Supercritical fluid chromatographic separation method of bakuchiol

Abstract

The invention relates to a supercritical fluid chromatographic separation method of bakuchiol, which comprises the following steps: dispersing a crude product in a solvent to prepare a test sample; carrying out supercritical fluid chromatographic separation on the test sample; the supercritical fluid chromatographic separation conditions are as follows: a mobile phase is a mixture of supercritical carbon dioxide and C1-C3 alcohol compounds in a volume ratio of (99.5-98.5): (0.5-1.5), and isocratic elution is performed; the stationary phase of the chromatographic column is octadecylsilane chemically bonded silica. The supercritical fluid chromatographic separation method of the bakuchiol makes up the research blank of analyzing and separating the bakuchiol by adopting the supercritical fluid chromatography in the traditional method, and can realize accurate detection and effective preparation of the bakuchiol. Meanwhile, the separation method is short in time consumption and good in separation effect.

Description

Technical Field

[0001] The invention relates to the technical field of chromatographic separation and preparation, and in particular to a supercritical fluid chromatographic separation method of bakuchiol. Background Art

[0002] Psoralea corylifolia L. is the dried mature fruit of the leguminous plant Psoralea corylifolia L., which has the functions of warming the kidney and promoting yang, absorbing qi and stopping diarrhea. The main chemical components of Psoralea corylifolia include coumarins, flavonoids, monoterpenoid phenols, lipids, volatile oils, resins, saponins, etc. In 1966, Mehta et al. isolated bakuchiol from ether extracts. Yao Xiangzhen et al. proposed that the toxicity of Psoralea corylifolia may be related to the content of bakuchiol based on the efficacy and toxicity studies of raw, old and new products of Psoralea corylifolia. In addition, studies have found that bakuchiol is an effective antioxidant that can significantly reduce the color difference caused by skin exposure to the external environment and has a soothing effect. In addition, it can also smooth fine lines and wrinkles. These advantages have made bakuchiol appear more and more in skin care products in recent years, and the latest research shows that these skin care products have unique benefits for all skin types when used topically on the skin. Therefore, it is of great significance to separate bakuchiol from the test sample in order to detect or prepare it.

[0003] Supercritical fluid is a state of matter above the critical pressure and critical temperature. It has the characteristics of low viscosity of gas, high density of liquid, and high diffusion coefficient between gas and liquid. Supercritical fluid chromatography (SFC) can make up for the shortcomings of gas chromatography (GC) and high performance liquid chromatography (HPLC). For example, SFC can analyze some substances that are not suitable for GC analysis, such as compounds with strong polarity, strong adsorption, poor thermal stability, and difficult volatility; SFC can analyze various compounds that are difficult to detect by HPLC, such as various natural products and high molecular polymers without UV absorption; compared with GC and HPLC, when processing a large number of samples, not only the analysis efficiency is significantly improved, but also the use of chemical reagents is greatly reduced. At the same time, CO2 resources are abundant, low in price, non-destructive to the analytical and purified compounds, low in reactivity and toxicity, and its critical temperature and pressure are compatible with chromatographs. Based on these technical advantages, SFC has gradually been recognized by the public and its coverage in the market has become increasingly larger. However, there is no method for separating and preparing bakuchiol using supercritical fluid chromatography. Summary of the invention

[0004] Based on this, the present invention provides a supercritical fluid chromatography separation method of bakuchiol which can effectively separate and prepare bakuchiol.

[0005] The specific technical solutions are as follows:

[0006] A supercritical fluid chromatography separation method for bakuchiol comprises the following steps:

[0007] The crude product is dispersed in a solvent to prepare the test sample;

[0008] Subjecting the test sample to supercritical fluid chromatography separation;

[0009] The conditions for the supercritical fluid chromatography separation include:

[0010] The mobile phase is a mixture of supercritical carbon dioxide and C1-C3 alcohol compounds in a volume ratio of (99.5-98.5):(0.5-1.5), and isocratic elution is performed;

[0011] The stationary phase of the chromatographic column is octadecylsilane bonded silica gel.

[0012] In one embodiment, the C1-C3 alcohol compound is methanol.

[0013] In one embodiment, the volume ratio of the supercritical carbon dioxide to the C1-C3 alcohol compound is 99:1.

[0014] In one embodiment, the temperature of the supercritical carbon dioxide when it is pumped in is 5°C to 10°C, and the density is 890kg / m 3 ~950kg / m 3 .

[0015] In one embodiment, the inner diameter of the chromatographic column is 10 mm and the length is 250 mm.

[0016] In one embodiment, the supercritical fluid chromatography separation conditions further include: column temperature is 38° C. to 40° C.; and / or,

[0017] The conditions for supercritical fluid chromatography separation also include: a flow rate of 10 mL / min to 12 mL / min.

[0018] In one embodiment, the supercritical fluid chromatography separation conditions also include: a detection wavelength of 260 nm.

[0019] In one embodiment, the supercritical fluid chromatography separation conditions also include: the system back pressure is 10Mpa~12Mpa.

[0020] In one embodiment, the solvent is n-hexane.

[0021] In one embodiment, the preparation method of the crude product comprises:

[0022] The medicinal material of Psoralea corylifolia was crushed and extracted with petroleum ether.

[0023] This application is based on supercritical fluid chromatography, using a mixture of supercritical carbon dioxide and C1-C3 alcohol compounds in a specific volume ratio as the mobile phase for isocratic elution, and a chromatographic column with an octadecylsilane bonded silica gel as the stationary phase, which can achieve effective separation and preparation of the bakuchiol peak in the test product, fill the research gap in the traditional method of using supercritical fluid chromatography for the analysis and separation of bakuchiol, and can achieve accurate detection and effective preparation of bakuchiol. At the same time, the separation method is short in time and has good separation effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is the separation spectrum of the bakuchiol obtained in Example 1 in a supercritical fluid chromatograph system;

[0025] Figure 2 This is the separation spectrum of the bakuchiol obtained in Comparative Example 1 in a supercritical fluid chromatography system. DETAILED DESCRIPTION

[0026] The supercritical fluid chromatographic separation method of bakuchiol of the present invention is further described in detail below in conjunction with specific examples. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the disclosure of the present invention more thoroughly understood.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0028] The optional scope of the terms "and / or", "or / and", and "and / or" used in this article includes any one of two or more related listed items, and also includes any and all combinations of the related listed items, said any and all combinations include any combination of any two related listed items, any more related listed items, or all related listed items.

[0029] In the present invention, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0030] In the present invention, when it comes to numerical ranges, unless otherwise specified, the above numerical ranges are deemed to be continuous and include the minimum and maximum values ​​of the range, as well as each value between such minimum and maximum values. Further, when a range refers to an integer, each integer between the minimum and maximum values ​​of the range is included. In addition, when multiple ranges are provided to describe features or characteristics, the ranges can be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges included therein.

[0031] The percentage content involved in the present invention, unless otherwise specified, refers to mass percentage for solid-liquid mixing and solid-solid mixing, and refers to volume percentage for liquid-liquid mixing.

[0032] The percentage concentrations involved in the present invention, unless otherwise specified, refer to the final concentration. The final concentration refers to the proportion of the added component in the system after the addition of the component.

[0033] The temperature parameters in the present invention, if not specifically limited, allow for both constant temperature treatment and treatment within a certain temperature range. The constant temperature treatment allows the temperature to fluctuate within the precision range controlled by the instrument.

[0034] The room temperature in the present invention generally refers to 4°C to 30°C, preferably 20±5°C.

[0035] In some examples of the present application, a supercritical fluid chromatography separation method of bakuchiol is provided, comprising the following steps:

[0036] The crude product is dispersed in a solvent to prepare the test sample;

[0037] Subjecting the test sample to supercritical fluid chromatography separation;

[0038] The conditions for the supercritical fluid chromatography separation include:

[0039] The mobile phase is a mixture of supercritical carbon dioxide and C1-C3 alcohol compounds in a volume ratio of (99.5-98.5):(0.5-1.5), and isocratic elution is performed; it can be understood that the time of isocratic elution can be set according to the peak of bakuchiol;

[0040] The stationary phase of the chromatographic column is octadecylsilane bonded silica gel.

[0041] In some of the examples, the C1-C3 alcohol compound is methanol.

[0042] Specifically, the volume ratio of the supercritical carbon dioxide to the C1-C3 alcohol compound includes but is not limited to: 99.5:0.5, 99.3:0.7, 99:1, 98.8:1.2, 98.5:1.5 or a range between any two of the foregoing. In some examples, the volume ratio of the supercritical carbon dioxide to the C1-C3 alcohol compound is 99:1.

[0043] In some of the examples, the temperature of the supercritical carbon dioxide when it is pumped in is 5°C to 10°C and the density is 890kg / m 3 ~950kg / m 3 .

[0044] In some examples, the chromatography column has an inner diameter of 10 mm and a length of 250 mm.

[0045] In some examples, the supercritical fluid chromatography separation condition further includes: a column temperature of 38° C. to 40° C. Specifically, the column temperature includes but is not limited to: 38° C., 38.5° C., 39° C., 39.5° C., 40° C., or a range between any two of the foregoing.

[0046] In some examples, the supercritical fluid chromatography separation condition also includes: a flow rate of 10 mL / min to 12 mL / min. Specifically, the flow rate includes but is not limited to: 10 mL / min, 11 mL / min, 12 mL / min, or a range between any two of the foregoing.

[0047] In some of the examples, the supercritical fluid chromatography separation conditions also include: a detection wavelength of 260 nm.

[0048] In some examples, the supercritical fluid chromatography separation condition also includes: a system back pressure of 10 MPa to 12 MPa. Specifically, the system back pressure includes but is not limited to: 10 MPa, 11 MPa, 12 MPa or a range between any two of the foregoing.

[0049] In some examples, the solvent is n-hexane. Optionally, the solvent is used to dilute the sample to be tested 3 to 7 times.

[0050] In some examples, the method for preparing the crude product includes:

[0051] The medicinal material of Psoralea corylifolia is crushed and extracted with petroleum ether. The crude extract of Bakuchiol obtained in this way is the crude product.

[0052] The following are examples. The experimental methods used in the examples are all conventional methods unless otherwise specified. The raw materials and reagents used can be obtained from commercial channels such as the market unless otherwise specified.

[0053] The test product used in the embodiment is a crude extract of bakuchiol obtained by crushing the psoralea corylifolia medicinal material and extracting it with petroleum ether.

[0054] Example 1

[0055] This embodiment is a method for separating bakuchiol using a supercritical fluid chromatography system. The chromatograph selected is a supercritical fluid chromatograph produced by Jiangsu Hanbang Technology Co., Ltd., and the chromatographic conditions are shown in Table 1 below.

[0056] Table 1

[0057]

[0058] The specific steps are:

[0059] (1) Preparation of test samples: Dilute the crude extract of bakuchiol by 5 times its volume with n-hexane and dissolve it with ultrasound to prepare the test sample;

[0060] (2) According to the chromatographic conditions in Table 1, ensure that the temperature of supercritical carbon dioxide entering the pump is 5℃~10℃ and the density is 890kg / m 3 ~950kg / m 3 , the test sample is separated, and the separation effect of the test sample is shown in the figure Figure 1 As shown, the purity of the collected bakuchiol was tested to be 99.89%.

[0061] Comparative Example 1

[0062] This comparative example is a method for separating bakuchiol using a supercritical fluid chromatography system. The chromatograph selected is a supercritical fluid chromatograph of Jiangsu Hanbang Technology Co., Ltd., and the chromatographic conditions are shown in Table 2 below.

[0063] Table 2

[0064]

[0065] The specific steps are the same as in Example 1 except for the chromatographic conditions. The separation effect diagram of the test sample is shown in FIG. Figure 2 As shown, it can be seen that some impurities have not been completely separated. The purity of the collected bakuchiol was tested to be 91.63%.

[0066] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0067] The above-described embodiments only express several implementation methods of the present invention, which are convenient for understanding the technical solutions of the present invention in detail, but they cannot be understood as limiting the scope of protection of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. It should be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided by the present invention are all within the protection scope of the claims attached to the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the contents of the attached claims, and the description can be used to interpret the contents of the claims.

Claims

1. A supercritical fluid chromatography separation method for bakuchiol, characterized in that: The steps include: The crude product is dispersed in a solvent to prepare the test sample; Subjecting the test sample to supercritical fluid chromatography separation; The conditions for the supercritical fluid chromatography separation include: The mobile phase is a mixture of supercritical carbon dioxide and C1-C3 alcohol compounds in a volume ratio of (99.5-98.5):(0.5-1.5), and isocratic elution is performed; The stationary phase of the chromatographic column is octadecylsilane bonded silica gel.

2. The supercritical fluid chromatography separation method of bakuchiol according to claim 1, characterized in that: The C1-C3 alcohol compound is methanol.

3. The supercritical fluid chromatography separation method of bakuchiol according to claim 1, characterized in that: The volume ratio of the supercritical carbon dioxide to the C1-C3 alcohol compound is 99:

1.

4. The supercritical fluid chromatography separation method of bakuchiol according to claim 1, characterized in that: The temperature of the supercritical carbon dioxide when it is pumped in is 5°C to 10°C, and the density is 890kg / m 3 ~950kg / m 3 .

5. The supercritical fluid chromatography separation method of bakuchiol according to claim 1, characterized in that: The inner diameter of the chromatographic column is 10 mm and the length is 250 mm.

6. The supercritical fluid chromatography separation method of bakuchiol according to any one of claims 1 to 5, characterized in that: The supercritical fluid chromatography separation conditions also include: column temperature of 38°C to 40°C; and / or, The conditions for supercritical fluid chromatography separation also include: a flow rate of 10 mL / min to 12 mL / min.

7. The supercritical fluid chromatography separation method of bakuchiol according to any one of claims 1 to 5, characterized in that: The conditions for supercritical fluid chromatography separation also include: the detection wavelength is 260nm.

8. The supercritical fluid chromatography separation method of bakuchiol according to any one of claims 1 to 5, characterized in that: The conditions for supercritical fluid chromatography separation also include: the system back pressure is 10Mpa~12Mpa.

9. The supercritical fluid chromatography separation method of bakuchiol according to any one of claims 1 to 5, characterized in that: The solvent is n-hexane.

10. The supercritical fluid chromatography separation method of bakuchiol according to any one of claims 1 to 5, characterized in that: The preparation method of the crude product comprises: The medicinal material of Psoralea corylifolia was crushed and extracted with petroleum ether.

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