Method for determining four mangnolia officinalis extracts in import and export cosmetics

Abstract

The invention discloses a method for determining four mangnolia officinalis extracts in import and export cosmetics, which is used for detecting magnolol, honokiol, magnoflorine and magnoliavine, and comprises the following steps: S1, uniformly mixing a cosmetic sample with a solvent; s2, after uniform mixing, carrying out vortex oscillation treatment on the sample; s3, after vortex oscillation treatment, carrying out ultrasonic extraction on the sample at room temperature to obtain an extracting solution; s4, mixing the extracting solution with an adsorbent to obtain a purified to-be-detected solution; and S5, filtering the to-be-detected liquid, and determining the to-be-detected liquid through LC-MS / MS to obtain a detection result. According to the present invention, the target object in the cosmetics with different matrix types can be effectively represented, the core functional components such as magnolol and honokiol in the cortex magnoliae officinalis extract can be detected, the qualitative and quantitative analysis can be further performed on the potential supervision risk substance, and the dual functions of quality evaluation and safety supervision can be provided for the cosmetics.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a method for determining four Magnolia officinalis extracts in imported and exported cosmetics. Background Technology

[0002] Magnolia officinalis extract is a natural active substance extracted from the bark, root bark, or branch bark of the Magnolia officinalis plant (Magnolia officinalis). Its main components include magnolol, honokiol, magnocuraneine, and magnoflorine. Magnolia officinalis extract possesses multiple biological activities and can be used in cosmetics. In terms of antibacterial activity, it exhibits significant broad-spectrum inhibitory effects against Gram-positive bacteria, Gram-negative bacteria, and fungi. In the anti-inflammatory and antioxidant fields, honokiol and honokiol demonstrate excellent free radical scavenging capabilities, with scavenging efficiency approaching that of ascorbic acid and significantly superior to the vitamin E analog Trolox, effectively reducing oxidative stress damage and alleviating chronic inflammation. Furthermore, the alkaloids in Magnolia officinalis extract have a significant muscle relaxant effect, inhibiting striated muscle contraction through a non-polarizing neuromuscular blocking mechanism. However, these alkaloids may cause neurotoxicity at high doses; therefore, the concentration of relevant alkaloids is strictly controlled (generally ≤0.2%) when Magnolia officinalis extract is used in leave-on cosmetics.

[0003] Magnolia officinalis extract faces multiple challenges in practical applications: on the one hand, alkaloids such as magnocurarine require strict dosage control due to their potential neurotoxicity; on the other hand, the qualitative and quantitative detection of magnololoids is crucial for product quality evaluation, yet a unified standard for multi-component detection methods of Magnolia officinalis extract has not been established, which to some extent limits product quality monitoring and safety assessment. Therefore, this invention aims to propose a method for the determination of Magnolia officinalis extract to address the aforementioned problems. Summary of the Invention

[0004] To address the aforementioned technical problems, the present invention aims to provide a method for determining four Magnolia officinalis extracts in imported and exported cosmetics. This method can effectively characterize target substances in cosmetics with various matrix types. It can not only detect the core functional components of Magnolia officinalis extract, magnolol and honokiol, but also perform qualitative and quantitative analysis on substances with potential regulatory risks. This method has the dual functions of quality evaluation and safety supervision for cosmetics.

[0005] To achieve the above-mentioned objectives, the technical solution adopted by the present invention is as follows:

[0006] A method for determining four Magnolia officinalis extracts in imported and exported cosmetics, targeting magnolol, honokiol, magnoflorine, and magnocuraneine, includes the following steps:

[0007] S1. Mix the cosmetic sample with the solvent until homogeneous;

[0008] S2. After mixing, the sample is subjected to vortex oscillation.

[0009] S3. After vortex oscillation treatment, the sample is subjected to ultrasonic extraction at room temperature to obtain the extract;

[0010] S4. Mix the extract with the adsorbent to obtain the purified test solution;

[0011] S5. After filtering the test solution, the results are obtained by LC-MS / MS.

[0012] Preferably, in step S1, the solvent includes one or more of methanol, acetonitrile, or ethanol. More preferably, the solvent is methanol, which has low toxicity and low cost while ensuring extraction efficiency.

[0013] Preferably, in step S2, the vortex oscillation processing time is 30 to 60 seconds.

[0014] Preferably, in step S3, the ultrasonic extraction time is 10–30 min. More preferably, the ultrasonic extraction time is 20 min.

[0015] Preferably, in step S4, the adsorbent includes one or more of n-hexane, silica gel, PSA, and C18.

[0016] More preferably, for cosmetic samples with cream, liquid or mask type matrix, in step S4, the extract is centrifuged to obtain supernatant, the supernatant is mixed with silica gel / PSA / C18 and vortexed, and then filtered to obtain the test solution.

[0017] More preferably, for cosmetic samples with cream-type magnolol, when determining the content of magnolol, magnolol, magnoflorine or magnocurane, the adsorbent is PSA.

[0018] More preferably, for cosmetic samples with an aqueous matrix, when determining the content of magnolol / and magnolol, the adsorbent is silica gel, and when determining the content of magnoflorine / magnolucurine, the adsorbent is C18.

[0019] More preferably, for cosmetic samples with a mask-type matrix, when determining the content of magnolol / and honokiol, the adsorbent is silica gel, and when determining the content of magnoflorine / magnocurane, the adsorbent is PSA.

[0020] More preferably, for cosmetic samples with an oil-based matrix, in step S4, the extract is mixed with n-hexane and vortexed, then frozen at -25°C in the dark to separate the n-hexane and methanol phases, the upper n-hexane phase is removed, and the lower methanol phase is filtered to obtain the test solution.

[0021] Beneficial effects:

[0022] This invention innovatively utilizes LC-MS / MS technology to effectively characterize target compounds in cosmetics with various matrix types. It can not only detect the core functional components magnolol and honokiol in Magnolia officinalis extract, but also perform qualitative and quantitative analysis on substances with potential regulatory risks (such as magnoflorine and magnocuraneine). The determination method of this invention has dual functions of quality evaluation and safety supervision for cosmetics. Moreover, it successfully overcomes the significant matrix effect by screening the four most widely used cosmetic matrices for Magnolia officinalis extract through purification methods, ensuring the broad applicability of the detection method. Attached Figure Description

[0023] Figure 1 The image shows the liquid chromatogram of Magnolia officinalis extract obtained from a C18 column. The left image contains magnolol and honokiol; the right image contains magnoflorine and magnocuraneine.

[0024] Figure 2 The figure shows the effect of different extraction times on the sample extraction rate. The top left figure shows the test results for magnolol, the top right figure shows the test results for magnolol, the bottom left figure shows the test results for magnoflorine, and the bottom right figure shows the test results for magnoflorine.

[0025] Figure 3 The following are the LC-MS / MS spectra of four Magnolia officinalis extracts: a is the LC-MS / MS spectrum of magnolol, b is the LC-MS / MS spectrum of magnolol, c is the LC-MS / MS spectrum of magnoflorine, and d is the LC-MS / MS spectrum of magnoflorine. Detailed Implementation

[0026] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the specific implementation methods of the present invention will be described below with reference to the accompanying drawings. Obviously, the drawings described below are merely some embodiments of the present invention. For those skilled in the art, other drawings and other implementation methods can be obtained based on these drawings without any creative effort.

[0027] This invention proposes a method for determining four Magnolia officinalis extracts in imported and exported cosmetics, specifically targeting the detection of magnolol, honokiol, magnoflorine, and magnocuraneine, comprising the following steps:

[0028] S1. Mix the cosmetic sample with the solvent until homogeneous;

[0029] S2. After mixing, the sample is subjected to vortex oscillation.

[0030] S3. After vortex oscillation treatment, the sample is subjected to ultrasonic extraction at room temperature to obtain the extract;

[0031] S4. Mix the extract with the adsorbent to obtain the purified test solution;

[0032] S5. After filtering the test solution, the results are obtained by liquid chromatography-tandem mass spectrometry (LC-MS / MS).

[0033] Preferably, in step S1, the solvent includes one or more of methanol, acetonitrile, or ethanol. More preferably, the solvent is methanol, which has low toxicity and low cost while ensuring extraction efficiency.

[0034] Preferably, in step S2, the vortex oscillation processing time is 30 to 60 seconds.

[0035] Preferably, in step S3, the ultrasonic extraction time is 10–30 min. More preferably, the ultrasonic extraction time is 20 min. This invention does not limit the ultrasonic extraction power; commonly used ultrasonic power in the art can be used.

[0036] Preferably, in step S4, the adsorbent includes one or more of n-hexane, silica gel, PSA, and C18.

[0037] More preferably, for cosmetic samples with cream, liquid or mask type matrix, in step S4, the extract is centrifuged to obtain supernatant, the supernatant is mixed with silica gel / PSA / C18 and vortexed, and then filtered to obtain the test solution.

[0038] More preferably, for cosmetic samples with cream-type magnolol, when determining the content of magnolol, magnolol, magnoflorine or magnocurane, the adsorbent is PSA.

[0039] More preferably, for cosmetic samples with an aqueous matrix, when determining the content of magnolol / and magnolol, the adsorbent is silica gel, and when determining the content of magnoflorine / magnolucurine, the adsorbent is C18.

[0040] More preferably, for cosmetic samples with a mask-type matrix, when determining the content of magnolol / and honokiol, the adsorbent is silica gel, and when determining the content of magnoflorine / magnocurane, the adsorbent is PSA.

[0041] More preferably, for cosmetic samples with an oil-based matrix, in step S4, the extract is mixed with n-hexane and vortexed, then frozen at -25°C in the dark to separate the n-hexane and methanol phases, the upper n-hexane phase is removed, and the lower methanol phase is filtered to obtain the test solution.

[0042] Preferably, in step S4, the mass spectrometry conditions are as follows:

[0043] Magnolol and magnolol were characterized using anionic mode, while magnoflorine and magnocuraneine were detected using cationic mode.

[0044] Other mass spectrometry conditions are as follows: ion source temperature: 550℃; ion spray voltage: 4000V; curtain gas flow: 170kPa; collision gas flow: 68kPa; auxiliary gas flow: 340kPa.

[0045] This invention innovatively utilizes LC-MS / MS technology to effectively characterize target substances in various cosmetics with different matrix types, such as oils, waters, creams, and masks. It can not only detect the core functional components magnolol and honokiol in Magnolia officinalis extract, but also perform qualitative and quantitative analysis of potentially regulatory risk substances (such as magnoflorine and magnocurarine). The assay method of this invention has dual functions of quality evaluation and safety supervision for cosmetics. Furthermore, by employing purification methods to screen the four most widely used cosmetic matrices for Magnolia officinalis extract, it successfully overcomes significant matrix effects, ensuring the broad applicability of the assay method.

[0046] This invention significantly improves analytical efficiency. Compared to existing detection methods that require over 1 hour per test (including approximately 38 minutes of scanning time and lengthy heating and cooling procedures), this invention, using LC-MS / MS analysis, reduces the scanning time to 5-10 minutes and eliminates the need for an additional temperature equilibration process. While improving analytical efficiency, this invention also significantly enhances detection sensitivity. The limit of quantitation (LOQ) of this invention reaches 0.01 μg / mL, compared to 10 μg / mL for existing detection methods, representing a 1000-fold increase in sensitivity.

[0047] The present invention significantly reduces operating costs. Existing detection methods are time-consuming, consume large amounts of solvent (100 mL of ethanol extract is required for a single test), and have high energy consumption of the instrument (gas chromatography). The present invention effectively overcomes these cost problems.

[0048] The technical solution of the present invention will be described in detail below with specific embodiments.

[0049] Sample determination method:

[0050] Weigh 0.200g of cosmetic (accurate to 0.001g) into a 50mL brown colorimetric tube, add 10.00mL of solvent, mix well, vortex for 30s, and then sonicate at room temperature (25.0±2.0℃) for 20min to obtain the extract.

[0051] For cosmetic samples with cream, liquid, or mask bases, the extract was centrifuged at 10000 rpm for 5 minutes to obtain the supernatant. 1.0 mL of the supernatant was then mixed with 50.0 mg of silica gel / PSA / C18, etc.

[0052] The QuEChERS adsorbent was mixed and vortexed for 15 min, then filtered through a microporous membrane (0.22 μm) to obtain the test solution.

[0053] For cosmetic samples with an oil-based matrix, in step S4, the extract is mixed with 10.00 mL of n-hexane and vortexed for 30 s. Then, it is frozen at -25 °C in the dark for 3 h to separate the n-hexane and methanol phases. The upper n-hexane phase is removed, and the lower methanol phase is filtered through a microporous membrane (0.22 μm) to obtain the test solution.

[0054] The test solution was analyzed by LC-MS / MS to obtain the detection results.

[0055] Screening of detection conditions:

[0056] Mass spectrometry condition screening:

[0057] The mass spectrometry conditions are shown in Table 1 below. Magnolol and magnolol were characterized using negative ion mode, while magnoflorine and magnocurarine were detected using cation mode. Ions marked with "*" in the table are quantitative ions. Other mass spectrometry conditions are as follows: ion source temperature: 550℃; ion spray voltage: 4000V; curtain gas flow: 170kPa; collision gas flow: 68kPa; auxiliary gas flow: 340kPa.

[0058] Table 1. Multiple reaction monitoring conditions for four Magnolia officinalis extracts using liquid chromatography-tandem mass spectrometry (LC-MS / MS).

[0059]

[0060] Liquid phase condition screening: such as Figure 1 As shown, the C18 column (Agilent Eclipse XDB-C18l) showed good separation and peak shape for magnolol and magnoflorine, while the C18 column (Agilent Extend-C18) showed good separation and peak shape for magnoflorine and magnoflorine-curane. Therefore, the corresponding C18 column was selected as the analytical column.

[0061] Specifically, under the conditions of acetonitrile-0.1% formic acid solution (volume ratio 90:10) at a flow rate of 0.5 mL / min, magnolol and magnocurane can be analyzed within 6 min; under the conditions of acetonitrile-0.1% formic acid + 5 mM ammonium formate solution (volume ratio 80:20) at a flow rate of 0.7 mL / min, magnoflorine and magnocurane can be analyzed within 6 min.

[0062] Sample pretreatment

[0063] Selection of extraction solvent: Based on the above sample determination methods, the effects of methanol, acetonitrile, and ethanol on the extraction efficiency of each component in four types of cosmetic bases were compared. As shown in Table 2, the recovery rate of ethanol was generally >105%. In contrast, the recovery rates of methanol and acetonitrile were closer to 100%. The extraction efficiency of methanol and acetonitrile as extraction solvents met the requirements. Considering that methanol is less toxic than acetonitrile and is cheaper, methanol was preferred as the extraction solvent.

[0064] Table 2 Spike recoveries of different extraction solvents (n=2)

[0065]

[0066] Selection of purification conditions: Based on the above sample determination methods, the purification effects of different adsorbents on the four Magnolia officinalis extracts in the four matrices were confirmed.

[0067] Spiking experiments were conducted using blank samples of aqueous solutions, creams, masks, and oils without Magnolia officinalis extract. Different treatments were used in the spiking experiments, including no purification, hexane liquid phase extraction (low temperature LLE), and purification using C18 (octadecyl bonded silica gel adsorbent) / PSA (N-propylethylenediamine adsorbent) / silica powder QuEChERS. The effects of different methods on the recovery rate of the target analyte were tested (with 100 μg / L methanol standard solution as a reference). The test results are shown in Table 3.

[0068] Using a 100 μg / L methanol standard solution as a reference, the recovery rate of Magnolia officinalis extract was poor without purification treatment, indicating that purification treatment is a necessary prerequisite for ensuring recovery. For magnolol and honokiol, PSA showed the best purification effect on cream matrices, while silica gel powder showed the best purification effect on aqueous and mask matrices. Based on the principle of "like dissolves like" and targeting the mass spectrometry matrix effect caused by lipid components, n-hexane adsorbent achieved the best purification effect on oil matrices. For magnoflorine and magnoflorine, PSA showed the best purification effect on cream and mask matrices, while C18 was most suitable for aqueous matrices.

[0069] Based on the test results, when determining magnolol and honokiol, PSA was used as the purifying agent for cream-type cosmetics, silica powder was used as the purifying agent for water-type and mask-type cosmetics, and n-hexane was used as the purifying agent for oil-type cosmetics; when determining magnoflorine and magnocurarine, PSA was used as the purifying agent for cream-type and mask-type cosmetics, C18 was used as the purifying agent for water-type cosmetics, and n-hexane was used as the purifying agent for oil-type cosmetics.

[0070] Table 3 Spike recoveries under different purification conditions (n=2)

[0071]

[0072] Selection of extraction time: Based on the above sample determination methods, the effect of ultrasonic extraction time on the extraction effect was investigated.

[0073] Spiking experiments were conducted using blank samples (aqueous, cream, mask, and oil formulations without Magnolia officinalis extract) to investigate the recoveries of each component of the extract after sonication for 10, 20, and 30 minutes. The test results are as follows: Figure 2 As shown.

[0074] When the ultrasonic extraction time was extended from 10 min to 20 min, the recovery rate of most samples of each component improved. When the ultrasonic extraction time was extended to 30 min, the recovery rate of most samples of each component no longer increased or decreased.

[0075] Based on the test results, the sample extraction time is 10 to 30 minutes, and preferably 20 minutes.

[0076] Matrix effect investigation: Blank samples (aqueous, cream, mask, and oil) without Magnolia officinalis extract were selected for matrix effect investigation experiments. Specifically, blank matrix solutions and methanol dilutions of standard substances at different concentrations were prepared. LC-MS / MS tests were used to obtain matrix working curves and standard working curves. The slopes of the matrix working curve (k1) and the standard working curve (k2) were substituted into the K-value calculation formula: K = (k1 / k2 - ... 1 The matrix effect K value is obtained by multiplying K by 100%. The K value is used as the evaluation index. If the K value is between -10% and 10%, it indicates that the matrix effect is weak and not obvious. If K < -10%, it shows a strong negative effect. If K > 10%, it shows a strong positive effect.

[0077] The results are shown in Table 4. For magnolol and magnoflorine, the matrix effect generally showed a significant positive effect (K > 10%) in all samples, while for magnoflorine and magnoflorine curare, the matrix effect generally showed a significant negative effect (K < -10%) in all samples. Therefore, this invention uses the external standard method based on the matrix standard working solution curve for quantitative calculation.

[0078] The method for preparing the standard substance blank matrix solution is as follows:

[0079] Accurately weigh 0.200 g of blank matrix cosmetic sample (accurate to 0.001 g) into a 50 mL brown colorimetric tube, add an appropriate amount of standard substance and 10.00 mL of methanol, mix well, vortex for 30 s, and then ultrasonically extract at room temperature (25.0 ± 2.0 ℃) for 20 min to obtain the extract.

[0080] For cosmetic samples with cream, liquid, or mask bases, the extract was centrifuged at 10,000 r / min for 5 min to obtain the supernatant. 1.0 mL of the supernatant was mixed with 50.0 mg of the appropriate QuEChERS adsorbent, such as silica gel / PSA / C18, and vortexed for 15 min. The mixture was then filtered through a microporous membrane (0.22 μm) to obtain the test solution.

[0081] For cosmetic samples with an oil-based matrix, the extract was mixed with 10.00 mL of n-hexane and vortexed for 30 s. Then, it was frozen at -25 °C in the dark for 3 h to separate the n-hexane and methanol phases. The upper n-hexane phase was removed, and the lower methanol phase was filtered through a microporous membrane (0.22 μm) to obtain the test solution.

[0082] Standard substance methanol dilution solution (standard solution):

[0083] Add appropriate amounts of standard substances (honokiol / magnoliol / magnoliol / magnolicurane) and 10.00 mL of methanol to a 50 mL brown colorimetric tube, mix well, vortex for 30 s, and then sonicate for 20 min at room temperature (25.0±2.0℃) to obtain the standard solution.

[0084] Table 4. Matrix effect K values ​​of Magnolia officinalis extract in different samples

[0085]

[0086] Stability Study of Standard Solutions: All four Magnolia officinalis extracts are phenolic compounds with multiple substituted active groups, which are easily oxidized under visible light excitation. Therefore, their standard substances and solutions must be stored away from light. To investigate the stability of the standard substances and the shelf life of the standard solutions, the standard solutions were stored under different temperature conditions and quantitatively analyzed at predetermined intervals. The results are shown in Table 5. Before each analysis, a mixed standard working solution containing the four Magnolia officinalis extracts at a concentration of 100 μg / L was prepared by fresh dilution of the standard solution for injection analysis. The test results showed no significant difference in the stability of the standard solutions under different temperature conditions.

[0087] Based on the test results, the preferred storage conditions for the standard solution and the diluted standard solution used in this invention are: 4°C protected from light, with a shelf life of 6 months.

[0088] Table 5. Experimental data of four Magnolia officinalis extracts in standard solutions under different storage temperatures (w: week, m: month)

[0089]

[0090] Confirmation of working curve and linear range: A series of matrix standard working solutions with different mass concentrations were injected sequentially, and a standard curve was plotted with peak area (y-axis) against mass concentration (x-axis). The concentration of the analyte compound and the corresponding peak area showed a good linear relationship, as shown in Table 6. The liquid chromatography of the standard working solutions is as follows: Figure 3 As shown.

[0091] Based on the test results, when the concentrations of the four Magnolia officinalis extracts in the sample exceed the corresponding linear range, the dilution factor of the sample can be appropriately increased; when the concentrations of the four Magnolia officinalis extracts in the sample are below the corresponding linear range, the purified solution can be appropriately concentrated. Preferably, in this invention, the sample concentration in the test solution is 10.0–200.0 μg / L.

[0092] Table 6 Standard Curve and Correlation Coefficient

[0093]

[0094] Confirmation of Limit of Detection (LOD) and Limit of Quantification (LOQ):

[0095] The detection limit of this invention is 3 times the signal-to-noise ratio (S / N = 3), corresponding to a concentration of 3.0 μg / L for all four Magnolia officinalis extracts. The limit of quantitation was confirmed by adding certified reference materials to the samples; this concentration level corresponded to a signal-to-noise ratio (S / N) greater than 10 times, and its accuracy and precision met the requirements of the method and regulations. The limit of quantitation for all four Magnolia officinalis extracts addressed in this invention is 10.0 μg / L.

[0096] Recovery and precision of the method:

[0097] Additive recovery experiments were conducted on four blank matrices: aqueous solution, cream, mask, and oil. Three different concentration levels of standard solution were added to each matrice, and the results were determined by LC-MS / MS. Six parallel samples were prepared for each concentration level. The measured values, average recovery rate, and relative standard deviation (RSD) of each additive level are shown in Tables 7 to 10.

[0098] Table 7. Spike recovery and precision results of magnolol

[0099]

[0100] Table 8 shows the spiked recoveries and precision results of magnolol.

[0101]

[0102]

[0103] Table 9. Spike recovery and precision results of magnoflorine.

[0104]

[0105] Table 10. Spike recovery and precision results of magnocurarine.

[0106]

[0107] Experimental results show that:

[0108] After spiked with magnolol, the recoveries of the four different matrix samples were as follows: water 101.0–105.8%, cream 99.3–100.2%, face mask 90.6–101.0%, and oil 91.4–104.2%. The RSD ranges were as follows: water 0.99–2.03%, cream 2.40–3.46%, face mask 0.71–2.96%, and oil 0.80–1.41%.

[0109] After spiked with magnolol, the recoveries of the four different matrix samples were as follows: water 100.0–105.5%, cream 91.7–106.0%, face mask 92.1–104.5%, and oil 102.2–105.5%. The RSD ranges were as follows: water 0.77–2.05%, cream 2.16–3.27%, face mask 1.02–2.69%, and oil 1.69–2.22%.

[0110] After spiked with magnoflorine in four different matrix samples, the recoveries ranged as follows: water 97.2–100.7%, cream 98.1–98.7%, face mask 95.1–101.8%, and oil 94.6–101.8%. The RSD ranges were as follows: water 0.68–2.00%, cream 0.80–1.70%, face mask 0.66–1.38%, and oil 0.49–1.21%.

[0111] The recoveries of magnocurarine spiked in four different matrix samples ranged as follows: water 98.2–102.0%, cream 94.0–97.1%, face mask 90.6–102.2%, and oil 91.4–104.2%. The RSD ranges were: water 0.37–1.61%, cream 0.80–1.99%, face mask 0.71–2.12%, and oil 0.80–1.41%.

[0112] By comparing and analyzing the verification results of various matrix liquid chromatography methods in the laboratory, the relative standard deviations were all less than 10%. The statistical results show that the verification results of this method meet the requirements of the relevant provisions of SN / T0001-2016 "Basic Provisions for the Compilation of Standards for Physicochemical Determination Methods of Export Food and Cosmetics". The determination method of this invention can effectively detect four Magnolia officinalis extracts in cosmetics.

[0113] Analysis of imported and exported cosmetic samples:

[0114] The determination method of this invention was used to determine the contents of magnolol, honokiol, magnoflorine and magnocurane in 20 imported and exported cosmetic samples (labeled as containing magnolia bark extract) and 3 magnolia bark extract cosmetic raw materials (oil). The determination data are shown in Table 11.

[0115] Table 11 shows that among the imported and exported cosmetic samples, 18 samples tested positive for magnolol, with contents ranging from 0.0127 to 645 μg / kg (some sample extracts were diluted or concentrated); 16 samples tested positive for honokiol, with contents ranging from 0.0170 to 570 μg / kg (some sample extracts were diluted or concentrated); 1 sample tested positive for magnoflorine, with a content of 0.0206 μg / kg (sample extract was concentrated); and 1 sample tested positive for magnocuronine, with a content of 0.0168 μg / kg (sample extract was concentrated). Among the cosmetic raw materials (oil form), 3 samples tested positive for honokiol, with contents ranging from 16300 to 71500 μg / kg (sample extract was diluted); 3 samples tested positive for honokiol, with contents ranging from 18700 to 91500 μg / kg (sample extract was diluted); magnocuronine and magnocuronine were not detected.

[0116] As can be seen, the determination method of the present invention can effectively detect four kinds of Magnolia officinalis extract in cosmetics with low detection limits. It not only detects the core functional components magnolol and honokiol in Magnolia officinalis extract, but also performs qualitative and quantitative analysis on substances with potential regulatory risks (such as magnoflorine and magnocurane), thus having the dual functions of quality evaluation and safety supervision.

[0117] Table 11 Sample Test Results

[0118]

[0119]

[0120] In summary, the limits of detection for magnolol, honokiol, magnoflorine, and magnocurarine in this invention are 0.15 mg / kg, and the limits of quantitation are 0.50 mg / kg. Within the spiking concentration range of 0.50 mg / kg to 10.0 mg / kg, the recoveries of magnolol in the four matrices were between 90.0% and 108.0%, with a relative standard deviation (RSD) of less than 3.46%; the recoveries of honokiol in the four matrices were between 90.0% and 109.0%, with an RSD of less than 3.27%; the recoveries of magnoflorine in the four matrices were between 93.5% and 103.0%, with an RSD of less than 2.00%; and the recoveries of magnocurarine in the four matrices were between 90.0% and 106.0%, with an RSD of less than 2.12%.

[0121] The embodiments provided by the present invention have been described in detail above. Specific examples have been used to illustrate the principles and implementation methods of the present invention, and the descriptions of the embodiments above are only for the purpose of helping to understand the core ideas of the present invention. It should be noted that those skilled in the art can make several improvements and modifications to the present invention without departing from the principles of the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.

Claims

1. A method for determining four Magnolia officinalis extracts in imported and exported cosmetics, characterized in that, The detection of magnolol, magnolol, magnoflorine, and magnocurane includes the following steps: S1. Mix the cosmetic sample with the solvent until homogeneous; S2. After mixing, the sample is subjected to vortex oscillation. S3. After vortex oscillation treatment, the sample is subjected to ultrasonic extraction at room temperature to obtain the extract; S4. Mix the extract with the adsorbent to obtain the purified test solution; S5. After filtering the test solution, the results are obtained by LC-MS / MS.

2. The determination method according to claim 1, characterized in that, In step S1, the solvent includes one or more of methanol, acetonitrile, or ethanol.

3. The determination method according to claim 1, characterized in that, In step S2, the vortex oscillation processing time is 30–60 s.

4. The determination method according to claim 1, characterized in that, In step S3, the ultrasonic extraction time is 10–30 min.

5. The determination method according to any one of claims 1-4, characterized in that, In step S4, the adsorbent includes one or more of n-hexane, silica gel, PSA, and C18.

6. The determination method according to claim 5, characterized in that, For cosmetic samples with cream, liquid, or mask-type bases, in step S4, the extract is centrifuged to obtain the supernatant, which is then mixed with silica gel / PSA / C18 and vortexed before being filtered to obtain the test solution.

7. The determination method according to claim 6, characterized in that, For cosmetic samples with cream-type magnolol, PSA is used as the adsorbent when determining the content of magnolol, magnolol, magnoflorine, or magnocurane.

8. The determination method according to claim 6, characterized in that, For cosmetic samples with aqueous matrix, silica gel is used as the adsorbent when determining the content of magnolol / magnolanine, and C18 is used as the adsorbent when determining the content of magnoflorine / magnocurane.

9. The determination method according to claim 6, characterized in that, For cosmetic samples with a mask-type matrix, silica gel was used as the adsorbent when determining the content of magnolol / honokiol, and PSA was used as the adsorbent when determining the content of magnoflorine / magnocurane.

10. The determination method according to claim 5, characterized in that, For cosmetic samples with an oil-based matrix, in step S4, the extract is mixed with n-hexane and vortexed, then frozen at -25°C in the dark to separate the n-hexane and methanol phases. The upper n-hexane phase is removed, and the lower methanol phase is filtered to obtain the test solution.

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