何传昌,关瑾,丁爽,阎峰,石爽,牛秋玲
HE Chuan-chang,GUAN Jin,DING Shuang,YAN Feng,SHI Shuang,NIU Qiu-ling

• 1:沈阳化工大学应用化学学院

摘要(Abstract)：

目的建立分离分析5-氨基乙酰丙酸及4种相关组分的反相离子对液相色谱法。方法考察了缓冲溶液的pH值、离子对的种类和浓度等色谱条件对分离分析的影响。以乙腈-20mmol·L-1磷酸二氢钾(pH值2.0,含2.0mmol·L-1辛烷磺酸钠)为流动相,在ZORBAX SB-C18色谱柱上,采用梯度洗脱,对5-氨基乙酰丙酸及4种相关组分进行分离分析。结果在优化色谱条件下,5-氨基乙酰丙酸及4种相关组分在15min内达到基线分离;5-氨基乙酰丙酸质量浓度在0.1~2.0g·L-1内与峰面积呈良好的线性关系,相关系数为0.999 5;检出限(信噪比为3)为3.0mg·L-1;样品的回收率为99.3%~100.5%,相对标准偏差为0.16%~0.92%。结论该方法操作简单、快速、准确可靠,可用于实际样品的分析,并获得了令人满意的结果。
Objective To develop a method for the separation and determination of 5-aminolevulinic acid(ALA)and related four substances by reversed-phase ion-pair high performance liquid chromatography(RPIP-HPLC).Methods The influences of pH of buffer,type and concentration of ion-pair were investigated by RPIP-HPLC on a ZORBAX SB-C18 column.The separation of the ALA and related four substances were studied using acetonitrile-20 mmol·L-1 potassium dihydrogen phosphate(containing 2.0 mmol·L-1 octanesulfonic acid sodium,pH2.0)as mobile phase with a gradient elution program and detection wavelength of 205 nm.Results Under the optimized condition,ALA and related four substances were baseline separated within 15 min.The calibration curve of ALA was good linear between the peak areas and the concentrations of 0.1-2.0 g·L-1 with correlation coefficients r2=0.999 5.The limit of detection(S/N=3)for ALA was 3.0 mg·L-1.The percentage recoveries were ranged from99.3% to 100.5% with acceptable relative standard deviations of 0.16%-0.92%.Conclusions The method is simple,sensitive,accurate and reliable for the determination of real samples.

关键词(KeyWords)： 5-氨基乙酰丙酸;反相离子对色谱法;含量测定
5-aminolevulinic acid;reversed-phase ion-pair high performance liquid chromatography;content determination

Abstract:

Keywords:

基金项目(Foundation): 辽宁省教育厅科学研究一般项目(L2012150)

作者(Author): 何传昌,关瑾,丁爽,阎峰,石爽,牛秋玲
HE Chuan-chang,GUAN Jin,DING Shuang,YAN Feng,SHI Shuang,NIU Qiu-ling

DOI: 10.14066/j.cnki.cn21-1349/r.2013.09.016

参考文献(References)：

• [1]汪良驹,姜卫兵,章镇,等.5-氨基乙酰丙酸的生物合成和生理活性及其在农业中的潜在应用[J].植物生理学通讯,2003,39(3):185-191.
• [2]张一宾.5-氨基乙酸丙酸(ALA)的功能与展开[J].世界农药,2006,28(4):14-17.
• [3]孙耀冉,陈涛,吕连营,等.新型绿色农药:5-氨基乙酰丙酸[J].农药,2008,47(10):710-711,725.
• [4]ZEITOUNI N C,OSEROFF A R,SHIEH S.Photodynamic therapy for nonmelanoma skin cancers.Current review and update[J].Mol Immunol,2003,39(17/18):1133-1136.
• [5]CHEN Hsin-ming,CHEN Chin-tin,YANG Hsiang,et al.Successful treatment of an extensive verrucous carcinoma with topical 5-aminolevulinic acid-mediated photodynamic therapy[J].J Oral Path Med,2005,34(4):253-256.
• [6]卢晓珣,郭坤元.光动力治疗药物5-氨基乙酰丙酸及其己脂的相关研究进展[J].实用医学杂志,2008,24(12):2035-2037.
• [7]赵春晖,穆江华,岑沛霖.化学法与生物转化合成5-氨基乙酰丙酸的研究进展[J].农药,2003,42(11):11-15.
• [8]MOENS L,COLO L.Synthesis of an acid addition salt ofδ-aminolevulinic acid from 5-bromolevulinic acid ester:US,5907058[P].1999-05-25.
• [9]文辉忠,范闽光.分光光度法测定δ-氨基乙酰丙酸的研究[J].广西化工,1997,26(2):47-50.
• [10]王凌健,倪迪安,叶叙,等.植物叶片中δ-氨基乙酸丙酸的测定[J].植物生理学通讯,1997,33(6):439-441.
• [11]BUNKE A,ZERBE O,SCHMID H,et al.Degradation mechanism and stability of 5-aminolevulinic acid[J].J Pharm Sci,2000,89(10):1335-1340.
• [12]BUNKE A,SCHMID H,BURMEISTER G,et al.Validation of a capillary electrophoresis method for determination of 5-aminolevulinic acid and degradation products[J].J Chromatogr A,2000,883(1/2):285-290.
• [13]范理,林建平,刘晓侠,等.反相高效液相色谱柱前衍生法测定发酵液中的5-氨基乙酰丙酸[J].分析化学,2006,34(7):937-938.
• [14]陈祝康,彭兴盛,吴伟.RP-HPLC测定盐酸氨基乙酰丙酸及其散剂的含量及有关物质[J].中国药学杂志,2007,42(24):1895-1897.