate of 20 DEG C/min with the programming rate of 10 DEG C/min, keeps 15min.Mass Spectrometry Conditions: transmission line temperature: 240 DEG C; Ion source temperature 230 DEG C; Solvent delay: 6min; Ionization mode: EI source; Ionizing energy: 70eV; Scan mode: Salbutamol Selected Ion Monitoring (SIM).

Retention time is compared in the present invention's employing, standard specimen addition method is qualitative.The retention time of 5 kinds of OIT compounds (MI, CMI, BIT, OI, DCOI) and internal standard compound matter and characteristic ion are in table 1.

Table 1 retention time and characteristic ion

Note: 1.-TMS is trimethyl silicone hydride product.

As the preferred embodiment of the present invention, adopt methylene chloride configuration standard solution (the OIT mixed standard solution of mixing OIT Standard Reserving Solution, the preparation of interior mark stock solution, variable concentrations gradient):

1) OIT Standard Reserving Solution is mixed: take 2-methyl-3 (2H)-OIT (MI), 5-chloro-2-methyl-2H-isothiazole-3-ketone (CMI), 1 respectively, 2-benzisothiazole base-3 (2H)-one (BIT), 2-octyl group-4-isothiazoline-3-ketone (OI), 4, the OIT standard specimen of the chloro-2-n-octyl of 5-bis--3-OIT (DCOI) 5 kinds of different qualities, in table 2, be accurate to 0.0001g, be placed in 50mL volumetric flask, add methylene chloride constant volume, shake up, obtain mixing OIT Standard Reserving Solution.

Table 2 mixes compound concentration in OIT Standard Reserving Solution

Sequence number	Compound title	Take quality (g)	Stock solution concentration (mg/mL)
				1	MI	0.1000	2.00
2	CMI	0.0250	0.50
				3	BIT	0.1000	2.00
4	OI	0.0807	1.61
				5	DCOI	0.0515	1.03

2) mark stock solution (inner mark solution) preparation in: accurately take mark 5-chloro-2-methyl-2H-isothiazole-3-ketone (CMI)-d3 in 0.005g, be accurate to 0.0001g, be placed in the volumetric flask of 25mL, add methylene chloride constant volume, obtain interior mark stock solution.

3) the OIT mixed standard solution of variable concentrations gradient: accurately pipette 5,10,20,50,100,200 μ L mixing OIT Standard Reserving Solution in 6 25mL volumetric flasks, add mark standard solution in 200 μ L respectively, use methylene chloride constant volume, shake up, obtain the OIT mixed standard solution of variable concentrations gradient, wherein, mark concentration is 1.6 μ g/mL.

Respectively the analysis of GC/MS choice ion pattern is carried out to the OIT mixed standard solution of 6 variable concentrations gradients, as shown in Figure 1 and Figure 2, with the peak area ratio (Y) of OIT compounds and internal standard compound to corresponding isothiazolinone compound concentration (X, μ g/mL) carry out linear regression analysis, obtain working curve regression equation and the related coefficient of 5 kinds of OIT compounds.By dilution least concentration mixed standard solution, OIT compounds produces and is not less than detection limit (LOD) and the quantitative limit (LOQ) that 3 times of signal to noise ratio (S/N ratio)s (S/N) and the analyte concentration corresponding to 10 times of signal to noise ratio (S/N ratio)s (S/N) count method respectively on instrument.Result is as shown in table 3.

The working curve of table 3 OIT compounds, related coefficient, detectability and quantitative limit

Adopt the recovery of Standard Addition Method for Determination detection method of the present invention, take a certain amount of smoke aqueous gel sample having detected 5 kinds of OIT content, divide 3 parts, above-mentioned hybrid standard sample is added respectively according to basic, normal, high 3 Pitch-based sphere, each Pitch-based sphere replication 6 times, the results are shown in Table 4.

Table 4 recovery and precision (n=6)

Detection method of the present invention can be carried out analysis at least 5 kinds of OIT compounds in smoke aqueous gel simultaneously be detected, and pre-treatment is easy, testing result is accurate and reproducible, highly sensitive, be applicable to the mensuration of 5 kinds of OIT compounds in smoke aqueous gel and other complex matrices.

Accompanying drawing explanation

Fig. 1 for (A) before derivative and derivative after mark in (B) and total ion current (TIC) figure of 5 kinds of OIT compounds.

In Fig. 1: 1.MI, 2.CMI-d3 (IS), 3.CMI, 4a.BIT, 4b.BIT derivant, 5.OI, 6.DCOI.

Fig. 2 is Selective ion mode stream (SIM) figure of standard model (C) and testing sample (D).

In Fig. 2: 1.MI, 2.CMI-d3 (IS), 3.CMI, 4a.BIT, 4b.BIT derivant, 5.OI, 6.DCOI.

Embodiment

Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.

Embodiment 1

Below by way of specific instantiation, embodiments of the present invention are described, those skilled in the art the content disclosed by this instructions can understand other advantages of the present invention and effect easily.The present invention can also be implemented or be applied by instantiation modes different in addition, and the every details in this instructions also can based on different viewpoints and application, carries out various modification or change not deviating under spirit of the present invention.

Embodiment 1

1) sample pre-treatments

Accurately take 8 kinds of dissimilar smoke aqueous gel sample 0.2g respectively in 50mL tool plug triangular flask, add extraction solvent methylene chloride 10mL and inner mark solution 80 μ L, ultrasonic 30min, leave standstill and get supernatant in 15mL centrifuge tube, the centrifugal 5min of 3000r/min, obtains supernatant;

2) get 1mL supernatant and cross 0.45 μm of organic phase filter membrane in chromatogram bottle, add 30 μ L BSTFA derivatization reagents, in 60 DEG C of water-baths, derivative 30min, is cooled to room temperature;

3) carry out quantitative test with gas chromatography-mass spectrum (GC-MS), testing result is as shown in table 5;

Wherein, GC-MS analysis condition is as follows:

Chromatographic condition: chromatographic column: DB-624 (30m × 0.25mm, 1.4 μm); Injector temperature: 250 DEG C; Carrier gas: high-purity helium, purity >=99.999%; Sample size: 2 μ L; Flow velocity: 1.2mL/min (constant current mode); Split ratio: 5:1; Heating schedule: initial temperature 80 DEG C keeps 2min, rises to 220 DEG C, then rise to 240 DEG C with the programming rate of 20 DEG C/min with the programming rate of 10 DEG C/min, keeps 15min.

Mass Spectrometry Conditions: transmission line temperature: 240 DEG C; Ion source temperature 230 DEG C; Solvent delay: 6min; Ionization mode: EI source; Ionizing energy: 70eV; Scan mode: Salbutamol Selected Ion Monitoring (SIM).

The content (μ g/g) of table 55 kinds of OIT compounds

Sample	MI	CMI	BIT	OI	DCOI
						1#	28.07	48.12	-	-	-
2#	23.94	55.36	-	-	-
						3#	19.43	54.13	-	-	-
4#	16.92	51.77	96.07	-	-
						5#	15.45	7.47	19.93	-	-

6#	70.18	10.45	-	-	-
						7#	24.59	75.70	-	-	-
8#	11.49	34.47	-	-	-

Note: "-": do not detect

Result shows, MI and CMI applies more Fungicidal compounds in smoke aqueous gel, and BIT application is relatively less, and OI and DCOI does not all detect in selected actual sample.

Those skilled in the art know, also not containing OI and DCOI in smoke aqueous gel conventional at present, but do not get rid of the appearance of the water base adhesive adding this compounds from now on, and adopt detection method of the present invention can detect five kinds of isothiazole phosphorus ketone compounds, for cigarette product quality safety provides technical support simultaneously.

Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.

Claims (9)

1. detect a method for OIT compounds in smoke aqueous gel, it is characterized in that, comprise the steps:

1) in smoke aqueous gel sample, extractant and inner mark solution is added, ultrasonic extraction, centrifugal, obtain supernatant;

2) by supernatant liquid filtering, add derivatization reagent and carry out derivative reaction, after reaction terminates, be cooled to room temperature;

3) GC-MS is utilized to step 2) OIT compounds carries out quantitative test in gained reactant liquor.

2. method according to claim 1, is characterized in that, step 1) in, described extraction solvent is selected from the one in methyl alcohol, normal hexane, methylene chloride, methyl tert-butyl ether, ethyl acetate.

3. method according to claim 2, is characterized in that, described extraction solvent is selected from methylene chloride.

4. method according to claim 1, is characterized in that, step 1) in, described inner mark solution is 5-chloro-2-methyl-3-isothiazolone-d3 solution, and concentration is 0.2mg/mL;

Described inner mark solution is 50-100 μ L:0.2g with the volume mass ratio of smoke aqueous gel sample.

5. method according to claim 1, is characterized in that, step 2) in, described derivatization reagent is two (TMS) trifluoroacetamide of N-methyl-N-(trimethyl silane) trifluoroacetamide or N, O-.

6. method according to claim 1, is characterized in that, step 2) in, described derivatization reagent is 10-60 μ L:0.2g with the volume mass ratio of smoke aqueous gel sample.

7. method according to claim 1, is characterized in that, step 2) in, described derivative reaction condition: temperature 40-80 DEG C, time 10-50min.

8. method according to claim 1, is characterized in that, step 3) in, in described GC-MS, chromatographic condition is as follows:

Chromatographic column: chromatographic column is selected from the one in nonpolar HP-5MS, the DB-624 of middle polarity and the HP-INNOWAX of strong polarity;

Injector temperature: 250 DEG C;

Carrier gas: high-purity helium, purity >=99.999%;

Sample size: 2 μ L;

Flow velocity: 1.2mL/min, constant current mode;

Split ratio: 5:1;

Heating schedule: initial temperature 80 DEG C keeps 2min, rises to 220 DEG C, then rise to 240 DEG C with the programming rate of 20 DEG C/min with the programming rate of 10 DEG C/min, keeps 15min;

Mass Spectrometry Conditions is as follows: transmission line temperature: 240 DEG C; Ion source temperature 230 DEG C; Solvent delay: 6min; Ionization mode: EI source; Ionizing energy: 70eV; Scan mode: Salbutamol Selected Ion Monitoring.

9. method according to claim 1, it is characterized in that, step 3) in, adopt methylene chloride to configure the OIT mixed standard solution of mixing OIT Standard Reserving Solution, the preparation of interior mark stock solution, variable concentrations gradient in described GC-MS, be specially:

1) OIT Standard Reserving Solution is mixed: take 2-methyl-3 (2H)-OIT, 5-chloro-2-methyl-2H-isothiazole-3-ketone, 1 respectively, 2-benzisothiazole base-3 (2H)-one, 2-octyl group-4-isothiazoline-3-ketone, 4, the OIT standard specimen of the chloro-2-n-octyl of 5-bis--3-OIT 5 kinds of different qualities, be placed in 50mL volumetric flask, add methylene chloride constant volume, shake up, obtain mixing OIT Standard Reserving Solution;

2) mark stock solution preparation in: accurately take mark 5-chloro-2-methyl-2H-isothiazole-3-ketone (CMI)-d3 in 0.005g, be placed in the volumetric flask of 25mL, add methylene chloride constant volume, obtain interior mark stock solution;

3) the OIT mixed standard solution of variable concentrations gradient: accurately pipette 5,10,20,50,100,200 μ L mixing OIT Standard Reserving Solution in 6 25mL volumetric flasks, add mark standard solution in 200 μ L respectively, use methylene chloride constant volume, shake up, obtain the OIT mixed standard solution of variable concentrations gradient, wherein, mark concentration is 1.6 μ g/mL.