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Objective To establish a method for the determination of biotin in formula food for special medical purposes by high performance liquid chromatography-post column derivatization. Methods The biotin in the sample was dissolved in water and extracted by enzymatic hydrolysis with amylase and papain at 60 ℃ for 1 hour in a water bath. Using Zorbax SB-AQ chromatographic column separation and entering the post column reaction device, the biotin was derived from fluorescein isothiocyanate labeled avidin. Derivatives were detected using a fluorescence detector with an excitation wavelength of 495 nm and an emission wavelength of 525 nm. The results were quantified by the external standard method. Results Under the optimized conditions, the biotin showed good (r2>0.999) linear relationships within the concentration range of 5.00‒75.00 ng/mL. The average recoveries were 97.5%‒100.1%, and the relative standard deviations were 0.86%‒2.9%. The limit of detection was 12 μg/kg, and the limit of quantification was 41 μg/kg. The relative standard deviations of the biotin standard solution and the sample solution to be tested within 24 h were 0.68% and 1.08%, respectively. There was no significant difference between the results of this method and GB 5009.256—2016 National food safety standard-Determination of biotin in food. Conclusion This method has simple pretreatment, high recovery, good sensitivity and precision, and can be used for the determination of biotin content in formula food for special medical purposes.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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