Codeofchina.com is in charge of this English translation. In case of any doubt about the English translation, the Chinese original shall be considered authoritative.
This standard is the revision of GB/T 14702-1993 Method for Determination of Vitamin B6 in Feed, it is more efficient, convenient and accurate with respect to the previous edition. This standard is developed by reference to the “Liquid Chromatographic Method for the Analysis of Ascorbic Acid, Niacinamide, Pyridoxol, Thiamine and Riboflavin in Compound Premix”, published by the Association of Analytical Communities, No. 67.
The technical deviations between this standard and the GB/T 14702-1993 are as follows: this standard specifies the determination of vitamin B6 in feed by high-performance liquid chromatography, and specifies the extraction conditions of specimen, chromatographic conditions and repeatability requirements; and specifies the scope of application is for the determination of vitamin B6 in compound premix and vitamin premix with vitamin B6 content greater than 30 mg/kg.
This standard was proposed by and is under the jurisdiction of the National Technical Committee 76 on Feed Industry of Standardization Administration of China.
The previous editions of this standard are as follows: GB/T 14702-1993
Determination of Vitamin B6 in Feeds — High-performance of Liquid Chromatography
1 Scope
This standard specifies the method for determination of vitamin B6 content in feeds by high-performance liquid chromatography.
This standard is applicable to the determination of vitamin B6 in compound premix and vitamin premix with vitamin B6 content greater than 30 mg/kg.
2 Normative References
The following normative documents contain provisions which, through reference in this text, constitute provisions of this standard. For dated references, subsequent amendments (excluding corrections), or revisions, of any of these publications do not apply to this part. However parties to agreements based on this standard are encouraged to investigate the possibility of applying the most recent editions of the normative documents indicated below. For undated references, the latest edition of the normative document referred to applies.
GB/T 6682 Water for Laboratory Use — Specifications and Test Methods
GB/T 14699.1 Animal Feeding Stuffs — Sampling
3 Principle
After ultrasonic extraction of the acid extract of vitamin B6 specimen, the test solution is injected into a high-performance liquid chromatography (HPLC) system for separation. Ultraviolet (or diode matrix detector) is used for detection. The content of vitamin B6 can be calculated by external standard method.
4 Reagent and solution
Unless otherwise specified, the reagents used shall be of analytical grade, the water used shall be distilled water, and the deionized water for chromatographic use shall meet the requirements of Grade 1 water specified in GB/T 6682.1.
4.1 Ethylenediamine tetraacetic acid disodium salt (EDTA).
4.2 Sodium 1-heptane sulfonate (PICB7), guaranteed reagent.
4.3 Glacial acetic acid, guaranteed reagent.
4.4 Triethylamine, chromatographically pure.
4.5 Methanol, chromatographically pure.
4.6 Hydrochloric acid solution: c(HCl) = 0.1 mol/L.
4.7 Extract: add 50 mg of EDTA (4.1) to a 1 000 mL volumetric bottle filled with about 700 mL/L deionized water, and then add 25 mL of glacial acetic acid (4.3) and 5 mL of triethylamine (4.4); dilute to the scale with deionized water and shake well. Take 860 mL of the solution, and mix with 140 mL of methanol.
4.8 Mobile phase: Weigh 50 mg (to the nearest 0.001 g) of EDTA (4.1) and 1.1 g (to the nearest 0.001 g) of sodium 1-heptane sulfonate (4.2) in a 1 000 mL volumetric bottle filled with about 700 mL deionized water, add 25 mL of glacial acetic acid (4.3) and 5 mL of triethylamine (4.4); after they are all dissolved, dilute to the scale with deionized water and shake well. Adjust the solution to pH 3.40 ± 0.02 with glacial acetic acid and triethylamine, and filter the solution with 0.45 μm filter membrane; Take 860 mL of the solution and mix with 140 mL of methanol (4.5); degassing by ultrasonic, and set aside.
4.9 Vitamin B6 standard solution:
a) Vitamin B6 standard stock solution: Accurately weigh 0.050 0 g of vitamin B6 in a 100 mL brown volumetric bottle, add hydrochloric acid solution (4.6) about two-thirds of the volume, and conduct ultrasonic treatment for 15 min. After the solution is completely dissolved, dilute to the scale with hydrochloric acid solution (4.6). The concentration of this solution is 500 μg/mL, it can be stored in a refrigerator at 4 ℃ and keep out of light for 3 months.
b) Vitamin B6 standard working solution A: accurately suck 2 mL of vitamin B6 standard stock solution [4.9 a)] and transfer it into a 50 mL brown volumetric flask, and dilute to the scale with mobile phase (4.8). The concentration of this standard working solution is 20 μg/mL
c) Vitamin B6 standard working solution B: accurately suck 5 mL of vitamin B6standard working solution A [4.9 b)] and transfer it into a 50 mL brown volumetric flask, and dilute to the scale with mobile phase (4.8). The concentration of this standard working solution is 2.0 μg/mL
Foreword II
1 Scope
2 Normative References
3 Principle
4 Reagent and solution
5 Apparatus
6 Preparation of Specimen
7 Analytical Procedure
8 Calculation of Results
9 Repeatability
Codeofchina.com is in charge of this English translation. In case of any doubt about the English translation, the Chinese original shall be considered authoritative.
This standard is the revision of GB/T 14702-1993 Method for Determination of Vitamin B6 in Feed, it is more efficient, convenient and accurate with respect to the previous edition. This standard is developed by reference to the “Liquid Chromatographic Method for the Analysis of Ascorbic Acid, Niacinamide, Pyridoxol, Thiamine and Riboflavin in Compound Premix”, published by the Association of Analytical Communities, No. 67.
The technical deviations between this standard and the GB/T 14702-1993 are as follows: this standard specifies the determination of vitamin B6 in feed by high-performance liquid chromatography, and specifies the extraction conditions of specimen, chromatographic conditions and repeatability requirements; and specifies the scope of application is for the determination of vitamin B6 in compound premix and vitamin premix with vitamin B6 content greater than 30 mg/kg.
This standard was proposed by and is under the jurisdiction of the National Technical Committee 76 on Feed Industry of Standardization Administration of China.
The previous editions of this standard are as follows: GB/T 14702-1993
Determination of Vitamin B6 in Feeds — High-performance of Liquid Chromatography
1 Scope
This standard specifies the method for determination of vitamin B6 content in feeds by high-performance liquid chromatography.
This standard is applicable to the determination of vitamin B6 in compound premix and vitamin premix with vitamin B6 content greater than 30 mg/kg.
2 Normative References
The following normative documents contain provisions which, through reference in this text, constitute provisions of this standard. For dated references, subsequent amendments (excluding corrections), or revisions, of any of these publications do not apply to this part. However parties to agreements based on this standard are encouraged to investigate the possibility of applying the most recent editions of the normative documents indicated below. For undated references, the latest edition of the normative document referred to applies.
GB/T 6682 Water for Laboratory Use — Specifications and Test Methods
GB/T 14699.1 Animal Feeding Stuffs — Sampling
3 Principle
After ultrasonic extraction of the acid extract of vitamin B6 specimen, the test solution is injected into a high-performance liquid chromatography (HPLC) system for separation. Ultraviolet (or diode matrix detector) is used for detection. The content of vitamin B6 can be calculated by external standard method.
4 Reagent and solution
Unless otherwise specified, the reagents used shall be of analytical grade, the water used shall be distilled water, and the deionized water for chromatographic use shall meet the requirements of Grade 1 water specified in GB/T 6682.1.
4.1 Ethylenediamine tetraacetic acid disodium salt (EDTA).
4.2 Sodium 1-heptane sulfonate (PICB7), guaranteed reagent.
4.3 Glacial acetic acid, guaranteed reagent.
4.4 Triethylamine, chromatographically pure.
4.5 Methanol, chromatographically pure.
4.6 Hydrochloric acid solution: c(HCl) = 0.1 mol/L.
4.7 Extract: add 50 mg of EDTA (4.1) to a 1 000 mL volumetric bottle filled with about 700 mL/L deionized water, and then add 25 mL of glacial acetic acid (4.3) and 5 mL of triethylamine (4.4); dilute to the scale with deionized water and shake well. Take 860 mL of the solution, and mix with 140 mL of methanol.
4.8 Mobile phase: Weigh 50 mg (to the nearest 0.001 g) of EDTA (4.1) and 1.1 g (to the nearest 0.001 g) of sodium 1-heptane sulfonate (4.2) in a 1 000 mL volumetric bottle filled with about 700 mL deionized water, add 25 mL of glacial acetic acid (4.3) and 5 mL of triethylamine (4.4); after they are all dissolved, dilute to the scale with deionized water and shake well. Adjust the solution to pH 3.40 ± 0.02 with glacial acetic acid and triethylamine, and filter the solution with 0.45 μm filter membrane; Take 860 mL of the solution and mix with 140 mL of methanol (4.5); degassing by ultrasonic, and set aside.
4.9 Vitamin B6 standard solution:
a) Vitamin B6 standard stock solution: Accurately weigh 0.050 0 g of vitamin B6 in a 100 mL brown volumetric bottle, add hydrochloric acid solution (4.6) about two-thirds of the volume, and conduct ultrasonic treatment for 15 min. After the solution is completely dissolved, dilute to the scale with hydrochloric acid solution (4.6). The concentration of this solution is 500 μg/mL, it can be stored in a refrigerator at 4 ℃ and keep out of light for 3 months.
b) Vitamin B6 standard working solution A: accurately suck 2 mL of vitamin B6 standard stock solution [4.9 a)] and transfer it into a 50 mL brown volumetric flask, and dilute to the scale with mobile phase (4.8). The concentration of this standard working solution is 20 μg/mL
c) Vitamin B6 standard working solution B: accurately suck 5 mL of vitamin B6standard working solution A [4.9 b)] and transfer it into a 50 mL brown volumetric flask, and dilute to the scale with mobile phase (4.8). The concentration of this standard working solution is 2.0 μg/mL
Contents of GB/T 14702-2002
Foreword II
1 Scope
2 Normative References
3 Principle
4 Reagent and solution
5 Apparatus
6 Preparation of Specimen
7 Analytical Procedure
8 Calculation of Results
9 Repeatability
