Microsoft Word 游景水-二校 4671.doc

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1 6 11 Vol. 6 No Journal of Food Safety and Quality Nov., 游景水 *, 王德伟 (, ) 摘要 : 目的 16 方法 6 mol/l, (OPA) 9- (FMOC) ; Agilent Hypersil AA-ODS ; (0.16% ) ; (FLD) ( : 340 nm, : 450 nm; 15 min, : 266 nm, : 305 nm) 结果 pmol/μl (r: , n=5), 91.6% 104.1%(n=9), (RSD) 0.9% 3.7%, pmol 结论,, 关键词 : ; ; ; ; ; 9- Determination of sixteen kinds of amino acids in the capsule of sheep placenta and medlar by online pre-column derivazation-reversed-phase high performance liquid chromatography YOU Jing-Shui *, WANG De-Wei (By-Health Co., Ltd., Zhuhai , China) ABSTRACT: Objective To establish a method for determination of sixteen kinds of amino acids in the capsule of sheep placenta and medlar by reversed-phase high performance liquid chromatography (RP-HPLC). Methods Proteins in the samples were hydrolyzed into amino acids in 6 mol/l hydrochloric acid solution, o-phthalaldehyde (OPA) and 9-fluorenylmethyl chloroformate (FMOC) were used as online pre-column derivative reagents for the primary amino acids and secondary amino acids respectively, and the chromatographic experiments were performed on an Agilent Hypersil AA-ODS column. The 0.16% sodium acetate solution was used as mobile phase. Signals were detected by fluorescence detection (FLD) (excitation wavelength: 340 nm, emission wavelength: 450 nm, excitation wavelength changed to 266 nm and emission wavelength changed to 305nm at the 15 min). Results The concentration of sixteen kinds of amino acids demonstrated a good linearity in the range of pmol/μl(r: , n=5). The recoveries were 91.6% 104.1% (n=9). The RSD were 0.9% 3.7%. The limits of detection were pmol. Conclusion This method has advantages of high sensitivity, precision and accuracy. It's suitable for the determination of sixteen kinds of amino acids in the capsule of sheep placenta and medlar. KEY WORDS: capsule of sheep placenta and medlar; online pre-column derivazation; amino acid; re- * 通讯作者 :,, @qq.com *Corresponding author: YOU Jing-Shui, Licensed Pharmacist, By-Health Co., Ltd., Zhuhai , China @qq.com

2 versed-phase high performance liquid chromatography; o-phthaldialdehyde; 9-fluorenylmethyl chloroformate 1 引言, [1], ( ), [2-8],, [9] (GB/T ) [2], ( (10 pmol) OPA FMOC,, (FLD) [10-15], 2 材料与方法 2.1 主要仪器 Agilent 1200 (, Agilent ), : Chem32; KQ-500E ( ); XW-80A ( ); DHG-9240A ( ); BPZ-6090Lc ( ) 2.2 试剂 OPA (, : BCBF1444); FMOC (, : BCBD9246); (, : BCBF0657); 250 pmol/μl (, : BCBD9914); (, ); (, ); (, ); (, ); (, ); (, ); (, ); (, ) 2.3 色谱条件 : Hypersil AA-ODS (200 mm 2.1 mm, 5 μm, ); : 0.45 ml/min; : 40 ; : 1 µl; : 30 min A(0.16% ): 1.6 g, 1000 ml,, 180 μl,, 2% ph 7.20±0.1, 3 ml,, 0.45 μm, ; B(0.16% ): 1.6 g, 200 ml,, 2% ph 7.20±0.1, 400 ml 400 ml,, 0.45 μm, 1 表 1 梯度洗脱比例 Table 1 Gradient elution ratio (min) (ml/min) A(%) B(%) : μl ; μl OPA ; 3 ( ) 0.0 μl ( ); X 1.0 μl ; 3 ( ) 0.0 μl ( ); 8 μl ;, 6 ; μl FMOC; 3 ( ) 0.0 μl ( ); 9 μl ;, 3 ; : (FLD) : 0 min, : 340 nm, : 450 nm, PTM : 11; 15 min, : 266 nm, : 305 nm, PTM : 对照品溶液的制备 250 pmol/μl, 0.1 mol/l : 5L pmol/μl

3 11, : 样品前处理 0.12 g, 25 ml, 6 mol/l 15 ml, 0.2 g,,,, 110 ±1, 22,,,, 50mL,,, 1 ml 10 ml,, ( ), 0.1 mol/l,, 0.45 μm, 3 结果与分析 3.1 线性试验和检出限试验 2.3, 5, (X: pmol/μl), (Y) (S/N)=3 (pmol) : pmol/μl (r: )(n=5), : pmol 表 2 16 种氨基酸的线性方程相关系数和检出限 Table 2 Linear equation, correlation coefficient and detection limit of the sixteen kinds of amino acids (pmol) Y = X Y = X Y = X Y = X Y = X Y = X Y = X Y = X Y = X Y = X Y = X Y = X Y = X Y = X Y =8.972 X Y = X 精密度试验 6,, 2.3,, 16 RSD 0.3% 2.0%, 3, 3, 3.3 加标回收试验 9, 3, 3, 250 nmol/μl μl,, 2.3, % 104.1%, RSD 0.9% 3.4% 4, 4, 3.4 方法专属性试验 16, 2.3, ,, 16, 16, 4 讨论 (OPA) 9- (FMOC), : (1), 16 (2), 16, [10,11],,, pmol, GB/T ( ) 10 pmol, 5 结论 16,,,

4 Table 3 表 3 精密度的试验 (n=6) The experiment of the precision (n=6) (mg/g) RSD(%) (mg/g) RSD(%) (mg/g) RSD(%) (mg/g) Table 4 表 4 加标回收率试验 (n=9) The experiment of standard addition recovery (n=9) (%) RSD(%) (%) RSD(%) (%) RSD(%) % % % % % % % % % % % % % % % % Fig. 1 1 The chromatogram of blank samples

5 11, : Fig The chromatogram of the sixteen kinds of amino acid standards Fig. 3 3 The chromatogram of samples 参考文献 [1],,,. [J]., 2007, 33(6): 1 4. Tan HP, Chen L, Ye SR, et al. Overview of amino acid determinations for tea [J]. China Meas Technol, 2007, 33(6): 1 4. [2] GB/T [S]. GB/T Determination of amino acids in foods [S]. [3],,. [J]., 2003, 31(5): Zhang GA, XU X, Zhang S. High throughput separation and identification for proteomics and recent development in proteome [J]. Anal Chem, 2003, 31(5): [4] NY/T [S]. NY/T Determination of amino acids in velvet antler [S]. [5] SN/T [S]. SN/T Method for the determination of total amino acid in import and export-amino acid automatic analyzer method [S]. [6] GB/T [S].

6 GB/T Laboratory animal-determination of amino acids for formula feeds [S]. [7] QB/T : [S]. QB/T Determination of free amino acids in Chinese rice wine: High performance liquid chromatography [S]. [8] QB/T [S]. QB/T Determination of amino acids in cosmetics [S]. [9] JY/T [S]. JY/T Gendral rules for amino acid analysis [S]. [10] GB/T : [S]. GB/T Inspection of germplasm for cultured fishes-part 11:Determination of amino acids content in muscle [S]. [11] NY/T [S]. NY/T Water-soluble fertilizers-determination of amino-acids content [S]. [12]. [J]., 1995, 2: Liu HW. Progress and comments on the analysis of amino acids in the column [J]. Amino Acids Biol Resour, 1995, 2: [13],,,. 4 [J]., 2009, 8: Gui L, Tian H, Zhen J, et al. Simultaneous determination of 4 kinds of amino acids neurotransmitters in brain tissue of mice by high performance liquid chromatography [J]. J Third Military Med Univ, 2009, 8: [14],. [J]. 2007, 2: Ji YF, Shi LF. Assay Amino Acid in Feed -Acid Hydrolytic Method Test and Research [J]. Grass-Feeding Livestock, 2007, 2: [15],. [J]., 2009, 5: Qu CL, Bai YP. Determination of amino acids in Panax ginseng by microwave hydrolysis and high performance liquid chromatography [J]. Anal Chem, 2009, 5: 作者简介 ( 责任编辑 : 白洪健 ) 游景水, 执业药师, 质量工程师, 主要研究方向为保健食品的质量检测 @qq.com

Microsoft Word 王德伟-二校

Microsoft Word 王德伟-二校 6 8 Vol. 6 No. 8 2015 8 Journal of Food Safety and Quality Aug., 2015 王德伟 *, 叶礼红, 杨祖伟 (, 519040) 摘要 : 目的方法, C 18, 0.68% : (V:V=82:18), 1.0 ml/min, 340 nm 结果 D- 0.052 1.045 mg/ml, 97.5%, RSD%=1.2(n=9)