Quantitative method for ultra-trace bisphenol compounds in wine

Abstract

The invention relates to a method for quantifying ultra-trace bisphenol compounds in wine, which comprises the following steps: 1, sample pretreatment: concentrating a to-be-detected wine sample to obtain a concentrated solution, carrying out constant volume treatment on the concentrated solution, carrying out refrigerated centrifugation treatment on the concentrated solution subjected to the constant volume treatment, and taking supernate obtained after the centrifugation treatment as a to-be-analyzed solution; wherein a constant volume solvent adopted in the constant volume treatment is methanol; and quantifying the sample: carrying out quantitative analysis on the to-be-analyzed solution by adopting a chromatography-mass spectrometry method. According to the research, a sample is pretreated by concentration; according to the method, the specific variety of bisphenol compounds are rapidly screened through chromatography-mass spectrometry, the method which is wide in variety coverage range, rapid, accurate and capable of quantifying the bisphenol compounds is established, and the method can be used for risk screening of ultra-trace bisphenol and derivatives thereof in Baijiu.

Description

Technical Field

[0001] This application belongs to the field of liquor detection technology, specifically relating to a quantitative method for ultra-trace bisphenol compounds in liquor. Background Technology

[0002] Bisphenol compounds have a basic structure consisting of two phenolic groups connected by bridging groups (such as isopropyl and sulfonyl groups). Bisphenol compounds and their derivatives are often used as initial raw materials, heat stabilizers and reinforcing agents for food contact plastic materials such as epoxy phenolic coatings and polyvinyl chloride organosol coatings, giving packaging materials properties such as transparency and impact resistance. However, they may leach out of plastics and migrate into food or water under acidic / alkaline environments or high temperatures. Summary of the Invention

[0003] To address the aforementioned technical problems, this application provides a method for quantifying trace amounts of bisphenol compounds in wine, comprising the following steps:

[0004] I. Sample pretreatment: The wine sample to be tested is concentrated to obtain a concentrated solution. The concentrated solution is then brought to a fixed volume. The concentrated solution after volume adjustment is then subjected to refrigerated centrifugation. The supernatant obtained after centrifugation is used as the solution to be analyzed.

[0005] The volume-fixing solvent used in the volume-fixing process is methanol;

[0006] II. Sample Quantification: The solution to be analyzed was quantitatively analyzed using chromatography-mass spectrometry.

[0007] In some specific embodiments, the concentration includes nitrogen blowing concentration.

[0008] In some specific embodiments, the nitrogen blowing concentration temperature is 50–70°C. In some specific embodiments, it can be 50°C, 51°C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C, 60°C, 61°C, 62°C, 63°C, 64°C, 65°C, 66°C, 67°C, 68°C, 69°C, 70°C, or any value between any two of the aforementioned values.

[0009] In some specific embodiments, the nitrogen blowing concentration temperature is 60°C.

[0010] In some specific embodiments, the needle velocity for nitrogen blowing concentration is 0.5 mm / min to 1.5 mm / min. In some specific embodiments, it can be 0.5 mm / min, 0.6 mm / min, 0.7 mm / min, 0.8 mm / min, 0.9 mm / min, 1.0 mm / min, 1.1 mm / min, 1.2 mm / min, 1.3 mm / min, 1.4 mm / min, 1.5 mm / min, or any value between any two of the aforementioned values.

[0011] In some specific embodiments, the pressure of the nitrogen blowing concentration is 2 psi to 4 psi; in some more specific embodiments, it can be 2 psi, 2.2 psi, 2.4 psi, 2.5 psi, 2.6 psi, 2.8 psi, 3.0 psi, 3.2 psi, 3.4 psi, 3.5 psi, 3.6 psi, 3.8 psi, 4.0 psi, or any value between any two of the aforementioned values.

[0012] In some specific embodiments, the concentration factor of the concentrate compared to the wine sample to be tested is 10 to 15; in some specific embodiments, it can be 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, or any value between any two of the aforementioned values.

[0013] In some specific embodiments, the concentrate is 10 times more concentrated than the wine sample to be tested.

[0014] In some specific embodiments, in step one, the volume of the sample to be tested is 5 ml, and the concentration factor of the concentrate compared to the wine sample to be tested is 10.

[0015] In some more specific embodiments, the volume of the concentrate after volume adjustment is 1 ml.

[0016] In some more specific embodiments, the ratio of the concentrate to the volume-fixing solvent in the concentrate after volume-fixing treatment is (1-2):(1-2); in some specific embodiments, it can be 1:1, 1:1.2, 1:1.5, 1:1.8, 1:2, 1.2:1, 1.2:1.2, 1.2:1.5, 1.2:1.8, 1.2:2, 1.5:1, 1.5:1.2, 1.5:1.5, 1.5:1.8, 1.5:2, 1.8:1, 1.8:1.2, 1.8:1.5, 1.8:1.8, 1.8:2, 2:1, 2:1.2, 2:1.5, 2:1.8, 2:2, and any ratio between any two of the aforementioned values.

[0017] In some specific embodiments, the chromatographic conditions include: mobile phase A being an aqueous solution and mobile phase B being methanol.

[0018] In some specific embodiments, the chromatographic conditions include: a PFP column (2.1 mm × 50 mm, 2.6 μm).

[0019] In some specific embodiments, the chromatographic conditions include: injection volume: 0.5–5.5 μL; in some specific embodiments, it can be 0.5 μL, 0.6 μL, 0.7 μL, 0.8 μL, 0.9 μL, 1.0 μL, 1.1 μL, 1.2 μL, 1.3 μL, 1.4 μL, 1.5 μL, 1.6 μL, 1.7 μL, 1.8 μL, 1.9 μL, 2.0 μL, 2.1 μL, 2.2 μL, 2.3 μL, 2.4 μL, 2.5 μL, 2.6 μL, 2.7 μL, ... 2.8 μL, 2.9 μL, 3.0 μL, 3.1 μL, 3.2 μL, 3.3 μL, 3.4 μL, 3.5 μL, 3.6 μL, 3.7 μL, 3.8 μL, 3.9 μL, 4.0 μL, 4.1 μL, 4.2 μL, 4.3 μL, 4.4 μL, 4.5 μL, 4.6 μL, 4.7 μL, 4.8 μL, 4.9 μL, 5.0 μL, 5.1 μL, 5.2 μL, 5.3 μL, 5.4 μL, 5.5 μL, and any value between any two of the aforementioned values.

[0020] In some specific embodiments, the chromatographic conditions include: injection volume: 1 μL. In some specific embodiments, the chromatographic conditions include: injection volume: 5 μL.

[0021] In some specific embodiments, the chromatographic conditions include: column temperature: 35℃~45℃; flow rate: 250μL / min~350μL / min; in some specific embodiments, the column temperature can be 35℃, 36℃, 37℃, 38℃, 39℃, 40℃, 41℃, 42℃, 43℃, 44℃, 45℃, or any value between any two of the aforementioned values. In some specific embodiments, the flow rate can be 250μL / min, 260μL / min, 270μL / min, 280μL / min, 290μL / min, 300μL / min, 310μL / min, 320μL / min, 330μL / min, 340μL / min, 350μL / min, or any value between any two of the aforementioned values.

[0022] In some specific embodiments, the chromatographic conditions include: column temperature: 40°C; flow rate: 300 μL / min.

[0023] In some specific embodiments, mass spectrometry employs a Q-Exactive quadrupole electrostatic field orbital trap high-resolution mass spectrometer.

[0024] In some specific embodiments, the mass spectrometer uses a heated HESI source with a capillary voltage of -3.7 kV to +3.7 kV; in some specific embodiments, the capillary temperature is 150℃ to 250℃; the sheath gas is 35 alb to 45 alb; the auxiliary gas is 10 alb to 20 alb; the ion transmission tube temperature is 300℃ to 400℃; the scanning mode is Full MS / ddMS2, and the scanning range is 150-600 m / z, with simultaneous acquisition of positive and negative ions. The capillary temperature of 150℃ to 250℃ can, in some specific embodiments, be 150℃, 160℃, 170℃, 180℃, 190℃, 200℃, 210℃, 220℃, 230℃, 240℃, 250℃, or any value between any two of the aforementioned values.

[0025] The sheath gas is 35arb to 45arb, and in some specific embodiments it can be 35arb, 36arb, 37arb, 38arb, 39arb, 40arb, 41arb, 42arb, 43arb, 44arb, 45arb, or any value between any two of the aforementioned values.

[0026] The auxiliary gas is 10arb to 20arb, and in some specific embodiments it can be 10arb, 11arb, 12arb, 13arb, 14arb, 15arb, 16arb, 17arb, 18arb, 19arb, 20arb, or any value between any two of the aforementioned values.

[0027] The temperature of the ion transmission tube is 300℃~400℃, and in some specific embodiments it can be 300℃, 310℃, 320℃, 330℃, 340℃, 350℃, 360℃, 370℃, 380℃, 390℃, 400℃, or any value between any two of the aforementioned values.

[0028] The first-level mass spectrometry resolution is 70,000, and the second-level mass spectrometry resolution is 17,500; the normalized collision energy (NCE) is 30, 45, and 60 eV.

[0029] Preferably, the mass spectrometry conditions include: a heated HESI source; capillary voltage: ±3.7 kV; capillary temperature: 200 °C; sheath gas: 40 arb; auxiliary gas: 15 arb; purge gas: 0 arb; ion transmission tube temperature: 350 °C; Full MS / ddMS2 scanning mode; scanning range: 150-600 m / z, simultaneous acquisition of positive and negative ions; primary mass spectrometry resolution of 70,000, secondary mass spectrometry resolution of 17,500; normalized collision energy (NCE) of 30, 45, and 60 eV.

[0030] In some specific embodiments, the bisphenol compounds include at least one of the following: bisphenol A, bisphenol B, bisphenol C, bisphenol F, bisphenol S, bisphenol Z, bisphenol AF, bisphenol AP, bisphenol P, 4,4-sulfonyl di(2-methylphenol), tetrachlorobisphenol A, tetrabromobisphenol A, bisphenol A diglycidyl ether, bisphenol A-(2-3-dihydroxypropyl)glycerol ether, bisphenol A-(2-3-dihydroxypropyl) ether, bisphenol A-(3-chloro-2-hydroxypropyl)glycerol ether, bisphenol A-(3-chloro-2-hydroxypropyl) ether, bisphenol A-(3-chloro-2-hydroxypropyl)(2-3-dihydroxypropyl) ether, bisphenol F diglycidyl ether, bisphenol F-(2-3-dihydroxypropyl) ether, and bisphenol F-(3-chloro-2-hydroxypropyl) ether.

[0031] Beneficial effects: This study employed concentration for sample pretreatment and methanol for volume adjustment. Chromatography-mass spectrometry (GC-MS) was used for rapid screening of specific types of bisphenol compounds, effectively reducing the false positive detection rate. A broad-coverage, rapid, and accurate method for the quantitative analysis of bisphenol compounds was established. This method can be used for risk screening of trace amounts of bisphenols and their derivatives in baijiu (Chinese liquor), providing a technical reference for the qualitative and quantitative analysis of bisphenols and their derivatives in baijiu. Attached Figure Description

[0032] Figure 1 shows the extraction chromatograms of 21 bisphenols and their derivatives;

[0033] Figure 2 The average spike recovery rate is given when water is used as the solvent in Example 3. Detailed Implementation

[0034] The following specific embodiments further illustrate the technical solution of the present invention. These specific embodiments do not represent a limitation on the scope of protection of the present invention. Non-essential modifications and adjustments made by others based on the concept of the present invention still fall within the scope of protection of the present invention.

[0035] Instruments and reagents

[0036] Purity ≥ 99%, Dr. Ehrenstorfer GmbH, Germany;

[0037] The methanol was LC-MS grade and purchased from Merck.

[0038] Anhydrous ethanol, HPLC grade, was purchased from TianDi Corporation, USA.

[0039] The mass spectrometer was a Q-Exactive quadrupole electrostatic field track trap high-resolution mass spectrometer, purchased from Thermo Fisher Scientific.

[0040] The XP205 electronic balance was purchased from Mettler.

[0041] The nitrogen blowing device was purchased from Auto EVA by Ruike.

[0042] The MILLI-Q pure water system was purchased from Millipore.

[0043] The high-speed refrigerated centrifuge was purchased from Hitachi, Japan.

[0044] To address the aforementioned technical problems, this application provides a method for quantifying trace amounts of bisphenol compounds in wine, comprising the following steps:

[0045] I. Sample pretreatment: The wine sample to be tested is concentrated to obtain a concentrated solution. The concentrated solution is then brought to a fixed volume. The concentrated solution after volume adjustment is then subjected to refrigerated centrifugation. The supernatant obtained after centrifugation is used as the solution to be analyzed.

[0046] The volume-fixing solvent used in the volume-fixing process is methanol;

[0047] II. Sample Quantification: The solution to be analyzed was quantitatively analyzed using chromatography-mass spectrometry.

[0048] In some specific embodiments, the concentration includes nitrogen blowing concentration.

[0049] In some specific embodiments, the nitrogen blowing concentration temperature is 50–70°C. In some specific embodiments, it can be 50°C, 51°C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C, 60°C, 61°C, 62°C, 63°C, 64°C, 65°C, 66°C, 67°C, 68°C, 69°C, 70°C, or any value between any two of the aforementioned values.

[0050] In some specific embodiments, the nitrogen blowing concentration temperature is 60°C.

[0051] In some specific embodiments, the needle velocity for nitrogen blowing concentration is 0.5 mm / min to 1.5 mm / min. In some specific embodiments, it can be 0.5 mm / min, 0.6 mm / min, 0.7 mm / min, 0.8 mm / min, 0.9 mm / min, 1.0 mm / min, 1.1 mm / min, 1.2 mm / min, 1.3 mm / min, 1.4 mm / min, 1.5 mm / min, or any value between any two of the aforementioned values.

[0052] In some specific embodiments, the pressure of the nitrogen blowing concentration is 2 psi to 4 psi; in some more specific embodiments, it can be 2 psi, 2.2 psi, 2.4 psi, 2.5 psi, 2.6 psi, 2.8 psi, 3.0 psi, 3.2 psi, 3.4 psi, 3.5 psi, 3.6 psi, 3.8 psi, 4.0 psi, or any value between any two of the aforementioned values.

[0053] In some specific embodiments, the concentration factor of the concentrate compared to the wine sample to be tested is 10 to 15; in some specific embodiments, it can be 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, or any value between any two of the aforementioned values.

[0054] In some specific embodiments, the concentrate is 10 times more concentrated than the wine sample to be tested.

[0055] In some specific embodiments, in step one, the volume of the sample to be tested is 5 ml, and the concentration factor of the concentrate compared to the wine sample to be tested is 10.

[0056] In some more specific embodiments, the volume of the concentrate after volume adjustment is 1 ml.

[0057] In some more specific embodiments, the ratio of the concentrate to the volume-fixing solvent in the concentrate after volume-fixing treatment is (1-2):(1-2); in some specific embodiments, it can be 1:1, 1:1.2, 1:1.5, 1:1.8, 1:2, 1.2:1, 1.2:1.2, 1.2:1.5, 1.2:1.8, 1.2:2, 1.5:1, 1.5:1.2, 1.5:1.5, 1.5:1.8, 1.5:2, 1.8:1, 1.8:1.2, 1.8:1.5, 1.8:1.8, 1.8:2, 2:1, 2:1.2, 2:1.5, 2:1.8, 2:2, and any ratio between any two of the aforementioned values.

[0058] In some specific embodiments, the chromatographic conditions include: mobile phase A being an aqueous solution and mobile phase B being methanol.

[0059] In some specific embodiments, the chromatographic conditions include: a PFP column (2.1 mm × 50 mm, 2.6 μm).

[0060] In some specific embodiments, the chromatographic conditions include: injection volume: 0.5–5.5 μL; in some specific embodiments, it can be 0.5 μL, 0.6 μL, 0.7 μL, 0.8 μL, 0.9 μL, 1.0 μL, 1.1 μL, 1.2 μL, 1.3 μL, 1.4 μL, 1.5 μL, 1.6 μL, 1.7 μL, 1.8 μL, 1.9 μL, 2.0 μL, 2.1 μL, 2.2 μL, 2.3 μL, 2.4 μL, 2.5 μL, 2.6 μL, 2.7 μL, ... 2.8 μL, 2.9 μL, 3.0 μL, 3.1 μL, 3.2 μL, 3.3 μL, 3.4 μL, 3.5 μL, 3.6 μL, 3.7 μL, 3.8 μL, 3.9 μL, 4.0 μL, 4.1 μL, 4.2 μL, 4.3 μL, 4.4 μL, 4.5 μL, 4.6 μL, 4.7 μL, 4.8 μL, 4.9 μL, 5.0 μL, 5.1 μL, 5.2 μL, 5.3 μL, 5.4 μL, 5.5 μL, and any value between any two of the aforementioned values.

[0061] In some specific embodiments, the chromatographic conditions include: injection volume: 1 μL. In some specific embodiments, the chromatographic conditions include: injection volume: 5 μL.

[0062] In some specific embodiments, the chromatographic conditions include: column temperature: 35℃~45℃; flow rate: 250μL / min~350μL / min; in some specific embodiments, the column temperature can be 35℃, 36℃, 37℃, 38℃, 39℃, 40℃, 41℃, 42℃, 43℃, 44℃, 45℃, or any value between any two of the aforementioned values. In some specific embodiments, the flow rate can be 250μL / min, 260μL / min, 270μL / min, 280μL / min, 290μL / min, 300μL / min, 310μL / min, 320μL / min, 330μL / min, 340μL / min, 350μL / min, or any value between any two of the aforementioned values.

[0063] In some specific embodiments, the chromatographic conditions include: column temperature: 40°C; flow rate: 300 μL / min.

[0064] In some specific embodiments, mass spectrometry employs a Q-Exactive quadrupole electrostatic field orbital trap high-resolution mass spectrometer.

[0065] In some specific embodiments, the mass spectrometer uses a heated HESI source with a capillary voltage of -3.7 kV to +3.7 kV; in some specific embodiments, the capillary temperature is 150℃ to 250℃; the sheath gas is 35 alb to 45 alb; the auxiliary gas is 10 alb to 20 alb; the ion transmission tube temperature is 300℃ to 400℃; the scanning mode is Full MS / ddMS2, and the scanning range is 150-600 m / z, with simultaneous acquisition of positive and negative ions. The capillary temperature of 150℃ to 250℃ can, in some specific embodiments, be 150℃, 160℃, 170℃, 180℃, 190℃, 200℃, 210℃, 220℃, 230℃, 240℃, 250℃, or any value between any two of the aforementioned values.

[0066] The sheath gas is 35arb to 45arb, and in some specific embodiments it can be 35arb, 36arb, 37arb, 38arb, 39arb, 40arb, 41arb, 42arb, 43arb, 44arb, 45arb, or any value between any two of the aforementioned values.

[0067] The auxiliary gas is 10arb to 20arb, and in some specific embodiments it can be 10arb, 11arb, 12arb, 13arb, 14arb, 15arb, 16arb, 17arb, 18arb, 19arb, 20arb, or any value between any two of the aforementioned values.

[0068] The temperature of the ion transmission tube is 300℃~400℃, and in some specific embodiments it can be 300℃, 310℃, 320℃, 330℃, 340℃, 350℃, 360℃, 370℃, 380℃, 390℃, 400℃, or any value between any two of the aforementioned values.

[0069] The first-level mass spectrometry resolution is 70,000, and the second-level mass spectrometry resolution is 17,500; the normalized collision energy (NCE) is 30, 45, and 60 eV.

[0070] Preferably, the mass spectrometry conditions include: a heated HESI source; capillary voltage: ±3.7 kV; capillary temperature: 200 °C; sheath gas: 40 arb; auxiliary gas: 15 arb; purge gas: 0 arb; ion transmission tube temperature: 350 °C; Full MS / ddMS2 scanning mode; scanning range: 150-600 m / z, simultaneous acquisition of positive and negative ions; primary mass spectrometry resolution of 70,000, secondary mass spectrometry resolution of 17,500; normalized collision energy (NCE) of 30, 45, and 60 eV.

[0071] In some specific embodiments, the bisphenol compounds include at least one of the following: bisphenol A, bisphenol B, bisphenol C, bisphenol F, bisphenol S, bisphenol Z, bisphenol AF, bisphenol AP, bisphenol P, 4,4-sulfonyl di(2-methylphenol), tetrachlorobisphenol A, tetrabromobisphenol A, bisphenol A diglycidyl ether, bisphenol A-(2-3-dihydroxypropyl)glycerol ether, bisphenol A-(2-3-dihydroxypropyl) ether, bisphenol A-(3-chloro-2-hydroxypropyl)glycerol ether, bisphenol A-(3-chloro-2-hydroxypropyl) ether, bisphenol A-(3-chloro-2-hydroxypropyl)(2-3-dihydroxypropyl) ether, bisphenol F diglycidyl ether, bisphenol F-(2-3-dihydroxypropyl) ether, and bisphenol F-(3-chloro-2-hydroxypropyl) ether.

[0072] Example 1.

[0073] I. Pre-processing

[0074] Thirty different types of baijiu were tested. 5 ml of each baijiu sample was placed in a 15 ml nitrogen blow-off tube. The nitrogen blow-off tube was set with the following parameters: gas pressure 3 psi, needle speed 1.0 mm / min, concentration temperature 60℃. After blowing the nitrogen down to 0.5 ml, the volume was adjusted to 1 ml with methanol, transferred to a glass centrifuge tube, and centrifuged at high speed for analysis.

[0075] II. Preparation of Standard Solutions

[0076] Accurately weigh 0.0500 g of each of the 21 bisphenols and their derivatives standards, dissolve them in methanol, and dilute to 50 mL in a brown volumetric flask. Shake well to obtain a standard stock solution with a concentration of 1000 mg / L, and store at 4°C for later use. Dilute with 50% methanol and water to obtain mixed standard working solutions of the 21 bisphenols and their derivatives with concentrations of 0.5, 1.0, 2.5, 5, 10, 25, and 50 μg / L, respectively.

[0077] III. Instrumental Analysis

[0078] Chromatographic conditions: Column: PFP (2.1 mm × 50 mm, 2.6 μm); Column temperature: 40 ℃; Injection volume: 1 μL; Flow rate: 300 μL / min; Mobile phase: A is aqueous solution, B is methanol; Gradient elution conditions: 0-2 min, 30% B, 2-8 min, 90% B, 8-10 min, 100% B, the entire analytical process takes 10 min.

[0079] A Q-Exactive quadrupole electrostatic field orbital trap high-resolution mass spectrometer was used. The mass spectrometry conditions were as follows: heated HESI source; capillary voltage: ±3.7 kV; capillary temperature: 200 °C; sheath gas: 40 arb (1 arb ≈ 0.3 L / min); auxiliary gas: 15 arb; purge gas: 0 arb; ion transmission tube temperature: 350 °C; Full MS / ddMS2 scan mode; scan range: 150-600 m / z; simultaneous acquisition of positive and negative ions; first-stage mass spectrometry resolution: 70,000; second-stage mass spectrometry resolution: 17,500; normalized collision energy (NCE): 30, 45, and 60 eV.

[0080] IV. Data Processing

[0081] Data acquisition and mass spectrometry analysis were performed using the instrument's built-in data analysis software Xcalibur 4.2, while the fragmentation pattern was studied using MassFrontier 7.0 software.

[0082] In this application, the precise mass number, ionization mode, retention time, qualitative ion, and detection limit of each standard are shown in Table 4 below.

[0083] Analysis revealed that among the aforementioned 30 samples, one sample tested positive for BADGE at a concentration of 1.5 μg / L, while the other products did not contain it.

[0084] Accuracy statement:

[0085] The mass spectrometry method described above in this application was used to analyze the aforementioned bisphenol A (BPA), bisphenol B (BPB), bisphenol C (BPC), bisphenol F (BPF), bisphenol S (BPS), bisphenol Z (BPZ), bisphenol AF (BPAF), bisphenol AP (BPAP), bisphenol P (BPP), 4,4-sulfonyl di(2-methylphenol) (DMBPS), tetrachlorobisphenol A (TCBPA), and tetrabromobisphenol A (TCBPA). A, TBBPA), Bisphenol A diglycidyl ether (Bisphenol A-diglycidyl, BADGE), Bisphenol A-(2,3-dihydroxypropyl)glycerol ether (BADGE-H2O), Bisphenol A-(2,3-dihydroxypropyl) ether (BADGE-2H2O), Bisphenol A-(3-chloro-2-hydroxypropyl)glycerol ether (BADGE-HCl), Bisphenol A-(3-chloro-2-hydroxypropyl) ether (BADGE-2HCl), Bisphenol A-(3-chloro-2-hydroxypropyl)(2,3-dihydroxypropyl) A series of mixed standard solutions of 21 bisphenols and their derivatives with standard mass concentrations ranging from 0.5 to 50 μg / L, including bisphenol F diglycidyl ether (BADGE-H2O-HCl), bisphenol F diglycidyl ether (BFDGE), bisphenol F-(2-3-dihydroxypropyl ether) (BFDGE-2H2O), and bisphenol F-(3-chloro-2-hydroxypropyl) ether (BFDGE-2HCl), were determined. Standard curves were plotted with the peak area y of the target component against the corresponding mass concentration x (μg / L), and the correlation coefficients were all greater than 0.9965. The detection limit was determined using actual standard samples. The linear equation, correlation coefficient, instrument detection limit, and method quantitation limit are shown in Table 1. The instrument detection limit is between 0.18 μg / L and 1.09 μg / L, and the quantitation limit applied for is between 0.03 μg / L and 0.69 μg / L (the quantitation limit in terms of the volume of the liquor before reduction and concentration). The sensitivity is improved by 12 times to 210 times.

[0086] Table 1. Linearity, Instrument Detection Limit, and Method Quantification Limit for 121 Target Substances

[0087]

[0088]

[0089] Spiking experiments were performed on blank samples, with three different mass concentrations of the target analyte added. Six parallel experiments were conducted for each spiking level. The average recovery and precision (expressed as relative standard deviation, RSD) were calculated, and the results are shown in Table 2. The recoveries of the target analyte ranged from 60.6% to 112.3%, with RSDs ranging from 0.6% to 15.7%.

[0090] Table 2. Spike recoveries and precision of 21 target analytes (n=6)

[0091]

[0092]

[0093]

[0094] Twenty-one bisphenol compounds and their derivatives were extracted. The target compounds showed good peak shapes and high responses, meeting the analytical requirements of the target compounds. The chromatograms of the target compounds extracted with quasi-molecular ion precise mass numbers are shown in Figure 1. As can be seen from Figure 1, each target compound can be effectively separated and quantified using the method of this application.

[0095] Example 2

[0096] The experimental method of Example 2 differs from that of Example 1 in that the same liquor sample was used, but the nitrogen blowing temperature was changed to 40℃ and 90℃ respectively. This example did not detect the content of the final sample, but the spiked recovery rate of the sample was detected under the same conditions as the example.

[0097] The specific testing method is as follows:

[0098] I. Pre-processing

[0099] The same baijiu sample was divided into multiple portions. 5 ml of each baijiu sample was placed in a 15 ml nitrogen blow-off tube. The nitrogen blow-off tube was set with the following parameters: gas pressure 3 psi, needle speed 1.0 mm / min, and concentration temperature 40℃ and 90℃, respectively. After blowing the nitrogen down to 0.5 ml, the volume was adjusted to 1 ml with methanol. The mixture was then transferred to a glass centrifuge tube, centrifuged at high speed, and then analyzed.

[0100] II. Preparation of Standard Solutions

[0101] Accurately weigh 0.0500 g of each of the 21 bisphenols and their derivatives standards, dissolve them in methanol, and dilute to 50 mL in a brown volumetric flask. Shake well to obtain a standard stock solution with a concentration of 1000 mg / L, and store at 4°C for later use. Dilute with 50% methanol and water to obtain mixed standard working solutions of the 21 bisphenols and their derivatives with concentrations of 0.5, 1.0, 2.5, 5, 10, 25, and 50 μg / L, respectively.

[0102] III. Instrumental Analysis

[0103] Chromatographic conditions: Column: PFP (2.1 mm × 50 mm, 2.6 μm); Column temperature: 40 ℃; Injection volume: 1 μL; Flow rate: 300 μL / min; Mobile phase: A is aqueous solution, B is methanol; Gradient elution conditions: 0-2 min, 30% B, 2-8 min, 90% B, 8-10 min, 100% B, the entire analytical process takes 10 min.

[0104] A Q-Exactive quadrupole electrostatic field orbital trap high-resolution mass spectrometer was used. The mass spectrometry conditions were as follows: heated HESI source; capillary voltage: ±3.7 kV; capillary temperature: 200 °C; sheath gas: 40 arb (1 arb ≈ 0.3 L / min); auxiliary gas: 15 arb; purge gas: 0 arb; ion transmission tube temperature: 350 °C; Full MS / ddMS2 scan mode; scan range: 150-600 m / z; simultaneous acquisition of positive and negative ions; first-stage mass spectrometry resolution: 70,000; second-stage mass spectrometry resolution: 17,500; normalized collision energy (NCE): 30, 45, and 60 eV.

[0105] IV. Data Processing

[0106] Data acquisition and mass spectrometry analysis were performed using the instrument's built-in data analysis software Xcalibur 4.2, while the fragmentation pattern was studied using MassFrontier 7.0 software.

[0107] The specific results obtained are as follows:

[0108] Using a recovery rate of 1 μg / L as the criterion, compared with the 60℃ in the examples, the results are shown in Table 3. The BPA recovery rate increased with increasing temperature, but at 40℃ and 90℃, the recovery rates were too low or too high, failing to meet the requirements of GB / T27404-2008 "Laboratory Quality Control Scope - Physicochemical Testing of Food". At a nitrogen blowing temperature of 90℃, the recovery rates of compounds such as BADGE and BADGE-HCl were too low, failing to meet the requirements. At a nitrogen blowing temperature of 60℃, the recovery rates of BPA, BPS, BADGE, BADGE-HCl, and BADGE-H2O were between 63.0% and 113.1%, meeting the requirements of GB / T27404-2008 "Laboratory Quality Control Scope - Physicochemical Testing of Food", therefore, a nitrogen blowing temperature of 60℃ was selected.

[0109] Table 3 Recovery rate (%) at different nitrogen blowing temperatures (1 μg / L spiked)

[0110] compound 40℃ 60℃ 90℃ BPA 44.6 113.1 141.8 BPS 71.1 70.7 100.9 BADGE 71.9 71.3 57.8 BADGE-HCL 63.7 63.0 55.5 <![CDATA[BADGE-H2O]]> 79.2 85.0 86.8

[0111] At the above different nitrogen blowing temperatures, it takes 23 minutes at 90℃ to reduce a 5ml wine sample to 0.5ml by nitrogen blowing, 45 minutes at 60℃, and 75 minutes at 40℃. It can be seen that the nitrogen blowing temperature of this application can achieve both rapid concentration and high recovery rate, thereby ensuring detection accuracy.

[0112] Example 3

[0113] The experimental method in Example 3 differs from that in Example 1 in that the same liquor sample was used, but the volumetric solvent was changed to water. This example did not perform content detection on the final sample, but rather detected the spiked recovery rate. The specific test method is as follows:

[0114] I. Pre-processing

[0115] The same baijiu sample was divided into multiple portions. 5 ml of each baijiu sample was placed in a 15 ml nitrogen blow-off tube. The nitrogen blow-off tube was set with the following parameters: gas pressure 3 psi, needle speed 1.0 mm / min, concentration temperature 60℃. Nitrogen was blown down to 0.5 ml, and the volume was adjusted to 1 ml with water. The mixture was then transferred to a glass centrifuge tube, centrifuged at high speed, and then analyzed.

[0116] II. Preparation of Standard Solutions

[0117] Accurately weigh 0.0500 g of each of the 21 bisphenols and their derivatives standards, dissolve them in methanol, and dilute to 50 mL in a brown volumetric flask. Shake well to obtain a standard stock solution with a concentration of 1000 mg / L, and store at 4°C for later use. Dilute with 50% methanol and water to obtain mixed standard working solutions of the 21 bisphenols and their derivatives with concentrations of 0.5, 1.0, 2.5, 5, 10, 25, and 50 μg / L, respectively.

[0118] III. Instrumental Analysis

[0119] Chromatographic conditions: Column: PFP (2.1 mm × 50 mm, 2.6 μm); Column temperature: 40 ℃; Injection volume: 1 μL; Flow rate: 300 μL / min; Mobile phase: A is aqueous solution, B is methanol; Gradient elution conditions: 0-2 min, 30% B, 2-8 min, 90% B, 8-10 min, 100% B, the entire analytical process takes 10 min.

[0120] A Q-Exactive quadrupole electrostatic field orbital trap high-resolution mass spectrometer was used. The mass spectrometry conditions were as follows: heated HESI source; capillary voltage: ±3.7 kV; capillary temperature: 200 °C; sheath gas: 40 arb (1 arb ≈ 0.3 L / min); auxiliary gas: 15 arb; purge gas: 0 arb; ion transmission tube temperature: 350 °C; Full MS / ddMS2 scan mode; scan range: 150-600 m / z; simultaneous acquisition of positive and negative ions; first-stage mass spectrometry resolution: 70,000; second-stage mass spectrometry resolution: 17,500; normalized collision energy (NCE): 30, 45, and 60 eV.

[0121] IV. Data Processing

[0122] Data acquisition and mass spectrometry analysis were performed using the instrument's built-in data analysis software Xcalibur 4.2, while the fragmentation pattern was studied using MassFrontier 7.0 software.

[0123] The specific results obtained are as follows Figure 2 As shown:

[0124] In Example 1, when methanol was used for volume adjustment, the average recoveries of the added standard (hereinafter referred to as the spiking concentration) at concentrations of 0.5 μg / L, 1.0 μg / L, and 2.5 μg / L were 68.3%, 78.2%, and 65.6%, respectively; while in Example 3, when water was used for volume adjustment, the average recoveries were 68.3%, 78.2%, and 65.6%, respectively. Figure 2 As shown, the recoveries at different spiking concentrations were all below 60%.

[0125] Comparative Example 1

[0126] The experimental method of Comparative Example 1 differs from that of Example 1 in that the same liquor sample was used, and 0.05% and 0.1% formic acid were added to mobile phase A. The analysis revealed that after adding the aforementioned two concentrations of formic acid, the response of bisphenol diglycidyl ethers such as BADGE and BFDGE and their derivatives in the HESI+ mode was not enhanced compared to Example 1, but bisphenols such as BPA, BPB, BPC, BPF, BPAP, BPP, and BPZ and their derivatives did not show peaks.

[0127] Example 4

[0128] The comparative sample was analyzed using an Eclipse XDB-C18 (2.1mm × 150mm, 1.8μm) with the other conditions being the same as in Example 1. The analysis time was found to have changed by 20 min.

[0129] Example 5

[0130] This comparative example used the same liquor sample as the test sample, with an injection volume of 5 μL for analysis. All other conditions were the same as in Example 1. The specific detection method is as follows:

[0131] I. Pre-processing

[0132] The same baijiu sample was divided into multiple portions. 5 ml of each baijiu sample was placed in a 15 ml nitrogen blow-off tube. The nitrogen blow-off tube was set with the following parameters: gas pressure 3 psi, needle speed 1.0 mm / min, and concentration temperature 60℃. After blowing the nitrogen down to 0.5 ml, the volume was adjusted to 1 ml with methanol. The mixture was then transferred to a glass centrifuge tube, centrifuged at high speed, and then analyzed.

[0133] II. Preparation of Standard Solutions

[0134] Accurately weigh 0.0500 g of each of the 21 bisphenols and their derivatives standards, dissolve them in methanol, and dilute to 50 mL in a brown volumetric flask. Shake well to obtain a standard stock solution with a concentration of 1000 mg / L, and store at 4°C for later use. Dilute with 50% methanol and water to obtain mixed standard working solutions of the 21 bisphenols and their derivatives with concentrations of 0.5, 1.0, 2.5, 5, 10, 25, and 50 μg / L, respectively.

[0135] III. Instrumental Analysis

[0136] Chromatographic conditions: Column: PFP (2.1 mm × 50 mm, 2.6 μm); Column temperature: 40 ℃; Injection volume: 5 μL; Flow rate: 300 μL / min; Mobile phase: A is aqueous solution, B is methanol; Gradient elution conditions: 0-2 min, 30% B, 2-8 min, 90% B, 8-10 min, 100% B, the entire analytical process takes 10 min.

[0137] A Q-Exactive quadrupole electrostatic field orbital trap high-resolution mass spectrometer was used. The mass spectrometry conditions were as follows: heated HESI source; capillary voltage: ±3.7 kV; capillary temperature: 200 °C; sheath gas: 40 arb (1 arb ≈ 0.3 L / min); auxiliary gas: 15 arb; purge gas: 0 arb; ion transmission tube temperature: 350 °C; Full MS / ddMS2 scan mode; scan range: 150-600 m / z; simultaneous acquisition of positive and negative ions; first-stage mass spectrometry resolution: 70,000; second-stage mass spectrometry resolution: 17,500; normalized collision energy (NCE): 30, 45, and 60 eV.

[0138] IV. Data Processing

[0139] Data acquisition and mass spectrometry analysis were performed using the instrument's built-in data analysis software Xcalibur 4.2, while the fragmentation pattern was studied using MassFrontier 7.0 software.

[0140] The detection limits compared with those of Example 1 are shown in Table 4 below.

[0141] Table 4

[0142]

[0143]

[0144] Increasing the sample injection volume from 1 μL to 5 μL and using the actual standard concentration as the basis for detection, the results showed that the instrument detection limits for 21 bisphenols and their derivatives were further reduced, with an average reduction of 18%.

[0145] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for quantifying trace amounts of bisphenol compounds in wine, characterized in that, Includes the following steps: I. Sample pretreatment: The wine sample to be tested is concentrated to obtain a concentrated solution. The concentrated solution is then brought to a fixed volume. The concentrated solution after volume adjustment is then subjected to refrigerated centrifugation. The supernatant obtained after centrifugation is used as the solution to be analyzed. The volume-fixing solvent used in the volume-fixing process is methanol; II. Sample Quantification: The solution to be analyzed was quantitatively analyzed using chromatography-mass spectrometry.

2. The quantitative method as described in claim 1, characterized in that, The concentration includes nitrogen blowing concentration.

3. The quantitative method as described in claim 2, characterized in that, The nitrogen blowing concentration temperature is 50–70°C.

4. The quantitative method as described in claim 1, characterized in that, The concentration of the concentrate is 10 to 15 times that of the wine sample to be tested.

5. The quantitative method as described in claim 1, characterized in that, The chromatographic conditions include: mobile phase A is an aqueous solution and mobile phase B is methanol.

6. The quantitative method as described in claim 1, characterized in that, The chromatographic conditions included a PFP column (2.1 mm × 50 mm, 2.6 μm).

7. The quantitative method as described in claim 1, characterized in that, The chromatographic conditions for the chromatography include: injection volume: 0.5–5.5 μL.

8. The quantitative method as described in claim 1, characterized in that, The chromatographic conditions include: column temperature: 35℃~45℃; flow rate: 250μL / min~350μL / min.

9. The quantitative method as described in claim 1, characterized in that, The mass spectrometry was performed using a Q-Exactive quadrupole electrostatic field track trap high-resolution mass spectrometer. Preferably, the mass spectrometry conditions include: a heated HESI source; capillary voltage: -3.7kV to +3.7kV; capillary temperature: 150℃ to 250℃; sheath gas: 35arb to 45arb; auxiliary gas: 10arb to 20arb; ion transmission tube temperature: 300℃ to 400℃; scanning mode: Full MS / ddMS2; scanning range: 150m / z to 600m / z.

10. The quantitative method as described in claim 1, characterized in that, The bisphenol compounds include at least one of the following: bisphenol A, bisphenol B, bisphenol C, bisphenol F, bisphenol S, bisphenol Z, bisphenol AF, bisphenol AP, bisphenol P, 4,4-sulfonyl di(2-methylphenol), tetrachlorobisphenol A, tetrabromobisphenol A, bisphenol A diglycidyl ether, bisphenol A-(2-3-dihydroxypropyl)glycerol ether, bisphenol A-(2-3-dihydroxypropyl) ether, bisphenol A-(3-chloro-2-hydroxypropyl)glycerol ether, bisphenol A-(3-chloro-2-hydroxypropyl) ether, bisphenol A-(3-chloro-2-hydroxypropyl)(2-3-dihydroxypropyl) ether, bisphenol F diglycidyl ether, bisphenol F-(2-3-dihydroxypropyl) ether, and bisphenol F-(3-chloro-2-hydroxypropyl) ether.

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