食品安全质量检测学报
食品安全質量檢測學報
식품안전질량검측학보
Journal of Food Safety & Quality
2015年
11期
4671-4676
,共6页
羊胎盘枸杞胶囊%柱前在线衍生%氨基酸%反相高效液相色谱法%邻苯二甲醛%9-芴甲基氯甲酸酯
羊胎盤枸杞膠囊%柱前在線衍生%氨基痠%反相高效液相色譜法%鄰苯二甲醛%9-芴甲基氯甲痠酯
양태반구기효낭%주전재선연생%안기산%반상고효액상색보법%린분이갑철%9-물갑기록갑산지
capsule of sheep placenta and medlar%online pre-column derivazation%amino acid%re-versed-phase high performance liquid chromatography%o-phthaldialdehyde%9-fluorenylmethyl chloroformate
目的：建立一种采用反相高效液相色谱法测定羊胎盘枸杞胶囊中16种氨基酸的含量。方法样品中的蛋白质在6 mol/L盐酸溶液中水解为氨基酸,其中一级氨基酸用邻苯二甲醛(OPA)、二级氨基酸用9-芴甲基氯甲酸酯(FMOC)进行柱前在线衍生；用Agilent Hypersil AA-ODS柱分离；用流动相(0.16%醋酸钠)进行梯度洗脱；经荧光检测器(FLD)检测(激发波长：340 nm,发射波长：450 nm；15 min时,激发波长：266 nm,发射波长：305 nm)。结果16种氨基酸在5～250 pmol/μL浓度范围内呈现良好的线性关系(r：0.99959～1.00000,n=5),平均加标回收率为91.6%～104.1%(n=9),相对标准偏差(RSD)为0.9%～3.7%,最低检测限为0.4～1.2 pmol。结论该方法灵敏度高,具有良好的精密度与准确度,可以有效地控制产品质量。
目的：建立一種採用反相高效液相色譜法測定羊胎盤枸杞膠囊中16種氨基痠的含量。方法樣品中的蛋白質在6 mol/L鹽痠溶液中水解為氨基痠,其中一級氨基痠用鄰苯二甲醛(OPA)、二級氨基痠用9-芴甲基氯甲痠酯(FMOC)進行柱前在線衍生；用Agilent Hypersil AA-ODS柱分離；用流動相(0.16%醋痠鈉)進行梯度洗脫；經熒光檢測器(FLD)檢測(激髮波長：340 nm,髮射波長：450 nm；15 min時,激髮波長：266 nm,髮射波長：305 nm)。結果16種氨基痠在5～250 pmol/μL濃度範圍內呈現良好的線性關繫(r：0.99959～1.00000,n=5),平均加標迴收率為91.6%～104.1%(n=9),相對標準偏差(RSD)為0.9%～3.7%,最低檢測限為0.4～1.2 pmol。結論該方法靈敏度高,具有良好的精密度與準確度,可以有效地控製產品質量。
목적：건립일충채용반상고효액상색보법측정양태반구기효낭중16충안기산적함량。방법양품중적단백질재6 mol/L염산용액중수해위안기산,기중일급안기산용린분이갑철(OPA)、이급안기산용9-물갑기록갑산지(FMOC)진행주전재선연생；용Agilent Hypersil AA-ODS주분리；용류동상(0.16%작산납)진행제도세탈；경형광검측기(FLD)검측(격발파장：340 nm,발사파장：450 nm；15 min시,격발파장：266 nm,발사파장：305 nm)。결과16충안기산재5～250 pmol/μL농도범위내정현량호적선성관계(r：0.99959～1.00000,n=5),평균가표회수솔위91.6%～104.1%(n=9),상대표준편차(RSD)위0.9%～3.7%,최저검측한위0.4～1.2 pmol。결론해방법령민도고,구유량호적정밀도여준학도,가이유효지공제산품질량。
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). MethodsProteins 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 deriva-tive 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). ResultsThe concentration of sixteen kinds of amino acids demonstrated a good linear-ity in the range of 5~250 pmol/μL(r: 0.99959~1.00000,n=5). The recoveries were 91.6%~104.1% (n=9). The RSD were 0.9%~3.7%. The limits of detection were 0.4~1.2 pmol.ConclusionThis method has ad-vantages 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.
