CN103798414B - A kind of edible oil comprising molecular marker and its preparation method and application - Google Patents
A kind of edible oil comprising molecular marker and its preparation method and application Download PDFInfo
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Abstract
The invention provides a kind of edible oil comprising molecular marker and its preparation method and application, this edible oil comprises the OPC that concentration is 1 ~ 80 μ g/ml.This edible oil can be used for conveniently, whether accurately detect it through culinary art and/or recuperation of heat process, and described detection method comprises the following steps: (1) measures actual procyanidin concentration (P in edible oil sample
t) and actual anthocyanin concentrations (A
t); (2) actual procyanidin concentration (P respectively
t) and theoretical procyanidin concentration (P
e), actual anthocyanin concentrations (A
t) and theoretical anthocyanin concentrations (A
e), and calculate actual procyanidin concentration (P respectively
t) and actual anthocyanin concentrations (A
t) ratio, theoretical procyanidin concentration (P
e) and theoretical anthocyanin concentrations (A
e) ratio, determine whether edible oil through culinary art and/or recuperation of heat process.<!--1-->
Description
Technical field
The invention belongs to Food Safety Analysis field, relate to a kind of edible oil comprising molecular marker and its preparation method and application, be specifically related to can be used for judging edible oil whether through edible oil and preparation method thereof of the molecular marker of culinary art and/or recuperation of heat process a kind of comprising, and a kind of method being detected the edible oil through culinary art and/recuperation of heat process by this molecular marker.
Background technology
In modern society, traditional food processing born by family kitchen and individual workship has fully achieved the meticulous division of labor of full industrial chain and industrialization, the large production of scale.This while drastically increasing Socialized Reading efficiency, lifting quality of the life, freeing general public from heavy housework, also a series of problem demanding prompt solution is created, particularly the subject under discussion of related food Health Administration is especially by social extensive concern, even rises to the height of food security.Wherein, through heat recovering process process or event that after culinary art, residual vegetable oil illegally flows into food processing field again make a very bad impression especially, not only threaten people ' s health, also cause social fear, the serious blow confidence of people for food security, will affect the sound development of relevant industries.
Because the raw-material source of this what is called " waste oil " and main production thereof are all produced very similar to normal edible vegetable oil, " waste oil " main component can carrying out in prior art detecting also obviously is not distinguished with normal edible vegetable oil, add different variety of raw material, various processes, and even in the normal edible vegetable oil of different production batch also there is larger difference in mentioned component content, therefore, expert and associated mechanisms still can not provide the mature technology means detecting this low-quality goods both at home and abroad at present.
Obviously, cause the basic reason of above-mentioned technical bottleneck to be: in vegetable oil residual after heat recovering process process or culinary art, to lack clear and definite, stable mark, cause utilizing merely traditional physics, chemistry and biochemical analysis technological means to carry out detection and lack reliable Testing index.
Summary of the invention
In order to overcome the above-mentioned defect of prior art, the present invention takes the lead in proposing the new thought of adding molecular marker in normal food is produced, and this molecular marker in culinary art or heat recycle process, altered chemical structure can occur.Therefore, as long as detect the structure change of this molecule, just clearly can understand this testing sample and whether experienced by culinary art or heat recycle process, this is by detecting mark content thus brand new technical scheme that is convenient, that monitor food quality and production technology exactly.
On the one hand, the invention provides a kind of edible oil, this edible oil comprises the OPC that concentration is 1 ~ 80 μ g/ml.Preferably, this edible oil comprises the OPC that concentration is 2 ~ 20 μ g/ml.
On the other hand, present invention also offers a kind of preparation method of above-mentioned edible oil, this preparation method comprises the following steps: in edible oil, add OPC and/or comprise the raw material of OPC, edible oil is comprised OPC that concentration is 1 ~ 80 μ g/ml.
Preferably, above-mentioned preparation method comprises the following steps: in edible oil, add OPC and/or comprise the raw material of OPC, edible oil is comprised OPC that concentration is 2 ~ 20 μ g/ml.
In above-mentioned preparation method, preferably, described OPC is food-grade OPC; The described raw material comprising OPC can be selected from the vegetable oil that comprises OPC and comprise in the food additives of OPC one or more, be preferably cold press grape-kernel oil.
Because relevant laws and regulations and product force the restriction of standard, in food and raw material thereof, add foreign substance must strictly follow specific technical requirement, as organism metabolism and safety evaluatio experiment, human-body safety intake and interpolation concentration foundation, the large-scale long-term intervention experiment of crowd etc. of additive.Obviously, by so vast and numerous screening and evaluation process, the risk of failure that the cost such as its human and material resources, financial resources, time is paid and born is very huge beyond doubt.Therefore, with less cost and in the short period of time, solve this important technical bottleneck according to above-mentioned thinking, optimal approach is exactly at existing food additives, nutrient, and even finds such molecular marker in natural food active ingredient.
The present inventor finds finally through large quantifier elimination and screening: OPC (proanthocyanidins, CASNo.4852-22-6, structure is shown in following formula I) as the efficient free radical scavenger of a large amount of existence natural in food plant and vegetable oil and antioxidant, and its catabolite anthocyanidin (anthocyanins, CASNo.528-58-5, structure is shown in Formula Il) be exactly such a pair desirable molecular marker.
Its basic technique principle is as follows: if containing a certain amount of OPC in edible vegetable oil, owing to being decomposed into anthocyanidin when OPC can experience heating environment in culinary art and the heat recovering process after discarding, therefore judge whether this detection sample have passed through above-mentioned culinary art, discards and heat recycle process by both mensuration content and ratio change thereof.
But, because most of normal edible vegetable oil procyanidins content is lower, and different variety of raw material, various processes, and even the content difference of different production batch product procyanidins is very large, therefore, is difficult to establish unified detection and criterion.
For this reason, one of technical essential of the present invention is: obtain product oil by normal edible vegetable oil production technology, and by one of following two kinds of modes or whole before dispatching from the factory, (this concentration is called initial procyanidin concentration P to make it contain certain density OPC
0):
(1) food grade OPC product or the esculent containing higher concentration OPC is directly added.
(2) add the specific edible vegetable oil being rich in OPC, as cold press grape pip wet goods, this is equal to the production technology of common edible mediation vegetable oil completely.
Initial procyanidin concentration (P
0) determination principle be:
(1) what meet existing analytical technology completely can detection range requirement.
(2) with the natural degradation speed of OPC in the edible vegetable oil term of validity for foundation, require for fully meeting above-mentioned 1st article in the whole term of validity and set a tolerance.
(3) production cost is considered.
The concrete numerical value of initial procyanidin concentration, can by the unified setting of government regulator; Also can by manufacturing enterprise according to different variety of raw material, various processes, so different production batch product is determined respectively, thus set up molecular marker system unique separately.
After this, owing to being decomposed into anthocyanidin when OPC can experience heating environment in culinary art and the heat recovering process after discarding, correlation technique detects department just by working sample procyanidins content and OPC and anthocyanidin content ratio, can judge whether this detection sample have passed through culinary art, discards and heat recycle process.
For avoiding endogenous anthocyanidin interference natural in product oil, initial anthocyanin concentrations (A can be measured if desired before product oil dispatches from the factory
0), and assess its annoyance level for above-mentioned testing result.
To sum up above factor, the present invention determines through experimental study, it is 1 ~ 80 μ g/ml that edible oil comprises concentration, and the OPC being preferably 2 ~ 20 μ g/ml can meet above-mentioned requirements, and this kind of edible oil can facilitated through culinary art and/or heat treatment recovery, effectively detecting.
As for the assay method of OPC and anthocyanidin, existing mature technology can be adopted, as: colorimetric method, thin-layer method, chromatography etc.; Also by preparing OPC and anthocyanidin antibody sets up immune analysis method, thus more high sensitivity, more simple and quick onthe technology of site test can be createed.
Owing to being all naturally occurring materials in food plant and vegetable oil as the OPC of molecular marker and anthocyanidin in the present invention, not only without the need to carrying out organism metabolism and safety evaluatio, human-body safety intake and adding concentration measuring and calculating, the large-scale long-term intervention experiment of crowd and corresponding report and rigister supervisor; And due to more than the antioxidation in its organism comparatively vitamin C and vitamin E high several times, effectively can prevent, block and stop the lipid peroxidation chain reaction that active oxygen radical causes, prevent the oxidative damage of human body, thus there is obvious anti-ageing effect, and having preventive and therapeutic effect to various diseases such as tumour, cerebrosclerosis, encephalatrophies, the nutrition and health care therefore also contributing to improving edible vegetable oil is worth.
So far, we just establish a set of discriminating through heat recovering process process or the molecular marker system of residual plant oil and corresponding technical method thereof after culinary art.And another significant advantage of the method is also: even if illegal fraud molecule has grasped the ins and outs of adding OPC in product oil, but because the anthocyanidin generated in a large number in recovered oil can not be removed, so the truth that also cannot cover up facts.Meanwhile, this system also can be used for the famous brand edible vegetable oil of differentiating that common non-recycled vegetable oil is palmed off.
Again on the one hand, the invention provides the method for a kind of detection through the above-mentioned edible oil of culinary art and/or recuperation of heat process, the method comprises the following steps:
(1) actual procyanidin concentration (P in edible oil is measured
t) and actual anthocyanin concentrations (A
t);
(2) actual procyanidin concentration (P respectively
t) and theoretical procyanidin concentration (P
e), actual anthocyanin concentrations (A
t) and theoretical anthocyanin concentrations (A
e), and calculate actual procyanidin concentration (P respectively
t) and actual anthocyanin concentrations (A
t) ratio, theoretical procyanidin concentration (P
e) and theoretical anthocyanin concentrations (A
e) ratio, judge described edible oil whether through culinary art and/or recuperation of heat process.
Concrete decision procedure is: first with the initial procyanidin concentration (P of product to be measured
0) and to detect the date be foundation to the real time difference indicated between the date of production, converts out the theoretical procyanidin concentration (P of this sample according to natural degradation speed
e) and theoretical anthocyanin concentrations (A
e).Then obtain actual procyanidin concentration (P by working sample
t) and actual anthocyanin concentrations (A
t).Concrete determinating mode is as follows:
Sequence number | OPC | Anthocyanidin | OPC/anthocyanidin | Judge conclusion |
1 | P T≈P E | A T≈A E | P T/A T≈P E/A E | Qualified certified products |
2 | P T≈P E | A T<A E | P T/A T>P E/A E | Qualified certified products |
3 | P T≈P E | A T>A E | P T/A T<P E/A E | Add the recovered oil of OPC |
4 | P T<P E | A T≈A E | P T/A T<P E/A E | Do not add the non-recycled oil of OPC |
5 | P T<P E | A T<A E | P T/A T≈P E/A E | Do not add the non-recycled oil of OPC |
6 | P T<P E | A T>A E | P T/A T<P E/A E | Do not add the recovered oil of OPC |
7 | P T>P E | A T≈A E | P T/A T>P E/A E | Add the non-recycled oil of OPC |
8 | P T>P E | A T<A E | P T/A T>P E/A E | Add the non-recycled oil of OPC |
9 | P T>P E | A T>A E | P T/A T≈P E/A E | Add the recovered oil of OPC |
In the above-mentioned methods, the assay method in described step (1) is colorimetric method, thin-layered chromatography or high performance liquid chromatography, is preferably high performance liquid chromatography.
Preferably, the condition of described high performance liquid chromatography comprises:
Chromatographic column is the reverse-phase chromatographic column adopting octadecylsilane chemically bonded silica filler; Column temperature is 35 DEG C; UV detect wavelength is 525nm; The mixed flow phase of the water of mobile phase to be volume ratio be 73:13:6:8, methyl alcohol, isopropyl alcohol and 10% aqueous formic acid; Flow rate of mobile phase is 1.0ml/ minute.
More preferably, described high performance liquid chromatography comprises the following steps:
(1) sample extraction
According to theoretical procyanidin concentration determination sample collection amount in determinand, make its theoretical content be 800 μ g, the sample to this determinand adds equivalent absolute methanol, fully centrifugal or stratification after concussion mixing, absorption methanol fractions is sample extracting solution, and sample extracting solution is settled to 10ml; If sample extracting solution volume is greater than 10ml, then concentrate by vacuum or heating;
(2) extraction recovery reference material
Get the determinand sample identical with above-mentioned collection capacity, throw in the OPC sterling suitable with theoretical anthocyanin concentrations with theoretical procyanidin concentration and anthocyanidin sterling respectively, using as extraction recovery reference material; And to process equally with above-mentioned sample, obtain extraction recovery with reference to liquid;
(3) concentration reference material is prepared
Procyanidin concentration reference material: get 7 pipe 10ml absolute methanols, add the methanol mother liquor of OPC sterling, makes its concentration be respectively 0,5,10,20,40,80,160 μ g/ml;
Anthocyanin concentrations reference material: get 7 pipe 10ml absolute methanols, add the methanol mother liquor of anthocyanidin sterling, makes its concentration be respectively 0,0.625,1.25,2.5,5,10,20 μ g/ml;
(4) OPC hydrolysis
Get the above-mentioned sample extracting solution of 1ml, extraction recovery with reference to liquid and procyanidin concentration reference material, add n-butanol and the mixed liquor of 12mol/L hydrochloric acid and the 2mol/L hydrochloric acid solution of 0.2ml2% ammonium ferric sulfate that 6.0ml volume ratio is 95:5 respectively, airtight be heated to 100 DEG C continue 45 minutes, cool for subsequent use;
(5) chromatographic condition
Chromatographic column is the reverse-phase chromatographic column adopting octadecylsilane chemically bonded silica filler; Column temperature is 35 DEG C; UV detect wavelength is 525nm; The mixed flow phase of the water of mobile phase to be volume ratio be 73:13:6:8, methyl alcohol, isopropyl alcohol and 10% aqueous formic acid; Flow rate of mobile phase is 1.0ml/ minute;
(6) detection and result calculate
A. sample extracting solution and extraction recovery are with reference to the total anthocyanin concentrations in liquid
With the procyanidin concentration reference material after said hydrolyzed step for standard, detect the sample extracting solution after said hydrolyzed step and extraction recovery reference liquid, obtain its total anthocyanin concentrations respectively;
B. sample extracting solution and extraction recovery are with reference to the anthocyanin concentrations in liquid
Be detect respectively after the formic acid of 1% without the sample extracting solution of said hydrolyzed step, extraction recovery with reference to adding final concentration in liquid and anthocyanin concentrations reference material, thus obtain its anthocyanin concentrations.If the measured value of extract or extraction recovery reference liquid exceedes reference material scope, measure again after diluting 5 ~ 10 times;
C. sample extracting solution and extraction recovery are with reference to the procyanidin concentration in liquid
Extract and reference liquid procyanidins concentration=total anthocyanin concentrations-anthocyanin concentrations
D. sample and extraction recovery reference material procyanidins apparent concentration and anthocyanidin apparent concentration
Apparent concentration=extract or with reference to concentration ÷ sample collection amount (ml) × 10 in liquid
E. OPC extraction recovery and anthocyanidin extraction recovery
The rate of recovery=(extraction recovery reference material apparent concentration-sample apparent concentration) ÷ throws in concentration × 100%
F. sample procyanidins actual concentrations and anthocyanidin actual concentrations
Actual concentrations=apparent concentration ÷ rate of recovery.
Detailed description of the invention
Describe the present invention in detail below by specific embodiment, should be appreciated that following embodiment is only for illustration of the present invention, and the scope do not limited the present invention in any way.
Embodiment 1
Finished product edible soybean oil adds certain proportion cold press grape-kernel oil, and it is 2.5 μ g/ml(P that high pressure lipuid chromatography (HPLC) records its procyanidin concentration
0); Meanwhile, recording anthocyanin concentrations is 0.03 μ g/ml(A
0), can think that this initial anthocyanin concentrations can be ignored for the annoyance level of testing result from now on accordingly.
Get the above-mentioned edible oil of part to decoct after stir-fry, discarded stacking three days, heating recovery, dehydration, filtration through high temperature and obtain recovered oil, record its procyanidin concentration 0.07 μ g/ml(P
t), anthocyanin concentrations 2.32 μ g/ml(A
t).
And the above-mentioned edible oil same period detected under normal storage conditions, find that its OPC and anthocyanin concentrations all do not have significant change.Therefore, P
e≈ P
0; A
e≈ A
0.
Therefore, P
t< P
e, A
t> A
e, P
t/ A
t< P
e/ A
e, according to the relevant determinating mode 6 in description of the present invention, this sample is successfully judged to be the recovered oil not adding OPC.
Wherein, the concentration determination principle of OPC and anthocyanidin and concrete grammar as follows:
1, principle: the OPC in sample extracts through methyl alcohol, generates peony anthocyanidin by heating under acid condition.
2, method
(1) sample extraction
According to theoretical procyanidin concentration determination sample collection amount in determinand, make its theoretical content about 800 μ g, this testing sample is added equivalent absolute methanol, fully centrifugal or stratification after concussion mixing, absorption methanol fractions is sample extracting solution.If sample extracting solution volume is greater than 10ml, then concentrate by vacuum or heating.Sample extracting solution is settled to 10ml.
(2) extraction recovery reference material
Get the testing sample identical with above-mentioned collection capacity, throw in the OPC sterling suitable with theoretical anthocyanin concentrations with theoretical procyanidin concentration and anthocyanidin sterling respectively, using as extraction recovery reference material; And to process equally with above-mentioned sample, obtain extraction recovery with reference to liquid.
(3) concentration reference material
Procyanidin concentration reference material:
Get 7 pipe 10ml absolute methanols, add the methanol mother liquor of OPC sterling, its concentration is respectively: 0,5,10,20,40,80,160 μ g/ml.
Anthocyanin concentrations reference material:
Get 7 pipe 10ml absolute methanols, add the methanol mother liquor of anthocyanidin sterling, its concentration is respectively: 0,0.625,1.25,2.5,5,10,20 μ g/ml.
(4) OPC hydrolysis
Get the above-mentioned sample extracting solution of 1ml, extraction recovery with reference to liquid and procyanidin concentration reference material, add the mixed liquor (volume ratio=95:5) of 6.0ml n-butanol and 12mol/L hydrochloric acid and hydrochloric acid (2mol/L) solution of 0.2ml2% ammonium ferric sulfate respectively, airtightly be heated to 100 DEG C and reach 45 minutes, cool for subsequent use.
(5) chromatographic condition
Chromatographic column: Shimadzu Shim-pakCLC-ODS4.6 × 150mm
Column temperature: 35 DEG C
Detector: UV-detector (525nm)
Mobile phase: water: methyl alcohol: isopropyl alcohol: 10% formic acid (volume ratio=73:13:6:8)
Flow velocity: 1.0ml/ minute
(6) detection and result calculate
A. sample extracting solution and extraction recovery are with reference to the total anthocyanin concentrations in liquid:
With the procyanidin concentration reference material after said hydrolyzed step for standard, detect the sample extracting solution after said hydrolyzed step and extraction recovery reference liquid, obtain its total anthocyanin concentrations (i.e. anthocyanidin and procyanidin concentration sum) respectively.
B. sample extracting solution and extraction recovery are with reference to the anthocyanin concentrations in liquid:
Be detect respectively after the formic acid of 1% without the sample extracting solution of said hydrolyzed step, extraction recovery with reference to adding final concentration in liquid and anthocyanin concentrations reference material, thus obtain its anthocyanin concentrations.If the measured value of extract or extraction recovery reference liquid exceedes reference material scope, measure again after 5 ~ 10 times can be diluted.
C. sample extracting solution and extraction recovery are with reference to the procyanidin concentration in liquid:
Extract and reference liquid procyanidins concentration=total anthocyanin concentrations-anthocyanin concentrations
D. sample and extraction recovery reference material procyanidins apparent concentration and anthocyanidin apparent concentration:
Apparent concentration=extract or with reference to concentration ÷ sample collection amount (ml) × 10 in liquid
E. OPC extraction recovery and anthocyanidin extraction recovery:
The rate of recovery=(extraction recovery reference material apparent concentration-sample apparent concentration) ÷ throws in concentration × 100%
F. sample procyanidins actual concentrations and anthocyanidin actual concentrations
Actual concentrations=apparent concentration ÷ rate of recovery.
Claims (6)
1. detect the method for edible oil through culinary art and/or recuperation of heat process, described edible oil comprises the OPC that concentration is 1 ~ 80 μ g/ml, and the method comprises the following steps:
(1) actual procyanidin concentration (P in edible oil is measured
t) and actual anthocyanin concentrations (A
t); With
(2) actual procyanidin concentration (P respectively
t) and theoretical procyanidin concentration (P
e), actual anthocyanin concentrations (A
t) and theoretical anthocyanin concentrations (A
e), and calculate actual procyanidin concentration (P respectively
t) and actual anthocyanin concentrations (A
t) ratio, theoretical procyanidin concentration (P
e) and theoretical anthocyanin concentrations (A
e) ratio, judge described edible oil whether through culinary art and/or recuperation of heat process;
Concrete determinating mode is as follows:
2. method according to claim 1, is characterized in that, described edible oil comprises the OPC that concentration is 2 ~ 20 μ g/ml.
3. method according to claim 1 and 2, is characterized in that, the assay method in described step (1) is colorimetric method, thin-layered chromatography or high performance liquid chromatography.
4. method according to claim 3, is characterized in that, described assay method is high performance liquid chromatography.
5. method according to claim 4, is characterized in that, the condition of described high performance liquid chromatography comprises:
Chromatographic column is the reverse-phase chromatographic column adopting octadecylsilane chemically bonded silica filler; Column temperature is 35 DEG C; UV detect wavelength is 525nm; The mixed flow phase of the water of mobile phase to be volume ratio be 73:13:6:8, methyl alcohol, isopropyl alcohol and 10% aqueous formic acid; Flow rate of mobile phase is 1.0ml/ minute.
6. method according to claim 4, is characterized in that, described high performance liquid chromatography comprises the following steps:
(1) sample extraction
According to theoretical procyanidin concentration determination sample collection amount in determinand, make its theoretical content be 800 μ g, add equivalent absolute methanol to this determinand sample, fully centrifugal or stratification after concussion mixing, absorption methanol fractions is sample extracting solution, and sample extracting solution is settled to 10ml; If sample extracting solution volume is greater than 10ml, then concentrate by vacuum or heating;
(2) extraction recovery reference material
Get the determinand sample identical with above-mentioned collection capacity, throw in the OPC sterling suitable with theoretical anthocyanin concentrations with theoretical procyanidin concentration and anthocyanidin sterling respectively, using as extraction recovery reference material; And to process equally with above-mentioned sample, obtain extraction recovery with reference to liquid;
(3) concentration reference material is prepared
Procyanidin concentration reference material: get 7 pipe 10ml absolute methanols, add the methanol mother liquor of OPC sterling, makes its concentration be respectively 0,5,10,20,40,80,160 μ g/ml;
Anthocyanin concentrations reference material: get 7 pipe 10ml absolute methanols, add the methanol mother liquor of anthocyanidin sterling, makes its concentration be respectively 0,0.625,1.25,2.5,5,10,20 μ g/ml;
(4) OPC hydrolysis
Get the above-mentioned sample extracting solution of 1ml, extraction recovery with reference to liquid and procyanidin concentration reference material, add n-butanol and the mixed liquor of 12mol/L hydrochloric acid and the 2mol/L hydrochloric acid solution of 0.2ml2% ammonium ferric sulfate that 6.0ml volume ratio is 95:5 respectively, airtight be heated to 100 DEG C continue 45 minutes, cool for subsequent use;
(5) chromatographic condition
Chromatographic column is the reverse-phase chromatographic column adopting octadecylsilane chemically bonded silica filler; Column temperature is 35 DEG C; UV detect wavelength is 525nm; The mixed flow phase of the water of mobile phase to be volume ratio be 73:13:6:8, methyl alcohol, isopropyl alcohol and 10% aqueous formic acid; Flow rate of mobile phase is 1.0ml/ minute;
(6) detection and result calculate
A. sample extracting solution and extraction recovery are with reference to the total anthocyanin concentrations in liquid
With the procyanidin concentration reference material after said hydrolyzed step for standard, detect the sample extracting solution after said hydrolyzed step and extraction recovery reference liquid, obtain its total anthocyanin concentrations respectively;
B. sample extracting solution and extraction recovery are with reference to the anthocyanin concentrations in liquid
Be detect respectively after the formic acid of 1% without the sample extracting solution of said hydrolyzed step, extraction recovery with reference to adding final concentration in liquid and anthocyanin concentrations reference material, thus obtain its anthocyanin concentrations; If the measured value of extract or extraction recovery reference liquid exceedes reference material scope, measure again after diluting 5 ~ 10 times;
C. sample extracting solution and extraction recovery are with reference to the procyanidin concentration in liquid
Extract and reference liquid procyanidins concentration=total anthocyanin concentrations-anthocyanin concentrations
D. sample and extraction recovery reference material procyanidins apparent concentration and anthocyanidin apparent concentration
Apparent concentration=extract or milliliter number × 10 with reference to concentration ÷ sample collection in liquid
E. OPC extraction recovery and anthocyanidin extraction recovery
The rate of recovery=(extraction recovery reference material apparent concentration-sample apparent concentration) ÷ throws in concentration × 100%
F. sample procyanidins actual concentrations and anthocyanidin actual concentrations
Actual concentrations=apparent concentration ÷ rate of recovery.
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