Preparation method and application of traditional Chinese medicine carbon quantum dots for improving the performance of mangiferin

Abstract

The invention discloses a method for preparing a Chinese medicine carbon quantum dot for improving the performance of mangiferin and its application. The molecular weight of the peony bark carbon quantum dot is greater than 3000Da. The preparation process includes: preparing peony bark charcoal: calcining the peony bark in a muffle furnace and cooling it naturally to room temperature to obtain peony bark charcoal; preparing a concentrated solution: crushing the peony bark charcoal, extracting the peony bark charcoal with deionized water to obtain a water extract, filtering the water extract with filter paper, and filtering the filtered water extract with a microporous filter membrane for a second time to obtain a concentrated solution; dialysis; centrifugation; taking the supernatant and freeze-drying to obtain the peony bark carbon quantum dot, and the finished product can be used for the preparation of mangiferin medicament. The invention aims to improve the water solubility and bioavailability of mangiferin by using carbon quantum dots in Chinese medicine charcoal medicine, and utilizes the advantages of excellent water solubility, low toxicity, environmental friendliness, wide source of raw materials, low cost, good biocompatibility, etc. to load mangiferin, thereby increasing its solubility and bioavailability in aqueous solution.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine carbon quantum dots, and particularly relates to a preparation method and application of traditional Chinese medicine carbon quantum dots for improving the performance of mangiferin. Background Art

[0002] Mangiferin, also known as Guan Zhimu Ning or mangoside, is a carbon ketoside of tetrahydroxypyrrolidone and a flavonoid compound of the bispyribac class. It is mainly derived from the dried rhizomes of the perennial herb Anemarrhena asphodeloides of the Liliaceae family, the leaves, fruits and barks of the plants almond and mango, and the roots of the Hainan Salacia Chemicalbook of the Pterospermaceae family. It has multiple pharmacological activities such as relieving cough and asthma, eliminating phlegm, immune anti-inflammatory and analgesic, protecting the liver and promoting bile secretion, resisting lipid peroxidation, anti-cancer, anti-diabetes, antibacterial and antiviral. Currently, the main clinical applications include mango cough tablets, mango cough capsules, mango glycoside tablets and mango glycoside pills.

[0003] It was found in research and practical applications that the solubility of mangiferin in water (0.111 mg / ml) is very small and it is almost insoluble in water, which makes it difficult to select the dosage form of mangiferin. At the same time, it also causes unsatisfactory absorption of mangiferin in the body and low relative bioavailability. Summary of the invention

[0004] In view of the above problems, the present invention provides a traditional Chinese medicine carbon quantum dot for improving the performance of mangiferin. The traditional Chinese medicine carbon quantum dot is peony bark carbon quantum dot, and the molecular weight of the peony bark carbon quantum dot is greater than 3000Da.

[0005] A further preferred technical solution is that the diameter of the peony bark carbon quantum dots is 0.5-10 nm.

[0006] Its further preferred technical solution is: including:

[0007] Preparation of peony bark charcoal: calcining peony bark in a muffle furnace and naturally cooling to room temperature to obtain peony bark charcoal;

[0008] Preparing a concentrated solution: crushing the peony bark charcoal, extracting the peony bark charcoal with deionized water to obtain a water extract, filtering the water extract with filter paper, and then filtering the filtered water extract with a microporous filter membrane for a second time to obtain a concentrated solution;

[0009] Dialysis: placing the concentrated solution into a dialysis bag for dialysis to obtain a dialysis bag solution;

[0010] Preparation of Paeonia suffruticosa carbon quantum dots: placing the liquid in the dialysis bag into a centrifuge for centrifugation, taking the supernatant and freeze-drying it to obtain the Paeonia suffruticosa carbon quantum dots.

[0011] A further preferred technical solution is: to prepare peony bark charcoal, the peony bark is placed in a muffle furnace, the temperature is raised to 65-75°C at a rate of 2-2.5°C / min, then the temperature is raised to 325-375°C at a rate of 4.5-5°C / min, and kept warm for 1 hour.

[0012] A further preferred technical solution is: to prepare peony bark charcoal, the peony bark is placed in a muffle furnace, the temperature is raised to 70°C at a rate of 2.25°C / min and kept warm for 20 minutes, then the temperature is raised to 350°C at a rate of 4.67°C / min and kept warm for 1 hour.

[0013] Its further preferred technical scheme is: to prepare the concentrated solution, the peony bark charcoal is crushed to obtain the peony bark charcoal powder, and the peony bark charcoal powder is extracted with deionized water, the mass-volume ratio of the deionized water to the peony bark charcoal powder is 1:5-20, the number of extractions is 2-4 times, and the extraction time is 1-3h.

[0014] A further preferred technical solution is: after obtaining a water extract and filtering the water extract with filter paper, the pore size of the microporous filter membrane is 0.22 or 0.45 μm.

[0015] A further preferred technical solution is: dialysis, which is to put the concentrated solution into a dialysis bag for dialysis, the dialysis time is 2-4 days, and the liquid outside the dialysis bag is replaced every 4-8 hours.

[0016] A further preferred technical solution is: centrifuging the liquid in the dialysis bag at 4000-6000 rpm for 10-30 minutes.

[0017] The invention discloses an application of traditional Chinese medicine carbon quantum dots for improving the performance of mangiferin. The traditional Chinese medicine carbon quantum dots can be used for the preparation of mangiferin medicament.

[0018] Beneficial effects of the present invention: The present invention aims to provide a method for improving the water solubility and bioavailability of mangiferin by using carbon quantum dots derived from traditional Chinese medicine charcoal medicine, and utilizes the advantages of excellent water solubility, low toxicity, environmental friendliness, wide source of raw materials, low cost, good biocompatibility, etc. to load mangiferin to increase its solubility and bioavailability in aqueous solution.

[0019] Other features and advantages of the present invention will be described in the following description, and partly become apparent from the description, or understood by practicing the present invention. The purpose and other advantages of the present invention can be realized and obtained by the structures pointed out in the description, claims and drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0021] Figure 1 HR-TEM images of Paeonia suffruticosa carbon quantum dots are shown;

[0022] Figure 2 The curve of the change of mangiferin content in rats over time is shown;

[0023] Figure 3 The graph shows the change curve of the content of mangiferin in rats over time in the presence of Paeonia suffruticosa carbon quantum dots prepared according to an embodiment of the present invention. DETAILED DESCRIPTION

[0024] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0025] Example 1

[0026] Preparation of Paeonia suffruticosa carbon quantum dots.

[0027] This drug is mainly composed of peony bark carbon quantum dots (MW>3000Da) with a diameter of 0.5-10nm. The preparation method is as follows:

[0028] 1. Raise the temperature of peony bark in a muffle furnace from 25°C to 70°C in 20 minutes and keep it for 20 minutes, then raise the temperature from 70°C to 350°C in 60 minutes and keep it at 350°C for 1 hour, and naturally cool to room temperature to calcine peony bark charcoal.

[0029] The peony bark charcoal calcined under this temperature control has a uniform calcination effect, the appearance of the peony charcoal turns black, the texture is crisp but no powder is produced, which meets the requirements of "frying charcoal and preserving its properties" and is easy to use without the need for multiple screening.

[0030] The peony bark charcoal was crushed into powder, extracted with 10 times the amount of deionized water for 3 times, each time for 2 hours, and the combined water extracts were first filtered with filter paper and then filtered through a 0.22 μm microporous filter membrane, and the water extracts were concentrated to 1 g / ml based on carbon powder.

[0031] Extracting peony bark charcoal powder with dequantized water can produce a purer finished product. Filtering through a 0.22μm microporous filter membrane can quickly and easily concentrate the water extract, and the appropriate pore size of the filter membrane ensures the retention of effective ingredients.

[0032] The concentrated solution of Paeonia suffruticosa charcoal water extract was placed in a dialysis bag (MW: 3000Da) and dialyzed for 3 days, with the water changed every 4 hours until the liquid outside the dialysis bag was colorless.

[0033] The liquid in the dialysis bag was centrifuged at 5000 rpm for 25 min, and the supernatant was freeze-dried to obtain Paeonia suffruticosa carbon quantum dots, the HR-TEM image of which is shown in Figure 1 As shown, the yield is 0.114%.

[0034] The peony bark carbon quantum dots provided by the present invention can be applied to the medical field and the cosmetic product development industry. The peony bark carbon quantum dots can be used as a carrier to load mangiferin, thereby increasing its solubility and bioavailability.

[0035] The solubility of mangiferin was studied by high performance liquid chromatography, and it was found that peony bark carbon quantum can increase the solubility of mangiferin.

[0036] Through pharmacokinetic experiments, SD rats were gavaged with mangoside or a mixed solution of mangoside and an equal amount of Paeonia suffruticosa carbon quantum dots. It was found that Paeonia suffruticosa carbon quantum dots increased the bioavailability of mangoside.

[0037] Example 2

[0038] Experiment on increasing the solubility of mangiferin by using carbon quantum dots from peony bark

[0039] Determination of the solubility of mangiferin in water

[0040] like Figure 1 As shown, mangiferin was dissolved in methanol to obtain mangiferin mother liquor, and the mother liquor was diluted 2, 4, 8, and 16 times respectively, and analyzed by high performance liquid chromatography to calculate the linear regression equation.

[0041] Separately, mangiferin was prepared into a saturated aqueous solution and diluted to 10, 20, 50, 100 times until completely dissolved. The content was determined by high performance liquid chromatography. The solubility of mangiferin in water was calculated to be 115.46 μg / ml using the above regression equation.

[0042] Study on the Solubility of Mangiferin from Paeonia Suffruticosa Carbon Quantum Dots

[0043] Take 30 mg of Paeonia suffruticosa carbon quantum dots and add water to make up to 10 ml to obtain 3000 μg / ml mother liquor, and dilute the mother liquor to 1500 μg / ml, 750 μg / ml, 375 μg / ml, 187.5 μg / ml, 93.5 μg / ml, 46.5 μg / ml, 23.3 μg / ml, 12.5 μg / ml, 6.35 μg / ml, 3.12 μg / ml and 1.06 μg / ml Paeonia suffruticosa carbon quantum dots solutions respectively.

[0044] Weigh 2.0 mg of mangiferin and place it in 12 2 ml centrifuge tubes, add 1 ml of the above-mentioned concentration of peony bark carbon quantum dot solution to each tube, ultrasonicate for 30 min, centrifuge at 10000 rpm for 10 min, take the supernatant and dilute 20 times for analysis by high performance liquid chromatography, and the solubility of mangiferin in peony bark carbon quantum dot solutions of different concentrations is shown in Table 1.

[0045] Table 1 Content of mangiferin in different concentrations of Paeonia suffruticosa carbon quantum dot solutions (μg / ml)

[0046]

[0047] A methodological investigation was conducted on this method, and the precision peak area RSD value was 0.09%, and the retention time RSD value was 0.22%; the stability peak area RSD value was 1.10%, and the retention time RSD value was 1.31%; the repeatability peak area RSD value was 2.92%, and the retention time RSD value was 2.49%; the sample recovery rate was between 100.68% and 102.78%, indicating that this method is better and the prepared samples are relatively stable.

[0048] The curve of the change of mangiferin content in rats over time is as follows Figure 2 As shown, 0.08 hours after administration, the mangiferin content in rats was 70-80 ng / L. 2.5 hours after administration, the mangiferin content in rats reached a peak of 160-180 ng / L. 2.5 hours after administration, the mangiferin content in rats entered the attenuation period. 12.5 hours after administration, the mangiferin content in rats decreased significantly and gradually decreased to zero.

[0049] The curve of the change of mangiferin content in rats with time in the presence of Paeonia suffruticosa carbon quantum dots is as follows: Figure 3As shown, it can be seen that 0.35 hours after administration, the content of mangoside in the rat body has reached a peak in the presence of peony bark carbon quantum dots. With the help of peony bark carbon quantum dots, the release rate of mangoside has been effectively improved, significantly shortening the onset time of mangoside. Between 0.35h and 2h, it can be seen that the content of mangoside in the rat body in the presence of peony bark carbon quantum dots has always maintained a high level, all exceeding the peak content of pure mangoside in rats, indicating that peony bark carbon quantum dots have a significant effect on the absorption of mangoside in the body. Peony bark carbon quantum dots can increase the solubility of mangoside in aqueous solution, and it has many advantages such as low toxicity, environmental friendliness, wide source of raw materials, low cost, and good biocompatibility.

[0050] Example 3

[0051] Pharmacokinetics of Paeonia suffruticosa carbon quantum dots in increasing the bioavailability of mangiferin

[0052] The purchased SD rats were divided into mangiferin group and moutan cortex carbon quantum dot group. The mangiferin group was intragastrically administered with 20 mg / kg mangiferin, and the moutan cortex quantum dot group was intragastrically administered with a mixed solution of 20 mg / kg mangiferin and 20 mg / kg moutan cortex carbon quantum dot. The rats were fasted but not watered before administration.

[0053] Before administration and 0.08h, 0.35h, 0.5h, 0.75h, 1h, 2h, 4h, 8h, 12h, and 24h after administration, 0.5ml of blood was collected from the canthus of the rats, placed in a centrifuge tube containing sodium heparin, and centrifuged at 4000rpm for 5min. The supernatant was taken to obtain plasma.

[0054] The protein in the plasma was removed with acetonitrile: glacial acetic acid = 9:1, vortexed, centrifuged at 10000 rpm for 10 min, and the supernatant was taken and dried with nitrogen. The nitrogen-dried sample was re-dissolved with 100 μl acetonitrile-0.4% glacial acetic acid (10:90), centrifuged at 10000 rpm for 10 min, and the operation was repeated twice, and the supernatant was taken for mass spectrometry analysis.

[0055] Preliminary experiments have found that the content of the sample is low after entering the blood, and mangoside cannot be detected by general high performance liquid chromatography or ultra-high performance liquid chromatography. In order to obtain a more intuitive comparison effect, this experiment uses UPLC-MS / MS, that is, ultra-high performance liquid chromatography tandem mass spectrometry. The ACQUITY UPLC BEH C18 column (2.1mm×100mm×1.7μm) was used, the column temperature was 30℃, the injection volume was 3μl, the flow rate was 0.2ml / min, and the mobile phase was 0.4% glacial acetic acid water (A)-acetonitrile (B) = 90:10 isocratic elution. The mass spectrometry mode was negative ion mode DP:-165V, CE:-32V, the parent ion peak was 421, and the daughter ion peak was 301.

[0056] As shown in Table 2 and Table 3, the addition of Paeonia suffruticosa carbon quantum dots promotes the absorption of mangiferin in the body, shortens the onset time of mangiferin, prolongs the half-life of mangiferin, and promotes mangiferin to reach organs from the blood, thereby improving its bioavailability.

[0057] Table 2 Pharmacokinetic parameters of mangiferin group

[0058]

[0059] Table 3. Pharmacokinetic parameters of Paeonia suffruticosa carbon quantum dots

[0060]

[0061]

[0062] Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent substitutions for some of the technical features therein; and these modifications or substitutions do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. An application of traditional Chinese medicine carbon quantum dots for improving the performance of mangiferin, characterized in that: The traditional Chinese medicine carbon quantum dots are used for the preparation of mangiferin medicaments. The traditional Chinese medicine carbon quantum dots are peony bark carbon quantum dots, and the molecular weight of the peony bark carbon quantum dots is greater than 3000Da.

2. The use of a Chinese medicine carbon quantum dots for improving the performance of mangiferin according to claim 1, characterized in that: The method for preparing the traditional Chinese medicine carbon quantum dots comprises: Preparation of peony bark charcoal: calcining peony bark in a muffle furnace and naturally cooling to room temperature to obtain peony bark charcoal; Preparing a concentrated solution: crushing the peony bark charcoal, extracting the peony bark charcoal with deionized water to obtain a water extract, filtering the water extract with filter paper, and then filtering the filtered water extract with a microporous filter membrane for a second time to obtain a concentrated solution; Dialysis: placing the concentrated solution into a dialysis bag for dialysis to obtain a dialysis bag solution; Preparation of Paeonia suffruticosa carbon quantum dots: placing the liquid in the dialysis bag into a centrifuge for centrifugation, taking the supernatant and freeze-drying it to obtain the Paeonia suffruticosa carbon quantum dots; The method for preparing peony bark charcoal comprises placing the peony bark in a muffle furnace, raising the temperature to 65-75°C at a rate of 2-2.5°C / min, then raising the temperature to 325-375°C at a rate of 4.5-5°C / min, and keeping the temperature for 1 hour.

3. The use of a Chinese medicine carbon quantum dots for improving the performance of mangiferin according to claim 2, characterized in that: The diameter of the Paeonia suffruticosa carbon quantum dots is 0.5-10 nm.

4. The use of traditional Chinese medicine carbon quantum dots for improving the performance of mangiferin according to claim 2, characterized in that: The method for preparing peony bark charcoal is to place the peony bark in a muffle furnace, raise the temperature to 70°C at a rate of 2.25°C / min and keep it warm for 20 minutes, then raise the temperature to 350°C at a rate of 4.67°C / min and keep it warm for 1 hour.

5. The use of traditional Chinese medicine carbon quantum dots for improving the performance of mangiferin according to claim 2, characterized in that: The preparation of the concentrated solution comprises crushing the peony bark charcoal to obtain peony bark charcoal powder, extracting the peony bark charcoal powder with deionized water, wherein the mass-volume ratio of the deionized water to the peony bark charcoal powder is 1:5-20, the extraction times are 2-4 times, and the extraction time is 1-3 hours.

6. The use of traditional Chinese medicine carbon quantum dots for improving the performance of mangiferin according to claim 2, characterized in that: The pore size of the microporous filter membrane is 0.22 or 0.45 μm.

7. The use of traditional Chinese medicine carbon quantum dots for improving the performance of mangiferin according to claim 2, characterized in that: The dialysis time is 2-4 days, and the external liquid of the dialysis bag is replaced every 4-8 hours.

8. The use of traditional Chinese medicine carbon quantum dots for improving the performance of mangiferin according to claim 2, characterized in that: The centrifugation is to centrifuge the liquid in the dialysis bag at 4000-6000 rpm for 10-30 minutes.

Images