CN101975837B - Method for determining content and purity of L-carnitine in milk powder - Google Patents
Method for determining content and purity of L-carnitine in milk powder Download PDFInfo
- Publication number
- CN101975837B CN101975837B CN2010102745735A CN201010274573A CN101975837B CN 101975837 B CN101975837 B CN 101975837B CN 2010102745735 A CN2010102745735 A CN 2010102745735A CN 201010274573 A CN201010274573 A CN 201010274573A CN 101975837 B CN101975837 B CN 101975837B
- Authority
- CN
- China
- Prior art keywords
- carnitine
- solution
- concentration
- sample
- gained
- 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.)
- Expired - Fee Related
Links
Images
Landscapes
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention discloses a method for determining content and purity of L-carnitine in milk powder, comprising the following steps in sequence: 1) taking the milk powder as a sample for sample treatment; 2) preparing standard solutions respectively by taking an L-carnitine standard substance and a D-carnitine standard substance; 3) correspondingly preparing an L-carnitine standard solution and a D-carnitine standard solution obtained the step 2) into an L-carnitine derivative solution containing gradient concentration and a D-carnitine solution containing gradient concentration; 4) carrying out derivatization reaction on the sample solutions; 5) carrying out high performance liquid chromatography on the products obtained in the step 3) and step 4); 6) acquiring the concentration cL of the L-carnitine and the concentration cD of the D-carnitine; and 7) acquiring the content and the purity of the L-carnitine in the milk powder sample. By adopting the method for determining the content and the purity of L-carnitine in the milk powder of the invention, the detection sensitivity can be greatly improved.
Description
Technical field
The present invention relates to the assay method of L-carnitine content and purity in a kind of milk powder; Be particularly related to the method for L-carnitine content and purity in a kind of column front derivation high effective liquid chromatography for measuring milk powder.
Background technology
The synthesis capability of L-carnitine in infants a little less than, must external source picked-up.Therefore existing in the world a plurality of countries and regions add the L-carnitine in baby's milk powder, domestic also have the baby's milk powder that adds the L-carnitine to go on the market.And carnitine has two kinds of existence forms (L type and D type) at nature, and there is negative interaction in the dextrorotation carnitine to health, and the l-cn of chemosynthesis generally is left-handed and dextrorotation carnitine potpourri, separates then to remove that the dextrorotation carnitine produces.
The detection of l-cn is general in the domestic health products adopts direct high performance liquid chromatography, and directly high performance liquid chromatography can not be distinguished left-handed and the dextrorotation carnitine.Cold peach blossoms etc. " D-and L-carnitine Enantiomers by Capillary Electrophoresis separate study " propose to separate the carnitine enantiomorph with CZE, and detect with UV-detector, and the Capillary Electrophoresis operation is had relatively high expectations, and are unfavorable for that the laboratory is universal; Use L-alanine-beta-naphthylamine to be derivative reagent among the Zhu Weixia etc. " rp-hplc determination of l-cn in the health products ", the HPLC UV-detector detects the content of l-cn in the health products; Use alpha-brominated Propiophenone to be derivative reagent among the Xu Juanjuan etc. " chiral hplc is measured the content of optical isomer in the l-cn ", the HPLC UV-detector detects the content of l-cn.
The context of detection of L-carnitine in milk powder, the analytical approach that the patent No. 02100582.6 " a kind of method of utilizing L-carnitine content in enzyme-colorimetric method for determining milk powder " proposes can not be distinguished the carnitine enantiomorph.
Method with L-carnitine in the column front derivation high effective liquid chromatography for measuring baby milk powder still is not reported.
Summary of the invention
The technical matters that the present invention will solve provides a kind of method of measuring L-carnitine content and purity in the milk powder.
In order to solve the problems of the technologies described above, the present invention provides the assay method of L-carnitine content and purity in a kind of milk powder, may further comprise the steps successively:
1), sample preparation:
Earlier milk powder 10～20g is settled to 100ml after with deionized water dissolving, removes the processing (promptly precipitating butter oil and lactoprotein with hydrochloric acid solution and sodium hydroxide solution respectively) of butter oil and lactoprotein more respectively, filtration, collected filtrating is sample solution;
2), preparation standard solution:
Take by weighing 50mg L-carnitine standard items,, and be settled to 10ml with deionized water dissolving; Gained solution obtains L-carnitine standard solution through 0.45 μ m membrane filtration;
Take by weighing 50mg D-carnitine standard items,, and be settled to 10ml with deionized water dissolving; Gained solution obtains D-carnitine standard solution through 0.45 μ m membrane filtration;
3) with step 2) the L-carnitine standard solution and the D-carnitine standard solution of gained operate respectively as follows:
The L-carnitine standard solution of getting 30 μ l is put in the volumetric flask, adds the 0.05M carbonic acid buffer of 80 μ l and the 1-of 80 μ l (9-fluorenyl)-ethyl chloroformate (i.e. (+)-FLEC reagent), the bottle stopper of volumetric flask beyond the Great Wall, and slowly rotated 3～5 seconds; Volumetric flask is put into 45 ℃ of water-baths place 1h, after taking out volumetric flask then and naturally cooling to room temperature, in volumetric flask, add deionized water and be settled to 10ml; Solution behind the constant volume is diluted with deionized water, obtain the series derivatives solution that L-carnitine concentration is respectively the L-carnitine of 0.5,1.0,1.5,2.0,2.5 μ g/ml;
In like manner, the D-carnitine standard solution of getting 30 μ l is put in the volumetric flask, adds the 0.05M carbonic acid buffer of 80 μ l and (+)-FLEC reagent of 80 μ l, the bottle stopper of volumetric flask beyond the Great Wall, and slowly rotated 3～5 seconds; Volumetric flask is put into 45 ℃ of water-baths place 1h, after taking out volumetric flask then and naturally cooling to room temperature, in volumetric flask, add deionized water and be settled to 10ml; Solution behind the constant volume is diluted with deionized water, obtain the series derivatives solution that D-carnitine concentration is respectively the D-carnitine of 0.5,1.0,1.5,2.0,2.5 μ g/ml;
The series derivatives solution of the L-carnitine of gained and the series derivatives solution of D-carnitine are used 0.45 μ m filtering with microporous membrane respectively; Get the L-carnitine derivative solution of gradient concentration and the D-carnitine derivative solution (being used for detecting of following step) of gradient concentration respectively with high performance liquid chromatography;
4), sample solution derivative reaction:
The sample solution of getting 60 μ l step 1) gained adds the 0.05M carbonic acid buffer of 30 μ l and (+)-FLEC reagent of 30 μ l again in volumetric flask, the bottle stopper of volumetric flask beyond the Great Wall, and slowly rotated 3～5 seconds; Volumetric flask is put into 45 ℃ of water-baths place 1h, after the taking-up volumetric flask naturally cools to room temperature from water-bath then, in volumetric flask, add deionized water and be settled to 10ml; Solution behind the constant volume with 0.45 μ m filtering with microporous membrane, is got sample introduction liquid;
5), high performance liquid chromatography detects:
The sample introduction liquid of the D-carnitine derivative solution of the L-carnitine derivative solution of the gradient concentration of step 3) gained and gradient concentration and step 4) gained is carried out high performance liquid chromatography respectively to be detected; Testing conditions is following:
Select the C18 chromatographic column for use, chromatographic condition is: moving phase is: the triethylamine phosphate buffer: the volume ratio of acetonitrile=73: 27; Flow velocity: 1ml/min; Detect wavelength: emission wavelength 269nm, detect wavelength 310nm; Sample size: 10 μ l;
Respectively the response peak area, the response peak area of sample introduction liquid of D-carnitine derivative solution of response peak area, gradient concentration of L-carnitine derivative solution of gradient concentration;
6), obtain the concentration c L of L-carnitine and the concentration c D of D-carnitine respectively:
Response peak area with the L-carnitine derivative solution of the gradient concentration of step 5) gained is mapped to concentration, the typical curve of L-carnitine;
Response peak area with the D-carnitine derivative solution of the gradient concentration of step 5) gained is mapped to concentration, the typical curve of D-carnitine;
With the response peak area of the sample introduction liquid of step 5) gained respectively with the typical curve contrast of the typical curve and the D-carnitine of above-mentioned L-carnitine; The result of gained multiply by the extension rate 10*1000/60 of sample solution in the derivatization reaction of step 4), thereby gets the concentration c L of L-carnitine in the sample solution and the concentration c D of D-carnitine respectively;
7), obtain L-carnitine content and purity in the powdered milk sample:
In the formula: cL---the concentration of L-carnitine in the sample solution, μ g/ml;
The quality of m---milk powder, g.
In the formula: cL---the concentration of L-carnitine in the sample solution, μ g/ml;
CD---the concentration of D-carnitine in the sample solution, μ g/ml.
Improvement as the assay method of L-carnitine content and purity in the milk powder of the present invention:
0.05M the preparation method of carbonic acid buffer is following: 338mg natrium carbonicum calcinatum and 152mg soda mint are dissolved in 100ml water;
The preparation method of triethylamine phosphate buffer solution is following: the triethylamine of 6.8ml is separated with 950ml is water-soluble, and using volume fraction then is that 84% phosphate aqueous solution is regulated pH to 2.5, and last water is settled to 1000ml.
Further improvement as the assay method of L-carnitine content and purity in the milk powder of the present invention: step 1) is:
Earlier with milk powder with deionized water dissolving after in 50 ℃ of water-bath 15min, ultrasonic Treatment 10min then is settled to 100ml with deionized water again;
The hydrochloric acid solution that adds 0.1mol/L is then regulated pH to 1.8～2.2, uses the nature filter paper filtering after leaving standstill 10min; The NaOH solution that in the filtrating of gained, adds 1mol/L is regulated pH to 5.8～6.2, carries out filtration second time with nature filter paper after leaving standstill 10min, and the filtrating that filter gained the second time is sample solution.
The present invention is with 1-(9-fluorenyl)-ethyl chloroformate, i.e. (+)-FLEC is a derivative reagent, on the carnitine enantiomorph, connects a fluorophor, thereby on chromatographic column, separates, and analyze with fluorescence detector, has improved detection sensitivity greatly.
In the present invention, agents useful for same is commercial product.For example, L-carnitine standard items, D-carnitine standard items, (+)-FLEC reagent can be available from Sigma company.
Method of the present invention has at first been got rid of the interference of fat and albumen in the milk powder; Then chiral resolution left-handed with dextrorotation carnitine enantiomter; And detect with high-efficient liquid phase chromatogram technique analysis, provide a kind of column front derivation high performance liquid chromatography to detect the method for L-carnitine content and purity in the milk powder.
The present invention utilizes the chirality derivatization method; The carnitine enantiomter is split; And carry out high performance liquid chromatography and detect, to measure the content and the purity of l-cn in the dispensed food for baby, significant to the quality control and the safety detection of dispensed food for baby and milk powder.
Adopt method of the present invention to measure the content and the purity of L-carnitine in the milk powder, have following advantage:
1, the present invention compares with the traditional analysis method, can chiral resolution L type and D type carnitine, reach the purpose of separating and detecting, and high performance liquid chromatograph uses in the laboratory more general;
2, the high performance liquid chromatography XRF accuracy of the present invention's employing is good, and separating power is good, and is highly sensitive, and detecting lower limit approximately is 1/10～1/100 of UV-detector, and instrumentation is simple;
3, can get rid of the interference of fat and albumen in the milk powder, effectively detect the content and the purity of L-carnitine.
Description of drawings
Do further explain below in conjunction with the accompanying drawing specific embodiments of the invention.
Fig. 1 is a L-carnitine typical curve;
Fig. 2 is a D-carnitine typical curve
Fig. 3 is the powdered milk sample chromatogram of embodiment 1.
Embodiment
Below in conjunction with the instance of L-carnitine content and purity in the column front derivation high effective liquid chromatography for measuring milk powder of the present invention, embodiment of the present invention is described.
The assay method of L-carnitine content and purity in embodiment 1, a kind of milk powder is characterized in that may further comprise the steps successively:
1), sample preparation:
Accurately take by weighing commercially available ordinary powdered milk sample 16.65g, be dissolved in the 30ml deionized water, 50 ℃ of water-bath 15min, ultrasound wave (frequency 30kHz) 10min fully dissolves milk powder, adds deionized water then and is settled to 100ml.
Regulate pH to 2 with the hydrochloric acid solution of 0.1mol/L then, use the nature filter paper filtering after leaving standstill 10min.Regulate pH to 6 the filtrating of gained with the NaOH solution of 1mol/L, carry out filtration second time with nature filter paper after leaving standstill 10min, the filtrating that filter gained the second time is sample solution.
2), preparation standard solution:
Take by weighing 50mg L-carnitine standard items,, and be settled to 10ml with deionized water dissolving; Gained solution obtains L-carnitine standard solution through 0.45 μ m membrane filtration;
Take by weighing 50mg D-carnitine standard items,, and be settled to 10ml with deionized water dissolving; Gained solution obtains D-carnitine standard solution through 0.45 μ m membrane filtration;
3), with step 2) the L-carnitine standard solution and the D-carnitine standard solution of gained operate respectively as follows:
The L-carnitine standard solution of getting 30 μ l is put in the volumetric flask, adds the 0.05M carbonic acid buffer of 80 μ l and (+)-FLEC reagent of 80 μ l, the bottle stopper of volumetric flask beyond the Great Wall, and slowly rotated 3～5 seconds; Volumetric flask is put into 45 ℃ of water-baths place 1h, after the taking-up volumetric flask naturally cools to room temperature from water-bath then, in volumetric flask, add deionized water and be settled to 10ml; Solution behind the constant volume is diluted with deionized water, obtain the series derivatives solution that L-carnitine concentration is respectively the L-carnitine of 0.5,1.0,1.5,2.0,2.5 μ g/ml;
In like manner, the D-carnitine standard solution of getting 30 μ l is put in the volumetric flask, adds the 0.05M carbonic acid buffer of 80 μ l and (+)-FLEC reagent of 80 μ l, the bottle stopper of volumetric flask beyond the Great Wall, and slowly rotated 3～5 seconds; Volumetric flask is put into 45 ℃ of water-baths place 1h, after the taking-up volumetric flask naturally cools to room temperature from water-bath then, in volumetric flask, add deionized water and be settled to 10ml; Solution behind the constant volume is diluted with deionized water, obtain the series derivatives solution that D-carnitine concentration is respectively the D-carnitine of 0.5,1.0,1.5,2.0,2.5 μ g/ml;
The L-carnitine series derivatives solution and the D-carnitine series derivatives solution of gained are used 0.45 μ m filtering with microporous membrane (can suitably pressurize) respectively; Get the L-carnitine derivative solution (totally 5 kinds) of gradient concentration and the D-carnitine derivative solution (totally 5 kinds) of gradient concentration respectively;
4), sample solution derivative reaction
Measure the preparation of reagent earlier:
The triethylamine of triethylamine phosphate buffer solution: 6.8ml is separated with 950ml is water-soluble, and the use volume fraction is its pH to 2.5 of phosphate aqueous solution adjusting of 84%, and last dilute with water is settled to 1000ml.
0.05M carbonic acid buffer: 338mg natrium carbonicum calcinatum and 152mg soda mint are dissolved in 100ml water.
The sample solution of getting 60 μ l step 1) gained adds the 0.05M carbonic acid buffer of 30 μ l and (+)-FLEC reagent of 30 μ l again in volumetric flask, the bottle stopper of volumetric flask beyond the Great Wall, and slowly rotated 3～5 seconds; Volumetric flask is put into 45 ℃ of water-baths place 1h, after the taking-up volumetric flask naturally cools to room temperature from water-bath then, in volumetric flask, add deionized water and be settled to 10ml; Solution behind the constant volume with 0.45 μ m filtering with microporous membrane (can suitably pressurize), is got sample introduction liquid.
5), high performance liquid chromatography detects
The sample introduction liquid of the D-carnitine derivative solution of the L-carnitine derivative solution of the gradient concentration of step 3) gained and gradient concentration and step 4) gained is carried out high performance liquid chromatography respectively to be detected; Testing conditions is following:
Select the C18 chromatographic column for use.Chromatographic condition is: moving phase is the triethylamine phosphate buffer: acetonitrile (73: 27, volume ratio); Flow velocity: 1ml/min; Detect wavelength: emission wavelength 269nm, detect wavelength 310nm; Sample size: 10 μ l.
Respectively the response peak area, the response peak area of sample introduction liquid of D-carnitine derivative solution of response peak area, gradient concentration of L-carnitine derivative solution of gradient concentration.
6), obtain L-carnitine content and purity:
Response peak area with the L-carnitine derivative solution of the gradient concentration of step 5) gained is mapped to concentration, the typical curve of L-carnitine; As shown in Figure 1: L-carnitine, c (μ g/ml)=2*10
-7A+0.0978; A is a peak area value in the formula;
Response peak area with the D-carnitine derivative solution of the gradient concentration of step 5) gained is mapped to concentration, the typical curve of D-carnitine; As shown in Figure 2: D-carnitine, c (μ g/ml)=2*10
-7A+0.1185; A is a peak area value in the formula.
With the response peak area of the sample introduction liquid of step 5) gained respectively with the typical curve contrast of the typical curve and the D-carnitine of above-mentioned L-carnitine; The result of gained multiply by the extension rate 10*1000/60 of sample solution in the derivatization reaction of step 4), thereby gets the concentration c L of L-carnitine in the sample solution and the concentration c D of D-carnitine respectively; Be specially:
The testing result figure of sample introduction liquid sees Fig. 3, and L-carnitine derivative peak area value is 158390, and D-carnitine derivative peak area is 0.Thereby L-carnitine concentration c L=(158390*2*10 in the sample solution
-7+ 0.0978) * 10*1000/60=21.58 μ g/ml; CD=0.
7), obtain L-carnitine content and purity in the powdered milk sample:
(annotate: in the formula: cL---the concentration of L-carnitine in the sample solution, μ g/ml;
The quality of m---milk powder, g.)
(annotate: in the formula: cL---the concentration of L-carnitine in the sample solution, μ g/ml;
CD---the concentration of D-carnitine in the sample solution, μ g/ml.)
Result of calculation is summed up as follows: the content of L-carnitine is 12.95mg/100g in the powdered milk sample, and the D-carnitine does not detect.
The assay method of L-carnitine content and purity in embodiment 2, a kind of milk powder is characterized in that may further comprise the steps successively:
1), sample preparation:
Accurately take by weighing commercially available baby formula milk powder I sample 14.95g, be dissolved in the 30m deionized water, 50 ℃ of water-bath 15min, ultrasound wave (frequency 30kHz) 10min, milk powder fully dissolves, and adds deionized water and is settled to 100ml.
Regulate pH to 1.9 with the hydrochloric acid solution of 0.1mol/L then, use the nature filter paper filtering after leaving standstill 10min.The filtrating of gained is regulated pH to 6.1 with the NaOH solution of 1mol/L, carry out the second time with nature filter paper and filter, the filtrating of filtering gained for the second time is sample solution.
Step 2)～step 6) all is equal to embodiment 1.
Thereby get the concentration c L=(812098*2*10 of L-carnitine in the sample solution respectively
-7+ 0.0978) * 10*1000/60=43.37 μ g/ml, the concentration c D=0 μ g/ml of D-carnitine;
7), obtain L-carnitine content and purity in the sample:
Computing formula is with embodiment 1.
Result of calculation is following: the content of L-carnitine is 29.01mg/100g in the baby formula milk powder I sample, and the D-carnitine does not detect.
Step 1)～step 6) all is equal to embodiment 2.
Thereby get the concentration c L=(848302*2*10 of L-carnitine in the sample solution respectively
-7+ 0.0978) * 10*1000/60=44.57 μ g/ml, the concentration c D=0 μ g/ml of D-carnitine;
7), obtain L-carnitine content and purity in the sample:
Computing formula is with embodiment 1.
The result is following: the content of L-carnitine is 28.52mg/100g in the baby formula milk powder II sample, and the D-carnitine does not detect.
In order to prove the correctness of conclusion of the present invention, utilize existing high performance liquid chromatography uv analysis method to detecting with the ordinary powdered milk sample among embodiment 1～embodiment 3, baby formula milk powder I sample, baby formula milk powder II sample fully: the result is following:
In the ordinary powdered milk sample: the content of L-carnitine is 13mg/100g; In the baby formula milk powder I sample: the content of L-carnitine is 29.1mg/100g; In the baby formula milk powder II sample: the content of L carnitine is 28.6mg/100g.Testing result is consistent with embodiment 1～embodiment 3 results.
At last, it is also to be noted that what more than enumerate only is several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.
Claims (2)
1. the assay method of L-carnitine content and purity in the milk powder is characterized in that may further comprise the steps successively:
1), sample preparation:
Earlier with powdered milk sample 10～20g with deionized water dissolving after in 50 ℃ of water-bath 15min, ultrasonic Treatment 10min then is settled to 100ml with deionized water again;
Remove the processing of butter oil and lactoprotein more respectively:
The hydrochloric acid solution that adds 0.1mol/L is regulated pH to 1.8～2.2, uses filter paper filtering after leaving standstill 10min; The NaOH solution that in the filtrating of gained, adds 1mol/L is regulated pH to 5.8～6.2, leaves standstill to carry out second time with filter paper behind the 10min and filter, and the filtrating that filter gained the second time is sample solution;
2), preparation standard solution:
Take by weighing 50mg L-carnitine standard items,, and be settled to 10ml with deionized water dissolving; Gained solution obtains L-carnitine standard solution through 0.45 μ m membrane filtration;
Take by weighing 50mg D-carnitine standard items,, and be settled to 10ml with deionized water dissolving; Gained solution obtains D-carnitine standard solution through 0.45 μ m membrane filtration;
3), with step 2) the L-carnitine standard solution and the D-carnitine standard solution of gained operate respectively as follows:
L-carnitine standard solution/D-carnitine standard solution of getting 30 μ l is put in the volumetric flask, adds the 0.05M carbonic acid buffer of 80 μ l and the 1-of 80 μ l (9-fluorenyl)-ethyl chloroformate; Volumetric flask is put into 45 ℃ of water-baths place 1h, after taking out volumetric flask then and naturally cooling to room temperature, in volumetric flask, add deionized water and be settled to 10ml; Solution behind the constant volume is diluted with deionized water, obtain L-carnitine series derivatives solution/D-carnitine series derivatives solution that L-carnitine/D-carnitine concentration is respectively 0.5,1.0,1.5,2.0,2.5 μ g/ml;
The L-carnitine series derivatives solution and the D-carnitine series derivatives solution of gained are used 0.45 μ m filtering with microporous membrane respectively; Get the L-carnitine derivative solution of gradient concentration and the D-carnitine derivative solution of gradient concentration respectively;
4), sample solution derivative reaction:
The sample solution of getting 60 μ l step 1) gained adds the 0.05M carbonic acid buffer of 30 μ l and the 1-of 30 μ l (9-fluorenyl)-ethyl chloroformate again in volumetric flask; Volumetric flask is put into 45 ℃ of water-baths place 1h, after the taking-up volumetric flask naturally cools to room temperature from water-bath then, in volumetric flask, add deionized water and be settled to 10ml; Solution behind the constant volume with 0.45 μ m filtering with microporous membrane, is got sample introduction liquid;
5), high performance liquid chromatography detects:
The sample introduction liquid of the D-carnitine derivative solution of the L-carnitine derivative solution of the gradient concentration of step 3) gained and gradient concentration and step 4) gained is carried out high performance liquid chromatography respectively to be detected; Testing conditions is following:
Select the C18 chromatographic column for use, chromatographic condition is: moving phase is: the triethylamine phosphate buffer: the volume ratio of acetonitrile=73: 27; Flow velocity: 1ml/min; Detect wavelength: emission wavelength 269nm, detect wavelength 310nm; Sample size: 10 μ l;
Respectively the response peak area, the response peak area of sample introduction liquid of D-carnitine derivative solution of response peak area, gradient concentration of L-carnitine derivative solution of gradient concentration;
6), obtain the concentration c L of L-carnitine and the concentration c D of D-carnitine:
Response peak area with the L-carnitine derivative solution of the gradient concentration of step 5) gained is mapped to concentration, the typical curve of L-carnitine;
Response peak area with the D-carnitine derivative solution of the gradient concentration of step 5) gained is mapped to concentration, the typical curve of D-carnitine;
With the response peak area of the sample introduction liquid of step 5) gained respectively with the typical curve contrast of the typical curve and the D-carnitine of above-mentioned L-carnitine; The result of gained multiply by the extension rate 10*1000/60 of sample solution in the derivatization reaction of step 4), thereby gets the concentration c L of L-carnitine in the sample solution and the concentration c D of D-carnitine respectively;
7), obtain L-carnitine content and purity in the powdered milk sample:
In the formula: cL---the concentration of L-carnitine in the sample solution, μ g/ml;
The quality of m---milk powder, g;
In the formula: cL---the concentration of L-carnitine in the sample solution, μ g/ml;
CD---the concentration of D-carnitine in the sample solution, μ g/ml.
2. the assay method of L-carnitine content and purity in the milk powder according to claim 1 is characterized in that:
0.05M the preparation method of carbonic acid buffer is following: 338mg natrium carbonicum calcinatum and 152mg soda mint are dissolved in 100ml water;
The preparation method of triethylamine phosphate buffer solution is following: the triethylamine of 6.8ml is separated with 950ml is water-soluble, and using volume fraction then is that 84% phosphate aqueous solution is regulated pH to 2.5, and last water is settled to 1000ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010102745735A CN101975837B (en) | 2010-09-07 | 2010-09-07 | Method for determining content and purity of L-carnitine in milk powder |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010102745735A CN101975837B (en) | 2010-09-07 | 2010-09-07 | Method for determining content and purity of L-carnitine in milk powder |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101975837A CN101975837A (en) | 2011-02-16 |
| CN101975837B true CN101975837B (en) | 2012-02-01 |
Family
ID=43575739
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010102745735A Expired - Fee Related CN101975837B (en) | 2010-09-07 | 2010-09-07 | Method for determining content and purity of L-carnitine in milk powder |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101975837B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104914183A (en) * | 2015-06-03 | 2015-09-16 | 沈阳化工大学 | Levocarnitine optical purity analysis method |
| CN106153773B (en) * | 2016-08-22 | 2019-12-03 | 河北三元食品有限公司 | Utilize the method for l-carnitine in ultra performance liquid chromatography tandem mass spectrum quantitative determination baby formula milk powder |
| CN108287205A (en) * | 2018-03-02 | 2018-07-17 | 常州兰陵制药有限公司 | Method in relation to substance in Levocarnitine determined by HPLC and its injection |
| CN112379016A (en) * | 2020-11-02 | 2021-02-19 | 江苏宝众宝达药业有限公司 | Method for measuring content of L-carnitine in 3-O-lauroyl-L-carnitine by high performance liquid chromatography |
| CN113219097B (en) * | 2021-01-15 | 2022-06-24 | 杭州海关技术中心 | Method for splitting and measuring carnitine enantiomer in infant formula milk powder |
| CN112684089A (en) * | 2021-01-15 | 2021-04-20 | 杭州海关技术中心 | Method for separating and determining carnitine enantiomer in health food based on ultra-high performance synthetic phase chromatography technology |
| CN113804790B (en) * | 2021-09-18 | 2023-03-14 | 四川新希望畜牧科技有限公司 | Method for simultaneously detecting additive, L-carnitine and D-carnitine |
-
2010
- 2010-09-07 CN CN2010102745735A patent/CN101975837B/en not_active Expired - Fee Related
Non-Patent Citations (4)
| Title |
|---|
| 徐娟娟等.手性高效液相色谱法测定左旋肉碱中光学异构体的含量.《现代食品科技》.2010,第26卷(第3期),311-313. * |
| 徐连明等.HPLC法测定减肥胶囊中左旋肉碱的含量.《海峡药学》.2009,第21卷(第11期),46-47. * |
| 朱筱玲等.减肥胶囊中左旋肉碱的HPLC测定.《食品工业科技》.2002,82-84. * |
| 赵亚明等.高效液相色谱法检测血浆左旋肉碱方法的建立.《中国临床神经科学》.2006,第14卷(第5期),528-532. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101975837A (en) | 2011-02-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101975837B (en) | Method for determining content and purity of L-carnitine in milk powder | |
| CN103713056B (en) | Method for simultaneously analyzing and detecting residual veterinary drug compositions in animal tissue | |
| JP5872768B2 (en) | Glycan sample preparation method | |
| CN102095715B (en) | Surface enhanced Raman spectrum detection method for antioxidant BHT (Butylated Hydroxytoluene) | |
| CN104569187B (en) | The detection method of whitening and speckle dispelling composition and device in cosmetics | |
| CN102539568A (en) | Ion chromatography method for detecting iodate and bromate in cosmetic | |
| CN108693273A (en) | Aflatoxin B1, the detection method of B2, G1, G2 in a kind of Chinese medicine | |
| TW201930874A (en) | Analysis method | |
| RU2532237C1 (en) | Method for chromatographic analysis of parabens (4-hydroxybenzoic acid esters) in food products, cosmetics, pharmaceutical preparations and biologically active additives | |
| CN106770116B (en) | The method of Se form in Solid Phase Extraction-AFS DETERMINATION honey and fruit juice | |
| CN103604674A (en) | Method for extracting and purifying ochratoxin A from yeast for making hard liquor | |
| CN110501438A (en) | The detection method of picosulfate sodium in a kind of slim tea | |
| CN108267515B (en) | Method for detecting content of D-amygdalin and/or L-amygdalin in substance to be detected | |
| CN107831231A (en) | A kind of method for detecting vitamin A impurity in multivitamin preparation | |
| CN101782557A (en) | Measuring method of xylooligosaccharide (counted by xylose) content in health food | |
| CN103257200A (en) | Method of determining content of sodium cyclamate in food through high performance liquid chromatography with ultraviolet detector | |
| CN112114061A (en) | Method for detecting taurine in formula food with special medical application | |
| CN104133032B (en) | A kind of efficient liquid phase detection method of inulin | |
| CN108614053B (en) | Rapid detection of preservatives and sweeteners in butter or cream | |
| CN108414640B (en) | High performance liquid chromatography determination method of ethyl vanillin beta-D-glucoside | |
| CN102818782A (en) | Determination method of total 2-(2-phenethyl) chromone compound content | |
| CN104280474A (en) | High performance liquid chromatography fluorescence detection method for simultaneously measuring four prohibited and limited additives in tobacco flavors and perfumes | |
| CN104297376A (en) | Method for detecting fructo-oligosaccharide in milk | |
| Zhong et al. | Multi-channel purge and trap system coupled with ion chromatography for the determination of alkylamines in cosmetics | |
| CN103472153B (en) | Method for detecting rhodamine B in pepper raw material and products made from pepper raw material |
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 | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120201 Termination date: 20120907 |