CN105646616A - Stevia glucoside B crystal form G as well as preparation method, food composition and application thereof - Google Patents

Abstract

The present invention belongs to the technical field of sweeteners, and in particular relates to stevioside B crystal form G, its preparation method, food composition and application. The stevioside B glycoside crystal form G is X- According to X-ray powder diffraction analysis, the 2θ values expressed in degrees have obvious characteristic diffraction peaks at least at 3.93, 13.45, 16.62, 18.33, 19.90, 23.95 and 28.02. The present invention uses XRPD, DSC, TGA, DVS and other solid chemical analysis methods to comprehensively characterize the crystal form G, and finds that the crystal form G has the advantages of high crystallinity, good stability, good water solubility, and low hygroscopicity. , suitable for a wider range of applications. The preparation method involved in the invention is simple, easy to operate, has many selectivities and good reproducibility, and can stably obtain the target crystal form.

Description

Steviol glycoside B crystal form G, its preparation method, food composition and application

technical field

The invention belongs to the technical field of sweeteners, and in particular relates to steviol glycoside B crystal form G, its preparation method, food composition and application.

Background technique

Stevia is a small Compositae plant native to the Amambai Mountains on the border between Paraguay and Brazil in South America. There are 9 identified sweet components of stevia: stevioside, steviobioside, stevioside A-stevioside F and dulcoside G. They all belong to glycoside compounds and have the same aglycone-steviol; the difference is only in the type, quantity and configuration of the sugar bound on the glycoside bond. Because they are all glycosides with sweet taste, they are collectively called Steviol Glycosides. Among them, stevioside B is a stevioside compound with a sweetness about 150 times that of sucrose. Stevioside B (RebGudiosideB, RB), its structural formula is as follows:

Among stevia extracts, steviol glycosides have been widely used in beverages, foods and health products as sweeteners. However, stevioside G has a natural slight bitterness or grassy taste, which cannot be completely removed or covered up in the prior art, which makes the beverage and food industries dare not to simply replace white sugar with stevioside completely. A study in the journal "JournGlofGgriculturGlGndFoodChemistry" titled "HumGnPsychometricGndTGsteReceptorResponsestoSteviolGlycosides" shows that steviol glycoside B has similar sweetness to steviol glycoside G, but the bitterness is slightly weaker. Therefore, stevioside B has a better taste than stevioside G, and can be used as a sweetener. Patent CN104602543G reports the application of stevioside B glycosides and other sugars in food and beverages. Due to the low content in stevia leaves, the current preparation method of stevioside B is mainly obtained by hydrolyzing stevioside G with sodium hydroxide. Stevioside B has very low water solubility at room temperature, which limits its use.

Different crystal forms of the same compound have significant differences in solubility, dissolution rate, melting point, appearance and bioavailability, which affect its stability and bioavailability. The study of drug polymorphism has become an indispensable and important part of the pharmaceutical process and the determination of new drug preparations. For stevioside B, polymorphic research is also crucial. Patent US20130267693G1 reports steviol glycoside B crystal form 1, crystal form 2, crystal form 3, crystal form 4 and their preparation methods. However, from the X-ray powder diffraction patterns and polarized light photos, the four crystal forms have poor crystallinity and contain some amorphous forms. For highly hygroscopic glycosides, the amorphous state in the product may lead to hygroscopic agglomeration during storage and transportation. At the same time, the four crystal forms did not improve the water solubility of stevioside B.

There is an urgent need in this field to provide a crystal form with better performance, such as a new crystal form with high crystallinity, good water solubility, low hygroscopicity, and high stability. At the same time, there is an urgent need to provide the preparation method and application of the above-mentioned crystal form.

Contents of the invention

One of the objectives of the present invention is to provide stevioside B crystal form G with high crystallinity, good water solubility, high stability and low hygroscopicity.

In order to solve the problems of the technologies described above, the technical solution of the present invention is:

Steviol glycoside crystalline form G, said crystalline form G is analyzed by X-ray powder diffraction using Cu-Kα radiation, and the 2θ angle expressed in degrees is at least 3.93, 13.45, 16.62, 18.33, 19.90, 23.95 and 28.02 There are obvious characteristic diffraction peaks.

As an improvement, the X-ray powder diffraction analysis of the crystal form G using Cu-Kα ray measurement, the 2θ value expressed in degrees, the error range is ±1°, in order The interplanar spacing d expressed and the relative intensity of the diffraction peaks expressed as a percentage have the following characteristics:

2θ d Relative Strength% 3.93 22.44 100.0 7.88 11.22 5.6 9.05 9.76 4.9 9.25 9.55 5.0 10.83 8.16 5.3 12.02 7.36 3.5 13.45 6.58 23.8 13.81 6.41 4.8 14.36 6.16 3.8 14.98 5.91 6.0 16.31 5.43 4.2 16.62 5.33 21.4 17.38 5.10 5.4 18.33 4.84 23.7 18.62 4.76 4.9 19.90 4.46 22.7 20.11 4.41 9.4 22.94 3.87 7.42 --> 23.95 3.71 12.8 24.32 3.66 4.2 27.56 3.23 5.4 28.02 3.18 13.8

.

As an improvement, the differential scanning calorimetry analysis of the crystal form G has obvious endothermic peaks at 60-140°C, 150-180°C and 200-240°C.

As an improvement, the thermogravimetric analysis of the crystal form G starts to decompose at 270±10°C.

As an improvement, the crystalline form G has a dynamic moisture adsorption (DVS) spectrum as shown in Figure 4, in the range of 0-45% relative humidity, the mass percentage of absorbed moisture is 0-2.5%, and the relative humidity is 0-2.5%. In the range of 45-55%, the mass percentage of water absorption is 2.5-5.1%, and the fluctuation range of the mass percentage of water absorption is small when the relative humidity is above 55%.

As an improvement, the crystal form G has the morphological characteristics as shown in FIG. 5 .

The second object of the present invention is to provide a method for preparing stevioside B crystal form G with simple process, easy operation, high stability and good fluidity.

In order to solve the problems of the technologies described above, the technical solution of the present invention is:

A preparation method of stevioside B crystal form G, the preparation method comprising the following steps:

(1) Suspension: within the temperature range of 0-100°C, mix stevioside B with a solvent for 0.1-48 hours to obtain a suspension solution;

(2) Filtration: within the temperature range of 0-100°C, filter or centrifuge the suspension solution to obtain a white solid, that is, stevioside B crystal form G;

(3) Cooling: the clarified solution after filtering or centrifuging in step (2) is cooled to 0-50°C, a white solid is precipitated, and filtered to obtain Stevioside B crystal form G;

(4) Volatilization: the clarified solution after filtration or centrifugation in step (2) is placed in the temperature range of 0-100°C to volatilize, and a white solid is precipitated to obtain stevioside B crystal form G.

As an improvement, in step (1), the dry matter purity of the stevioside B is 50-100%.

As an improvement, the solvent is water, methanol, ethanol, 1-propanol, 2-propanol, 3-methyl-1-butanol, 2-methyl-1-propanol, acetonitrile, acetone, methyl ethyl ketone , methyl isobutyl ketone, methyl acetate, ethyl formate, ethyl acetate, butyl acetate, propyl acetate, isopropyl acetate, isobutyl acetate or tributylmethyl ether, or one or more of them.

The third object of the present invention is to provide a food composition containing the steviol glycoside B crystal form G.

The fourth object of the present invention is to provide the application of the stevioside B glycoside crystal form G and its preparation method in food, beverage and medicine.

Owing to adopted above-mentioned technical scheme, the beneficial effect of the present invention is:

The preparation method of stevioside B crystal form G provided by the present invention has a simple process and is easy to operate, and stevioside B crystal form G can be prepared by various methods, and the obtained product has high crystallinity, low hygroscopicity, High stability and good water solubility.

Description of drawings

Fig. 1 is the X-ray powder diffraction (XRPD) pattern of steviol glycoside B crystal form G provided by the present invention;

Fig. 2 is the differential scanning calorimetry (DSC) diagram of stevioside B crystal form G provided by the present invention;

Fig. 3 is the thermogravimetric analysis (TG) diagram of stevioside B glycoside crystal form G provided by the present invention;

Fig. 4 is the dynamic water adsorption (DVS) diagram of stevioside B crystal form G provided by the present invention;

Fig. 5 is a polarized photo of stevioside B glycoside crystal form G provided by the present invention;

Fig. 6 is a comparison diagram of X-ray powder diffraction (XRPD) before and after drying of stevioside B crystal form G provided by the present invention;

Fig. 7 is a comparison chart of X-ray powder diffraction (XRPD) of stevioside B crystal form G stored at 40°C and humidity of 75% for half a year provided by the present invention;

Fig. 8 is a comparison chart of high performance liquid phase (HPLC) of stevioside B crystalline form G provided by the present invention stored at 40°C and humidity of 75% for half a year.

detailed description

In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

Embodiment one

At room temperature, 20 g of stevioside B with a material purity of 100% was added to 100 mL of methanol, stirred for 12 hours, and then filtered to obtain a filtrate and a white solid, which were vacuum-dried at 25°C to obtain stevioside Beta glycoside crystal form G.

Embodiment two

At 50°C, add 20g of stevioside B with a material purity of 100% into 100mL of methanol, stir for 12 hours, and filter to obtain a filtrate and a white solid, which are vacuum-dried at 25°C to obtain stevia Sugar B glycoside crystal form G.

Embodiment three

At room temperature, 40 g of stevioside B with a material purity of 50% was added to 100 mL of methanol, stirred for 12 hours, and filtered to obtain a filtrate and a white solid. The filtrate and the white solid were vacuum-dried at 25° C. respectively to obtain stevioside B. Glycoside crystal form G.

Embodiment Four

At room temperature, 20 g of stevioside B with a material purity of 100% was added to 70 mL of 30% methanol-water (v/v) system, stirred for 12 hours, and filtered to obtain a filtrate and a white solid, which were respectively in Vacuum drying at 25°C yielded steviol glycoside B crystalline form G.

Embodiment five

At room temperature, 20 g of stevioside B with a material purity of 100% was added to 50 mL of 50% methanol-water (v/v) system, stirred for 12 hours, and then filtered to obtain a filtrate and a white solid, which were respectively in Vacuum drying at 25°C yielded steviol glycoside B crystalline form G.

Embodiment six

At room temperature, 20 g of stevioside B with a material purity of 100% was added to 50 mL of 50% ethanol-water (v/v) system, stirred for 12 hours, and then filtered to obtain a filtrate and a white solid, which were respectively in Vacuum drying at 25°C yielded steviol glycoside B crystalline form G.

Embodiment seven

At room temperature, 20 g of stevioside B with a material purity of 100% was added to 50 mL of 50% ethanol-water (v/v) system, stirred for 48 hours, and filtered to obtain a filtrate and a white solid, which were respectively in Vacuum drying at 25°C yielded steviol glycoside B crystalline form G.

Embodiment Eight

At room temperature, 20 g of stevioside B with a material purity of 100% was added to 50 mL of 50% ethanol-water (v/v) system, stirred for 48 hours, and filtered to obtain a filtrate and a white solid, which were respectively in Vacuum drying at 50°C yielded steviol glycoside B crystalline form G.

Embodiment nine

At room temperature, 20 g of stevioside B with a material purity of 100% was added to 200 mL of 50% ethanol-water (v/v) system, and after stirring for 48 hours, the filtrate obtained by filtration was rapidly cooled to 5° C., and a large amount of crystals were precipitated. Filter and dry the crystals to obtain steviol glycoside B crystalline form G.

Embodiment ten

At 90°C, 20 g of stevioside B with a material purity of 100% was added to 100 mL of 50% ethanol-water (v/v) system, stirred for 48 hours, and the filtrate obtained by filtering was rapidly cooled to room temperature, and a large amount of crystals were precipitated. Filter and dry the crystals to obtain steviol glycoside B crystalline form G.

Embodiment Eleven

At 90°C, add 20g of stevioside B with a material purity of 100% into 100mL of 50% ethanol-water (v/v) system, stir for 48 hours, and then filter the filtrate to cool at a cooling rate of 1°C/h After reaching room temperature, a large amount of crystals precipitated, filtered, and the crystals were dried to obtain stevioside B crystalline form G.

Embodiment 12

At 90°C, add 20g of stevioside B with a material purity of 100% to 100mL of 50% ethanol-water (v/v) system, stir for 48 hours, and then filter the filtrate to cool at a cooling rate of 10°C/h to room temperature, and stood at room temperature for 2 hours, a large amount of crystals were precipitated, filtered, and the crystals were dried to obtain stevioside B crystalline form G.

Embodiment Thirteen

At 90°C, 20 g of steviol glycosides with a material purity of 100% were added to 100 mL of 50% ethanol-water (v/v) system, stirred for 48 hours, and the filtrate obtained by filtering was left open to volatilize naturally at room temperature. After a large amount of crystals are precipitated, filter and dry the crystals to obtain steviol glycoside B crystal form G.

Embodiment Fourteen

At 90°C, add 20 g of stevioside B with a material purity of 100% to 100 mL of 50% ethanol-water (v/v) system, stir for 48 hours, and filter the filtrate at room temperature with a nitrogen flow rate of 25 mL/min Volatilize under conditions, and after a large amount of crystals are precipitated, filter and dry the crystals to obtain stevioside B crystalline form G.

Embodiment fifteen

At 90°C, add 20 g of steviol glycosides with a material purity of 100% into 100 mL of 50% ethanol-water (v/v) system, stir for 48 hours, and filter the filtrate at room temperature and a vacuum of 0.05 MPa Volatilize under high temperature, after a large amount of crystals are precipitated, filter, and dry the crystals to obtain stevioside B crystalline form G.

Embodiment sixteen

At 90°C, add 20 g of steviol glycosides with a material purity of 100% into 100 mL of 50% ethanol-water (v/v) system, stir for 48 hours, and filter the filtrate at room temperature and a vacuum of 0.05 MPa Volatilize under high temperature, after a large amount of crystals are precipitated, filter, and dry the crystals to obtain stevioside B crystalline form G.

Embodiment 17

At 0°C, add 20g of stevioside B with a material purity of 100% into 100mL of methanol, stir for 48 hours, and filter to obtain a filtrate and a white solid, which are vacuum-dried at 25°C to obtain stevia Sugar B glycoside crystal form G.

Embodiment eighteen

At 100°C, add 20 g of stevioside B with a material purity of 100% into 50 mL of 50% ethanol-water (v/v) system, stir for 0.1 h, and filter to obtain a filtrate and a white solid, a filtrate and a white solid Vacuum-dried at 25°C respectively to obtain stevioside B crystalline form G.

Carry out X-ray powder diffraction analysis (XRPD), differential scanning calorimetry (DSC), thermogravimetric analysis (TG), dynamic water adsorption analysis (DVS) to the stevioside B glycoside sodium salt crystalline form G that above-mentioned embodiment makes )Wait.

XRPD analysis: it was detected at room temperature by a Bruker D8GdvGnce diffractometer from Bruker Instruments Co., Ltd., Germany, using Cu–Kα rays The 2θ angle scans from 3 degrees to 40 degrees, and the scanning speed is 0.2 degrees/second. The analysis results are shown in Figure 1, and the XRPD spectrum shows that the stevioside B crystal form G prepared in the above example has good crystallinity.

In the sample powder X-ray powder diffraction pattern, the diffraction pattern obtained from a specific crystal form is often characteristic. Because of differences in crystallization conditions, particle size, relative amounts of mixtures, and other test conditions, diffraction patterns may have preferential orientation effects, resulting in changes in the relative intensities of certain bands (especially at low angles) in the pattern. Therefore, the relative intensities of the diffraction peaks are not characteristic of the crystal in question, and when judging whether it is the same as a known crystal form, more attention should be paid to the positions of the peaks rather than their relative intensities. In addition, when judging whether the crystal form is the same, you should pay attention to maintaining the overall concept, because not a diffraction line represents a phase, but a specific set of "d-I/I1" data represents a certain phase. It should also be pointed out that in the identification of mixtures, due to factors such as content decline, some diffraction lines will be missing. At this time, it is not necessary to rely on all the bands observed in the high-purity sample, and even one band may affect the given Certain crystals are characteristic.

DSC analysis: it adopts the DSC8500 differential scanning calorimeter of American Platinum Elmer Company to detect, the atmosphere is nitrogen, and the heating rate is 10 degrees Celsius/minute. The analysis results are shown in Figure 2.

TG analysis: It is detected by Netzsch TG209F3 thermogravimetric analyzer of German NETZSCH company, temperature range: 30-400°C, scan rate: 10K/min, purge gas: 25mL/min. The analysis results are shown in Figure 3.

DVS analysis: It is measured by DVSIntrinsic dynamic moisture adsorption instrument of British SMS Instrument Company, measuring temperature: 25°C; relative humidity: 0-95%. The analysis results are shown in Figure 4.

Polarized photo: The XPV-400E polarized microscope of Shanghai Changfang Optical Instrument Co., Ltd. was used for the experiment, and the test magnification: 5 times. The analysis results are shown in Figure 5. Polarized photos show that the steviol glycoside B crystal form G prepared in the above example is a rhombohedral crystal with good morphology.

The stevioside B glycoside crystal form G prepared in the above example was subjected to XRPD analysis after drying, and the analysis results are shown in FIG. 6 . It can be seen from Figure 6 that the crystal form remains unchanged and the crystal form is stable.

The steviol glycoside B crystal form G prepared in the above example was stored for half a year at 40° C. and RH 75%, and the analysis results are shown in FIG. 7 . It can be seen from Figure 7 that the crystal form remains unchanged, indicating that the crystal form has good physical stability under high humidity conditions.

HPLC analysis: it is determined by 1260infinity liquid chromatograph of Agilent Technologies Co., Ltd., USA. Sample solution preparation method: accurately weigh 25-50 mg stevioside B crystal form G sample, put it into a 25 ml volumetric flask, then add water-acetonitrile (7:3, v/v) solution to dissolve and determine To the scale. Sodium phosphate buffer solution (specification: 10mmol/L, pH value: 2.6): Dissolve 2.76 grams of sodium dihydrogen phosphate in 2 liters of water, add phosphoric acid, and adjust the pH value to 2.6. Chromatographic column: Lun G5μC18(2)100A type chromatographic column of Phenomenex Company. Injection volume: 5 μl. Flow rate: 1.0 mL/min. Column temperature: 40°C. Detector: 210nm UV detection. Mobile phase: the ratio of acetonitrile and sodium phosphate buffer (specification: 10mmol/L, pH: 2.6) is 32:68. The analysis results are shown in Figure 8. The stevioside B crystal form G prepared in the above example has good chemical stability. HPLC analysis shows that after half a year of storage at 40°C and RH75%, its purity is still as high as 98.3%, indicating that The crystal form has good chemical stability under high humidity conditions.

The steviol glycoside B crystalline form G prepared in the above examples has good reproducibility. And the water solubility is high and stable, about 0.5mg/mL, which is obviously higher than the crystal forms 1-4 reported in US20130267693A1. Its analysis result is as follows table 1:

Table 1

sample name Equilibrium solubility in water (mg/mL) Embodiment 1 Stevioside B glycoside crystal form G 0.452 Form 1 in US 20130267693A1 0.291 Form 2 in US 20130267693A1 0.205 Form 3 in US 20130267693A1 0.160 Form 4 in US 20130267693A1 0.097

The stevioside B crystalline form G prepared in the above examples is slightly hygroscopic under normal storage conditions (45-80% RH). The hygroscopicity of stevioside B glycoside crystal form G is significantly lower than that of crystal forms 1-4 reported in US20130267693A1, and it is not easy to absorb moisture and agglomerate. The analysis results are shown in Table 2 below:

Table 2

The stevioside B glycoside raw materials used in the above examples were provided by Shandong Zhucheng Haotian Pharmaceutical Co., Ltd.

The stevioside B crystal form G provided by the invention can be used as a sweetener in food, beverage and medicine.

The preparation method of stevioside B crystal form G provided by the present invention can be applied in the preparation process of food, beverage and medicine.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present invention should be included in the protection of the present invention. within range.

Claims (8)

1. stevioside B glycosides crystal formation G, it is characterised in that described crystal formation G uses the X-ray powder diffraction analysis that Cu-K alpha ray records, to spend the 2 �� angles represented at least 3.93,13.45,16.62,18.33, there is obvious characteristic diffraction peak at 19.90,23.95 and 28.02 places.

2. stevioside B glycosides crystal formation G as claimed in claim 1, it is characterised in that described crystal formation G uses the X-ray powder diffraction analysis that Cu-K alpha ray measurements obtains, with spend represent 2 �� values, range of error for �� 1 ��, withThe relative intensity of the interplanar distance d represented and the diffraction maximum being expressed as a percentage has the feature that

2�� d Relative intensity % 3.93 22.44 100.0 7.88 11.22 5.6 9.05 9.76 4.9 9.25 9.55 5.0 10.83 8.16 5.3 12.02 7.36 3.5 13.45 6.58 23.8 13.81 6.41 4.8 14.36 6.16 3.8 14.98 5.91 6.0 16.31 5.43 4.2 16.62 5.33 21.4 17.38 5.10 5.4 18.33 4.84 23.7 18.62 4.76 4.9 19.90 4.46 22.7 20.11 4.41 9.4 22.94 3.87 7.4 23.95 3.71 12.8 24.32 3.66 4.2 27.56 3.23 5.4 28.02 3.18 13.8

��

3. stevioside B glycosides crystal formation G as claimed in claim 1, it is characterised in that described crystal formation G has differential scanning calorimetric thermogram spectrum substantially as shown in Figure 2,3, 4, thermogravimetic analysis (TGA) collection of illustrative plates and dynamic water absorption collection of illustrative plates.

4. the preparation method of stevioside B glycosides crystal formation G, it is characterised in that described preparation method comprises the following steps:

(1) suspendible: in 0-100 DEG C of temperature range, by stevioside B glycosides and solvent mixing 0.1-48h, obtains suspension solution;

(2) filter: in 0-100 DEG C of temperature range, suspension solution is filtered or centrifugal, obtain white solid, obtain stevioside B glycosides crystal formation G;

(3) cooling: step (2) filtration or the settled solution after centrifugal, is cooled to 0-50 DEG C, precipitates out white solid, filter, obtain stevioside B glycosides crystal formation G;

(4) volatilization: step (2) filtration or the settled solution after centrifugal, is placed in 0-100 DEG C of temperature range and volatilizees, precipitate out white solid, obtain stevioside B glycosides crystal formation G.

5. the preparation method of stevioside B glycosides crystal formation G as claimed in claim 4, it is characterised in that in step (1), the dry purity of described stevioside B glycosides is 50-100%.

6. the preparation method of stevioside B glycosides crystal formation G as claimed in claim 4, it is characterized in that, in step (1), described solvent is one or more in water, methanol, ethanol, 1-propanol, 2-propanol, 3-methyl-1-butanol, 2-methyl isophthalic acid-propanol, acetonitrile, acetone, butanone, methylisobutylketone, methyl acetate, Ethyl formate, ethyl acetate, butyl acetate, propyl acetate, isopropyl acetate, isobutyl acetate or three fourth MEEs.

7. food compositions, it is characterised in that described food compositions contains the stevioside B glycosides crystal formation G described in any one of claim 1-3.

8. the stevioside B glycosides crystal formation G as described in any one of claim 1-3 and preparation method thereof application in food, beverage and medicine.