CN102778517B - Method for detecting cocoa powder adulteration based on lipid clustering analysis - Google Patents
Method for detecting cocoa powder adulteration based on lipid clustering analysis Download PDFInfo
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Abstract
The invention discloses a method for detecting cocoa powder adulteration based on lipid clustering analysis. Cocoa butter is extracted from the cocoa powder by a Soxhlet extraction method; a reversed-phase high-performance liquid chromatography (RP-HPLC) separation-evaporative light scattering detector (ELSD) is utilized to detect, and a lipid chromatograph finger-print of the cocoa powder is constructed; and a cocoa powder adulteration sample which simulates respectively mixing with chestnut shell powder and longan shell powder and respectively mixing with a cocoa butter substitute and palm oil is subjected to lipid fingerprint analysis and adulteration distinguishing test by applying a hierarchical cluster analysis (HCA) process according to a relative peak area of a common characteristic peak of the fingerprint. The result shows that the method is capable of verifying the adulterated cocoa powder sample mixed with 1.2% or more of the cocoa butter substitute or the palm oil based on the percentage composition of the sample mass. The method is more convenient and sensitive and better in repeatability, thereby being utilized to detect the cocoa powder mixed with a forged and fake sample without lipid or low-lipid impurities, wherein the cocoa butter substitute or the palm oil and the like is artificially mixed so that the lipid content reaches the standard.
Description
Technical field
The present invention relates to a kind of cocoa power based on finger-print and mingle detection method, particularly a kind of by making up the method for glyceride finger-print to judge whether cocoa power is mingled.
Background technology
Natural cocoa butter (cocoa butter or cacao butter, CB) be cocoa bean through the grease with specific function that milling process makes, abound with in the torrid zone (mainly in Africa), be the milk yellow solid under the normal temperature, outer appearnce is Chinese wax seemingly, has aromatic odor.Cocoa power has the unique perfume of strong fragrance, contains various nutrient elements and active components such as abundant polyphenol, protein, carbohydrates, cocoa butter, is to make chocolate important source material, also for the production of one of the world today's three big hobby property beverages.Eighties of last century is since the mid-90, expansion day by day along with the cocoa products range of application, the expansion of foreign trade, China's cocoa processing industry development is swift and violent, meanwhile, a large amount of inferior or personation cocoa power floods market on the home market, not only serious harm food security, also influenced the international trade prestige of China and the interests of regular cocoa processing producer.Gold immortality etc. has been carried out exploratory development to the PCR detection method of mixing the plant-derived compositions of several external sources such as big dregs of beans, sesame seed meal, peanut shell and chestnut shell in the cocoa power, the method accuracy of setting up is subject to impurity effect, can not differentiate this main endogenous adulterant of cocoa shell powder; In addition, the plant-derived composition of the external source that may add is of a great variety, is difficult to expect, can not carry out PCR one by one to the plant-derived composition of external source that institute might add in the practical operation and detect, so this method has very big limitation, is difficult to popularization.For this reason, this paper attempts setting up cocoa power from lipid fingerprint analysis angle and mingles detection method, namely with the cocoa butter in the cocoa power sample of 11 batches of separate sources of soxhlet extraction extraction, detect the lipid chromatographic fingerprinting that obtains cocoa power by reversed-phase high-performance liquid chromatography (HPLC) separation-evaporative light-scattering detector (ELSD), relative peak area according to its common characteristic peak, application system cluster analysis (HCA) method is mixed chestnut shell powder and longan shell powder respectively and is mixed substitute of cocoa fat respectively and palmitic cocoa power is mingled sample and carried out the lipid fingerprint analysis simulation.
The present invention can be the quality determining method of China's cocoa power and perfect provide scientific basis and reference are provided in the lifting of quality standard, thereby promotes the quality of China's cocoa power product to improve and stablize, and consumers' rights and interests are safeguarded in standard cocoa power market better.
Summary of the invention
The technical problem to be solved in the present invention provides accurately and reliably a kind of and easier, can be applied to the detection method of mingling of cocoa power.
Technical scheme of the present invention is:
At first, make up cocoa power glyceride standard finger-print, comprise extraction, the fingerprint analysis of high performance liquid chromatography and the determining of standard finger-print thereof of cocoa power lipid;
Then, according to the assay method that cocoa power glyceride is formed, measure the lipid glyceride composition that sample is mingled in simulation;
At last, according to cocoa power sample and the matrix of simulating the relative peak area composition of mingling sample lipid glyceride component, carry out cluster analysis.
Concrete steps of the present invention are:
1, the method for building up of cocoa power lipid glyceride finger-print comprises the following steps:
(1) extraction of cocoa power lipid: take by weighing 2.50g left and right sides cocoa power, place filtration paper cylinder, put into the extraction tube of Soxhlet extractor, and in extraction flask, add about 2/3 sherwood oil, 47 ℃ of water-baths, 6h.Take out extraction flask, use Rotary Evaporators, be concentrated into dried.Then with extraction flask at 105 ℃ of dry 2h, take out and to be positioned over cooling in the exsiccator, weigh, repeat this and be operated to constant weight.Accurately take by weighing the lipid that a certain amount of extraction obtains, be dissolved in methenyl choloride, making its concentration is about 5mg/mL.
(2) efficient liquid phase chromatographic analysis: chromatographic column: symmetry C18 (250mm * 4.6mm, 5 μ m).Phase: the A that flows is acetonitrile/chloroform (70:30), and B is acetonitrile/chloroform (30:70).Binary gradient elution program: 0min, 100%A; 5min, 70%A; 25min, 0%A; 25.1 ~ 30min, 100%A.Column temperature: 40 ℃.Flow velocity: 0.8mL/min.The ELSD drift tube temperature: 85 ℃, N2 flow velocity: 2.5L/min.Distinguish the sample introduction analysis with this understanding, obtain the chromatographic fingerprinting of cocoa power glyceride.Be reference with No. 11 peaks, the relative retention time at each peak in the calculation sample.
(3) foundation of standard finger-print: by glyceride high-efficient liquid phase chromatogram in the cocoa power sample in 11 different places of production is compared, determine the common characteristic peak, and chromatographic data is imported the standard colour chart finger-print that the finger-print special software obtains the cocoa power glyceride that is made of its common characteristic peak.Testing sample can contrast with standard finger-print, distinguishes its similarities and differences.
The common characteristic peak has 14, and the relative standard deviation RSD of their relative retention time RT (be reference with No. 11 peak POS) is all less than 1%; That is:
No. 1 average RT in peak is that 0.411, RSD is 0.18%;
No. 2 average RT in peak are that 0.465, RSD is 0%;
No. 3 average RT in peak are that 0.703, RSD is 0.15%;
No. 4 average RT in peak are that 0.743, RSD is 0.22%;
No. 5 average RT in peak are that 0.786, RSD is 0.07%;
No. 6 average RT in peak are that 0.848, RSD is 0.12%;
No. 7 average RT in peak are that 0.878, RSD is 0.02%;
No. 8 average RT in peak are that 0.899, RSD is 0%;
No. 9 average RT in peak are that 0.944, RSD is 0.03%;
No. 10 average RT in peak are that 0.977, RSD is 0.06%;
No. 11 average RT in peak are that 1.000, RSD is 0%;
No. 12 average RT in peak are that 1.107, RSD is 0%;
No. 13 average RT in peak are that 1.183, RSD is 0.34%;
No. 14 average RT in peak are that 1.215, RSD is 0.34%;
The fingerprint peaks that wherein surpasses total peak area 3% has 3, is respectively: No. 8 peaks, relative peak area 10.848% ~ 12.403%; No. 11 peaks, relative peak area 51.518% ~ 53.078%; No. 12 peaks, relative peak area 30.463% ~ 32.677%.
2, cluster analysis
The present invention select the Ward method as clustering method, Chebychev distance as the measuring distance method, average is 1 as standardization, carries out cluster analysis.No. 11 total fingerprint peaks is more stable, and peak area number percent is about 50%, and separates better with adjacent peaks, and retention time is more placed in the middle, therefore sets No. 11 peaks for reference to the peak.Then, use the matrix that the relative peak area (with No. 11 peaks as the reference peak) at 14 common characteristic peaks forms and carry out cluster analysis.
The present invention has set up the HPLC finger-print of the lipid glyceride composition of cocoa power, and its similarity is all more than 0.995.The matrix that the relative peak area at 14 common characteristic peaks of the HPLC finger-print that application cocoa power lipid glyceride is formed is formed, birds of the same feather flock together method to mingling cocoa power analysis with system, mix the percentage composition that 1.2%(accounts for sample quality) and above substitute of cocoa fat or the palmitic cocoa power sample of mingling can be differentiated.This method is reliable and stable, and good reproducibility can be applicable to the detection of mingling of cocoa power.
Description of drawings
Fig. 1: 11 HPLC finger-prints that place of production cocoa power lipid glyceride is formed;
Each peak retention time (min) is respectively: No. 1 peak-7.291; No. 2 peaks-8.245; No. 3 peaks-12.469; No. 4 peaks-13.184; No. 5 peaks-13.943; No. 6 peaks-15.038; No. 7 peaks-15.571; No. 8 peaks-15.947; No. 9 peaks-16.745; No. 10 peaks-17.331; No. 11 peaks-17.739; No. 12 peaks-19.635; No. 13 peaks-20.978; No. 14 peaks-21.561.
Fig. 2: cocoa power sample lipid chromatogram is mingled in simulation;
Fig. 3: simulation is mixed the cocoa power sample clustering of mingling of chestnut shell and substitute of cocoa fat and is analyzed dendrogram;
Fig. 4: longan shell is mixed in simulation and the palmitic cocoa power sample clustering of mingling is analyzed dendrogram.
Embodiment
The present invention will be further described below in conjunction with embodiment, and following embodiment only is used for explanation the present invention but not limitation of the present invention.
Embodiment 1: mix chestnut shell and substitute of cocoa fat cocoa power mingle detection method research
1, instrument, sample and reagent
1.1 sample
The cocoa power sample of 11 batches of separate sources and mingle lipid substitute of cocoa fat (BS2000): gather by Wuxi City product quality supervision and testing institute office;
Material chestnut shell powder is mingled in simulation: made by the Chinese chestnut that sends out the supermarket available from the big profit of Wuxi City;
1.2 reagent
Sherwood oil, methenyl choloride: analyze pure, (Chemical Reagent Co., Ltd., Sinopharm Group); Acetonitrile: chromatographically pure, (Chemical Reagent Co., Ltd., Sinopharm Group);
1.3 instrument
High performance liquid chromatograph (comprising the G1311A quaternary pump, G1313A automatic sampler, the online degasser of G1379A, G1316A column oven, Agilent1100 chem workstation): Agilent1100, U.S. Agilent company;
Evaporative light-scattering detector: Alltech2000, U.S. Agilent company;
BUCHI Soxhlet extractor: B-811, Switzerland BUCHI Labortechnik AG company limited; DLSB-cryogenic liquid ebullator: DLSB-5/20, Zhengzhou Greatwall Scientific Industrial ﹠ Trading Co., Ltd.;
Rotary evaporator: RE52-2, Shanghai Hu Xi analytical instrument Co., Ltd., Factory;
Recirculated water is used vacuum pump more: SHZ-3, Shanghai Hu Xi analytical instrument Co., Ltd., Factory.Nitrile, methyl alcohol are chromatographically pure.
2, the extraction of cocoa power lipid
Take by weighing 2.50g left and right sides cocoa power, place filtration paper cylinder, put into the extraction tube of Soxhlet extractor, and in extraction flask, add about 2/3 sherwood oil, 47 ℃ of water-baths, 6h.Take out extraction flask, use Rotary Evaporators, be concentrated into dried.Then with extraction flask at 105 ℃ of dry 2h, take out and to be positioned over cooling in the exsiccator, weigh, repeat this and be operated to constant weight.Accurately take by weighing the lipid that a certain amount of extraction obtains, be dissolved in methenyl choloride, making its concentration is about 5mg/mL.
3, efficient liquid phase chromatographic analysis
Chromatographic column: symmetry C18 (250mm * 4.6mm, 5 μ m).Phase: the A that flows is acetonitrile/chloroform (70:30), and B is acetonitrile/chloroform (30:70).Binary gradient elution program: 0min, 100%A; 5min, 70%A; 25min, 0%A; 25.1 ~ 30min, 100%A.Column temperature: 40 ℃.Flow velocity: 0.8mL/min.The ELSD drift tube temperature: 85 ℃, N2 flow velocity: 2.5L/min.
Sample introduction analysis respectively with this understanding obtains the fingerprint spectrogram that cocoa power sample lipid glyceride is formed, and sees accompanying drawing 1.Be reference with No. 11 peaks, the relative retention time at each peak in the calculation sample.
4, the foundation of standard finger-print and total fingerprint peaks feature
High-performance liquid chromatogram determination by the composition of the lipid glyceride in the cocoa power sample in 11 different places of production, compare its chromatogram, determine 14 at common characteristic peak, the relative standard deviation RSD of the relative retention time RT at total peak (be reference with No. 11 peaks) is all less than 1%; The fingerprint peaks that wherein surpasses total peak area 3% has 3, is respectively: No. 8 peaks, relative peak area 10.848% ~ 12.403%; No. 11 peaks, relative peak area 51.518% ~ 53.078%:12 peak, relative peak area 30.463% ~ 32.677%.
5, cluster analysis
Hierarchial-cluster analysis is derived from taxonomy, is based on the similarity of sample.Its basic thought is to regard each sample of sample set to be clustered as a class separately, carries out cluster after the distance between regulation or definition sample and sample or the distance between similarity measurement and class and the class then.When cluster begins, between each self-forming one class of each sample, class and class and the distance between sample and the sample be identical, chosen distance minimum a pair of is that the pair of sample of similarity maximum is merged into a new class; Calculate the distance between this new class and other all classes again; Relatively after each distance, will be merged into another new class apart from two classes of minimum again.Sample in all sample sets is classified as till the class.Whole cluster process has carried out the n(number of samples) operation of-1 time the new class of merging, obtain n-1 and class distance.This n-1 merging process also can be represented with dendrogram.Just can draw the relevant information of sample class according to the dendrogram that provides.
Measure the composition of the triglyceride of the cocoa power that mixes chestnut shell and substitute of cocoa fat according to 2 and 3 method, and select the Ward method as clustering method, Chebychev distance as the measuring distance method, average is 1 as standardization, carries out cluster analysis.No. 11 total fingerprint peaks is more stable, and peak area is bigger, and separates better with adjacent peaks, and retention time is more placed in the middle, and therefore setting No. 11 peaks is with reference to the peak.Then, the matrix of forming with the relative peak area at common characteristic peak and non-total peak (with No. 11 peak POS as the reference peak) carries out cluster analysis.
Because being substantially free of lipid in the chestnut shell, so when mixing chestnut shell in the S11 cocoa power, then the amount of cocoa butter content in the cocoa power can reduce, up to standard for making amount of cocoa butter content, illegal producer mixes a certain amount of substitute of cocoa fat usually simultaneously.Therefore this experimental simulation prepares the samples (total lipid content all reaches 10% behind the admixture substitute of cocoa fat) of the different amount of 8 admixtures substitute of cocoa fat, see Table 1(after measured in the S11 cocoa power original total lipid content be 9.96%, namely be approximately 10%.）。Extract lipid in these samples according to the method for step 2, and (Fig. 2 a) to carry out the HPLC fingerprint analysis according to step 3.The matrix of forming according to the relative peak area of chromatographic peak carries out cluster analysis, by the cluster analysis dendrogram (Fig. 3) of gained as seen: sample A1 can not (distinguish the S1～S11) from pure cocoa power sample, A2～A8 then all can distinguish over S1～S11, namely mix 1.2% and the sample of above substitute of cocoa fat can be differentiated.
Table 1: simulation is mixed the cocoa power of mingling of chestnut shell and substitute of cocoa fat and is formed
6, stratographic analysis precision
The cocoa butter that the S11 cocoa power extracts through Soxhlet is configured to the sample solution of 5mg/mL, and continuous sample introduction 5 times is added up relative retention time (retention time with No. 11 peaks is reference) and the relative peak area at each total peak.The result shows, the relative retention time at each total peak and the relative standard deviation of relative peak area (RSD)＜2% show that the chromatographic fingerprint analytical precision test of cocoa power lipid meets the requirements.
7, method repeatability
Take by weighing 5 parts in S11 cocoa power sample, extract lipid and carry out the HPLC analysis by 1.2 and 1.3 respectively, respectively relative retention time and the relative peak area at total peak are added up, their relative standard deviation (RSD) is all less than 3%.
8, method stability test
The S11 cocoa power is configured to the sample solution of 5mg/mL through the cocoa butter of Soxhlet extraction, at room temperature (about 18 ℃) are preserved, respectively at 0,5,10,15,20, the 25h sample introduction is analyzed, and relative retention time (retention time with No. 11 peaks is reference) and the relative peak area at each total peak are added up.The result shows that the relative retention time at each total peak and relative peak area RSD are all less than 5%.
More than test shows, above-mentioned finger print measuring method precision, stable, reliable.
Embodiment 2: that mixes longan shell and palmitic cocoa power mingles detection method research
1, instrument, sample and reagent
1.1 sample
The cocoa power sample of 11 batches of separate sources and mingle lipid palm oil sample: gather by Wuxi City product quality supervision and testing institute office;
Material longan shell powder is mingled in simulation: made by the longan of sending out the supermarket available from the big profit of Wuxi City;
1.2 reagent
Sherwood oil, methenyl choloride: analyze pure, (Chemical Reagent Co., Ltd., Sinopharm Group); Acetonitrile: chromatographically pure, (Chemical Reagent Co., Ltd., Sinopharm Group);
1.3 instrument
High performance liquid chromatograph (comprising the G1311A quaternary pump, G1313A automatic sampler, the online degasser of G1379A, G1316A column oven, Agilent1100 chem workstation): Agilent1100, U.S. Agilent company;
Evaporative light-scattering detector: Alltech2000, U.S. Agilent company;
BUCHI Soxhlet extractor: B-811, Switzerland BUCHI Labortechnik AG company limited; DLSB-cryogenic liquid ebullator: DLSB-5/20, Zhengzhou Greatwall Scientific Industrial ﹠ Trading Co., Ltd.;
Rotary evaporator: RE52-2, Shanghai Hu Xi analytical instrument Co., Ltd., Factory;
Recirculated water is used vacuum pump more: SHZ-3, Shanghai Hu Xi analytical instrument Co., Ltd., Factory.Nitrile, methyl alcohol are chromatographically pure.
2, the extraction of cocoa power lipid
Take by weighing 2.50g left and right sides cocoa power, place filtration paper cylinder, put into the extraction tube of Soxhlet extractor, and in extraction flask, add about 2/3 sherwood oil, 47 ℃ of water-baths, 6h.Take out extraction flask, use Rotary Evaporators, be concentrated into dried.Then with extraction flask at 105 ℃ of dry 2h, take out and to be positioned over cooling in the exsiccator, weigh, repeat this and be operated to constant weight.Accurately take by weighing the lipid that a certain amount of extraction obtains, be dissolved in methenyl choloride, making its concentration is about 5mg/mL.
3, efficient liquid phase chromatographic analysis
Chromatographic column: symmetry C18 (250mm * 4.6mm, 5 μ m).Phase: the A that flows is acetonitrile/chloroform (70:30), and B is acetonitrile/chloroform (30:70).Binary gradient elution program: 0min, 100%A; 5min, 70%A; 25min, 0%A; 25.1 ~ 30min, 100%A.Column temperature: 40 ℃.Flow velocity: 0.8mL/min.The ELSD drift tube temperature: 85 ℃, N2 flow velocity: 2.5L/min.
Sample introduction analysis respectively with this understanding obtains the fingerprint spectrogram that cocoa power sample lipid glyceride is formed, and sees accompanying drawing 1.Be reference with No. 11 peaks, the relative retention time at each peak in the calculation sample.
4, the foundation of standard finger-print and total fingerprint peaks feature
High-performance liquid chromatogram determination by the composition of the lipid glyceride in the cocoa power sample in 11 different places of production, compare its chromatogram, determine 14 at common characteristic peak, the relative standard deviation RSD of the relative retention time RT at total peak (be reference with No. 11 peaks) is all less than 1%; The fingerprint peaks that wherein surpasses total peak area 3% has 3, is respectively: No. 8 peaks, relative peak area 10.848% ~ 12.403%; No. 11 peaks, relative peak area 51.518% ~ 53.078%; No. 12 peaks, relative peak area 30.463% ~ 32.677%.
5, cluster analysis
Measure the composition of the triglyceride that mixes longan shell and palmitic cocoa power according to 2 and 3 method,, and select the Ward method as clustering method, Chebychev distance as the measuring distance method, average is 1 as standardization, carries out cluster analysis.No. 11 total fingerprint peaks is more stable, and peak area is bigger, and separates better with adjacent peaks, and retention time is more placed in the middle, and therefore setting No. 11 peaks is with reference to the peak.Then, the matrix of forming with the relative peak area at common characteristic peak and non-total peak (with No. 11 peaks as the reference peak) carries out cluster analysis.
Also be substantially free of lipid in the longan shell powder, in like manner make the sample (total lipid content also all reaches 10% behind the admixture substitute of cocoa fat) of 8 admixture longan shell powder and different amount substitute of cocoa fat, see Table 2.Extract lipid in these samples according to the method for step 2, and carry out HPLC fingerprint analysis (Fig. 2 b) according to step 3.The matrix of forming according to the relative peak area of chromatographic peak carries out cluster analysis, by the cluster analysis dendrogram (Fig. 4) of gained as seen: sample B1 can not (distinguish the S1～S11) from pure cocoa power sample, B2～B8 then all can distinguish over S1～S11, namely for the cocoa power sample of admixture longan shell powder, when mixing 1.2% and can be differentiated during above substitute of cocoa fat.
Table 2: longan shell is mixed in simulation and the palmitic cocoa power of mingling is formed
6, stratographic analysis precision
The cocoa butter that the S11 cocoa power extracts through Soxhlet is mixed with the sample solution of 5mg/mL, and continuous sample introduction 5 times is added up relative retention time (retention time with No. 11 peaks is reference) and the relative peak area at each total peak.The result shows, the relative retention time at each total peak and the relative standard deviation of relative peak area (RSD)＜2% show that the chromatographic fingerprint analytical precision test of cocoa power lipid meets the requirements.
7, method repeatability
Take by weighing 5 parts in S11 cocoa power sample, extract lipid and carry out the HPLC analysis by 1.2 and 1.3 respectively, respectively relative retention time and the relative peak area at total peak are added up, their relative standard deviation (RSD) is all less than 3%.
8, method stability test
The S11 cocoa power is configured to the sample solution of 5mg/mL through the cocoa butter of Soxhlet extraction, at room temperature (about 18 ℃) are preserved, respectively at 0,5,10,15,20, the 25h sample introduction is analyzed, and relative retention time (retention time with No. 11 peaks is reference) and the relative peak area at each total peak are added up.The result shows that the relative retention time at each total peak and relative peak area RSD are all less than 5%.
More than test shows, above-mentioned finger print measuring method precision, stable, reliable.
Claims (2)
1. the cocoa power based on the lipid cluster analysis is mingled detection method, it is characterized in that, comprises the steps:
1) make up cocoa power glyceride standard finger-print, carry out the high performance liquid chromatography fingerprint analysis that glyceride is formed behind the lipid of the authentic cocoa power sample of some batches of separate sources of extraction, and definite common characteristic peak; Specifically comprise the steps:
A) described cocoa power lipid extracting method is: take by weighing 2.50g left and right sides cocoa power, place filtration paper cylinder, put into the extraction flask of Soxhlet extractor, and add about 2/3 sherwood oil in extraction flask, 47 ℃ of water-baths, 6h; Take out extraction flask, use Rotary Evaporators, be concentrated into dried; Then with extraction flask at 105 ℃ of dry 2h, take out and to be positioned over cooling in the exsiccator, weigh, repeat this and be operated to constant weight; Accurately take by weighing and extract the lipid that obtains, be dissolved in methenyl choloride, making its concentration is about 5mg/mL;
B) described HPLC analytical method is: chromatographic column: symmetry C18,250mm * 4.6mm, 5 μ m; Phase: A flows: acetonitrile and chloroform volume ratio are 70:30, B: acetonitrile and chloroform volume ratio are 30:70; Binary gradient elution program: 0min, 100%A; 5min, 70%A; 25min, 0%A; 25.1～30min, 100%A; Column temperature: 40 ℃; Flow velocity: 0.8mL/min; The ELSD drift tube temperature: 85 ℃, N2 flow velocity: 2.5L/min; The sample that a) obtains of sample introduction analytical procedure with this understanding obtains the efficient liquid-phase chromatograph finger print atlas of cocoa power glyceride;
C) definite method of described standard finger-print is: by extracting the high-performance liquid chromatogram determination of the lipid glyceride composition that obtains in the cocoa power sample in 11 different places of production, compare its chromatogram, determined that standard finger-print has 14 characteristic peaks, described common characteristic peak with the relative standard deviation RSD of No. 11 peak POS relative retention time RT that is reference all less than 1%; Wherein
No. 1 average RT in peak is that 0.411, RSD is 0.18%;
No. 2 average RT in peak are that 0.465, RSD is 0%;
No. 3 average RT in peak are that 0.703, RSD is 0.15%;
No. 4 average RT in peak are that 0.743, RSD is 0.22%;
No. 5 average RT in peak are that 0.786, RSD is 0.07%;
No. 6 average RT in peak are that 0.848, RSD is 0.12%;
No. 7 average RT in peak are that 0.878, RSD is 0.02%;
No. 8 average RT in peak are that 0.899, RSD is 0%;
No. 9 average RT in peak are that 0.944, RSD is 0.03%;
No. 10 average RT in peak are that 0.977, RSD is 0.06%;
No. 11 average RT in peak are that 1.000, RSD is 0%;
No. 12 average RT in peak are that 1.107, RSD is 0%;
No. 13 average RT in peak are that 1.183, RSD is 0.34%;
No. 14 average RT in peak are that 1.215, RSD is 0.34%;
2) preparation method of the cocoa power glyceride standard finger-print in the employing step 1) is measured the glyceride of cocoa power unknown sample and is formed;
3) with step 2) chromatogram of the unknown sample that obtains compares with the standard finger-print that step 1) obtains, and the matrix according to their relative peak areas are formed carries out cluster analysis, and whether the judgement cocoa power is mingled.
2. the method for claim 1, it is characterized in that, in the described step 3) cluster analysis select the Ward method as clustering method, Chebychev distance as the measuring distance method, average is 1 as standardization, carry out cluster analysis, the matrix of forming with the relative peak area at 14 common characteristic peaks carries out cluster analysis, and wherein No. 11 peaks are as the reference peak, its average RT is that 1.000, RSD is 0%.
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| CN104215711A (en) * | 2014-09-15 | 2014-12-17 | 湖北中烟工业有限责任公司 | Fingerprint determination method of active ingredients in cocoa powder |
| CN112195270A (en) * | 2020-11-17 | 2021-01-08 | 谱尼测试集团北京检验认证科学研究院有限公司 | Primer probe and kit for specific detection of cocoa plant-derived components |
| CN112782293B (en) * | 2020-12-17 | 2023-01-13 | 谱尼测试集团北京检验认证科学研究院有限公司 | Quantitative detection method for cocoa shell doped in cocoa powder |
| CN113624885A (en) * | 2021-08-19 | 2021-11-09 | 宁波市疾病预防控制中心 | Method for rapidly identifying adulterated wheat flour in chestnut flour |
| CN114139643B (en) * | 2021-12-07 | 2022-11-29 | 佳力士添加剂（海安）有限公司 | Monoglyceride quality detection method and system based on machine vision |
| CN115078606B (en) * | 2022-06-14 | 2023-06-06 | 浙江工商大学 | Method for identifying varieties of minced shrimp products based on lipidomic |
| CN116359420B (en) * | 2023-04-11 | 2023-08-18 | 烟台国工智能科技有限公司 | Chromatographic data impurity qualitative analysis method based on clustering algorithm and application |
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