CN103115887B - Measurement method of total saponins in health food using propolis as raw material - Google Patents
Measurement method of total saponins in health food using propolis as raw material Download PDFInfo
- Publication number
- CN103115887B CN103115887B CN201310035767.3A CN201310035767A CN103115887B CN 103115887 B CN103115887 B CN 103115887B CN 201310035767 A CN201310035767 A CN 201310035767A CN 103115887 B CN103115887 B CN 103115887B
- Authority
- CN
- China
- Prior art keywords
- health food
- propolis
- raw material
- water
- total saposins
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Jellies, Jams, And Syrups (AREA)
Abstract
The invention relates to a measurement method of total saponins in health food, and particularly relates to a measurement method of total saponins in health food using propolis as a raw material. The measurement method provided by the invention comprises a standard curve making process, a sample treatment process, a test liquid column chromatography isolation process, a color developing process and a total saponins calculating process. In the standard curve making process, a standard curve is obtained by using an absorbance value and a mass of ginsenoside as curves; in the sample treatment process, liquid to be tested is obtained by taking 0.3g of the health food using the propolis as the raw material; in the test liquid column chromatography isolation process, column chromatography isolation is carried out on the liquid to be tested by using a chromatographic column, so as to obtain an evaporation pan with the liquid to be tested which is evaporated to be dry; in the color developing process, the absorbance value of the liquid to be tested is obtained; and in the total saponins calculating process, the content of the total saponins in the health food using the propolis as the raw material can be obtained. The measurement method provided by the invention has the advantages of simple extraction method, high recovery rate, good accuracy, simplicity, convenience and rapidness.
Description
Technical field
The present invention relates to the assay method of total saposins in a kind of health food, especially relating to a kind of take propolis as the assay method of total saposins in the health food of raw material.
Background technology
Propolis is the resin of honeybee herborization, and mixed with the tackiness material of self secretion.Propolis can qi-restoratives weak, change turbid fat, only quench one's thirst; External application removing toxicity for detumescence, convergence myogenic.Propolis is widely used as healthy food material.
Complicated components in propolis, water insoluble, the propolis after purification is that brown is block, has both been inconvenient to take, and is also unfavorable for absorbing, and often needs to add auxiliary material and ensure that propolis can with the dispersion of the state of fine-powder in the product when being therefore processed into health food.
Be in the health food of raw material with propolis, can be furnished with the raw material containing saponins compound such as American Ginseng, ginseng, general flavone and total saposins are often defined as the functional component of product by this kind of health food simultaneously.The total saposins assay method of bibliographical information has: vanillin assay, thin-layered chromatography, GC method, HPLC method, LC-MS method, CE method etc.; The panaxoside in health products is measured as Shen Xianghong, Ren Yiping, Chen Yi .HPLC method and vanillin assay. Chinese Journal of Health Laboratory Technology, 2004, disclose vanillin assay and HPLC method in 14 (6): 681-683.The assay method of total saposins in the health food provided with technical manual (version in 2003) evaluated by health food; it is the classical assay method of total saposins in current health food; when measuring the health food containing propolis raw material by this method; due to the protection by supplies such as propolis, disintegrant, filling agents; in product, total saposins is difficult to be extracted efficiently; total saponin content measurement result is not often inconsistent with actual formula ratio, poor accuracy.
Also there is now other the method for measuring total saponin content, if publication date is on October 31st, 2012, publication number is in the Chinese patent of CN102759514A, disclose the assay method of the content of general ginsenoside in a kind of injection qi-tonifying and pulse-restoring preparation, the method comprises the following steps: 1) preparation of reference substance respectively precision take general ginsenoside reference substance, add methyl alcohol to dissolve, obtained solution, 2) injection Yiqi and vein recovery drug powder is got in the preparation of need testing solution, accurately weighed, use water-soluble solution, cross resin column, after rinsing with methanol-water, with methanol-eluted fractions, eluent filters, obtained solution, 3) assay method is accurate respectively draws need testing solution and reference substance solution, the content of general ginsenoside in determined by ultraviolet spectrophotometry sample.The method can not measure with propolis the total saposins in the health food being raw material.
In sum, also do not have a kind of extracting method simple at present, the recovery is high, and accuracy is good, is the assay method of total saposins in the health food of raw material simply and rapidly with propolis.
Summary of the invention
The object of the invention is to overcome above shortcomings in prior art, and provide a kind of extracting method simple, the recovery is high, and accuracy is good, is the assay method of total saposins in the health food of raw material simply and rapidly with propolis.
The present invention's adopted technical scheme that solves the problem is: the assay method that should be with propolis total saposins in the health food of raw material, is characterized in that: this assay method comprises standard curve making operation, sample preparation operation, test solution cross post operation, colour developing operation and total saposins calculation process;
In described standard curve making operation, draw 0ml respectively, 10ml, 20ml, 40ml, 60ml, 80ml, ginsenoside Re's titer of 100ml and 120ml concentration known is placed in eight evaporating dishes, after ginsenoside Re's titer drying in ware to be evaporated, first add the sodium hydroxide solution that 5ml volumetric molar concentration is 0.1M, add water 20ml again, shaking up rear salt acid for adjusting pH value is 6.5-7.0, thus obtain eight parts of standard mother liquors, eight parts of standard mother liquors are placed in the ultrasonic 20-40min breakdown of emulsion of water-bath of 45-55 DEG C, be cooled to normal temperature, adopt the volumetric flask water constant volume of 50ml to 50ml respectively, shake up rear filtration, discard 5ml just filtrate, get 1.0ml subsequent filtrate as standard solution, thus obtain eight parts of standard solutions, adopt chromatographic column to carry out column operation to every part of standard solution, first wash post with the ethanol that 25mL percent by volume is 70%, discard eluent, wash post with 25mL again, discard eluent, accurately add 1.0mL standard solution, cross post, with the washing post of 25mL, to wash away the water-solubility impurities such as sugared part, discard eluent, with the ethanol elution ginsenoside that 25mL percent by volume is 70%, collect eluent in evaporating dish, volatilize in the bath that discharges water, thus obtain the evaporating dish after eight volatilizations, the vanillic aldehyde glacial acetic acid solution that 0.2mL percent by volume is 5% is accurately added in the evaporating dish volatilized, rotate evaporating dish, residue is all dissolved, then adds 0.8mL perchloric acid, move in 5mL band plug graduated centrifuge tube after mixing, be placed in the water-bath of less than 60 DEG C and heat 10min, take out, after ice bath cooling, accurately add glacial acetic acid 5.0mL, after shaking up, carry out colorimetric estimation in 560nm wavelength place with 1cm cuvette together with standard pipe, thus obtain eight absorbances, an absorbance is corresponding with the quality of the ginsenoside contained in an evaporating dish, adopts the quality of absorbance and ginsenoside to do curve, obtains typical curve,
In described sample preparation operation, getting with propolis is that the health food 0.3g of raw material is placed in 50ml volumetric flask, add 5ml volumetric molar concentration be the sodium hydroxide solution of 0.1M to dissolve health food, after health food dissolves, add water 20ml, shaking up rear salt acid for adjusting pH value is 6.5-7.0, and in the water-bath of 45-55 DEG C, ultrasonic 20-40min breakdown of emulsion, is cooled to normal temperature, with water constant volume to 50ml, shake up rear filtration, discard 5ml just filtrate, get 1.0ml subsequent filtrate as test solution to be measured;
Described test solution is crossed in post operation, adopts chromatographic column to treat test fluid and carries out column operation, first wash post with the ethanol that 25mL percent by volume is 70%, discard eluent, then wash post with 25mL, discard eluent, accurately add 1.0mL test solution to be measured, cross post, with the washing post of 25mL, to wash away the water-solubility impurities such as sugared part, discarding eluent, is the ethanol elution ginsenoside of 70% by 25mL percent by volume, collects eluent in evaporating dish, discharge water in bathing and volatilize, obtain test solution to be measured and volatilize evaporating dish;
In described colour developing operation, volatilize in evaporating dish at test solution to be measured and accurately add the vanillic aldehyde glacial acetic acid solution that 0.2mL percent by volume is 5%, rotate evaporating dish, residue is all dissolved, add 0.8mL perchloric acid again, move in 5mL band plug graduated centrifuge tube after mixing, be placed in the water-bath of less than 60 DEG C and heat 10min, take out, after ice bath cooling, accurately add glacial acetic acid 5.0mL, after shaking up, colorimetric estimation is carried out in 560nm wavelength place together with standard pipe with 1cm cuvette, thus obtain the absorbance of test solution to be measured, this absorbance is corresponding with the health food taking propolis as raw material,
In described total saposins calculation process, the absorbance of test solution to be measured is substituted in typical curve, obtains the quality of the ginsenoside contained in test solution to be measured, thus obtain with propolis be raw material health food in the content of total saposins.
The present invention adopts sodium hydrate aqueous solution to dissolve with propolis to be the health food of raw material, discharge by the total saposins composition of propolis, disintegrant and filling agent embedding, make it soluble in water, then use salt acid for adjusting pH value to 6.5-7.0, ensure the stability of total saposins in water, and propolis is separated out again, eliminate the interference of propolis, thus be effectively extracted total saposins.The total saposins in the sample containing propolis, disintegrant, filling agent is measured by this method, there is extracting method simple, the recovery is high, accuracy is good, simple and rapid feature, may be used for taking propolis as raw material take total saposins as the quality control of the health food of functional component, also may be used for the mensuration of the total saposins in general health food.
As preferably, the present invention in standard curve making operation and test solution cross in post operation, the long 20cm-25cm of chromatographic column used, internal diameter 0.6cm-0.8cm, top connects the storage liquid funnel of 30ml; The Amberlite-XAD-2 macroreticular resin that the in-built 6cm of chromatographic column is high, above increases the neutral alumina of 1cm.
As preferably, the present invention, in sample preparation operation, when using sodium hydroxide solution to dissolve health food, adopts the mode of the mode of jolting dispersion or ultrasonic wave added dispersion to dissolve.
As preferably, the present invention, in sample preparation operation, adds 20ml water and after shaking up in the health food dissolved, the hydrochloric acid for adjust ph to be percent by volume be 10% hydrochloric acid.
As preferably, in standard curve making operation of the present invention, the regression equation of typical curve is y=ax+b, and wherein, y represents absorbance, and x represents the quality of panaxoside, and a is positive number, and b is Arbitrary Digit.
As preferably, in standard curve making operation of the present invention, the regression equation of typical curve is y=0.0036x-0.0043, represents that the unit of the y of absorbance is A, represents that the unit of the x of the quality of panaxoside is mg.
As preferably, the coefficient R of regression equation of the present invention
2=0.9937, illustrate and present good linear relationship within the scope of 20mg-240mg.
As preferably, in sample preparation operation of the present invention, be that the health food of raw material is grinds powder with propolis.Make the present invention can dissolving health food more quickly and effectively thus.
As preferably, the concentration of ginsenoside Re's titer of the present invention is 2.0mg/mL.
The present invention compared with prior art, have the following advantages and effect: the dissolving of employing sodium hydrate aqueous solution take propolis as the health food of raw material, discharge by the total saposins composition of propolis, disintegrant and filling agent embedding, make it soluble in water, then use salt acid for adjusting pH value to 6.5-7.0, ensure the stability of total saposins in water, and propolis is separated out again, eliminate the interference of propolis, thus be effectively extracted total saposins.The total saposins in the sample containing propolis, disintegrant, filling agent is measured by this method, there is extracting method simple, the recovery is high, accuracy is good, simple and rapid feature, may be used for taking propolis as raw material take total saposins as the quality control of the health food of functional component, also may be used for the mensuration of the total saposins in general health food.
The present invention has made innovation in the selection of extracting reagent, aldehyde radical in phenolic hydroxyl group in saponin(e structure and vanillic aldehyde structure carries out aldol reaction and generates red condensation product, the phenolic hydroxyl group of glucoside unit is oxidized to carboxyl, with aldehyde condensation reaction, carbohydrate, flavone compound are because of the interference measurement containing phenolic hydroxyl group.The present invention once adopted methyl alcohol to extract total saposins, because propolis is soluble in methyl alcohol, so with methyl alcohol as extraction agent, total saposins can be extracted completely, but sample is while extracting total saposins with methyl alcohol, also propolis is extracted, in order to methyl alcohol need dry up by the interference getting rid of propolis, then the total saposins that is dissolved in water, because propolis is still present in methanolic extract, and firmly total saposins is embedded, so by shipwreck with stripping total saposins, cause Lower result.The present invention also once adopted hot water return method to extract, although more better than methyl alcohol method, the recovery increases, but it is still on the low side, and assay is unstable, this may be because heating can increase the solubleness of disintegrant, filling agent, but can not improve the solubleness of propolis in water, in addition, unstable result is relevant with factors such as sample thickness, Extracting temperature, times.Analyze the character of propolis and total saposins, the present invention adopts sodium hydrate aqueous solution to extract total saposins, propolis is soluble in alkaline aqueous solution, total saposins is able to be released in alkaline aqueous solution from propolis, adjust ph is to acid again, and propolis is precipitated, and total saposins is still stayed in water, extract total saposins with sodium hydrate aqueous solution, the recovery can reach 90.3%-109.7%.
The present invention has made innovation in the selection of extract concentration, total saposins is unstable in alkaline aqueous solution, optimum pH is 6.03, on the books during the ginseng liquid body preparation TSPG PHm that this " Chinese Journal of Modern Applied Pharmacy magazine " the 15th published in August, 1998 is studied by Xu Wenxiang in volume supplementary issue measures.Propolis in sample contains a large amount of acid ingredient, and first extraction agent sodium hydrate aqueous solution carries out neutralization reaction with the acidic materials in propolis, thus dissolves propolis, and the amount of propolis determines the consumption of extraction agent.If extract concentration is too high, can accelerate the hydrolysis of saponin(e, concentration is too low, then propolis is not dissolved, and saponin constituent can not fully stripping.The present invention is under the prerequisite of lot of experiments, final high spot reviews volumetric molar concentration is these three kinds of concentration of 0.1M, 0.5M and 1M, and these three kinds of consumptions of 2mL, 3mL and 5mL are to the dissolving situation of sample, find that sample weighting amount is when 0.3g, use NaOH that 5ml volumetric molar concentration is 0.1M can sample dissolution well, during adjust ph, the consumption of hydrochloric acid be minimum.
The present invention has made innovation in the selection of extracting pH value, and sample needs adjust ph after being dissolved by sodium hydroxide solution, ensures that total saposins is fully extracted and is not hydrolyzed.The present invention is under the prerequisite of lot of experiments, and when final primary study pH value is 5,6,7, the recovery of total saposins, finds that the recovery when pH value is 7 is best, possible cause is due in acid condition, has embedded caused by part saponin(e when propolis is separated out again.
Embodiment
Below by embodiment, the present invention is described in further detail, and following examples are explanation of the invention and the present invention is not limited to following examples.
Embodiment.
Be that the assay method of total saposins in the health food of raw material comprises standard curve making operation, sample preparation operation, test solution cross post operation, colour developing operation and total saposins calculation process with propolis in the present embodiment.
In standard curve making operation, draw 0ml respectively, 10ml, 20ml, 40ml, 60ml, 80ml, 100ml and 120ml concentration is that ginsenoside Re's titer of 2.0mg/mL is placed in eight evaporating dishes, after ginsenoside Re's titer drying in ware to be evaporated, first add the sodium hydroxide solution that 5ml volumetric molar concentration is 0.1M, add water 20ml again, shaking up rear salt acid for adjusting pH value is 6.5-7.0, thus obtain eight parts of standard mother liquors, eight parts of standard mother liquors are placed in the ultrasonic 30min breakdown of emulsion of water-bath of 50 DEG C, be cooled to normal temperature, adopt the volumetric flask water constant volume of 50ml to 50ml respectively, shake up rear filtration, discard 5ml just filtrate, get 1.0ml subsequent filtrate as standard solution, thus obtain eight parts of standard solutions.
Adopt chromatographic column to carry out column operation to every part of standard solution, the long 20cm-25cm of chromatographic column used, internal diameter 0.6cm-0.8cm, top connects the storage liquid funnel of 30ml; The Amberlite-XAD-2 macroreticular resin that the in-built 6cm of chromatographic column is high, above increases the neutral alumina of 1cm.First wash post with the ethanol that 25mL percent by volume is 70%, discard eluent, then wash post with 25mL, discard eluent, accurately add 1.0mL standard solution, cross post, with the washing post of 25mL, to wash away the water-solubility impurities such as sugared part, discarding eluent, is the ethanol elution ginsenoside of 70% by 25mL percent by volume, collects eluent in evaporating dish, discharge water in bathing and volatilize, thus obtain the evaporating dish after eight volatilizations.
The vanillic aldehyde glacial acetic acid solution that 0.2mL percent by volume is 5% is accurately added in the evaporating dish volatilized, rotate evaporating dish, residue is all dissolved, then adds 0.8mL perchloric acid, move in 5mL band plug graduated centrifuge tube after mixing, be placed in the water-bath of less than 60 DEG C and heat 10min, take out, after ice bath cooling, accurately add glacial acetic acid 5.0mL, after shaking up, carry out colorimetric estimation in 560nm wavelength place with 1cm cuvette together with standard pipe, thus obtain eight absorbances.An absorbance is corresponding with the quality of the ginsenoside contained in an evaporating dish, the quality of absorbance and ginsenoside is adopted to do curve, obtain typical curve, adopt EXCEL software, the regression equation of trying to achieve this typical curve is y=0.0036x-0.0043, wherein, y represents absorbance, and the unit of y is the quality that A, x represent panaxoside, the unit of x is the coefficient R of mg, this regression equation
2=0.9937, illustrate and present good linear relationship within the scope of 20mg-240mg.The regression equation of the typical curve in the present invention can be y=ax+b, and wherein, y represents absorbance, and x represents the quality of panaxoside, and a is positive number, and b is Arbitrary Digit.
In sample preparation operation, getting with propolis is that the health food 0.3g of raw material is placed in 50ml volumetric flask, add 5ml volumetric molar concentration be the sodium hydroxide solution of 0.1M to dissolve health food, after health food dissolves, add water 20ml, shaking up rear salt acid for adjusting pH value is 6.5-7.0, and in the water-bath of 50 DEG C, ultrasonic 30min breakdown of emulsion, is cooled to normal temperature, with water constant volume to 50ml, shake up rear filtration, discard 5ml just filtrate, get 1.0ml subsequent filtrate as test solution to be measured.The present invention is when using sodium hydroxide solution to dissolve health food, and the mode that jolting can be adopted to disperse or the mode adopting ultrasonic wave added to disperse are dissolved; Hydrochloric acid for adjust ph can be the hydrochloric acid of 10% for percent by volume; Take propolis as the health food normally grinds powder of raw material.
Cross in post operation at test solution, adopt chromatographic column to treat test fluid and carried out column operation, the long 20cm-25cm of chromatographic column used, internal diameter 0.6cm-0.8cm, top connects the storage liquid funnel of 30ml; The Amberlite-XAD-2 macroreticular resin that the in-built 6cm of chromatographic column is high, above increases the neutral alumina of 1cm.First wash post with the ethanol that 25mL percent by volume is 70%, discard eluent, then wash post with 25mL, discard eluent, accurately add 1.0mL test solution to be measured, cross post, with the washing post of 25mL, to wash away the water-solubility impurities such as sugared part, discarding eluent, is the ethanol elution ginsenoside of 70% by 25mL percent by volume, collects eluent in evaporating dish, discharge water in bathing and volatilize, obtain test solution to be measured and volatilize evaporating dish.
In colour developing operation, volatilize in evaporating dish at test solution to be measured and accurately add the vanillic aldehyde glacial acetic acid solution that 0.2mL percent by volume is 5%, rotate evaporating dish, residue is all dissolved, add 0.8mL perchloric acid again, move in 5mL band plug graduated centrifuge tube after mixing, be placed in the water-bath of less than 60 DEG C and heat 10min, take out, after ice bath cooling, accurately add glacial acetic acid 5.0mL, after shaking up, colorimetric estimation is carried out in 560nm wavelength place together with standard pipe with 1cm cuvette, thus obtain the absorbance of test solution to be measured, this absorbance is corresponding with the health food taking propolis as raw material.
In total saposins calculation process, the absorbance of test solution to be measured is substituted in the regression equation of typical curve, obtains the quality of the ginsenoside contained in test solution to be measured, thus obtain with propolis be raw material health food in the content of total saposins.
The present invention can adopt the TU-1810SPC ultraviolet-visible pectrophotometer manufactured by Beijing Puxi General Instrument Co., Ltd to measure absorbance, Sartorius BS224S analytical balance can be adopted to take sample, pH precision test paper can be adopted to survey pH value.The tablet being main material production with propolis, American ginseng extract in the present invention is provided by Hangzhou Words Bee Apiculture Co., Ltd; Ginsenoside Re's standard items used are Nat'l Pharmaceutical & Biological Products Control Institute, and lot number is 110754-200822, and mass percentage is 88.8%; It is pure that normal butyl alcohol used, NaOH, hydrochloric acid, methyl alcohol, ethanol, glacial acetic acid, perchloric acid are analysis; Neutral alumina is that chromatography is used; Water can be dual distilled water.
Below the range of linearity of the assay method in the present invention, detectability, the recovery and precision are analyzed.
Linear relationship: be that ginsenoside Re standard solution 0ml, 10ml, 20ml, 40ml, 60ml, 80ml, 100ml and 120ml of 2.0mg/mL is placed in evaporating dish by concentration, be placed in the water-bath lower than 60 DEG C and volatilize, or hot blast drying (it is overheated not make).Represent absorbance (A) with y, x represents ginsenoside Re quality (mg), uses EXCEL software, and the regression equation of trying to achieve method is y=0.0036x-0.0043, coefficient R
2=0.9937, illustrate and present good linear relationship in the scope of 20mg-240mg.
Method detectability: to reagent blank METHOD FOR CONTINUOUS DETERMINATION 6 times (see table 1), calculates to detect with 3 of its absorbance times of standard deviations and is limited to 9.9mg.
Precision test: take 6 parts of tablet sample said method ungrease treatments and measure, measurement result is in table 3.Its relative standard deviation is 3.65%.
Recovery test: (lot number is 110801 to take the same batch sample of known content, content 2.51g/100g) 6 parts, every part of about 0.12g, accurately weighed, be placed in 50ml volumetric flask, add the ginsenoside Re reference substance of 2.925mg respectively, carry out process and the mensuration of sample by this assay method, calculate the recovery, the results are shown in Table 4, recovery of standard addition is at 90.3%-109.7%, and relative standard deviation is 6.05%.
The accuracy in the present embodiment with propolis being the assay method of total saposins in the health food of raw material is high, and the mode below by contrast is described.Adopt the assay method (being called for short Ministry of Public Health's method) of total saposins in health food evaluation and technical manual (version in 2003) health food and this assay method to process respectively to 3 batches of American ginseng extracts and 3 batches of products, measurement result is respectively in table 5 and table 6.As seen from table, American ginseng extract adopts two kinds of methods not have significant difference, and 3 batches of product differentiation are extremely remarkable, and adopts this assay method measured value consistent with formula ratio, therefore the accuracy of this assay method is high.
The present invention adopts sodium hydrate aqueous solution to dissolve propolis, discharge by the total saposins composition of propolis, disintegrant, filling agent embedding, make it soluble in water, then use salt acid for adjusting pH value to 6.5-7.0, ensure the stability of total saposins in water, and propolis is separated out again, eliminate the interference of propolis, thus be effectively extracted total saposins.The total saposins in the sample containing propolis, disintegrant, filling agent is measured with this assay method, have that extracting method is simple, the recovery is high, accuracy is good, simple and rapid feature, may be used for taking propolis as raw material take total saposins as the quality control of the health food of functional component, also can be used for the mensuration of the total saposins in general health food.
Although the present invention with embodiment openly as above; but it is also not used to limit protection scope of the present invention; any technician being familiar with this technology, not departing from the change and retouching done in the spirit and scope of the present invention, all should belong to protection scope of the present invention.
Claims (8)
1. be an assay method for total saposins in the health food of raw material with propolis, it is characterized in that: this assay method comprises standard curve making operation, post operation crossed by sample preparation operation, test solution, colour developing operation and total saposins calculation process;
In described standard curve making operation, draw 0ml respectively, 10ml, 20ml, 40ml, 60ml, 80ml, ginsenoside Re's titer of 100ml and 120ml concentration known is placed in eight evaporating dishes, after ginsenoside Re's titer drying in ware to be evaporated, first add the sodium hydroxide solution for extracting total saposins that 5ml volumetric molar concentration is 0.1M, add water 20ml again, shaking up rear salt acid for adjusting pH value is 6.5-7.0, thus obtain eight parts of standard mother liquors, eight parts of standard mother liquors are placed in the ultrasonic 20-40min breakdown of emulsion of water-bath of 45-55 DEG C, be cooled to normal temperature, adopt the volumetric flask water constant volume of 50ml to 50ml respectively, shake up rear filtration, discard 5ml just filtrate, get 1.0ml subsequent filtrate as standard solution, thus obtain eight parts of standard solutions, adopt chromatographic column to carry out column operation to every part of standard solution, first wash post with the ethanol that 25mL percent by volume is 70%, discard eluent, wash post with 25mL again, discard eluent, accurately add 1.0mL standard solution, cross post, with the washing post of 25mL, to wash away the water-solubility impurities such as sugared part, discard eluent, with the ethanol elution ginsenoside that 25mL percent by volume is 70%, collect eluent in evaporating dish, volatilize in the bath that discharges water, thus obtain the evaporating dish after eight volatilizations, the vanillic aldehyde glacial acetic acid solution that 0.2mL percent by volume is 5% is accurately added in the evaporating dish volatilized, rotate evaporating dish, residue is all dissolved, then adds 0.8mL perchloric acid, move in 5mL band plug graduated centrifuge tube after mixing, be placed in the water-bath of less than 60 DEG C and heat 10min, take out, after ice bath cooling, accurately add glacial acetic acid 5.0mL, after shaking up, carry out colorimetric estimation in 560nm wavelength place with 1cm cuvette together with standard pipe, thus obtain eight absorbances, an absorbance is corresponding with the quality of the ginsenoside contained in an evaporating dish, adopts the quality of absorbance and ginsenoside to do curve, obtains typical curve,
In described sample preparation operation, getting with propolis is that the health food 0.3g of raw material is placed in 50ml volumetric flask, add 5ml volumetric molar concentration be 0.1M for extracting the sodium hydroxide solution of total saposins to dissolve health food, after health food dissolves, add water 20ml, shaking up rear salt acid for adjusting pH value is 6.5-7.0, and in the water-bath of 45-55 DEG C, ultrasonic 20-40min breakdown of emulsion, is cooled to normal temperature, with water constant volume to 50ml, shake up rear filtration, discard 5ml just filtrate, get 1.0ml subsequent filtrate as test solution to be measured; When using sodium hydroxide solution to dissolve health food, the mode of the mode of jolting dispersion or ultrasonic wave added dispersion is adopted to dissolve;
Described test solution is crossed in post operation, adopts chromatographic column to treat test fluid and carries out column operation, first wash post with the ethanol that 25mL percent by volume is 70%, discard eluent, then wash post with 25mL, discard eluent, accurately add 1.0mL test solution to be measured, cross post, with the washing post of 25mL, to wash away the water-solubility impurities such as sugared part, discarding eluent, is the ethanol elution ginsenoside of 70% by 25mL percent by volume, collects eluent in evaporating dish, discharge water in bathing and volatilize, obtain test solution to be measured and volatilize evaporating dish;
In described colour developing operation, volatilize in evaporating dish at test solution to be measured and accurately add the vanillic aldehyde glacial acetic acid solution that 0.2mL percent by volume is 5%, rotate evaporating dish, residue is all dissolved, add 0.8mL perchloric acid again, move in 5mL band plug graduated centrifuge tube after mixing, be placed in the water-bath of less than 60 DEG C and heat 10min, take out, after ice bath cooling, accurately add glacial acetic acid 5.0mL, after shaking up, colorimetric estimation is carried out in 560nm wavelength place together with standard pipe with 1cm cuvette, thus obtain the absorbance of test solution to be measured, this absorbance is corresponding with the health food taking propolis as raw material,
In described total saposins calculation process, the absorbance of test solution to be measured is substituted in typical curve, obtains the quality of the ginsenoside contained in test solution to be measured, thus obtain with propolis be raw material health food in the content of total saposins.
2. according to claim 1 take propolis as the assay method of total saposins in the health food of raw material, it is characterized in that: cross in post operation with test solution in standard curve making operation, the long 20cm-25cm of chromatographic column used, internal diameter 0.6cm-0.8cm, top connects the storage liquid funnel of 30ml; The Amberlite-XAD-2 macroreticular resin that the in-built 6cm of chromatographic column is high, above increases the neutral alumina of 1cm.
3. according to claim 1 take propolis as the assay method of total saposins in the health food of raw material, it is characterized in that: in sample preparation operation, in the health food dissolved, add 20ml water and after shaking up, the hydrochloric acid for adjust ph to be percent by volume be 10% hydrochloric acid.
4. according to claim 1 take propolis as the assay method of total saposins in the health food of raw material, it is characterized in that: in described standard curve making operation, the regression equation of typical curve is y=ax+b, wherein, y represents absorbance, x represents the quality of panaxoside, and a is positive number, and b is Arbitrary Digit.
5. according to claim 4 take propolis as the assay method of total saposins in the health food of raw material, it is characterized in that: in described standard curve making operation, the regression equation of typical curve is y=0.0036x-0.0043, the unit of the y of expression absorbance is A, represents that the unit of the x of the quality of panaxoside is mg.
6. according to claim 5 take propolis as the assay method of total saposins in the health food of raw material, it is characterized in that: the coefficient R of described regression equation
2=0.9937, illustrate and present good linear relationship within the scope of 20mg-240mg.
7. according to claim 1 take propolis as the assay method of total saposins in the health food of raw material, and it is characterized in that: in described sample preparation operation, is that the health food of raw material is grinds powder with propolis.
8. according to claim 1 take propolis as the assay method of total saposins in the health food of raw material, it is characterized in that: the concentration of described ginsenoside Re's titer is 2.0mg/mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310035767.3A CN103115887B (en) | 2013-01-30 | 2013-01-30 | Measurement method of total saponins in health food using propolis as raw material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310035767.3A CN103115887B (en) | 2013-01-30 | 2013-01-30 | Measurement method of total saponins in health food using propolis as raw material |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103115887A CN103115887A (en) | 2013-05-22 |
| CN103115887B true CN103115887B (en) | 2015-06-03 |
Family
ID=48414312
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310035767.3A Active CN103115887B (en) | 2013-01-30 | 2013-01-30 | Measurement method of total saponins in health food using propolis as raw material |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103115887B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104777248B (en) * | 2015-04-02 | 2016-10-26 | 上海交通大学 | In beverage total saponin content detection purification and detection method |
| CN112098551A (en) * | 2020-09-04 | 2020-12-18 | 广东省中药研究所 | Method for detecting ginsenoside in health food |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5899732A (en) * | 1981-12-08 | 1983-06-14 | Rooto Seiyaku Kk | Analysis of dammarane saponin |
| JPS60219556A (en) * | 1984-04-16 | 1985-11-02 | Shin Nippon Yakuhin Kk | Quantitative determination of saponin of gynostemma pentaphyllum |
| CN101354382A (en) * | 2007-07-24 | 2009-01-28 | 中国药科大学 | Simultaneous quantitative analysis method of five active components in pseudo-ginseng saponin body |
| CN102048778A (en) * | 2009-11-05 | 2011-05-11 | 天津天士力现代中药资源有限公司 | Method for detecting ginseng extract |
| CN102058647A (en) * | 2009-11-17 | 2011-05-18 | 天津天士力制药股份有限公司 | Ginseng glycoside H extract and preparation method thereof |
| CN102058642A (en) * | 2009-11-17 | 2011-05-18 | 天津天士力制药股份有限公司 | Ginseng saponin extract and preparation method thereof |
| CN102058644A (en) * | 2009-11-17 | 2011-05-18 | 天津天士力制药股份有限公司 | Ginseng saponin H extract and preparation method thereof |
| CN102058645A (en) * | 2009-11-17 | 2011-05-18 | 天津天士力制药股份有限公司 | Ginseng glycoside H extractive and preparation method thereof |
| CN102058643A (en) * | 2009-11-17 | 2011-05-18 | 天津天士力制药股份有限公司 | Ginseng saponin H extract and preparation method thereof |
-
2013
- 2013-01-30 CN CN201310035767.3A patent/CN103115887B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5899732A (en) * | 1981-12-08 | 1983-06-14 | Rooto Seiyaku Kk | Analysis of dammarane saponin |
| JPS60219556A (en) * | 1984-04-16 | 1985-11-02 | Shin Nippon Yakuhin Kk | Quantitative determination of saponin of gynostemma pentaphyllum |
| CN101354382A (en) * | 2007-07-24 | 2009-01-28 | 中国药科大学 | Simultaneous quantitative analysis method of five active components in pseudo-ginseng saponin body |
| CN102048778A (en) * | 2009-11-05 | 2011-05-11 | 天津天士力现代中药资源有限公司 | Method for detecting ginseng extract |
| CN102058647A (en) * | 2009-11-17 | 2011-05-18 | 天津天士力制药股份有限公司 | Ginseng glycoside H extract and preparation method thereof |
| CN102058642A (en) * | 2009-11-17 | 2011-05-18 | 天津天士力制药股份有限公司 | Ginseng saponin extract and preparation method thereof |
| CN102058644A (en) * | 2009-11-17 | 2011-05-18 | 天津天士力制药股份有限公司 | Ginseng saponin H extract and preparation method thereof |
| CN102058645A (en) * | 2009-11-17 | 2011-05-18 | 天津天士力制药股份有限公司 | Ginseng glycoside H extractive and preparation method thereof |
| CN102058643A (en) * | 2009-11-17 | 2011-05-18 | 天津天士力制药股份有限公司 | Ginseng saponin H extract and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 中华人民共和国卫生部.保健食品中总皂苷的测定.《保健食品评价与技术规范(2003年版)》.2003, * |
| 高效液相色谱串联质谱测定蜂胶中氯霉素药物残留量;周萍等;《中国蜂业》;20100901;第61卷(第9期);第42页右栏第1段 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103115887A (en) | 2013-05-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102954943B (en) | A kind of detect the detection method of tea polysaccharide in Folium camelliae assamicae or Pu'er tea | |
| CN102466658B (en) | Measurement method of content of 5-hydroxymethylfurfural (5-HMF) in injection | |
| CN102297926B (en) | Rapid TLC identification and three-component simultaneous quantitative determination method for antitussive tablets | |
| CN102854281B (en) | Detection method of sugar-free strong loquat syrup | |
| CN104161847B (en) | A kind of quality determining method of the Chinese medicine composition treating diabetic retinopathy | |
| CN102269751B (en) | Detection method of Liuweinengxiao preparation | |
| Yang et al. | Simultaneous determination of 15 flavonoids in Scutellaria barbata–Hedyotis diffusa Herb Pair by HPLC Q-TOF MS | |
| CN103115887B (en) | Measurement method of total saponins in health food using propolis as raw material | |
| CN109549966A (en) | A kind of glycyrrhiza glabra extract and the preparation method and application thereof | |
| CN102028773B (en) | Method for identifying radix rosae gigantea extract | |
| CN105911154A (en) | Method for determination of chlorogenic acid, galuteolin and total flavone content of honeysuckle | |
| CN103432196A (en) | Desmodium styracifolium total flavonoids and preparation method of Desmodium styracifolium total flavonoids and total polysaccharides | |
| CN107727763A (en) | Seven kinds of benzyl carbinol glycosides composition fast qualitatives and quantitative detecting method in saline cistanche medicinal material | |
| Arayne et al. | Validated spectrophotometric method for quantitative determination of simvastatin in pharmaceutical formulations and human serum | |
| CN101332287B (en) | Quality control method of Danle capsule | |
| CN103822888B (en) | Catch up with the quality determining method of yellow grass | |
| CN102331467B (en) | Method for detecting quality of south isatis root granules | |
| CN104820026A (en) | Litchi seed dripping pill quality detection method | |
| CN103983735B (en) | A kind of detection method preparing medical capsule for treating pelvic inflammatory disease | |
| CN108645951B (en) | A kind of thin-layer identification method of fructus crataegi medicinal material and medicine materical crude slice | |
| CN103091437B (en) | Determination method of hesperidin content | |
| CN103940942B (en) | A kind of detection method of CHANGYANNING preparation | |
| CN106706835A (en) | Quality detection method of trollius chinensis bunge effervescent tablets | |
| CN105616946A (en) | Preparation for treating cough, preparation method and quality control method thereof | |
| CN1853674B (en) | Quality controlling method of Xingdan injection |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |