Method for determining linalool in blast bead essence

Abstract

The invention relates to the technical field of analysis of linalool in essence, and discloses a method for determining linalool in bead blasting essence, which comprises the following steps: S1, preparing bead blasting essence extracting solution; s2, preparing a standard working solution; s3, drawing a standard working curve of the linalool: performing liquid chromatography-mass spectrometry (UPLC-MS / MS) analysis on the standard working solution of the linalool, and drawing the standard working curve of the linalool; s4, determination of a blasting bead essence extracting solution: performing UPLC-MS / MS analysis on linalool in the blasting bead essence extracting solution under the conditions in the step S3; and S5, calculating the content of linalool in the blast bead essence extracting solution according to the standard working curve of linalool. According to the method, the content of linalool in the blast bead essence can be simply, quickly and accurately determined, and false positive detection results are almost avoided through monitoring / detection of double ion pairs of a tandem mass spectrometry detector.

Description

Technical Field

[0001] The present invention relates to the technical field of linalool analysis in flavors, and particularly to a method for determining linalool in flavors. Background Art

[0002] Linalool is a chain-like terpene alcohol compound. It is a colorless liquid at room temperature with a lily of the valley scent. As one of the most commonly used and largest-volume spices in the fragrance industry, linalool occupies an important position in the international market. However, studies have shown that linalool may cause adverse reactions in some people, such as low mood, depression, and even affect respiratory health. In addition, due to its potential allergenicity, it is included in the list of 26 fragrance ingredients that may cause allergic reactions specified in the EU Cosmetics Directive. Explosive bead flavors, also known as fragrance pills or brittle capsules, are a sealed capsule formed by encapsulating the fragrance liquid with a polymer, which is commonly found in food and fast-moving consumer goods. This product is popular with consumers because of its unique fragrance-adding method, and research on it is also increasing. Explosive bead flavors are composed of two parts: core material and wall material. The core material contains flavors and fragrances and an appropriate amount of solvent, while the wall material is generally composed of gelatin, modified starch, carrageenan and other substances. When the wall material breaks under certain conditions, the fragrance components in the core material will be released, thereby enhancing the fragrance.

[0003] Since the contents of the popping bead flavors are affected by many factors, such as the origin of raw materials and processing technology, as well as the quality stability between batches, it is necessary to prevent exogenous linalool contamination or accidental addition without the knowledge of multiple parties. Therefore, in order to ensure product quality, it is particularly important to develop a method that can accurately measure the content of linalool. Its main function is to screen the risk transmission from the supplier to the downstream user without knowing it, and to conduct risk investigation when the supply and demand parties are about to finalize the product.

[0004] At present, the linalool content in popping pearl essence can be analyzed by gas chromatography (GC), gas chromatography-mass spectrometry (GC-MS) and other technologies. However, these methods have some challenges. For example, caprylic decanoic acid glyceride in popping pearl essence may remain on the gas chromatography column, which requires high temperature (over 300°C) treatment for dozens of minutes after each analysis to remove the residue, which not only prolongs the detection time (usually more than one hour), but may also shorten the life of the chromatographic column. Secondly, in order to avoid the occurrence of false positives, it is urgent to develop an HPC-MS / MS method that can be mutually verified with other methods. For example, ISO17234-1 "Leather-Chemical Test-Determination of Certain Azo Dyes in Leather" and EU EN 14362-1 "Determination of Certain Aromatic Amines Derived from Azo Dyes in Textiles" clearly require: If one method is used to detect the presence of aromatic amines, it is necessary to confirm it with another one or two different methods to avoid misjudgment. Moreover, the standard also stipulates that only when both methods give positive results can it be determined that the sample contains banned azo dyes. It can be seen that a single detection method of linalool has its drawbacks. Given the importance of this component, all stakeholders have the need to establish another mutual verification method. Summary of the invention

[0005] In view of this, the purpose of the present invention is to provide a method for determining linalool in popping bead essence, which can simply, quickly and accurately determine the content of linalool in popping bead essence, and almost avoid the occurrence of false positive detection results through monitoring / detection of dual ion pairs of tandem mass spectrometry detector.

[0006] The present invention solves the above technical problems by the following technical means:

[0007] A method for determining linalool in blasting bead flavor, including the following steps:

[0008] S1. Preparation of the extract of the popping pearl essence: After the popping pearl essence is preliminarily treated, a solvent is added, and then N-propylethylenediamine is added as a dispersed solid phase extractant, and vortexed for 30 seconds to 1 minute to form a mixed solution, and the mixed solution is centrifuged to separate the supernatant and the precipitate, and the supernatant is filtered, and the filtered liquid is the popping pearl essence extract;

[0009] S2. Preparation of standard working solutions: Prepare linaol standard working solutions;

[0010] S3. Drawing of standard working curves of linalool: Liquid chromatography-mass spectrometry combined with UPLC-MS / MS to draw the standard working curve of linalool;

[0011] S4. Determination of the blasting bead flavor extract: UPLC-MS / MS analysis of linalool in the blasting bead flavor extract is performed using the conditions in step S3;

[0012] S5. Referring to the standard working curve of linalool, the content of linalool in the blasting bead flavor extract was calculated.

[0013] A further preferred method for the preliminary treatment of the bursting bead flavor in step S1 is to cut ordinary qualitative filter paper into small pieces, put the small pieces of filter paper into an oven for high-temperature impurity removal, weigh the bursting bead flavor, wrap the bursting bead flavor with filter paper, break it, and place it in a centrifuge tube together with the filter paper.

[0014] Preferably, the solvent in step S1 is one of methanol and acetonitrile.

[0015] Preferably, the centrifugal treatment process parameter of the step S1 is that the centrifugal separation temperature is 20-25°C, the rotation speed is 10000-15000 rpm, and the time is 5-8 min.

[0016] Preferably, the filtration of step S1 is filtered using a 0.22 µm organic filter membrane.

[0017] Preferably, the concentration of the linalool standard working solution in step S1 is 0 μg / mL, 5 μg / mL, 10 μg / mL, 20 μg / mL, and 50 μg / mL.

[0018] Preferably, the chromatography condition in the liquid chromatography-mass spectrometry combined with UPLC-MS / MS analysis conditions in the step S3 is,

[0019] The column is a C18 bonded silicon ball filler liquid chromatography column (100×2.1 mm, 2.7 μm) or equivalent column;

[0020] The mobile phases were pure water and acetonitrile;

[0021] Gradient elution procedure: 0-10 min, the ratio of acetonitrile to water linearly increased from 20:80 to 100% acetonitrile;

[0022] 10-15 min, 100% acetonitrile, 15-16 min, reduced from 100% acetonitrile to 20% acetonitrile, 16-20 min, 20% acetonitrile; the running time is 20 min in total;

[0023] Column temperature: 20-40°C; flow rate: 0.3-0.6 mL / min; injection volume: 1-5 µL.

[0024] Preferably, the chromatographic conditions are: the column temperature is 30°C, the flow rate is 0.6 mL / min, and the injection volume is 2 µL.

[0025] Preferably, the mass spectrometry condition in the liquid chromatography-mass spectrometry combined with UPLC-MS / MS analysis conditions in the step S4 is,

[0026] The mass number of precursor ion detection is 155.2±1, the mass number of sub-ion detection is 137.0±1, 81.0±1; the decluster voltage (DP) is 15-50 V, and the collision energy (CE) is 15-40 V.

[0027] Preferably, the drawing of the standard curve in step S4 is to establish a linear equation with the concentration of linalool standard working solution as the horizontal coordinate and the mass spectrometry response peak area as the vertical coordinate.

[0028] Preferably, the calculation formula for linalool content in the blasting bead extract of step S5 is:

[0029]

[0030] Where:

[0031] C—The concentration of linalool in the flavor and flavor sample obtained from the standard curve in micrograms per milliliter (μg / mL);

[0032] C0—The concentration of linalool in the blank sample derived from the standard curve in micrograms per milliliter (μg / mL);

[0033] V—volume of the extract, in milliliters (mL);

[0034] k—dilution multiple;

[0035] m—sample mass, in grams (g).

[0036] The average value of the two parallel measurements was used as the final measurement result, and the accuracy was as high as 0.1 mg / kg.

[0037] Beneficial effects of the present invention:

[0038] (1) Ultra-high performance liquid chromatography was used as an instrumental analysis method, and a corresponding method for processing the popping essence was established to obtain a clean solution for the machine and reduce the interference of the matrix on the target object. The monitoring / detection of the dual ion pair of the tandem mass spectrometer detector almost avoided the occurrence of false positive test results.

[0039] (2) The interference of non-target substances in the matrix is ​​greatly reduced by adding N-propylethylenediamine (PSA) bonded silica gel as a dispersed solid-phase extractant and a certain proportion of solvent dilution.

[0040] (3) According to the characteristics of the target compound, a new rapid analysis method was established through the optimization of UPLC chromatography conditions, and the detection limit of the method was 0.5 μg / mL. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1It is a chromatogram of a typical standard solution under the chromatographic conditions of Example 2 of the present invention.

[0042] Figure 2 It is a chromatogram of the Explosive Bead Fragrance B under the chromatographic conditions of Example 2 of the present invention. DETAILED DESCRIPTION

[0043] The following will be combined with the specific embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention:

[0044] Example 1

[0045] A method for determining linalool in blasting bead flavor, including the following steps:

[0046] S1. Preparation of the extract of the popping pearl essence: cut the ordinary qualitative filter paper into small pieces, put the small pieces of filter paper into the oven for high-temperature impurity removal, weigh 0.1 g of the popping pearl essence, wrap it with filter paper, crush it, put it into a centrifuge tube together with the filter paper, add 5 mL of methanol, and then add 100 mg of N-propylethylenediamine dispersed solid phase extractant, vortex mix for 30 seconds to form a mixed solution, centrifuge the mixed solution, and the centrifugal process parameters are centrifugal separation temperature of 20°C, speed of 10000 rpm, and time of 5 min. Separate the supernatant and the precipitate, take the supernatant and filter it through a 0.22 µm organic filter membrane, and the filtered liquid is the popping pearl essence extract;

[0047] S2. Preparation of standard working solutions: Prepare 0 μg / mL, 5 μg / mL, 10 μg / mL, 20 μg / mL, 50 μg / mL standard working solutions of linaol;

[0048] S3. Drawing of the standard working curve of linalool: The linalool standard working solution was analyzed by liquid chromatography-mass spectrometry coupled with UPLC-MS / MS. The concentration of the linalool standard working solution was used as the horizontal axis and the mass spectrometry response peak area was used as the vertical axis. A linear equation was established to draw the standard working curve of linalool.

[0049] The chromatographic conditions are:

[0050] The chromatographic column is a C18 bonded silicon ball filler liquid chromatography column, 100×2.1 mm, 2.7 μm;

[0051] The mobile phases were pure water and acetonitrile;

[0052] Gradient elution procedure: 0-10 min, the ratio of acetonitrile to water linearly increased from 20:80 to 100% acetonitrile;

[0053] 10-15 min, 100% acetonitrile, 15-16 min, reduced from 100% acetonitrile to 20% acetonitrile, 16-20 min, 20% acetonitrile; the running time is 20 min in total;

[0054] Column temperature: 20°C; flow rate: 0.3 mL / min; injection volume: 1 µL.

[0055] The mass spectrometry conditions were:

[0056] The mass number of precursor ion detection is 154.2, the mass number of sub-ion detection is 136.0 and 80.0; the decluster voltage (DP) is 15 V, collision energy (CE) is 15 V.

[0057] S4. Determination of the blasting bead flavor extract: UPLC-MS / MS analysis of linalool in the blasting bead flavor extract is performed using the conditions in step S3;

[0058] S5. Referring to the standard working curve of linalool, the content of linalool in the blasting bead flavor extract was calculated.

[0059] The calculation formula for linalool content in the burlap flavor extract is:

[0060]

[0061] Where:

[0062] C—The concentration of linalool in the blasting bead flavor obtained from the standard curve, in micrograms per milliliter (μg / mL);

[0063] C0—The concentration of linalool in the blank sample derived from the standard curve in micrograms per milliliter (μg / mL);

[0064] V—volume of the extract, in milliliters (mL);

[0065] k—dilution multiple;

[0066] m—sample mass, in grams (g).

[0067] The average value of the two parallel measurements was used as the final measurement result, and the accuracy was as high as 0.1 mg / kg.

[0068] Example 2

[0069] A method for determining linalool in blasting bead flavor, including the following steps:

[0070] S1. Preparation of the extract of the popping pearl essence: cut the ordinary qualitative filter paper into small pieces, put the small pieces of filter paper into the oven for high-temperature impurity removal, weigh 0.1 g of the popping pearl essence, wrap it with filter paper, crush it, put it into a centrifuge tube together with the filter paper, add 5 mL of acetonitrile, and then add 100 mg of N-propylethylenediamine dispersed solid phase extractant, vortex mix for 45 seconds to form a mixed solution, centrifuge the mixed solution, and the centrifugal process parameters are centrifugal separation temperature of 22.5℃, speed of 12500 rpm, and time of 6.5 min. Separate the supernatant and the precipitate, take the supernatant and filter it through a 0.22 µm organic filter membrane, and the filtered liquid is the popping pearl essence extract;

[0071] S2. Preparation of standard working solutions: Prepare 0 μg / mL, 5 μg / mL, 10 μg / mL, 20 μg / mL, 50 μg / mL standard working solutions of linaol;

[0072] S3. Drawing of the standard working curve of linalool: The linalool standard working solution was analyzed by liquid chromatography-mass spectrometry coupled with UPLC-MS / MS. The concentration of the linalool standard working solution was used as the horizontal axis and the mass spectrometry response peak area was used as the vertical axis. A linear equation was established to draw the standard working curve of linalool.

[0073] The chromatographic conditions are:

[0074] The chromatographic column is a C18 bonded silicon ball filler liquid chromatography column, 100×2.1 mm, 2.7 μm;

[0075] The mobile phases were pure water and acetonitrile;

[0076] Gradient elution procedure: 0-10 min, the ratio of acetonitrile to water linearly increased from 20:80 to 100% acetonitrile;

[0077] 10-15 min, 100% acetonitrile, 15-16 min, reduced from 100% acetonitrile to 20% acetonitrile, 16-20 min, 20% acetonitrile; the running time is 20 min in total;

[0078] The column temperature was 30°C, the flow rate was 0.6 mL / min, and the injection volume was 2 µL.

[0079] The mass spectrometry conditions were:

[0080] The mass number of precursor ion detection is 155.2, the mass number of sub-ion detection is 137.0 and 81.0; the decluster voltage (DP) is 32.5 V, collision energy (CE) is 27.5 V

[0081] S4. Determination of the blasting bead flavor extract: UPLC-MS / MS analysis of linalool in the blasting bead flavor extract is performed using the conditions in step S3;

[0082] S5. Referring to the standard working curve of linalool, the content of linalool in the blasting bead flavor extract was calculated.

[0083] The calculation formula for linalool content in the burlap flavor extract is:

[0084]

[0085] Where:

[0086] C—The concentration of linalool in the blasting bead flavor obtained from the standard curve, in micrograms per milliliter (μg / mL);

[0087] C0—The concentration of linalool in the blank sample derived from the standard curve in micrograms per milliliter (μg / mL);

[0088] V—volume of the extract, in milliliters (mL);

[0089] k—dilution multiple;

[0090] m—sample mass, in grams (g).

[0091] The average value of the two parallel measurements was used as the final measurement result, and the accuracy was as high as 0.1 mg / kg.

[0092] Example 3

[0093] A method for determining linalool in blasting bead flavor, including the following steps:

[0094] S1. Preparation of the extract of the popping pearl essence: cut the ordinary qualitative filter paper into small pieces, put the small pieces of filter paper into the oven for high-temperature impurity removal, weigh 0.1 g of the popping pearl essence, wrap it with filter paper, crush it, put it into a centrifuge tube together with the filter paper, add 5 mL of methanol, and then add 100 mg of N-propylethylenediamine dispersed solid phase extractant, vortex mix for 1 minute to form a mixed solution, centrifuge the mixed solution, and the centrifugal process parameters are centrifugal separation temperature of 25°C, speed of 15000 rpm, and time of 8 min. Separate the supernatant and the precipitate, take the supernatant and filter it through a 0.22 µm organic filter membrane, and the filtered liquid is the popping pearl essence extract;

[0095] S2. Preparation of standard working solutions: Prepare 0 μg / mL, 5 μg / mL, 10 μg / mL, 20 μg / mL, 50 μg / mL standard working solutions of linaol;

[0096] S3. Drawing of the standard working curve of linalool: The linalool standard working solution was analyzed by liquid chromatography-mass spectrometry coupled with UPLC-MS / MS. The concentration of the linalool standard working solution was used as the horizontal axis and the mass spectrometry response peak area was used as the vertical axis. A linear equation was established to draw the standard working curve of linalool.

[0097] The chromatographic conditions are:

[0098] The chromatographic column is a C18 bonded silicon ball filler liquid chromatography column, 100×2.1 mm, 2.7 μm;

[0099] The mobile phases were pure water and acetonitrile;

[0100] Gradient elution procedure: 0-10 min, the ratio of acetonitrile to water linearly increased from 20:80 to 100% acetonitrile;

[0101] 10-15 min, 100% acetonitrile, 15-16 min, reduced from 100% acetonitrile to 20% acetonitrile, 16-20 min, 20% acetonitrile; the running time is 20 min in total;

[0102] Column temperature: 40°C; flow rate: 0.6 mL / min; injection volume: 5 µL.

[0103] The mass spectrometry conditions were:

[0104] The mass number of precursor ion detection is 156.2, the mass number of sub-ion detection is 138.0 and 82.0; the decluster voltage (DP) is 50 V, and the collision energy (CE) is 40 V.

[0105] S4. Determination of the blasting bead flavor extract: UPLC-MS analysis of linalool in the blasting bead flavor extract using the conditions in step S3;

[0106] S5. Referring to the standard working curve of linalool, the content of linalool in the blasting bead flavor extract was calculated.

[0107] The calculation formula for linalool content in the burlap flavor extract is:

[0108]

[0109] Where:

[0110] C—The concentration of linalool in the blasting bead flavor obtained from the standard curve, in micrograms per milliliter (μg / mL);

[0111] C0—The concentration of linalool in the blank sample derived from the standard curve in micrograms per milliliter (μg / mL);

[0112] V—volume of the extract, in milliliters (mL);

[0113] k—dilution multiple;

[0114] m—sample mass, in grams (g).

[0115] The average value of the two parallel measurements was used as the final measurement result, and the accuracy was as high as 0.1 mg / kg.

[0116] Linalool was measured in Example 2, and Table 1 data was obtained based on the calculation formula for linalool content.

[0117] Table 1 Linalool content in samples

[0118]

[0119] Recovery and reproducibility of the invented method

[0120] The standard addition method was used to determine the recovery of the method. Weigh 0.1 g of the popping pearl flavor, add a mixed standard solution with the same content, extract according to the pretreatment conditions of this method, repeat 6 times, and determine the spiked recovery and repeatability. The results are shown in Table 2. The average recovery range is 94.5%, and the repeatability experiment RSD is 6.5%, which can meet the needs of quantitative analysis.

[0121] Table 2 Recovery and repeatability of the method

[0122]

[0123] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions of the present invention, which should be included in the scope of the claims of the present invention. The techniques, shapes, and structural parts not described in detail in the present invention are all known technologies.

Claims

1. A method for determining linalool in popping pearl essence, characterized in that: The following steps are involved: S1. Preparation of the extract of the popping pearl essence: After the popping pearl essence is preliminarily treated, a solvent is added, and then N-propylethylenediamine is added as a dispersed solid phase extractant, and vortexed for 30 seconds to 1 minute to form a mixed solution, and the mixed solution is centrifuged to separate the supernatant and the precipitate, and the supernatant is filtered, and the filtered liquid is the popping pearl essence extract; S2. Preparation of standard working solution: prepare linalool standard working solution; S3. Preparation of the standard working curve of linalool: The linalool standard working solution was analyzed by liquid chromatography-mass spectrometry coupled with UPLC-MS / MS to prepare the standard working curve of linalool; S4. Determination of the extract of popping pearl essence: UPLC-MS / MS analysis of linalool in the extract of popping pearl essence was performed using the conditions of step S3; S5. Calculate the content of linalool in the extract of the explosive pearl essence by referring to the standard working curve of linalool.

2. The method for determining linalool in a popping pearl flavor according to claim 1, characterized in that: The solvent in step S1 is one of methanol and acetonitrile.

3. The method for determining linalool in a popping pearl essence according to claim 1, characterized in that: The centrifugal treatment process parameters of step S1 are as follows: centrifugal separation temperature is 20-25° C., rotation speed is 10000-15000 rpm, and time is 5-8 min.

4. The method for determining linalool in a popping pearl flavor according to claim 1, characterized in that: The filtration in step S1 is performed using a 0.22 µm organic filter membrane.

5. The method for determining linalool in a popping pearl flavor according to claim 1, characterized in that: The concentrations of the linalool standard working solution in step S1 are 0 μg / mL, 5 μg / mL, 10 μg / mL, 20 μg / mL, and 50 μg / mL.

6. The method for determining linalool in a popping bead flavor according to claim 1, characterized in that: The chromatographic conditions in the UPLC-MS / MS analysis conditions of step S3 are: The chromatographic column is a C18 bonded silica ball filler liquid chromatography column (100×2.1 mm, 2.7 μm) or an equivalent column; The mobile phases were pure water and acetonitrile; Gradient elution program: 0-10 min, the ratio of acetonitrile to water increased linearly from 20:80 to 100% acetonitrile; 10-15 min, 100% acetonitrile, 15-16 min, from 100% acetonitrile to 20% acetonitrile, 16-20 min, 20% acetonitrile; total running time 20 min; Column temperature: 20-40℃; flow rate: 0.3-0.6 mL / min; injection volume: 1-5 µL.

7. The method for determining linalool in a popping bead flavor according to claim 5, characterized in that: The chromatographic conditions are as follows: column temperature of 30°C, flow rate of 0.6 mL / min, and injection volume of 2 µL.

8. The method for determining linalool in a popping bead flavor according to claim 1, characterized in that: The mass spectrometry conditions in the UPLC-MS / MS analysis conditions of step S3 are: The mass number of the parent ion detection was 155.2±1, and the mass numbers of the daughter ion detection were 137.0±1 and 81.0±1; the declustering voltage (DP) was 15-50 V, and the collision energy (CE) was 15-40 V.

9. The method for determining linalool in a popping pearl essence according to claim 1, characterized in that: The drawing of the standard curve in step S4 is specifically to establish a linear equation with the concentration of the linalool standard working solution as the abscissa and the mass spectrometry response peak area as the ordinate.

10. The method for determining linalool in a popping bead flavor according to claim 1, characterized in that: The calculation formula for the linalool content in the popping pearl essence extract in step S5 is: Where: C—the concentration of linalool in the popping pearl flavor obtained from the standard curve, in micrograms per milliliter (μg / mL); C0—linalool concentration in the blank sample obtained from the standard curve, in micrograms per milliliter (μg / mL); V—volume of the extract, in milliliters (mL); k—dilution multiple; m—sample mass, in grams (g); The average of two parallel measurements was taken as the final measurement result, accurate to 0.1 mg / kg.

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