CN117629903A - Quality evaluation method of ginseng oyster paste - Google Patents
Quality evaluation method of ginseng oyster paste Download PDFInfo
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- CN117629903A CN117629903A CN202311383971.4A CN202311383971A CN117629903A CN 117629903 A CN117629903 A CN 117629903A CN 202311383971 A CN202311383971 A CN 202311383971A CN 117629903 A CN117629903 A CN 117629903A
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Abstract
The invention relates to a quality evaluation method of ginseng oyster paste, which belongs to the technical field of traditional Chinese medicine, and comprises the following test items: comprising the following steps: s1, performing sensory property evaluation experiments; and at least two of the following test items: s2, measuring the polyphenol content of the ginseng oyster paste; s3, measuring the content of total saponins in the ginseng oyster paste; s4, measuring the polysaccharide content of the ginseng oyster paste; s5, measuring the content of total flavonoids in the ginseng oyster paste. The evaluation method of the invention performs quality control on the ginseng oyster cream by screening proper evaluation indexes, and better plays the role of health care and health preservation of the ginseng oyster cream.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicines, in particular to a quality evaluation method of ginseng oyster paste, which is used for controlling the quality of ginseng oyster paste by screening proper evaluation indexes and better playing the health care and health preserving effects of the ginseng oyster paste.
Background
Especially, the modern society has fast life rhythm and high working pressure, and more men face the problems of qi and blood deficiency and kidney qi deficiency, so that the male reproductive health problem is caused, and serious troubles are caused to the life quality and physical and mental health. At present, the market lacks special medicines aiming at the diseases, and most of health-care products are promoted to be completely inconsistent with the components which actually exert the medicine effect.
The hormone components are added in part of health care products in violation, and although the strong nerve excitation stimulation effect is caused in a short time, after the drug effect is resolved, the physical burden of a patient is increased, the health state is further deteriorated, a large amount of the drugs have to be repeatedly used, and finally irreversible damage is caused.
The ginseng oyster paste is mainly prepared from traditional Chinese medicinal materials such as ginseng, oyster and the like, has the effects of tonifying qi and blood, calming heart and soothing nerves, and is used for relieving uncomfortable symptoms caused by deficiency of qi and blood and malnutrition of heart spirit. The ginseng oyster paste has good conditioning and health care effects on patients with kidney qi deficiency and body deficiency, can radically regulate eight extra channels, relax the channels and activate collaterals, and eliminate sub-health problems caused by kidney qi deficiency and body deficiency.
Meanwhile, the ginseng oyster paste belongs to the homology of medicine and food, can be directly eaten, is not limited by environmental conditions, and is deeply favored by wide health-preserving lovers. In addition, the ginseng oyster paste can be matched with foods such as red dates, longan, medlar and the like in the eating process, so that the ginseng oyster paste can supplement needed nutrition for the body.
The quality research of the ginseng oyster paste in the prior art is less, a large number of inferior products which are filled in a second time exist on the market, the quality of the products is difficult to distinguish by non-professionals, and general consumers are often subjected to false marketing and even sales fraud of illegal vendors.
Although the Chinese pharmacopoeia provides some detection methods for the Chinese medicinal ointment, most of the detection methods are general detection and analysis methods, have no pertinence, and cannot effectively distinguish the substantial quality of the ginseng oyster ointment.
Disclosure of Invention
Aiming at the problems that the quality evaluation method of the ginseng oyster paste is rough and cannot effectively evaluate the actual quality expression of the ginseng oyster paste in the prior art, the invention provides the quality evaluation method of the ginseng oyster paste, the quality control of the ginseng oyster paste can be realized by the method, and the formula product which can exert the health care and health preservation effects most can be screened.
In order to achieve the above object, the following technical solutions are provided:
a quality evaluation method of Ginseng radix Concha Ostreae paste comprises:
s1, performing sensory property evaluation experiments;
and at least two of the following test items:
s2, measuring the polyphenol content of the ginseng oyster paste;
s3, measuring the content of total saponins in the ginseng oyster paste;
s4, measuring the polysaccharide content of the ginseng oyster paste;
s5, measuring the content of total flavonoids in the ginseng oyster paste.
According to the method for evaluating the quality of the ginseng oyster paste, a plurality of detection and analysis indexes are designed by combining the characteristics of the ginseng oyster paste product, the quality performance of the product can be accurately researched and analyzed, the effective evaluation and analysis of the part of the active ingredients in various traditional Chinese medicinal material ingredients, which are converted into the oyster paste, are ensured, and the quality of the ginseng oyster paste researched by the evaluation method meets the expected target.
According to the ginseng oyster paste quality evaluation method, the test items are not distinguished in sequence, and the test sequence can be adjusted according to actual conditions.
Further, S1, sensory property evaluation experiment: the test items include: mouthfeel, morphology and color.
Wherein, the taste score accounts for 40%, the morphology score accounts for 30%, and the color score accounts for 30%.
Further, the scoring criteria for sensory trait evaluation were as follows:
further, S2, measuring polyphenol content of the ginseng oyster paste: adopting gallic acid as a standard component, diluting to prepare a reference substance solution with gradient concentration, adding a Fu Lin Fen test solution and a sodium carbonate solution, fully shaking uniformly, and fixing the volume by using distilled water; the absorbance was measured at 768nm and a standard curve was drawn.
Then, the ginseng oyster paste is taken and dissolved in ethanol solution, and the supernatant is taken, and the ginseng oyster paste is detected under the same detection condition.
Further, S2, measuring polyphenol content of the ginseng oyster paste: the gallic acid is adopted as a standard component, diluted into a control solution with the concentration of 0.05mg/mL, 0, 0.2, 0.4, 0.6, 0.8 and 1.0mL are respectively absorbed into test tubes with plugs, 0.5mL of Fu Lin Fen test solution is respectively added, 2mL of 7.5 percent sodium carbonate solution is added within 5min, the mixture is fully and evenly shaken, and distilled water is used for fixing the volume to 10mL; after being fully and uniformly mixed, the mixture is transferred to a light-shielding environment for reaction for 120min, the absorbance is measured at 768nm, and 768nm is the maximum absorption wavelength determined by scanning. And drawing a standard curve by taking the concentration of the reference substance solution as an abscissa and the absorbance as an ordinate.
Then, 2.00g of ginseng oyster cream is taken and dissolved in 60v% ethanol solution and fixed in a 50mL volumetric flask, 0.2mL of the ginseng oyster cream is taken and added into a 25mL volumetric flask, and the ginseng oyster cream is detected under the same detection condition.
Further, S3, measuring the content of total saponins of the ginseng oyster paste: dissolving diosgenin reference substance in methanol to obtain reference substance solution with gradient concentration; volatilizing methanol, adding vanillin acetic acid solution, adding perchloric acid under ice water bath condition for reaction, measuring absorbance at 550nm, and drawing to obtain a standard curve equation of diosgenin;
adding ethanol into the ginseng oyster paste for ultrasonic dissolution; centrifuging and collecting supernatant; volatilizing ethanol; adding vanillin acetic acid solution, adding perchloric acid under ice water bath condition for reaction, measuring absorbance at 550nm, and calculating to obtain sapogenin content.
Further, S3, measuring the content of total saponins of the ginseng oyster paste: placing 3.0mg of diosgenin reference substance in 25mL volumetric flask, and adding methanol to constant volume to scale mark; taking 0.3, 0.6, 0.9, 1.2, 1.5 and 1.8mL of the mother solution by a pipette, placing the mother solutions in test tubes with plugs, respectively marking, volatilizing methanol at 60 ℃, and adding 0.2mL of the prepared vanillin acetic acid solution; adding 0.8mL of perchloric acid under the ice water bath condition, shaking uniformly, immediately carrying out water bath at 60 ℃ for 15min, and taking out the immediately ice water bath for 2min; taking out, adding 5mL glacial acetic acid, shaking, standing for 15min, measuring absorbance at 550nm, and drawing to obtain the standard curve equation of diosgenin.
Weighing appropriate amount of Ginseng radix Concha Ostreae paste, placing in 50mL volumetric flask, adding 80% ethanol, and dissolving with ultrasound; centrifuging to collect 0.2mL of supernatant, adding 10mL test tube, volatilizing ethanol, and measuring absorbance according to the same method as that of the diosgenin standard; and carrying out a standard curve equation of the diosgenin, and calculating to obtain the sapogenin content.
Further, S4, measuring the polysaccharide content of the ginseng oyster paste: 17.5mg of anhydrous glucose reference substance is precisely weighed and dissolved in a 100mL volumetric flask to prepare a glucose reference substance solution of 0.175 mg/mL. Precisely measuring 0.1, 0.2, 0.3, 0.4 and 0.5mL of reference substance solution, placing the reference substance solutions into test tubes with plugs, respectively adding 1.5mL of 4.5% phenol solution, uniformly mixing, rapidly adding 6.5mL of sulfuric acid, shaking uniformly, preserving heat in a water bath at 40 ℃ for 30 minutes, taking out, placing in an ice water bath for 5 minutes, taking out, taking the corresponding reagent as a blank, measuring absorbance at a wavelength of 490nm according to an ultraviolet-visible spectrophotometry, taking absorbance as an ordinate, taking concentration as an abscissa, and drawing a standard curve.
Precisely weighing 0.2g of ginseng oyster extract, dissolving in a 50mL volumetric flask, fixing the volume, and performing absorbance of a ginseng oyster extract sample solution according to an experimental method drawn by a glucose standard curve; and carrying the measured absorbance value into a glucose concentration absorbance regression equation, and calculating to obtain the polysaccharide concentration of the ginseng oyster extract sample.
Further, S5, measuring the content of total flavonoids in the ginseng oyster paste: weighing rutin control 3.1mg, adding appropriate amount of 60v% CH 3 CH 2 Micro-dissolving in OH water bath, placing in 5mL volumetric flask, and fixing volume to obtain rutin control solution of 0.64 mg/mL; precisely taking 1.5, 2.0, 2.5, 3.0, 3.5mL and 0.1mol/mL AlCl with 2mL concentration respectively in 10mL volumetric flask 3 3mL of CH with a concentration of 1mol/mL 3 COOK solution, 60v% CH 3 CH 2 OH is fixed in volume, shaken uniformly, kept stand for 30min, then scanned by a corresponding blank reagent according to an ultraviolet-visible spectrophotometry, absorbance is measured at a wavelength of 424nm, and a standard curve is drawn.
Weighing 0.0802g of ginseng oyster paste, dissolving in 2mL centrifuge tube, adding 60v% CH 3 CH 2 Fully dissolving OH, centrifuging, filtering to obtain supernatant, precisely measuring 1mL of supernatant in a 10mL volumetric flask; and (3) measuring absorbance according to the same test method of the rutin standard substance, and carrying out calculation to obtain a total flavone content test result by a rutin concentration absorbance regression equation of the rutin standard substance brought into a standard curve.
Through the technical scheme, compared with the prior art, the invention has the following technical effects:
1. according to the method for evaluating the quality of the ginseng oyster paste, a plurality of detection and analysis indexes are designed by combining the characteristics of the ginseng oyster paste product, the quality performance of the product can be accurately researched and analyzed, the effective evaluation and analysis of the part of the active ingredients in various traditional Chinese medicinal material ingredients, which are converted into the oyster paste, are ensured, and the quality of the ginseng oyster paste researched by the evaluation method meets the expected target.
2. The evaluation method of the invention performs quality control on the ginseng oyster cream by screening proper evaluation indexes, and better plays the role of health care and health preservation of the ginseng oyster cream.
Description of the drawings:
FIG. 1 is an absorbance curve of taurine standard solution.
FIG. 2 is a standard curve drawn for a polyphenol standard solution.
FIG. 3 is a standard curve of diosgenin.
FIG. 4 is a glucose standard curve.
Fig. 5 is a rutin control standard curve.
FIG. 6 is a software operation interface in the process of drawing a standard curve according to the rutin content test result.
Detailed Description
In order to more clearly describe the objects, technical solutions and advantages of the present invention in the specific embodiments, the following detailed description will be made with reference to the specific embodiments. The following specific examples relate to the specific technical solutions only for the purpose of clearly and completely describing the innovative technical solution of the present invention, and are themselves only some, but not all, examples of the specific embodiments that the present invention may employ and are not to be construed as limiting the innovative solutions of the present invention. Any solution employing the same inventive concept should be included in the scope of protection of the present invention.
For those skilled in the art, reference may be made to the conventional technical manual in the art for an understanding of the solutions described in the specific examples of the present invention, and appropriate understanding or adjustments may be made with reference to the places where the above terms appear, so that the same or similar technical solutions are deduced without the inventive effort.
The ethanol solutions which are not specifically described in the invention are all ethanol water solutions, wherein the percentage value is the volume percentage of ethanol in the ethanol water solution. The rest solid reagent preparation solutions are prepared by weight percent, a certain amount of solution is firstly measured, and then corresponding solute is added according to the weight percent to prepare corresponding solution for use.
Example 1
Preparation process of ginseng oyster paste
The raw materials of the ginseng oyster paste are prepared according to the following formula: 30 g of mulberry, 30 g of oyster, 3 g of ginseng (less than 5 years old), 10 g of polygonatum, 20g of medlar, 20g of raspberry, 15 g of poria cocos, 15 g of Chinese yam, 15 g of fructus alpiniae oxyphyllae, 20g of isomaltooligosaccharide and 3 g of liquorice.
1. Oyster extraction
Adding water into oyster according to a feed liquid ratio of 1:10 immersing, adding 3000U/g enzyme, and performing enzymolysis for 6h, and adjusting various proteases to perform enzymolysis test under the conditions of optimal enzymolysis temperature and the same pH value, wherein the specific protease type selection and enzyme adding U/g, temperature, time, pH value and other conditions are shown in table 1.
As can be seen from the results in table 1, the cellulase: the highest yield of taurine extracted by trypsin (mixing ratio) (1:1) through enzymolysis is 1.256 percent. Thus, alkaline protease is used as a preferred enzyme for the extraction of oyster taurine. And measuring taurine content by adopting a pharmacopoeia method, bringing the taurine content into the taurine standard solution absorbance curve shown in figure 1, and calculating to obtain different protease enzymolysis conditions and taurine yield.
TABLE 1 enzymatic hydrolysis conditions of different proteases and taurine yield
2. Extracting other medicinal materials
The extraction process of the rest materials (except oyster) of the ginseng oyster paste is as follows. The rest materials are dried to constant weight, crushed into powder, sieved by a 60-mesh sieve, 20g of powder is weighed into two parts, the powder is refluxed for 4 hours at 80 ℃ with 95v% ethanol, and residues are dried for standby. One part is added into a flash extractor, and the other part is decocted with conventional water, and water is used as an extraction solvent. After the extraction is finished, carrying out suction filtration, concentrating the filtrate under reduced pressure to 50mL, regulating the volume fraction of ethanol in the polysaccharide solution to 80%, precipitating with ethanol at 4 ℃ overnight, centrifugally collecting precipitate, drying until the precipitate has no alcohol smell, dissolving with a proper amount of distilled water, and freeze-drying to obtain crude polysaccharide (2 parts).
TABLE 2 polysaccharide content and yield (wt%) extracted in two different ways
| Extraction mode | Temperature (. Degree. C.) | Time (h) | Number of extractions | Feed-to-liquid ratio | Polysaccharide yield | Polysaccharide content |
| Decocting in water | 100 | 4 | 2 | 1：30 | 9.3％ | 40.2％ |
| Flash extraction | 60 | 0.5 | 1 | 1：30 | 15.2％ | 60.33％ |
3. Concentrating
Concentrating the extractive solution obtained by extracting Concha Ostreae and other materials to density of 1.35g/mL by membrane permeation concentration technology; the membrane permeation concentration time is quick, the active ingredients are well reserved, and the energy is saved.
The membrane permeation concentration is a non-heating concentration process technology, and is often divided into reverse osmosis, nanofiltration, ultrafiltration and microfiltration, the principle is that the technology for separating, classifying, purifying and enriching two-component or multi-component gas or liquid by means of concentration difference, chemical potential difference or external force through the permeation action of a specific membrane is one of the most advanced technologies in the technical field of modern separation. The method is characterized in that the efficient concentration and enrichment or separation of substances are realized under the normal-temperature physical environment, the impurities are effectively removed, and the whole process has low energy consumption and no phase change.
3. Honey for training
Isomaltooligosaccharides (edible for diabetics): adding appropriate amount of water, heating with small fire, stirring, dissolving, removing floating foam, filtering with No. 4 sieve, heating with small fire, stirring, and refining to obtain golden yellow thick liquid.
The relative density of refined honey is 1.37g/mL.
3. Ointment for collecting
Concentrating to obtain concentrated extract with density of 1.35g/mL, adding 3 times of isomaltooligosaccharide, decocting to pasty, heating with slow fire, mixing, and stirring to prevent bottoming or gelatinization. Sticking bamboo chip to paste, and stopping heating to obtain Ginseng radix Concha Ostreae paste.
The ginseng oyster paste used in the following examples is the ginseng oyster paste product prepared by the method of example 1, and the specific oyster extraction method is a method for selecting serial number 5 (cellulase: trypsin, mixing ratio 1:1, and enzyme addition amount is 2000U/g); extracting the other materials except Concha Ostreae with flash extractor.
Comparative example 1
Preparation process of ginseng oyster paste
The raw materials of the ginseng oyster paste were prepared according to the same raw material formulation as in example 1.
The extraction process is similar to that of example 1, except that the rotary evaporator is used for reduced pressure evaporation concentration in the concentration process, and the membrane permeation concentration technology is not used, so that the reduced pressure evaporation concentration takes 3 hours, and the equipment operation energy consumption is relatively high. Concentrating under reduced pressure to obtain concentrated extract with feed liquid density of 1.35g/mL, adding isomaltooligosaccharide, decocting to pasty state, heating with slow fire, mixing, and stirring until the material is thread-shaped to obtain Ginseng radix Concha Ostreae extract.
Example 2
Sensory trait evaluation experiment
The sensory shape of the ginseng oyster paste prepared in example 1 was evaluated, and the experimental method was as follows.
The sensory evaluation method is a main food texture evaluation means, has strong subjectivity and poor reliability. To improve the reliability of sensory evaluation, 10 panelists consisted of 5 men and 5 women. The food with heavier smell is evaluated about one to two hours after meals, the food with heavier smell is not eaten within half an hour before tasting, the paste is firstly rinsed with purified water and then evaluated, the evaluated place is bright and odorless, the light judgment and the smell interference are not influenced, and the sensory evaluation criteria are shown in the following table.
Sensory scoring criteria
The scores obtained will be statistically demonstrated, and the final experimental results are shown in the following table.
| Sequence number | Sensory trait score |
| 1 | 81.38 |
| 2 | 81.25 |
| 3 | 80.87 |
| 4 | 80.12 |
| 5 | 71.82 |
| 6 | 74.36 |
| 7 | 79.25 |
| 8 | 72.28 |
| 9 | 73.81 |
| 10 | 74.92 |
The average score is 77.01, the overall sensory evaluation score of the ginseng oyster paste prepared in example 1 is high, the quality is good, and the ginseng oyster paste prepared in example 1 is subjected to chemical analysis in the following examples to evaluate the content of active ingredients.
Example 3
Ginseng oyster extract polyphenol content determination
1. Standard curve drawing
Precisely weighing 5.0mg of gallic acid, dissolving with distilled water, placing into a 100mL volumetric flask to fix volume, preparing into 0.05mg/mL reference substance solution, respectively sucking 0, 0.2, 0.4, 0.6, 0.8 and 1.0mL into test tubes with plugs, respectively adding 0.5mL of Fu Lin Fen test solution, adding 2mL of 7.5% sodium carbonate solution within 5min, shaking completely, and fixing volume to 10mL with distilled water. After being fully and uniformly mixed, the mixture is transferred to a light-shielding environment for reaction for 120min, the absorbance is measured at 768nm, and 768nm is the maximum absorption wavelength determined by scanning. And drawing a standard curve by taking the concentration of the reference substance solution as an abscissa and the absorbance as an ordinate.
Standard equation y=113.65x+0.0638, r 2 = 0.9978, the result is shown in fig. 2.
The most important thing to measure the total content with an ultraviolet spectrophotometer is to draw a standard curve, and the amounts of the reference substance and the color-developing agent must be strictly controlled in the process, and the used container must be washed clean. And (3) carrying out gradient dilution on the standard solution to obtain a plurality of concentration gradients for measurement, drawing an accurate standard curve by using the test results of the standard solutions with different concentrations, and improving the test precision.
2. Content determination
2.00g of the ginseng oyster paste prepared in example 1 was dissolved in 60v% ethanol solution and fixed in a 50mL volumetric flask, 0.2mL was added to the 25mL volumetric flask, and the experiment was performed as described above.
3. Experimental results
Dilution factor: 2/50×0.2/25=0.32 mg/mL
Absorbance: 0.5362 the corresponding concentration=0.0042 mg/mL is obtained by equation y=113.65x+0.0638.
Therefore, the polyphenol content in the sample was 0.0042++0.32X100% =1.31%.
Comparative example 1
Ginseng oyster extract polyphenol content determination
The polyphenol content of the ginseng oyster shell cream prepared in example 1 was measured, and the experimental method was as follows.
1. Standard curve drawing
The standard curve was plotted as in step 1 of example 3.
2. Content determination
2.00g of the ginseng oyster paste prepared in the example 1 is taken, distilled water is added for dissolution, stirring is carried out for 5min, the mixture is dissolved in a volumetric flask of 100mL, 0.2mL of the mixture is taken and added into the volumetric flask of 25mL, the operation is carried out by the same method according to the standard curve drawing, and the polyphenol content is determined experimentally.
3. Experimental results
The polyphenol content of the ginseng oyster extract sample is 0.87% which is lower than the test result of the example 3, and the test result of the example 3 is correct by adopting the national standard method for correction and inspection, and the test result of the comparative example distilled water extraction has deviation.
Test results show that although distilled water is adopted to dissolve the sample of the ginseng oyster cream, and a larger volumetric flask is adopted to dissolve the sample, the distilled water is fully stirred, but the distilled water is insufficient in dissolving polyphenol in the sample, so that the test results are inaccurate, and finally ethanol is selected to dissolve and disperse the sample, so that the polyphenol content in the ginseng oyster cream is tested.
Example 4
Content determination of total saponins of Ginseng radix Concha Ostreae extract
1. Preparation of experiments
1. Drawing standard curve of diosgenin
(1) Taking 3.0mg of diosgenin reference substance, placing in 25mL volumetric flask, adding methanol to constant volume until the scale mark is reached.
(2) Pipette 0.3, 0.6, 0.9, 1.2, 1.5 and 1.8mL of the mother solution are placed in a test tube with a stopper, respectively marked, methanol is volatilized at 60 ℃, and 0.2mL of the prepared vanillin acetic acid solution is added.
(3) Under the ice water bath condition, 0.8mL of perchloric acid is added, the mixture is uniformly shaken, the water bath is immediately carried out at 60 ℃ for 15min, and the immediate ice water bath is taken out for 2min.
(4) Taken out, 5mL of glacial acetic acid is added and shaken well, and the mixture is kept stand for 15min, and the absorbance is measured at 550 nm.
Drawing to obtain a standard curve equation of the diosgenin: y=16.916x+0.1159, r 2 = 0.9996, the result is shown in fig. 3.
2. Preparation of an object to be measured
(1) Weight reduction method A proper amount of the ginseng oyster paste prepared in example 1 (4.5304 g is weighed in practical experiment) is put into a 50mL volumetric flask, and is added with 80v% ethanol for ultrasonic dissolution
(2) Centrifuging the above solution to collect supernatant
(3) Collecting supernatant 0.2mL in 10mL test tube, volatilizing ethanol, and measuring absorbance of total saponin extract of Ginseng radix and Concha Ostreae according to the preparation method of diosgenin standard curve
2. Experimental results
Measuring absorbance of total saponin extract of the ginseng oyster extract sample: 0.3226, carrying into a standard curve equation, and calculating to obtain the total saponin content of the ginseng oyster shell cream: 1.62%.
Example 5
Ginseng oyster extract polysaccharide content determination
1. Drawing a standard curve:
17.5mg of anhydrous glucose reference substance is precisely weighed and dissolved in a 100mL volumetric flask to prepare a glucose reference substance solution of 0.175 mg/mL. Precisely measuring 0.1, 0.2, 0.3, 0.4 and 0.5mL of reference substance solution, placing the reference substance solutions into test tubes with plugs, respectively adding 1.5mL of 4.5% phenol solution, uniformly mixing, rapidly adding 6.5mL of sulfuric acid, shaking uniformly, preserving heat in a water bath at 40 ℃ for 30 minutes, taking out, placing in an ice water bath for 5 minutes, taking out, taking the corresponding reagent as a blank, measuring absorbance at a wavelength of 490nm according to an ultraviolet-visible spectrophotometry, taking absorbance as an ordinate, taking concentration as an abscissa, and drawing a standard curve.
The glucose standard curve is plotted in FIG. 4, where the abscissa indicates concentration in μg/mL and the ordinate indicates absorbance (A). And fitting according to the test result to obtain a glucose concentration absorbance regression equation.
Y＝0.0627X-0.0307，R 2 ＝0.9988。
2. And (3) content measurement:
0.2g of the ginseng oyster paste prepared in example 1 is precisely weighed and dissolved in a 50mL volumetric flask to fix the volume, 1.5mL, 1.0mL and 2.0mL are respectively taken, and the absorbance of the ginseng oyster paste sample solution is carried out according to the experimental method drawn by the glucose standard curve from the step of adding 1.5mL of the 4.5% phenol solution respectively. And carrying the absorbance value obtained by the test into a glucose concentration absorbance regression equation, and calculating to obtain the polysaccharide concentration of the ginseng oyster extract sample.
The polysaccharide content test process of the ginseng oyster shell cream sample is repeated three times, and the sample measurement data are shown in the following table.
| Sequence number | 490.0nm | 490.0nm | 490.0nm | Average value of | Concentration (μg/mL) | Content of |
| 1 | 0.3933 | 0.3933 | 0.3933 | 0.3933 | 6.76331738 | 42.2％ |
| 2 | 0.7901 | 0.7901 | 0.7901 | 0.7901 | 13.0909091 | 40.8％ |
| 3 | 2.0489 | 2.0493 | 2.049 | —— | —— | —— |
Wherein, the third test data are invalid, and the average value is calculated according to the 1 st to 2 nd measurement results, so that the polysaccharide content in the ginseng oyster cream is 41.5%.
According to the component list of the ginseng oyster paste, mulberry, fragrant solomonseal rhizome and siberian solomonseal rhizome belong to the same genus plant of the same family and are main components with higher proportion. Taking rhizoma polygonati as a center point, consulting 20 edition pharmacopoeia, taking anhydrous glucose as a reference substance to prepare a glucose reference substance solution of 0.6mg/mL, and respectively taking 1.0, 1.5, 2.0, 2.5 and 3.0mL of the reference substance solution to dilute in a 50mL test tube with a plug. According to the actual test result, finally transferring 0.2, 0.6, 0.8 and 1.0mL of solution to be diluted according to the modification of the law color development index, and operating to find that the absorbance value of the measured solution shows an ascending trend, and the concentration of 0.4mL reaches 0.768 and is higher; adjusting the concentration of the reference substance solution to 0.3mg/mL, precisely removing 0.3, 0.5, 0.7, 0.8 and 0.9mL, diluting and measuring, wherein the absorbance of the measured result is lower than 0.2, the concentration of the solution is lower, continuously adjusting the concentration to be constantly and precisely absorbed by 1.0, 1.5, 2.0, 2.5 and 3.0mL, and measuring according to a law, wherein the absorbance of the measured result is lower than 0.2; and (3) adjusting the scheme, searching a regression literature, inquiring a teacher and the like, finally adjusting the glucose concentration to 0.175mg/mL, precisely removing 0.1, 0.2, 0.3, 0.4 and 0.5mL, directly adding distilled water to 2mL, and performing legal measurement.
The display agent is prepared at present and a 4.5% phenol solution is selected. If the phenol solution is left for too long, phenol will oxidize and cause errors in the measurement results.
Example 6
Determination of total flavone content in ginseng oyster extract
1. Drawing a standard curve:
accurately weighing rutin control 3.1mg, adding 60% CH 3 CH 2 And (3) carrying out micro-dissolution on the OH water bath, placing the solution into a 5mL volumetric flask for constant volume, and preparing a rutin control solution with the concentration of 0.64 mg/mL. Precisely taking 1.5, 2.0, 2.5, 3.0, 3.5mL and 0.1mol/mL AlCl with 2mL concentration respectively in 10mL volumetric flask 3 3mL of CH with a concentration of 1mol/mL 3 COOK solution, 60v% CH 3 CH 2 OH is fixed in volume, shaken uniformly, kept stand for 30min, and then the corresponding blank reagent is used for scanning the wavelength according to an ultraviolet-visible spectrophotometry, so that the absorption of the solution at 424nm is known, namely, the absorbance is measured at the wavelength of 424nm, the absorbance is taken as an ordinate, the concentration is taken as an abscissa, and a standard curve is drawn.
The rutin standard curve is shown in FIG. 5, wherein the abscissa represents concentration, the unit of μg/mL, and the ordinate represents absorbance (A). And fitting according to the test result to obtain a rutin concentration absorbance regression equation.
Regression equation y= 0.0284X-0.03, r 2 ＝0.9997。
The abscissa X is the rutin concentration, and the unit mu g/mL; the ordinate Y is absorbance (a).
The software operation interface in the process of drawing the standard curve according to the rutin content test result is shown in figure 6. In the original measurement process, each group of data is found to be in an unstable state, and analysis determines that the first two groups of data are unstable measured data values due to the fact that the concentration of the solution is not too low between 0.2 and 0.8, and the later groups of data values are all between 0.2 and 0.8. Meanwhile, examination of the solution found that white flocs appeared at the bottom of the solution, presumably due to flocs at the bottom of the solution.
The color reagent used for the total flavone determination is AlCl with the concentration of 2mL and 0.1mol/mL 3 3mL of CH with a concentration of 1mol/mL 3 COOK solutions, wherein the main effect is Al 3+ The substances reacted with the flavonoids absorb ultraviolet light, and act as pH regulator (buffer solution) by adding CH 3 COOK can promote Al by gradually increasing pH of originally acidic solution 3+ Is reacted with a flavonoid compound to obtain the product,however, when the solution is too alkaline, al in the solution 3+ Will be with OH + Reaction to Al (OH) 3 The white precipitate, i.e. the bottom floc of the volumetric flask, was adjusted to reduce and avoid this during the calibration curve drawing.
2. And (3) content measurement:
accurately weighing 0.0802g of ginseng oyster paste prepared in example 1, dissolving in 2mL centrifuge tube, adding 60% CH 3 CH 2 OH was dissolved well, the supernatant was collected by centrifugation, 1mL of the supernatant was precisely measured in a 10mL volumetric flask from "2 mL of 0.1mol/mL AlCl, respectively 3 And (3) measuring absorbance according to a law, carrying out a rutin concentration absorbance regression equation of a standard curve, and calculating to obtain a total flavone content test result.
The test was repeated twice to obtain total flavone content data of ginseng oyster extract samples as shown in the following table.
| Sequence number | 424.0nm | 424.0nm | 424.0nm | Average value of | Concentration (μg/mL) | Content of |
| 1 | 0.5004 | 0.5005 | 0.5005 | 0.5005 | 18.6795775 | 0.38％ |
| 2 | 0.4075 | 0.4076 | 0.4077 | 0.4076 | 15.4084507 | 0.37％ |
According to the two measurement results, calculating the average value to obtain the total flavone content of the ginseng oyster cream of 0.375%.
Claims (10)
1. A quality evaluation method of ginseng oyster is characterized by comprising the following steps:
s1, performing sensory property evaluation experiments;
and at least two of the following test items:
s2, measuring the polyphenol content of the ginseng oyster paste;
s3, measuring the content of total saponins in the ginseng oyster paste;
s4, measuring the polysaccharide content of the ginseng oyster paste;
s5, measuring the content of total flavonoids in the ginseng oyster paste.
2. The method for evaluating the quality of ginseng oyster according to claim 1, wherein S1, sensory property evaluation experiment: the test items include: mouthfeel, morphology and color.
3. The method for evaluating the quality of ginseng oyster according to claim 2, wherein the taste score is 40%, the morphology score is 30% and the color score is 30%.
4. A method for evaluating the quality of oysters according to claim 3, wherein the evaluation of sensory properties is based on the following scoring criteria:
5. the method for evaluating the quality of ginseng and oyster according to claim 1, wherein gallic acid is adopted as a standard component, a reference substance solution with gradient concentration is diluted and prepared, a Fu Lin Fen test solution and a sodium carbonate solution are added, the mixture is fully and uniformly shaken, and distilled water is used for constant volume; measuring absorbance at 768nm, and drawing a standard curve;
then, the ginseng oyster paste is taken and dissolved in ethanol solution, and the supernatant is taken, and the ginseng oyster paste is detected under the same detection condition.
6. The method for evaluating the quality of ginseng oyster according to claim 5, wherein the content of polyphenols in the ginseng oyster extract is determined by S2: the gallic acid is adopted as a standard component, diluted into a control solution with the concentration of 0.05mg/mL, 0, 0.2, 0.4, 0.6, 0.8 and 1.0mL are respectively absorbed into test tubes with plugs, 0.5mL of Fu Lin Fen test solution is respectively added, 2mL of 7.5 percent sodium carbonate solution is added within 5min, the mixture is fully and evenly shaken, and distilled water is used for fixing the volume to 10mL; fully and uniformly mixing, transferring to a light-shielding environment for reaction for 120min, measuring absorbance at 768nm, wherein 768nm is the maximum absorption wavelength determined by scanning; drawing a standard curve by taking the concentration of the reference substance solution as an abscissa and the absorbance as an ordinate;
then, 2.00g of ginseng oyster cream is taken and dissolved in 60v% ethanol solution and fixed in a 50mL volumetric flask, 0.2mL of the ginseng oyster cream is taken and added into a 25mL volumetric flask, and the ginseng oyster cream is detected under the same detection condition.
7. The method for evaluating the quality of ginseng oyster according to claim 1, wherein the content of total saponins of the ginseng oyster cream is determined by S3: dissolving diosgenin reference substance in methanol to obtain reference substance solution with gradient concentration; volatilizing methanol, adding vanillin acetic acid solution, adding perchloric acid under ice water bath condition for reaction, measuring absorbance at 550nm, and drawing to obtain a standard curve equation of diosgenin;
adding ethanol into the ginseng oyster paste for ultrasonic dissolution; centrifuging and collecting supernatant; volatilizing ethanol; adding vanillin acetic acid solution, adding perchloric acid under ice water bath condition for reaction, measuring absorbance at 550nm, and calculating to obtain sapogenin content.
8. The method for evaluating the quality of ginseng oyster according to claim 7, wherein the content of total saponins of the ginseng oyster cream is determined by the following steps: placing 3.0mg of diosgenin reference substance in 25mL volumetric flask, and adding methanol to constant volume to scale mark; taking 0.3, 0.6, 0.9, 1.2, 1.5 and 1.8mL of the mother solution by a pipette, placing the mother solutions in test tubes with plugs, respectively marking, volatilizing methanol at 60 ℃, and adding 0.2mL of the prepared vanillin acetic acid solution; adding 0.8mL of perchloric acid under the ice water bath condition, shaking uniformly, immediately carrying out water bath at 60 ℃ for 15min, and taking out the immediately ice water bath for 2min; taking out, adding 5mL glacial acetic acid, shaking, standing for 15min, measuring absorbance at 550nm, and drawing to obtain a standard curve equation of diosgenin;
weighing appropriate amount of Ginseng radix Concha Ostreae paste, placing in 50mL volumetric flask, adding 80% ethanol, and dissolving with ultrasound; centrifuging to collect 0.2mL of supernatant, adding 10mL test tube, volatilizing ethanol, and measuring absorbance according to the same method as that of the diosgenin standard; bringing the standard curve equation of the diosgenin into the standard curve equation of the diosgenin, and obtaining the sapogenin content by the technology.
9. The method for evaluating the quality of ginseng oyster according to claim 1, wherein the content of the polysaccharide in the S4 ginseng oyster paste is determined: accurately weighing 17.5mg of anhydrous glucose reference substance, dissolving in a 100mL volumetric flask, and preparing into 0.175mg/mL glucose reference substance solution; precisely measuring 0.1, 0.2, 0.3, 0.4 and 0.5mL of reference substance solution, placing the reference substance solutions into test tubes with plugs, respectively adding 1.5mL of 4.5% phenol solution, uniformly mixing, rapidly adding 6.5mL of sulfuric acid, shaking uniformly, preserving heat in a water bath at 40 ℃ for 30 minutes, taking out, placing in an ice water bath for 5 minutes, taking out, taking the corresponding reagent as a blank, measuring absorbance at a wavelength of 490nm according to an ultraviolet-visible spectrophotometry, taking absorbance as an ordinate and concentration as an abscissa, and drawing a standard curve;
precisely weighing 0.2g of ginseng oyster extract, dissolving in a 50mL volumetric flask, fixing the volume, and performing absorbance of a ginseng oyster extract sample solution according to an experimental method drawn by a glucose standard curve; and carrying the measured absorbance value into a glucose concentration absorbance regression equation, and calculating to obtain the polysaccharide concentration of the ginseng oyster extract sample.
10. The method for evaluating the quality of ginseng oyster according to claim 1, wherein the content of total flavonoids in the ginseng oyster cream is determined by S5: weighing rutin control 3.1mg, adding appropriate amount of 60v% CH 3 CH 2 Micro-dissolving in OH water bath, placing in 5mL volumetric flask, and fixing volume to obtain rutin control solution of 0.64 mg/mL; precisely taking 1.5, 2.0, 2.5, 3.0, 3.5mL and 0.1mol/mL AlCl with 2mL concentration respectively in 10mL volumetric flask 3 3mL of CH with a concentration of 1mol/mL 3 COOK solution, 60v% CH 3 CH 2 OH is subjected to constant volume, shaking is carried out, standing is carried out for 30min, the corresponding blank reagent is used for scanning the wavelength according to an ultraviolet-visible spectrophotometry, the absorbance is measured at the wavelength of 424nm, and a standard curve is drawn;
weighing 0.0802g of ginseng oyster paste, dissolving in 2mL centrifuge tube, adding 60v% CH 3 CH 2 Fully dissolving OH, centrifuging, filtering to obtain supernatant, precisely measuring 1mL of supernatant in a 10mL volumetric flask; and (3) measuring absorbance according to the same test method of the rutin standard substance, and carrying out calculation to obtain a total flavone content test result by a rutin concentration absorbance regression equation of the rutin standard substance brought into a standard curve.
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