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Determination of L-carnitine in infant foods and dairy products
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GB 29989-2013: Determination of L-carnitine in infant foods and dairy products
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GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National Food Safety Standard -
Determination of Levocarnitine in Foods and
Milk Products for Infants and Young Children
Issued on. NOVEMBER 29, 2013
Implemented on. JUNE 1, 2014
Issued by. National Health and Family Planning Commission of the
People's Republic of China
Table of Contents
1 Scope... 3
2 Principle... 3
3 Reagents and Materials... 3
4 Apparatuses and devices... 5
5 Analytical Procedures... 5
6 Expression of analytical result... 6
7 Precision... 6
8 Other... 6
National Food Safety Standard -
Determination of Levocarnitine in Foods and Milk
Products for Infants and Young Children
1 Scope
This Standard specifies the method for determination of levocarnitine in foods and
milk products for infants and young children.
This Standard is applicable to the determination of levocarnitine in foods and milk
products for infants and young children.
2 Principle
Extract the specimen with water; use perchloric acid to precipitate the protein; then
filtrate it. After the filtrate is gone through alkaline saponification, the combined-state
levocarnitine in solution is dissociated. Under the catalysis of carnitine acetyl
transferase, levocarnitine reacts with acetyl coenzyme A to generate acetylcarnitine
and free coenzyme A. Free coenzyme A reacts with 2-nitrobenzoic acid to generate
yellow substance; the color of which is proportional to the content of free coenzyme A.
As the free coenzyme A and levocarnitine are of same mole reaction, the content of
levocarnitine in specimen may be calculated indirectly.
3 Reagents and Materials
Note. Unless otherwise specified, all reagents adopted in this method are analytical reagents
and the water is Grade 3 specified in GB/T 6682.
3.1 Reagents
3.1.1 Perchloric acid (HClO4).
3.1.6 Ethylenediamine tetraacetic acid disodium (C10H14N2Na2·2H2O).
3.1.7 Acetyl coenzyme A (AcetylCoA). kept under 2~8°C.
3.1.8 Carnitine acetyl transferase (CAT). kept under 2~8°C.
3.2 Reagent preparation
3.2.1 Perchloric acid solution (13%). dilute 13mL of perchloric acid to 100mL.
3.2.2 Sodium hydroxide solution (10mol/L). dissolve 40g of sodium hydroxide with
water; dilute to 100mL after cooling down.
3.2.3 Potassium hydroxide solution (4.0mol/L). dissolve 22.4g with water; dilute to
100mL after cooling down.
3.2.6 Acetyl coenzyme A (AcetylCoA) solution. dissolve 20.0mg of acetyl coenzyme
A into 2.0mL water. Prepare it just before use.
3.2.7 Carnitine acetyl transferase (CAT) solution. pipet 100μL of carnitine acetyl
transferase suspension; centrifuge it for 10min with rotation speed of 1500r/min;
discard the supernatant liquid and dissolve the sediment into 2mL of water. Prepare it
just before use.
3.3 Standard product
Standard levocarnitine product (C7H15NO3). purity ≥98%.
3.4 Preparation of standard solution
4 Apparatuses and devices
4.1 Analytical balance. with a sensitivity of 0.1mg.
4.2 pH meter. with a precision of 0.01.
4.5 Spectrophotometer.
5 Analytical Procedures
5.1 Specimen treatment
Accurately weigh 5g (accurate to 0.0001g) of well-mixed specimen; place it into a
beaker; dissolve it with 30mL of 40°C warm water; transfer it into 100mL volumetric
flask. Add 10mL of 13% perchloric acid solution into it; keep static for 20min after
mixed uniformly. Fix the volume to the scale with distilled water; filter it with
quantitative filter paper.
5.2 Drawing of standard curve
Pipet 2.0mL of standard levocarnitine working solution into a 1cm cuvette; add 0.8mL
of color rendering working solution and 100μL of acetyl coenzyme A solution into it;
then cover it; after mixing it uniformly, put it in spectrophotometer;
5.3 Specimen determination
Take 2.0mL of specimen treatment solution; determine its absorbance according to
the procedures specified in Section 5.2.Obtain the concentration of specimen
solution to be tested through standard curve.
6 Expression of analytical result
The content X of levocarnitine in specimen is expressed by mass fraction mg/100g
and calculated according to Formula (1).
7 Precision
The absolute deviation of two independent determination results under repeatability
conditions shall not exceed 10% of arithmetic mean.
8 Other
Method detection limit is 0.6mg/100g; quantitation limit is 2mg/100g.
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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