CN104330484A - Detection method of vitamin B12 content in vitamin complex additive - Google Patents
Detection method of vitamin B12 content in vitamin complex additive Download PDFInfo
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- CN104330484A CN104330484A CN201410573261.2A CN201410573261A CN104330484A CN 104330484 A CN104330484 A CN 104330484A CN 201410573261 A CN201410573261 A CN 201410573261A CN 104330484 A CN104330484 A CN 104330484A
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Abstract
The invention relates to a detection method of the vitamin B12 content in a vitamin complex additive, which belongs to the fields of food additives and detection methods of the food additives. The detection method comprises the following steps: (1) preparing a vitamin B12 stock solution with the concentration of 1 mg/ml, then preparing serial solutions with the normal concentrations of 0.5, 1.0, 2.0, 3.0, 4.0 and 5.0mu g/ml, putting on an instrument to measure under certain chromatographic conditions, and calculating a regression equation and correlation coefficients; and (2) weighing a test sample, putting on the instrument to measure under the same chromatographic conditions, and calculating the concentration of the test sample according to the peak height of a standard sample, the concentration of the standard sample and the peak height of the test sample. According to the detection method, the measuring precision and accuracy can meet the requirement of dairy product analysis, and the detection method has the characteristics of high accuracy, high repeatability, simplicity and convenience in operation, low detection limit and the like and can be used as a conventional detection method to be applied.
Description
Technical field
The present invention relates to the detection method of cobalamin content in a kind of compound vitamin additive, belong to food additives and detection method field thereof.
Background technology
Cobalamin is the maximum and the most complicated one of all vitamins Middle molecule, but perhaps human body is one minimum in all vitamins to its requirement, about only needing 3 emblems gram every day.Excessive cobalamin also can have side effects, and a large amount of cobalamin also can cause folic acid deficiency.Cobalamin can improve the utilization factor of folic acid, if be deficient in vitamin B12, then the utilization of folic acid is affected.Vitamin B12 deficiency also can cause peripheral neuritis.The Early manifestation that child lacks cobalamin is spiritual abnormal feeling, have a dull expression on one's face, cry less less noisy, slow in reacting, like that sleep, brothers' unintentional motion or head and limbs tremble, finally cause anaemia.The people of expert to 323 B12 that are deficient in vitamin of the Columbia University of USA New York and the University of Colorado of Denver investigates, and finds wherein the neurotrosis that degree is different and psychiatric symptoms to have been appearred in the people (totally 141 people) of nearly half.Symptom comprises dementia, depression, memory disorders, equilibrium function imbalance etc., and what have also has schizoid performance.After doctor gives these people's replenishing vitamins B12, the psychiatric symptoms of many patients disappears completely.
According to the data reported at present, measure cobalamin and mainly contain four kinds of methods: colourimetry, atomic absorption spectrophotometry, high performance liquid chromatography and microbial method.Microbial method measures the universal method as food vitamins B12, and this method sensitivity is high especially, can determine the content of 0.01ng/ml in sample liquid.But due to this method complex operation, round of visits is long, technical difficulty is large, the many and poor reproducibility of influence factor, high in cost of production problem and can not widespread use.Particularly measure the high sample of the cobalamin content such as adjuvant by this method, because extension rate is comparatively large and be difficult to grasp etc. and make error at measurment comparatively large, not bery suitable; And first three methods is mostly for sample (general content mg/g level) that drug class equal size is higher.And the cobalamin content in milk powder specific complex vitamin(e) supplement is generally 10 μ g/g or lower (in national regulations baby formula milk powder, cobalamin content is >=0.8 μ g/100g).Therefore, existing detection method is generally difficult to detect cobalamin content in baby formula milk powder.
Therefore study that a kind of accuracy is high, the low detection method being applicable to cobalamin content in baby formula milk powder of high, easy and simple to handle, the detection limit of repeatability is necessary.
Summary of the invention
The present invention aims to provide high, repeated high, easy and simple to handle, the low detection method being applicable to cobalamin content in baby formula milk powder of detection limit of a kind of accuracy.
The detection method of cobalamin content in described compound vitamin additive, comprises the following steps:
(1) preparing cobalamin concentration is the storing solution of 1mg/ml; Then be mixed with the solution that concentration is respectively the series standard concentration of 0.5,1.0,2.0,3.0,4.0,5.0 μ g/ml, sample size is respectively 20 μ L.On under certain chromatographic condition, machine measures and calculates regression equation and related coefficient.
(2) take test sample 10g to be put in 125ml triangular flask.Ultrasound wave is put into after adding the distilled water dissolving of 30m130-50 DEG C, l0 divides quiet rear taking-up, proceed in 50ml volumetric flask and add 10% solution of zinc sulfate 5ml constant volume, mix rear Filter paper filtering, filtrate by after the filter membrane of 0.45 μm under identical chromatographic condition on machine measure.Calculate according to standard sample peak height, standard sample concentration and test sample peak height and test sample concentration.
Preferably, chromatographic condition of the present invention is chromatographic column is ODS chromatographic column, and chromatographic column theoretical cam curve is 2216; Column temperature is room temperature; Flow rate of mobile phase 1ml/min; Measure wavelength 361nm; Mobile phase is sodium dihydrogen phosphate (10g/L pH=3.5): the mixed liquor of methyl alcohol=70:30.
Detection method of the present invention, the typical curve obtained is:
Y=-0.1340+0.0009592X，R=0.9976。Result shows: be proportionate between B12 concentration and peak height in measurement range, and correlativity is good, can be used for external standard method and carries out quantitative measurement.
Detection method of the present invention, the computing formula of vitamin B2 content is:
Vitamin B2 content (μ g/100g)=(h1 × V × C)/(h2 × m) × 100, wherein h1 is test specimens peak height (mm), h2 is standard sample peak height (mm), V is test specimens liquid long-pending (mL), m is test specimens quality (g), and C is cobalamin concentration of standard solution (μ g/ml).
In detection method of the present invention, adopt cobalamin content in high effective liquid chromatography for measuring compound vitamin additive, owing to often making carrier with organism in sample, even if so also there will be muddy phenomenon by filter membrane after directly dissolving with distilled water, and can effectively organic principle be precipitated when making precipitation agent with zinc sulfate, liquid clarification to be measured, is more conducive to machine and measures.After measured, the method recovery and Precision test result better, the recovery is between 90-102%, average out to 96.4%, the coefficient of variation that sample replication is 12 times is 0.54%, the method detects and is limited to 4ng, can be calculated detecting of cobalamin in the special vitamin(e) supplement of baby formula milk powder and is limited to 1 μ g/g, can meet the mensuration of cobalamin in the special vitamin(e) supplement of any milk powder.
Detection method of the present invention, adopt HPLC method to measure its mensuration preci-sion and accuracy of cobalamin in the special vitamin(e) supplement of baby formula milk powder and can meet the requirement of dairy products analysis, there is the features such as accuracy is high, repeatability is high, easy and simple to handle, detection limit is low, can be used as common detection methods application.
Embodiment
Embodiment one:
(1) preparing cobalamin concentration is the storing solution of 1mg/ml; Then be mixed with the solution that concentration is respectively the series standard concentration of 0.5,1.0,2.0,3.0,4.0,5.0 μ g/ml, sample size is respectively 20 μ L.Under certain chromatographic condition, (chromatographic column is ODS chromatographic column, and chromatographic column theoretical cam curve is 2216; Column temperature is room temperature; Flow rate of mobile phase 1ml/min; Measure wavelength 361nm; Mobile phase is sodium dihydrogen phosphate (10g/L pH=3.5): the mixed liquor of methyl alcohol=70:30) upper machine measures and calculates regression equation and related coefficient.The typical curve obtained is:
Y=-0.1340+0.0009592X，R=0.9976。Result shows: be proportionate between B12 concentration and peak height in measurement range, and correlativity is good, can be used for external standard method and carries out quantitative measurement.
(2) take test sample 10g to be put in 125ml triangular flask.Ultrasound wave is put into after adding the distilled water dissolving of 30m130-50 DEG C, l0 divides quiet rear taking-up, proceed in 50ml volumetric flask and add 10% solution of zinc sulfate 5ml constant volume, mix rear Filter paper filtering, filtrate by after the filter membrane of 0.45 μm under identical chromatographic condition on machine measure.Calculate according to standard sample peak height, standard sample concentration and test sample peak height and test sample concentration.
The computing formula of vitamin B2 content is:
Vitamin B2 content (μ g/100g)=(h1 × V × C)/(h2 × m) × 100, wherein h1 is test specimens peak height (mm), h2 is standard sample peak height (mm), V is test specimens liquid long-pending (mL), m is test specimens quality (g), and C is cobalamin concentration of standard solution (μ g/ml).
Embodiment two: replica test, recovery test and detection limit are tested
After measured, the method recovery and Precision test result better, the recovery is between 90-102%, average out to 96.4%, the coefficient of variation that sample replication is 12 times is 0.54%, the method detects and is limited to 4ng, can be calculated detecting of cobalamin in the special vitamin(e) supplement of baby formula milk powder and is limited to 1 μ g/g, can meet the mensuration of cobalamin in the special vitamin(e) supplement of any milk powder.
Claims (2)
1. the detection method of cobalamin content in compound vitamin additive, comprises the following steps:
(1) preparing cobalamin concentration is the storing solution of 1mg/ml; Then be mixed with the solution that concentration is respectively the series standard concentration of 0.5,1.0,2.0,3.0,4.0,5.0 μ g/ml, sample size is respectively 20 μ L, and under certain chromatographic condition, machine measures and calculates regression equation and related coefficient;
(2) taking test sample 10g is put in 125ml triangular flask;
Ultrasound wave is put into after adding the distilled water dissolving of 30m130-50 DEG C, l0 divides quiet rear taking-up, proceed in 50ml volumetric flask and add 10% solution of zinc sulfate 5ml constant volume, mix rear Filter paper filtering, filtrate by after the filter membrane of 0.45 μm under identical chromatographic condition on machine measure, to calculate with test sample peak height according to standard sample peak height, standard sample concentration and test sample concentration.
2. the detection method of cobalamin content in compound vitamin additive as claimed in claim 1, it is characterized in that described chromatographic condition be chromatographic column is ODS chromatographic column, chromatographic column theoretical cam curve is 2216; Column temperature is room temperature; Flow rate of mobile phase 1ml/min; Measure wavelength 361nm; Mobile phase is the sodium dihydrogen phosphate of 10g/L, pH=3.5: the mixed liquor of methyl alcohol=70:30.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107860847A (en) * | 2017-11-14 | 2018-03-30 | 济南维瑞医药科技开发有限公司 | Vitamin B in one kind detection multivitamin preparation12Method about material |
| CN108445092A (en) * | 2018-01-30 | 2018-08-24 | 上海信谊万象药业股份有限公司 | A kind of lysine vitamin B12The analysis method that drug content measures in granule |
| CN109917037A (en) * | 2019-03-26 | 2019-06-21 | 广西壮族自治区食品药品检验所 | Gentamicin procaine ties up the HPLC detection method of vitamin B12 content in B12 particle/capsule |
-
2014
- 2014-10-23 CN CN201410573261.2A patent/CN104330484A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107860847A (en) * | 2017-11-14 | 2018-03-30 | 济南维瑞医药科技开发有限公司 | Vitamin B in one kind detection multivitamin preparation12Method about material |
| CN108445092A (en) * | 2018-01-30 | 2018-08-24 | 上海信谊万象药业股份有限公司 | A kind of lysine vitamin B12The analysis method that drug content measures in granule |
| CN109917037A (en) * | 2019-03-26 | 2019-06-21 | 广西壮族自治区食品药品检验所 | Gentamicin procaine ties up the HPLC detection method of vitamin B12 content in B12 particle/capsule |
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Application publication date: 20150204 |