Microsoft Word 蒋孟虹-二校897

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1 7 3 Vol. 7 No Journal of Food Safety and Quality Mar., 2016 B 12 * 蒋孟虹, 许苹, 秦峰, 刘浩, 杨美成 (, ) 摘要 : 目的 B 12 方法, B 12, (36±1) 19~20 h, 结果 B ~0.010 ng/ml, B μg/100 g 结论 B 12 关键词 : B 12 ; ; Determination of trace vitamin B 12 in enteral nutrition powder by microbiological method JIANG Meng-Hong, XU Ping, QIN Feng, LIU Hao, YANG Mei-Cheng * (Shanghai Institute for Food and Drug Control, Shanghai , China) ABSTRACT: Objective To establish a method for the determination of trace vitamin B 12 in enteral nutrition powder by microbiological method. Methods The test organism was Lactobacillus leichmamii. The medium without vitamin B 12 was used as assay medium. The sample were incubated for 19~20 h at the temperature of (36±1) and measured by microbial turbidimetric method. Results Vitamin B 12 had a good linear relationship in the range of 0.001~0.010 ng/ml, and the content of vitamin B 12 in enteral nutrition powder was 2.9 μg/100 g. Conclusion This method is accurate and reliable, and suitable for the assay of trace vitamin B 12 in enteral nutrition powder. KEY WORDS: vitamin B 12 ; enteral nutrition powder; microbiological method 1 引言 B 12, B,,, B 12, DNA, [1], B 12, B 12,,, [2-4], B 12, B 12 [5-11] [12] [13] * 通讯作者 :,,, yangmeicheng@vip.sina.com *Corresponding author: YANG Mei-Cheng, Ph.D., Chief Pharmacist, Shanghai Institute for Food and Drug Control, Shanghai , China. yangmeicheng@vip.sina.com

2 894 7 [14] [15-16],, B 12,, B 12, [16],, B 12 (Lactobacillus leichmannii) B 12, B 12,,, B 12,,, B 12 2 材料与方法 2.1 材料与试剂 (CP225D, Sartorius); ph (Seven easy, Mettler Toledo); (MS 3, IKA); (HVA-85, Hirayama); (KB240, Binder); (UV2450, ) (ATCC 7830, American Type Culture Collection); B 12 ( 96.2%, : 203B, ); ( : ); ( BD ); ( BD ); B 12 ( ); (, ) 2.2 检测方法测试菌液的制备,, (36±1) 24 h, 2~3, (36±1) 18~24 h, 2000 r/min 2~3 min,, 10 ml,, 2 10 ml, 10 ml,, 550 nm,,, 60%~80% 标准曲线的制作 (1) B 12 (10 μg/ml): 20 mg B 12, ( 25%) B μg/ml B 12 (100 ng/ml): 5.0 ml B ml B 12 (1 ng/ml): 5.0 ml B ml : B 12, 0.02 ng/ml, 0.01 ng/ml (2) 1 4, B 12, 表 1 标准曲线的制备 Table 1 Preparation of standard curve 0.01 ng/ml (ml) 0.02 ng/ml (ml) (ml) (ml) S S S S S S S S S S

3 3, : B 试样的处理 1.3 g, 1.0 g, ( 1 )1.2 g, 100 ml ( ), 10 ml, 150 ml,, min,, B 12, ph 4.5±0.2, 250 ml, 5 ml 100 ml, 20~30 ml, ph 6.8±0.2, B 12 ph, 待测溶液的制备 2 4, B 12, 表 2 待测溶液的制备 Table 2 Preparation of samples (ml) (ml) (ml) 灭菌接种及培养 min, μl ( S1 ),,, (36±1) 19~20 h 测定, S1,,,, B 12,, B 12, B 12 3 结果与分析 3.1 测试菌液的制备测试菌液透光率的确定,,,, 60%~80%, 测试菌液加入量的考察,, μl 4,,, 50 μl, 3.2 标准曲线的制作 10 ml, 5 ml,, 10 ml, ng/ml,,,, B 12, B 12, B 12,,,,,, ng/ml 3.3 待测液的制备 10 ml, 5 ml, ml, 10 ml, B ~0.02 ng/ml ( g), 2 g,

4 896 7 ( ), 3.4 培养及测定时间的考察, (36±1) 19~20 h,,, 18 h, 1 h 20 h,,,, 30%,, 3.5 标准曲线的绘制 B 12, B 12, ( 1) B ~0.010 ng/ml, Y= X , r= 结论 B 12,,, B 12, 参考文献 [1],. B 12 [J]., 2012, 24(4): Lv YJ, Huang JM. Progress on the studies of vitamin B 12 [J]. Chin Food Hyg, 2012, 24(4): [2] Suarez L, Hendricks K, Felkner M, et al. Maternal serum B 12 levels and risk for neural tube defects in a Texas-Mexico border population [J].Ann Epidemiol, 2003, 13(2): [3] Molloy AM, Kirke PN, Troendle JF, et al. Maternal vitamin B 12 status and risk of neural tube defects in a population with high neural tube defect prevalence and no folic acid fortification [J]. Pediatrics, 2009, 123(3): [4],,. B 12 [J]., 2003, 15(3): Lu J, Yang DJ, Wang ZT. Review of chemical methods for determining cyanocobalamin [J]. Chin Food Hyg, 2003, 15(3): [5] Guggisberg D, Risse MC, Hadorn R. Determination of vitamin B 12 in meat products by RP-HPLC after enrichment and purification on an immunoaffinity column [J]. Meat Sci, 2012, 90(2): [6],,. HPLC B 12 [J]., 2004, (2): Hao YP, Jiang JD, Fang YG. Research on the determination 1 B 12 Fig. 1 Standard curve of vitamin B 样品测定,, B 12, ±15%,, B 12, B μg/100 g method of vitamin B 12 in composite vitamin additives for milk powder and formula by HPLC [J]. China Food Addit, 2004, (2): [7],,,. - B 12 [J]. ( ), 2013, 39(2): Chu XJ, Tao BH, Lai SY, et al. Quantification of vitamin B 12 in infant formula by dimensional liquid chromatography with solid phase extraction [J]. J Zhejiang Univ (Agric Life Sci), 2013, 39(2): [8],,,. / B 12 [J]., 2013, 32(3):

3, : B 12 897 389 392. Wu N, Jiang JD, Tao DL, et al. Determination of vitamin B 12 content in milk powder by immunoaffinity column purification -HPLC method [J].

Simultaneous determination of eight water-soluble vitamins in baby milk powder by high performance liquid chromatography [J]. J Instrumental Anal, 2008, 27(4): 408 411. [10],,,. 9 [J].

5 3, : B Wu N, Jiang JD, Tao DL, et al. Determination of vitamin B 12 content in milk powder by immunoaffinity column purification -HPLC method [J]. J Instrumental Anal, 2013, 32(3): [9],,,. 8 [J]., 2008, 27(4): Liu N, Chen DZ, Tang H, et al. Simultaneous determination of eight water-soluble vitamins in baby milk powder by high performance liquid chromatography [J]. J Instrumental Anal, 2008, 27(4): [10],,,. 9 [J]., 2014, 5(3): Ren DD, Xie YF, Liu JJ, et al. Simultaneous determination of nine water-soluble vitamins in foods by high performance liquid chromatography [J]. J Food Saf Qual, 2014, 5(3): [11] GB/T B 12 [S]. GB/T Determination of vitamin B 12 in health foods [S]. [12] Karmi O, Zayed A, Suheir Baraghethi, et al. Measurement of vitamin B 12 concentration: a review on available methods [J]. IIOAB J, 2011, 2(2): [13] Chen JH, Jiang SJ. Determination of cobalamin in nutritive supplements and chlorella foods by capillary electrophoresis-inductively coupled plasma mass spectrometry [J]. J Agric Food Chem, 2008, 56(4): [14] L Sagaya SK, Thakur MS. Competitive immunoassay for analysis of vitamin B 12 [J]. Anal Biochem, 2011, 418(2): [15],,,. B 12 [J]., 2014, 148: Wu H, Liu DH, Zhang H, et al. Determination of vitamin B 12 content in milk powder by immunoaffinity column purification HPLC method [J]. China Dairy, 2014, 148: [16] GB B 12 [S]. GB Food safety national standard-determination of vitamin B 12 in food for infants and young children, milk and milk products [S]. 作者简介 ( 责任编辑 : 杨翠娜 ) 蒋孟虹, 硕士, 药师, 主要研究方向为药品食品检验 jmh1122ow@163.com 杨美成, 博士, 主任药师, 主要研究方向为实验室质量管理药物分析与微生物学检验 yangmeicheng@vip.sina.com

Microsoft Word - 3098-3102 20150611007-蔡伟江-二校

Microsoft Word - 3098-3102 20150611007-蔡伟江-二校 6 8 Vol. 6 No. 8 2015 8 Journal of Food Safety and Quality Aug., 2015 蔡伟江 *, 邓鹏飞, 刘光兰, 李丹 (, 519040) 摘要 : 目的, 方法 (4.6 mm 250 mm, 5.0 μm) ; : : : =70: 16: 14(V:V:V),, : 1.0 ml/min; : 40 ; : 10