阅读说明：本技术 一种软糖中胭脂红酸的提取纯化方法 (Method for extracting and purifying carminic acid in soft sweets ) 是由 杨祖伟 苏杜威 陈绮梦 魏鲜娥 李珍 黄进丽 吴小翠 苏昭伦 叶少文 于 2019-11-15 设计创作，主要内容包括：本发明公开了一种软糖中胭脂红酸的提取纯化方法,包括：(1)将样品加入水,在70～80℃条件下加热,使样品溶解,依次加入饱和硫酸钠溶液和稀酸溶液,混匀,得到样品溶液；(2)将样品溶液离心,离心液加入到装有聚酰胺的层析柱中；(3)用洗涤液洗脱层析柱,再用水洗脱层析柱至洗脱液呈中性,弃去洗脱液；(4)再用解析液分多次洗脱层析柱,收集洗脱液,加入磷酸溶液,即得。本发明针对软糖类产品,将胭脂红酸在酸性环境下用聚酰胺粉吸附除杂,用解析液洗脱后,可将糖类等复杂基质与胭脂红酸有效分离,得到纯度高的胭脂红酸提取液,解决了软糖中胭脂红酸含量较低导致的检测不准确的问题,从而实现软糖中胭脂红酸含量的准确检测。(The invention discloses a method for extracting and purifying carminic acid in soft sweets, which comprises the following steps: (1) adding water into a sample, heating at 70-80 ℃ to dissolve the sample, sequentially adding a saturated sodium sulfate solution and a dilute acid solution, and uniformly mixing to obtain a sample solution; (2) centrifuging the sample solution, and adding the centrifugate into a chromatographic column filled with polyamide; (3) eluting the chromatographic column with a washing solution, eluting the chromatographic column with water until the eluent is neutral, and discarding the eluent; (4) eluting the chromatographic column with resolving liquid for multiple times, collecting eluate, and adding phosphoric acid solution. Aiming at the soft sugar products, the carminic acid is adsorbed by polyamide powder to remove impurities in an acidic environment, and after the adsorption and the elution are carried out by using the analytic liquid, the complex matrixes such as the sugar and the like can be effectively separated from the carminic acid to obtain the carminic acid extracting solution with high purity, so that the problem of inaccurate detection caused by low content of the carminic acid in the soft sweets is solved, and the accurate detection of the content of the carminic acid in the soft sweets is realized.)

1. A method for extracting and purifying carminic acid in soft sweets is characterized by comprising the following steps:

(1) adding water into a soft sweet sample, heating at 70-80 ℃ to dissolve the sample, sequentially adding a saturated sodium sulfate solution and a dilute acid solution, and uniformly mixing to obtain a sample solution; the dilute acid is citric acid or formic acid;

(2) centrifuging the sample solution, and adding the centrifugate into a chromatographic column filled with polyamide;

(3) eluting the chromatographic column with a washing solution, eluting the chromatographic column with water until the eluent is neutral, and discarding the eluent; the washing liquid contains (0-4) by volume: (1-3): (18-30) a mixed aqueous solution of a saturated sodium sulfate solution, a citric acid solution and water;

(4) eluting the chromatographic column for multiple times by using the resolving liquid, collecting the eluent, and adding a phosphoric acid solution to obtain a carminic acid extracting solution; the analysis solution contains (0.8-1.2) by volume: (8-12): (36.8-41.2) a mixed aqueous solution of ammonia water, ethanol and water.

2. The method for extracting and purifying carminic acid from soft sweets according to claim 1, wherein in the step (1), the ratio of the soft sweets sample, the water saturated sodium sulfate solution and the dilute acid solution is (0.1-5) g: (8-30) ml: (0-4) ml: (0.4-3) ml.

3. The method for extracting and purifying carminic acid from soft sweets according to claim 1, wherein in the step (1) and the step (2), the mass concentration of the citric acid is 0.8% -1.2%.

4. The method for extracting and purifying carminic acid from soft sweets according to claim 1, wherein in the step (2), the rotation speed of the centrifugation is 6000-8000 r/min, and the centrifugation time is 3-5 min.

5. The method for extracting and purifying carminic acid from soft sweets according to claim 1, wherein in the step (2), the sample solution is dissolved by absolute ethyl alcohol and then centrifuged to obtain a centrifugate and a residue, and the centrifugate is added into a chromatographic column filled with polyamide for elution.

6. The method for extracting and purifying carminic acid from soft sweets according to claim 5, wherein the residues are washed with ethanol with a volume concentration of 60-70%, the centrifugate and the ethanol washing solution are combined, the mixture is concentrated under reduced pressure at 60-80 ℃ to obtain a concentrated solution, and the concentrated solution is transferred to a chromatographic column filled with polyamide by using a citric acid solution for elution.

7. The method for extracting and purifying carminic acid from soft sweets as claimed in claim 1, wherein in step (2), the particle size of the polyamide is 100-200 mesh.

8. The method for extracting and purifying carminic acid from soft sweets according to claim 1, wherein in the step (4), the volume concentration of the phosphoric acid solution is 3% to 4%.

Technical Field

The invention belongs to the technical field of health-care products and foods, and particularly relates to a method for extracting and purifying carminic acid in soft sweets.

Background

Cochineal red is a natural pigment extracted from cochineal bodies, is pink to purple, is an anthraquinone natural pigment, is different from other natural pigments, has stable physicochemical properties, is regarded as the safest natural pigment, and can be used as a colorant for drinks such as wine, fruit pulp and cold drinks, candies, cakes, meats, sausages and the like. GB 2760 supplement 2014 food safety national standard food additive use standard specifies the addition amount of cochineal red, and the addition amount needs to be calculated by carminic acid.

The currently reported detection methods of carminic acid mainly include spectrophotometry, thin layer chromatography, high performance liquid chromatography and the like. In the detection process of the spectrophotometry, the total absorbance at a single wavelength is required, so that the condition that non-carminic acid also absorbs at the wavelength cannot be eliminated, the accuracy of the detection result is influenced, and the analysis and the determination are time-consuming. And the high performance liquid chromatography is easily interfered by complex matrixes in food and is limited in application. The conventional high performance liquid chromatography only simply processes a sample, and the sample facing complex matrixes such as saccharides and the like in actual detection cannot effectively separate carminic acid, is difficult to characterize and easily blocks a chromatographic column, so that the service life of the chromatographic column is greatly reduced. Meanwhile, the content of carminic acid in the soft candy sample is very low, and the carminic acid cannot be effectively detected only by simple treatment, so that the aim of controlling the carminic acid is difficult to achieve.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide a method for extracting and purifying carminic acid from soft sweets, which can effectively separate complex matrixes such as saccharides and the like from carminic acid and improve the accuracy of detecting the content of carminic acid.

The invention is realized by the following technical scheme:

a method for extracting and purifying carminic acid in soft sweets comprises the following steps:

(1) adding water into a soft sweet sample, heating at 70-80 ℃ to dissolve the sample, sequentially adding a saturated sodium sulfate solution and a dilute acid solution, and uniformly mixing to obtain a sample solution; the dilute acid is citric acid or formic acid;

(2) centrifuging the sample solution, and adding the centrifugate into a chromatographic column filled with polyamide;

(3) eluting the chromatographic column with a washing solution, eluting the chromatographic column with water until the eluent is neutral, and discarding the eluent; the washing liquid contains (0-4) by volume: (1-3): (18-30) a mixed aqueous solution of a saturated sodium sulfate solution, a citric acid solution and water;

(4) eluting the chromatographic column for multiple times by using the resolving liquid, collecting the eluent, and adding a phosphoric acid solution to obtain a carminic acid extracting solution; the analysis solution contains (0.8-1.2) by volume: (8-12): (36.8-41.2) a mixed aqueous solution of ammonia water, ethanol and water.

Preferably, in the step (1), the ratio of the soft candy sample, water, saturated sodium sulfate solution and dilute acid solution is (0.1-5) g: (8-30) ml: (0-4) ml: (0.4-3) ml.

Preferably, in the step (1) and the step (2), the mass concentration of the citric acid is 0.8-1.2%.

Preferably, in the step (2), the rotation speed of the centrifugation is 6000 to 8000r/min, and the centrifugation time is 3 to 5 min.

Preferably, in the step (1), after two solvents, namely a saturated sodium sulfate solution and a dilute acid solution, are sequentially added, the sample solution needs to be ensured not to generate solidification and gel substances, and after centrifugation, the centrifugate is clear and has good fluidity and can smoothly pass through a chromatographic column filled with polyamide powder (the flow rate is more than 0.5 drop/S); if the requirement cannot be met, in the step (2), the sample solution is dissolved by absolute ethyl alcohol and then centrifuged to obtain centrifugate and residue, and the centrifugate is added into a chromatographic column filled with polyamide for elution.

Further preferably, the residue is washed for 1-2 times by using ethanol with the volume concentration of 60-70%, the centrifugate and the ethanol washing liquid are combined and concentrated under reduced pressure at the temperature of 60-80 ℃ to obtain a concentrated solution, and the concentrated solution is transferred to a chromatographic column filled with polyamide by using a citric acid solution for elution.

Preferably, in the step (2), the particle size of the polyamide is 100-200 meshes, the specification of the chromatographic column is 10mm multiplied by 250mm, a cock is arranged, and the polyamide powder is dispersed by a small amount of 1% citric acid solution and transferred to the chromatographic column.

Preferably, in the step (4), the volume concentration of the phosphoric acid solution is 3-4%.

Compared with the prior art, the invention has the following beneficial effects:

the invention provides a method for extracting and purifying carminic acid in soft sweets aiming at soft sugar products, wherein the carminic acid is adsorbed by polyamide powder to remove impurities in an acidic environment, and after being eluted by analytic liquid, complex matrixes such as sugar and the like can be effectively separated from the carminic acid to obtain a high-purity carminic acid extracting solution, so that the problem of inaccurate detection caused by low content of the carminic acid in the soft sweets is solved, and the accurate detection of the content of the carminic acid in the soft sweets is realized.

Detailed Description

The present invention is further illustrated by the following specific embodiments, which are not intended to limit the scope of the invention.

The raw materials of the invention are all commercially available.