GB 7300.209-2025 Feed additives—Part 2：Vitamins, provitamins and chemically well-defined substances having similar effect—Cyanocobalamin （Vitamin B12） English, Anglais, Englisch, Inglés, えいご
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ICS
CCS H
National Standard of the People's Republic of China
GB 7300.209-2025
Feed additives - Part 2: Vitamins, provitamins and chemically well-defined substances having similar effect - Cyanocobalamin （Vitamin B12）
饲料添加剂 第2部分：维生素及类维生素 氰钴胺（维生素B12）
Issue date: 2025-12-31 Implementation date: 2027-01-01
Issued by the General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China
the Standardization Administration of the People's Republic of China
Contents
Foreword
Introduction
1 Scope
2 Normative References
3 Terms and Definitions
4 Common Name, Molecular Formula, Relative Molecular Mass, and Chemical Structural Formula
5 Technical Requirements
6 Sampling
7 Test Methods
8 Inspection Rules
9 Labeling, Packaging, Transportation, Storage, and Shelf Life
Introduction
Feed additives refer to substances added in small or trace amounts during the processing, manufacturing, and use of feed, including nutritional feed additives and general feed additives. For ease of use, and according to product categories, GB 7300 "Feed Additives" is divided into the following 13 major categories:
------ Amino acids, amino acid salts, and analogues;
------ Vitamins and vitamin-like substances;
------ Mineral elements and their complexes (chelates);
------ Enzyme preparations;
------ Microorganisms;
------ Non-protein nitrogen;
------ Antioxidants;
------ Preservatives, mold inhibitors, and acidity regulators;
------ Coloring agents;
------ Flavoring and appetite-stimulating substances;
------ Binders, anti-caking agents, stabilizers, and emulsifiers;
------ Polysaccharides and oligosaccharides;
------ Others.
The product cyanocobalamin (Vitamin B₁₂) covered by this document belongs to the second major category: Vitamins and vitamin-like substances. As cyanocobalamin (Vitamin B₁₂) is the ninth product standard published under this major category, this document is assigned the number GB 7300.209 and serves as Part 209 of GB 7300.
Feed Additives — Part 2: Vitamins and Vitamin-Like Substances — Cyanocobalamin (Vitamin B₁₂)
1 Scope
This document provides the common name, molecular formula, relative molecular mass, and chemical structural formula of cyanocobalamin (Vitamin B₁₂). It specifies the technical requirements, inspection rules, labeling, packaging, transportation, storage, and shelf life of the feed additive cyanocobalamin (Vitamin B₁₂), and describes the sampling and test methods.
This document applies to the feed additive cyanocobalamin obtained by purification and crystallization from a fermentation broth containing cobalamins produced by microbial fermentation.
2 Normative References
The contents of the following documents constitute essential provisions of this document through normative reference. For dated references, only the edition corresponding to the date applies. For undated references, the latest edition (including any amendments) applies.
GB/T 603 Chemical Reagents — Preparations of reagents and solutions for use in test methods
GB/T 6435-2014 Determination of Moisture in Feeds
GB/T 6682 Water for Analytical Laboratory Use — Specification and Test Methods
GB/T 8170 Rules of Rounding Off for Numerical Values & Expression and Judgment of Limiting Values
GB 10648 Feed Labeling
GB/T 13079 Determination of Total Arsenic in Feeds
GB/T 13080 Determination of Lead in Feeds — Atomic Absorption Spectrometry
GB/T 13082 Determination of Cadmium in Feeds
GB/T 14699 Feeding Stuffs — Sampling
3 Terms and Definitions
No terms and definitions are stipulated in this document.
4 Common Name, Molecular Formula, Relative Molecular Mass, and Chemical Structural Formula
4.1 Common Name
Cyanocobalamin (Vitamin B₁₂).
4.2 Molecular Formula
C₆₃H₈₈CoN₁₄O₁₄P.
4.3 Relative Molecular Mass
1355.38 (calculated based on the 2024 international relative atomic masses).
4.4 Chemical Structural Formula
The chemical structural formula of cyanocobalamin (Vitamin B₁₂) is shown in Figure 1.
5 Technical Requirements
5.1 Appearance and Properties
Shall be dark red crystals or crystalline powder.
5.2 Identification
5.2.1 Ultraviolet-Visible Absorption Spectrum Identification
The sample solution shall have maximum absorption at wavelengths of 278 nm ± 1 nm, 361 nm ± 2 nm, and 550 nm ± 2 nm. The ratio of the absorbance at 361 nm to that at 278 nm shall be 1.70 to 1.90. The ratio of the absorbance at 361 nm to that at 550 nm shall be 3.15 to 3.40.
5.2.2 Infrared Spectrum Identification
The infrared spectrum of the sample shall be consistent with that of the cyanocobalamin (Vitamin B₁₂) reference standard.
5.3 Physicochemical Indicators
Shall comply with the requirements specified in Table 1.
5.4 Hygiene Indicators
Shall comply with the requirements specified in Table 2.
6 Sampling
Carry out according to GB/T 14699.
7 Test Methods
Warning: Some reagents used in the test methods are corrosive. Exercise caution during operation and take appropriate safety and protective measures.
7.1 General Provisions
Unless otherwise specified, all reagents used in this document shall be of analytical grade. Water for chromatographic analysis shall be Grade 1 water as specified in GB/T 6682; water for other uses shall be Grade 3 water as specified in GB/T 6682. All reagents or solutions shall be prepared according to the provisions of GB/T 603.
7.2 Appearance and Properties
Place an appropriate amount of the sample on a clean, dry white porcelain plate. Observe its color and form under natural light.
7.3 Identification Tests
7.3.1 Ultraviolet-Visible Absorption Spectrum Identification
7.3.1.1 Apparatus
7.3.1.1.1 Ultraviolet-visible spectrophotometer: equipped with 1 cm quartz cuvettes.
7.3.1.1.2 Analytical balance: accuracy of 0.01 mg.
7.3.1.2 Test Procedure
Accurately weigh an appropriate amount of the sample (accurate to 0.01 mg), dissolve in water and dilute quantitatively to prepare a solution containing approximately 25 μg/ml of cyanocobalamin. Measure using an ultraviolet-visible spectrophotometer. The absorption spectrum shall show maximum peaks at (278 ± 1) nm, (361 ± 2) nm, and (550 ± 2) nm. The ratio of the absorbance at 361 nm to that at 278 nm shall be 1.70 to 1.90. The ratio of the absorbance at 361 nm to that at 550 nm shall be 3.15 to 3.40.
7.3.2 Infrared Spectrum Identification
7.3.2.1 Reagents or Materials
Potassium bromide: spectroscopic grade.
7.3.2.2 Apparatus
7.3.2.2.1 Infrared spectrophotometer.
7.3.2.2.2 Analytical balance: accuracy of 0.01 mg.
7.3.2.3 Test Procedure
Operate protected from light. Weigh an appropriate amount of the sample, grind it uniformly with potassium bromide (7.3.2.1). The approximate ratio of sample to potassium bromide is 1:200. Prepare a pellet and record the infrared spectrum of the sample using an infrared spectrophotometer in the wavenumber range of 4000 cm⁻¹ to 400 cm⁻¹. The infrared spectrum of the sample shall be consistent with that of the cyanocobalamin (Vitamin B₁₂) reference standard (see Annex A).
7.4 Cyanocobalamin
7.4.1 Principle
Cyanocobalamin (Vitamin B₁₂) has maximum absorption at 361 nm, and the solution concentration and absorbance comply with the Lambert-Beer law. Accordingly, the absorbance of the sample solution can be measured using an ultraviolet-visible spectrophotometer, and the cyanocobalamin content in the sample is calculated based on the molar absorptivity coefficient.
