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[1]吕铎,邢光南,盖钧镒.大豆种子中嘌呤含量的HPLC 测定方法[J].大豆科学,2016,35(04):655-659.[doi:10.11861/j.issn.1000-9841.2016.04.0655]
　LYU Duo,XING Guang-nan,GAI Jun-yi.HPLC Method for Determining Purine Content in Soybean Seeds[J].Soybean Science,2016,35(04):655-659.[doi:10.11861/j.issn.1000-9841.2016.04.0655]

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大豆种子中嘌呤含量的HPLC 测定方法

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷期数: 2016年04期页码: 655-659 栏目: 出版日期: 2016-07-20

Title:: HPLC Method for Determining Purine Content in Soybean Seeds

作者:: 吕铎; 邢光南; 盖钧镒; ( 南京农业大学大豆研究所/国家大豆改良中心/农业部大豆生物学与遗传育种重点实验室( 综合) /作物遗传与种质创新国家重点实验室，江苏南京210095)

Author(s):: LYU Duo; XING Guang-nan; GAI Jun-yi; Soybean Ｒesearch Institute of Nanjing Agricultural University / National Center for Soybean Improvement / Key Laboratory for Biology and Genetic Improvementof Soybean ( General) ，Ministry of Agriculture / National Key Laboratory for Crop Genetic and Germplasm Enhancement，Nanjing 210095，China

关键词:: 大豆种子; 嘌呤含量; 高效液相色谱分析

Keywords:: Soybean seed; Purine Content; High performance liquid chromatography ( HPLC)

分类号:: S565. 1

DOI:: 10.11861/j.issn.1000-9841.2016.04.0655

文献标志码:: A

摘要:: 根据检测不同高氯酸浓度和水解温度对大豆种子的水解效果，检测不同流动相对鸟嘌呤、腺嘌呤、次黄嘌呤、黄嘌呤4 种成分的分离效果，建立了稳定准确的大豆种子中多组分嘌呤含量的高效液相色谱分析方法。结果表明:在100℃条件下， 35% ( v /v) 的高氯酸处理60 min 时豆粉水解效果最佳; 使用Welch Ultimate Polar ＲP18 ( 4. 6 mm ×250 mm，5. 0 μm) 色谱柱以0. 02 mol·L － 1 磷酸二氢钾缓冲液( pH2. 9) 作为流动相时，4种嘌呤在0. 05 ～ 25 mg·L － 1浓度范围内均可完全分离且响应峰面积与浓度呈良好线性相关( r ＞ 0. 999 9) ，相对标准偏差为2. 97% ～ 6. 25%，加标回收率为89. 19% ～ 110. 97%。

Abstract:: A stable and accurate HPLC ( high performance liquid chromatography) procedure for measuring multiple componentsof purines in soybean seeds was established based on choosing an appropriate HClO4 content and temperature for hydrolysisof mashed soybean specimen and choosing an appropriate buffer solution mobile phase. The optimized hydrolysis conditionwas hydrolyzing mashed soybean seed with 35% ( v /v) of HClO4 for 60 min at 100℃. The optimized chromatography conditionwas using Welch Ultimate Polar ＲP18 ( 4. 6 mm × 250 mm，5. 0 μm) as chromatographic column with 0. 02 mol·L － 1KH2 PO4buffer solution ( pH2. 9) as mobile phase at flow rate of 1. 0 mL·min － 1. Under this system，the ademine，guanine，hypoxanthineand xanthine within 0. 05 － 25 mg·L － 1 could be fully separated and the response peak area was linearly correlated withpurine content perfectly ( r ＞ 0. 999 9) with the range of relative standard deviation ( ＲSD or CV) as 2. 97% － 6. 25% and therange of spiked recovery rate as 89. 19% － 110. 97%.

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备注/Memo

基金项目: 国家重点基础研究发展计划“973 计划”( 2011CB1093 ) ; 农业部公益性行业专项( 201203026-4 ) ; 国家自然科学基金( 31571694) ; 教育部111 项目( B08025 ) ; 教育部长江学者和创新团队( PCSＲT13073 ) ; 中央高校基本科研业务费项目( KYZ201202-8) ; 国家现代农业产业技术体系( CAＲS-04) ; 江苏省优势学科建设工程专项; 江苏省JCIC-MCP 项目。第一作者简介: 吕铎( 1990-) ，男，硕士，主要从事大豆遗传育种研究。E-mail: 18298345890@163. com。通讯作者: 盖钧镒( 1936-) ，男，教授，博导，主要从事大豆遗传育种和植物数量遗传学研究。E-mail: sri@ njau. edu. cn。

更新日期/Last Update: 2016-08-23