GB/T 5009.185-2003 Determination of patulin in apple and hawthorn products

This standard specifies the determination method of patulin in apple and hawthorn products. This standard is applicable to the determination of patulin in apple and hawthorn products. The detection limit of this standard is: 3μg/L. GB/T 5009.185-2003 Determination of patulin in apple and hawthorn products GB/T5009.185-2003 Standard download decompression password: www.bzxz.net

ICS67.040
National Standard of the People's Republic of China
GB/T5009.185--2003
Partially replaces GB14974-1994
Determination of patulin in apple and hawthorn products2003-08-11Promulgated
Ministry of Health of the People's Republic of China
Standardization Administration of the People's Republic of China
2004-01-01Implementation
GB/T5009.185—2003
This standard replaces the test method in Chapter 3 of GB14974—1994 "Hygienic Standard for Limit of Patulin in Apple and Hawthorn Products". This standard is proposed and managed by the Ministry of Health of the People's Republic of China. This standard is drafted by the Institute of Nutrition and Food Hygiene, Chinese Academy of Preventive Medicine. The main drafters of this standard are Wu Nan, Liu Yong and Liu Xingjie. 484
1 Scope
Determination of patulin in apple and hawthorn products This standard specifies the determination method of patulin in apple and hawthorn products. This standard is applicable to the determination of patulin in apple and hawthorn products. The detection limit of this standard is: 3 μg/L.
2 Principle
GB/T5009.185-—2003bzxz.net
After the patulin in the sample is extracted, purified, concentrated and developed by thin layer, it is quantified by ultraviolet reflected light scanning using a thin layer scanner. 3 Reagents
3.1 Silica gel GF254.
3.2 Thin layer chromatography developing solvent Horizontal: chloroform-acetone (30+1.5), vertical: toluene-ethyl acetate-formic acid (50+15+1). 3.3 Patulin standard.
3.4 ​​Ethyl acetate.
3.5 1.5% sodium carbonate solution.
3.6 Anhydrous sodium sulfate.
3.7 Chloroform.
3.8 Display agent: Dissolve 0.1g MBTH·HCl·H.O3-methyl-2-benzothiazolone hydrochloride in 20mL distilled water, store in a refrigerator, and re-prepare every 3 days. 4 Instruments
Thin layer scanner.
4.2 Chromatographic tank (standard tank with an inner diameter of 11.5cm and a height of 20cm). 4.3 Glass plate 10cm×10cm.
4.4 Ultraviolet light.
5 Operation method
5.1 Extraction
5.1.1 Fruit juice, fruit wine: Measure 25mL of fruit juice, place it in a separatory funnel, add an equal volume of ethyl acetate, shake for 2 minutes, let stand to separate, repeat the above steps twice, combine the organic phases, add 2.5mL of 1.5% sodium carbonate, shake for 1 minute, let stand to separate, discard the sodium carbonate layer, and treat with sodium carbonate again in the same steps as above. Filter the extract into a 100mL pear-shaped bottle, concentrate to nearly 40% by vacuum decompression in a 40℃ water bath, wash the bottle wall with a little trinitroform, concentrate to dryness, add 0.4mL of trinitroform to make up the volume, and use it for thin layer chromatography determination. 5.1.2 Jam: Weigh 25 g of the sample and place it in a mortar. Add an appropriate amount of anhydrous sodium sulfate and grind it. Then weigh it into a conical flask and add 80 mL of ethyl acetate to soak for 30 min. Oscillate for 30 min, filter, and take 50 mL of the filtrate. The following operations are the same as 5.1.1. 5.2 Determination
5.2.1 Preparation of thin layer plate: Take 55 g of silica gel GF2, add 15 mL of water, and coat it on a 10 cm × 10 cm glass plate. Coat 5 pieces at a time with a thin layer thickness of 0.3 mm. After drying in the shade, bake at 105°C for 2 h and put it in a desiccator for later use. 5.2.2 Sample spotting: Take a thin plate, and drip 10μL of 1.0μg/mL paucidine standard solution with a microsyringe at 10cm from the bottom and right, and drip 10μL of sample solution at 4cm from the left. On the same vertical line of the sample point, 2cm from the top, spot 20ng of standard 485
GB/T5009.185-2003
solution as the position reference point.
5.2.3 Development: After horizontal development to the top, take out the volatiles, and carry out longitudinal development. After reaching the top, take out the volatiles and observe under 254nm ultraviolet light. If a bright absorption point appears, the sample is positive. Scanning quantitative determination is carried out. 5.2.4 Thin layer chromatography scanning determination: Instrument operating conditions: measurement wavelength 270nm reference wavelength 310nm; reflected light determination; scanning speed 40nm/min; recorder paper speed 20nm/min, determination of paucidine peak area in standard and sample. 5.2.5 Confirmation of positive samples: Spray the TLC plate of the positive sample with MBTH color developer, bake at 130℃ for 15min, cool to room temperature, and observe under 365nm ultraviolet light. The chloranthene should appear as orange-yellow spots. 5.2.6 Calculation: The calculation of the chloranthene content in juice and fruit is shown in formula (1) and formula (2). Juice X=c×
Jam: X=CX
Where:
×V×Dx
Content of chloroquine, in micrograms per milliliter (μg/mL); Concentration of chloroquine standard solution, in micrograms per milliliter (μg/mL); Peak area of ​​chloroquine in sample solution;
Peak area of ​​chloroquine in standard solution:
Volume of chloroform added to make up to volume, in milliliters (mL); D dilution multiple of sample solution;
6Precision
Mass of the sample, in grams (g);
Volume of the liquid sample, in liters (mL). 8
The absolute difference between two independent measurement results obtained under repeatability conditions shall not exceed 10% of the mean value. 486
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