Microsoft Word 苏昭仑-二校 3114

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1 6 8 Vol. 6 No Journal of Food Safety and Quality Aug., 2015 苏昭仑 *, 黄康惠, 张喜金, 陈彩云, 陈宏壁, 林贤邦 (, ) 摘要 : 目的方法, ; : (SUPELCO, C 18, 4.6 mm 150 mm, 5 μm); : 1.0 ml /min; : 40 ; : 20 μl; : 294 nm; :, ; α- γ- δ- ; 结果 0.01~0.20 mg/ml, r>0.9990, 93.5%~103.8%, RSD<4.0%, 0.02 μg/kg, 0.1 μg/kg 结论,,, 关键词 : ; ; ; E Determination of mixed tocopherols in oil health food by reversed-phase high performance liquid chromatography SU Zhao-Lun *, HUANG Kang-Hui, ZHANG Xi-Jin, CHEN Cai-Yun, CHEN Hong-Bi, LIN Xian-Bang (By-Health Co., Ltd., Zhuhai , China) ABSTRACT: Objective To establish a method for determination of mixed tocopherols in oil health foods by reversed-phase high performance liquid chromatography. Methods The samples were saponified by potassium hydroxide methanol, and neutralized by hydrochloric acid methanol. The extracts were separated by RP-HPLC with the column of SUPELCO, C 18 (4.6 mm 150 mm, 5 μm), flow rate of 1.0 ml/min, column temperature of 40, injection volume of 20 μl, wavelength of 294 nm, mobile phase of water and methanol, and with gradient elution to separate α-, γ-and δ-tocopherol; then were determined by external standard method. Results Tocopherols standard curve had a good linear relationship within 0.01~0.20 mg/ml range (r>0.9990), the recovery was 93.5%~103.8%, RSD 4.0%, limit of determination was 0.02 μg/kg, and the limit of quantity was 0.1 μg/kg. Conclusion The method is accurate and reliable, saving pretreatment time and reagents, can reduce the test cost; it is suitable for the determination of mixed tocopherol in oil health food. KEY WORDS: mixed tocopherols; health foods; reversed-phase high performance liquid chromatography; vitamin E * 通讯作者 :,, suwheel@126.com *Corresponding author: SU Zhao-Lun, Licensed Pharmacist, By-Health Co., Ltd., Zhuhai , China. suwheel@126.com

2 8, : 引言 E,, d-α- 8 E, α- β- γ- δ- α- β- γ- δ- E d-α- ; ; d-α- d-α-,, [1-4] [5-6] [7-8] [9-15],, ;, β- γ-, ;,,, ;, β- γ-,, [15],, 2 材料与方法 2.1 仪器与试剂 : ( :, : 1260, VWD); ( : Mettler Toledo, : XP205) : 3322TG ( ) : (, CNW); (, CNW); (, ); (, ); (, ); ; d-α- ( : Dr, : D0050, : 99.8%); d-γ- ( : SUPELCO, : LB95755V, : 99%); d-δ- ( : SUPELCO, : LB94736V, : 99%) 2.2 仪器条件 : SUPELCO, C 18, mm, 5 μm; : 1.0 ml /min; : 40 ; : 20 μl; : 294 nm; : A, B, 1 表 1 梯度洗脱比例 Table 1 Gradient elution ratio (min) A(%) B(%) 标准溶液的制备标准储备液的配制 d-α-, d-γ-, d-δ- 50 mg 50 ml, ( 1.0 mg/ml), 标准储备液浓度校正 d-α-, d-γ-, d-δ ml, 10 ml,, 294 nm d-α- 298 nm d-γ-, 298 nm d-δ- C=A/E 100 C ( mg/ ml), A, E 1% (d-α- 71, d-γ- 92.8, d-δ- 91.2) 标准工作溶液配制 d-α-, d-γ-, d-δ ml 50 ml,,, mg/ml 2.4 样品前处理 1.0 g 50 ml, 0.2 g, ml (0.5

3 mol/l), min,,, ml (0.5 mol/l), 25 ml, 3 结果与讨论 3.1 样品前处理方法的优化 [15], (1+1), 2, ( 3 h),, (0.5 mol/l), (0.5 mol/l),,, ( 45 min) 3.2 色谱条件的选择色谱柱的选择 150 mm C 18 α- γ- δ-, 150 mm, α- γ- δ-, 250 mm 流动相的选择, β- γ-, γ- β- 3.3 方法的线性范围确认,,, α- γ- δ- 0.01~0.20 mg/ml, r 方法的回收率实验 6, 1.00 ml, %~103.8%, RSD 4.0%, 检出限, (S/N) 3, 0.02 μg/kg, (S/N) 10, 0.1 μg/kg, 表 2 结果对比 (n=10) Table 2 Results comparison (n=10) (mg/100 g) RSD(%) (mg/100 g) RSD(%) 94.3~ ~ ~ ~ ~ ~ TG 16.7~ ~ Fig. 1 1 The chart of mixed standard solution

4 8, : 3117 Table 3 表 3 加标回收率和相对标准偏差 (n=6) The recovery and relative standard deviation (n=6) (%) RSD(%) α- 95.4~ γ- 94.2~ δ- 96.1~ α- 95.1~ γ- 93.6~ δ- 94.4~ α- 96.9~ γ- 93.5~ δ- 96.4~ α- 95.1~ TG γ- 94.5~ δ- 95.3~ Fig. 2 2 The chromatogram of blank samples 3.6 样品的测定, 3322TG, 40,, 4, 3~6 4 结论 [15] [e],,,, [e], ;, ( 3 h),,,,, ( 45 min),, 表 4 总生育酚的含量 (n=10) Table 4 The content of tocopherols (n=10) (mg/100 g) (mg/100 g) 102.4~ ~ ~ TG 17.6~

5 Fig. 3 3 The chromatogram of auxiliary lowering soft capsule sample Fig. 4 4 The chromatogram of conjugated linoleic acid soft capsule sample Fig. 5 5 The chromatogram of salmon fish oil capsules sample

8, : 3119 Fig.6 6 3322TG The chromatogram of fish oil 3322TG sample 参考文献 [1] Tǜtem E, Apak R, Gǜnayd E, et al.

6 8, : 3119 Fig TG The chromatogram of fish oil 3322TG sample 参考文献 [1] Tǜtem E, Apak R, Gǜnayd E, et al. Spectrophotom etric determ ination of vitamin E (α-tocopherol) using copper (II) neocuproine reagent [J]. Talanta, 1997, 44: [2] Proetp P, Pineda M, Aguilar M. Spectrophotometric quantitation of antioxidant capacity through the form ation of a phosphomolybdenum complex: specific application to the determination of vitamin E [J]. Anal Biochem, 1999, 269: [3] Amin AS. Colorimetric determination of tocopheryl acetate (Vitamin E) in pure form and in multi-vitamin capsules [J]. Eur J Pharm Biopharm, 2001, 51: [4] Jadoon S, Waseem A, Yaqoob M, et al. Flow injection spectrophotometric determination of vitamin E in pharmaceuticals milk powder and blood serum using potassium ferricyanide-fe (III) detection system [J]. Chin Chem Lett, 2010, 21: [5] [M]., Pharmacopoeia of the People's Republic of China 2010 edition [M]. Chinese Medical Science and Technology Press, [6] GB E [S]. GB Food additive-natural vitamin E [S]. [7] Kamal A, Gorgen S, Pettersson J, et al. Normal-phase high performance liquid chromatography of tocopherols and tocotrienols comparison of different chromatographic columns [J]. J Chromatogr A, 2000, 881: [8] Nagy K, Courtet MC, Holst B, et al. Comprehensive analysis of vitamin e constituents in human plasma by liquid chromatography-mass spectrometry [J]. Anal Chem, 2007, 79: [9] GB/T E [S]. GB/T Determination of vitamin E in feeds high-performance liquid chromatography [S]. [10],,. HPLC A E [J]., 2006, 6(5): Wang H, Liu HH, Yin JW. Determination of vitamin A and vitamin E by HPLC in foods [J]. China Tropical Med, 2006, 6(5): [11] Paliakov EM, Crow BS, Bishop MJ, et al. Rapid quantitative determination of fat-soluble vitamins and coenzyme Q-10 in human serum by reversed phase ultra-high pressure liquid chromatography with UV detection [J]. J Chromatogr B, 2009, 877: [12] Gliszczynska AS, Sikorska E. Simple reversed-phase liquid chromatography method for determination of tocopherols in edible plant oils [J]. J Chromatogr A, 2004, 1048: [13] Tasioula MM, Okogeri O. Simultaneous determination of phenolic compounds and tocopherols in virgin olive oil using HPLC and UV detection [J]. Food Chem, 2001, 74: [14] Gimeno E, Calero E, Castellote AI, et al. Simultaneous determination of α-tocopheroland β carotene in olive oil by reversed-phase high-performance liquid chromatography [J]. J Chromatogr A, 2000, 881: [15] GB/T A E [S]. GB/T Determination of retinol and tocopherol in foods [S]. 作者简介 ( 责任编辑 : 白洪健 ) 苏昭仑, 执业中药师, 主要研究方向为中药和保健食品安全检测 suwheel@126.com

g ml 10% ph 色谱条件 kinetex C μm 100 A 4. 6 mm 150 mm 25 5 μl A B 10 mmol /L ml /min

g ml 10% ph 色谱条件 kinetex C μm 100 A 4. 6 mm 150 mm 25 5 μl A B 10 mmol /L ml /min 25 4 2013 8 监测技术 5 1 2 1 ( 1. 江苏省疾病预防控制中心, 江苏南京 210009; 2. 南京医科大学第二附属医院, 江苏南京 210029) 5 Phenomenex kinetex C 18 2. 6 μm 4. 6 mm 150 mm 10 mmol /L ph 2. 5 + 60 40 DAD 280 nm 5 20. 0 mg /L ~ 100 mg /L 0.