This standard specifies the method for the determination of melatonin in health foods packaged in capsules or tablets with melatonin as the active ingredient. This standard is applicable to the determination of melatonin in health foods packaged in capsules or tablets with melatonin as the active ingredient. The detection limit of the first method of this standard, high performance liquid chromatography-ultraviolet detection method, is 0.5ng; when the sampling volume is 0.5, the detection concentration is 0.07mg/kg. The detection limit of the second method, high performance liquid chromatography-fluorescence method, is 30pg, and the linear range is 0.05ng~0.50ng. GB/T 5009.170-2003 Determination of melatonin content in health foods GB/T5009.170-2003 Standard download decompression password: www.bzxz.net

ECS67.040
National Standard of the People's Republic of China
GB/T5009.170-—2003
Determination of melatonin in health foods foods2003-08-11 Issued
Ministry of Health of the People's Republic of China
Standardization Administration of China
2004-01-01 Implementation
This standard was proposed and managed by the Ministry of Health of the People's Republic of China, and the first unit of the standard is the Food and Drug Administration of the Ministry of Health. The second unit of this standard is the Institute of Nutrition and Food Hygiene of the Chinese Academy of Preventive Medicine, the Food Hygiene Inspection Institute of the Ministry of Health, and the Beijing Municipal Health and Epidemic Prevention Station. The first drafters of this standard are Yang Dajin, Wang Zhutian, Fang Congrong, Wu Guohua, Li Jianwu, Ji Ping, Wu Chengning, and Wu Guohua. 4us
GB/T5009.170—2003
Latnin (M=latnnin) is a kind of indole hormonal substance in vivo, its chemical structure is N-ethyl-4-methoxytryptamine, the nucleus has a wide range of physiological effects. Such products have been widely used in the world, and in recent years, my country has also begun to mass produce and sell such products. In order to improve the quality of melanin products and protect the health of consumers, the determination method of melanin in chain food is established.
1 Dong Mo
Determination of melanin content in health food
GE/T5009.170—2003
This standard specifies the determination method of melanin in health food with melanin as the effective ingredient in preservative packaging. This standard is applicable to the determination of melanin in health food with super fruit capsule as the effective ingredient in plastic or tablet packaging. The detection limit of this standard is 0.5ng when the dosage is 9.5, and the total concentration is 1.n7mg/kg. The second method is high-efficiency immunochromatographic fluorescence method. The particle limit is 30 μg. The linear range is 0.ng~0.50. The first method is high-efficiency color - external detection method
2 The black potential in the fiber is filtered and then detected by high-performance liquid chromatography with ultraviolet detector. The retention time of the chromatogram is determined and the external quantification is performed.
3 Trial production
Except for instructions, the reagents and distilled water or deionized water or methanol with linearity should be determined in the analytical instrument. 3.1: chromatographic purity.
3.2 Water: high quality
3.3 Three kinds of acetic acid, high purity,
3.4 ​​High performance liquid chromatography: single + water + three bottles of acetic acid 145 + 5 + 0.5C5.3.5 Cabinet of meletonin standard product, 3.6 Fu without plate push to prepare
Hold the weighing plate 30 to prepare the standard product 105. Add 0.0% alcohol and dilute to the system. Accurately draw 2ml of the above liquid into a 10L container, add flowing water (8.4) and dilute to the standard concentration, the sieve concentration is 0.00mg/mI. 4 Collection equipment
4 High performance phase chromatograph: with UV detector. 4.2 Ultrasonic sedimentation apparatus,
4.3 Centrifugation.
5 Analysis steps
5. Sample treatment: Grind the tablets into powder using a grinder to make them uniform. 5.2 Accurately weigh about 1 tablet or 100g of mass in a 1.5L volumetric flask. Add 703% ethanol to the mark. Use an ultrasonic precipitator to extract for 0 min. Roll the extract to a high temperature until it drops to a certain level. Accurately add 30ml of water to a 30L volumetric flask, add the mixture to the mark with a constant volume, filter and separate by 4-way separation. 5.3 DeterminationwwW.bzxz.Net
5.3.! 5.3.1.1 Chromatographic column i[indaFakC, 4.6am×250tim.5.3.1.2 UV detection, detection line length 22m, 5.3.1.3 Speed ​​0.a nLmn
5.3.14, hanging bottle: room temperature.
5.3.2 Chromatographic analysis
CR/T5009.170—2003
Inject the standard solution and sample purification solution into the commercial liquid chromatography, qualitatively by retention time, quantitatively by peak selection or peak area compared with the standard,
5. 3. 3 Chromatogram See Figure 1, Calculation of the chromatogram results: Calculate according to the formula: The above xrx10x number, x: The content of the flammable element in the sample is in mg/kg, the peak quotient or peak area of ​​the sample is in mg/L (mL), and the peak quotient or peak area of ​​the standard is in g/L (mL). The sample quality is in g (mL). The results are calculated by retaining two validation coefficients. In order to ensure precision, the difference between the results of two independent determinations under repeatability conditions shall not exceed 5% of the arithmetic mean. High performance liquid chromatography fluorescence method
The second method
F source avoidance
The irradiated substance in the sample is repeatedly taken from the lungs, prepared into a kind of acetylene, diluted in a certain ratio, injected into the corresponding high-temperature chamber, separated by zone chromatography, and detected by fluorescence detection. The qualitative analysis is carried out according to the retention time and the peak area comparison with the standard product. 9 Reagents
9.1 Acetone, monohydrate,
9.2 Standard: The maximum purity is 99.7% 92.1 Standard: Accurately weigh the required standard product (0.10-1% alcohol and ether) and store it at -20%.
9.2.2 Use of liquid: Accurately measure the quantitative standard before use, and use the standard product with a kind of acetylene as required, and use the standard product with a kind of acetylene.
1 Instruments required
13 High-speed liquid chromatography equipment: with fluorescence rectifier and microprocessor. +8
0.2 High-speed centrifuge.
10.3 Superwave cleaning machine.
11 Sample preparation
Mix the sample into a granule and set aside; the contents of the capsule are mixed and set aside. 12. Separation
12.1 Extraction
GB/T5009.170—2Q03
Requirements Weigh 100 ml of sample: add about 3 ml of methanol to the sample in a mol volume. Ultrasonicate for 13 minutes. Centrifuge and take the supernatant 10 ml. Add about 3 ml of methanol to a volume of 1 ml. Repeat the above method twice, add the supernatant, and add the mixture until the sample is stable. Take an appropriate amount of this solution, dilute it with water and make it to volume to prepare the sample solution. 12.2 Determination
12.2.1 High-efficiency oil-phase chromatography conditions
12.2.1.1 Chromatographic medium: A:]rimaC-4.5mruX250rmfn non-ferrous metal 12.2.1.2 Mobile phase: ALC-4.5mruX250rmfn non-ferrous metal 12.2.1.3 Flow rate: 1 ml./nis.
12.2, 1, 4 Flow rate: 10 L-
12.2.1.5 Detector: fluorescence detection Excitation wavelength: 286nm Emission wavelength: 352nm 12.2.2 Chromatographic analysis
After adjusting the instrument to the maximum state, add 10% standard liquid and the purified sample to the solution, and add the sample height or area to the standard solution for a certain period of time. The standard surface of this method is 0.5 ng~.50 ng. 12.2.3 Chromatographic diagram
The color section is shown in Figure 2.
13 Calculation of results
Calculate using formula (2):
Where:
X-MXVIUO
mX V,x1 000
The melanin content in the sample, in milligrams per gram (mg/kg); The melanin content in the prepared solution, in nanograms (g); The mass of the sample, in grams
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