CN110220960B - 一种l-精氨酸的检测方法及传感器 - Google Patents
一种l-精氨酸的检测方法及传感器 Download PDFInfo
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- CN110220960B CN110220960B CN201910604859.6A CN201910604859A CN110220960B CN 110220960 B CN110220960 B CN 110220960B CN 201910604859 A CN201910604859 A CN 201910604859A CN 110220960 B CN110220960 B CN 110220960B
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
本发明公开了一种L‑精氨酸(L‑Arg)的检测方法及传感器,所述方法包括合成二茂铁功能化十六肽二硫代环戊烷(FC‑P16 Peptide)、制备FC‑P16 Peptide/AuE多肽复合膜修饰电极和L‑精氨酸检测等步骤。结果表明,FC‑P16 Peptide/AuE多肽复合膜修饰电极对L‑Arg表现出良好的电化学响应特性。在10 mmol/L磷酸盐缓冲溶液(PBS,pH=7.4)中,该多肽复合膜修饰电极的DPV响应峰电流与L‑Arg的浓度在1.0×10‑13~1.0×10‑7 mol/L范围内存在良好的线性关系,检测限为1.0×10‑13 mol/L。该修饰电极具有良好的重现性、重复性与选择性,在生命科学与营养健康方面有潜在的应用价值。
Description
技术领域
本发明属于化学/生物传感技术领域,具体涉及一种L-精氨酸的检测方法及传感器。
背景技术
L-精氨酸(L-arginine,L-Arg)又称蛋白氨基酸,是维持婴幼儿生长发育必不可少的氨基酸。它是鸟氨酸循环的中间代谢物,能促使氨转变成为尿素,从而降低血氨含量。它有帮助改善免疫系统健康和抵御疾病的作用。在身体受伤情况下,身体免疫系统处于最佳状态,可以加快身体疗伤速度。总的来说,精氨酸在伤口愈合、细胞分裂、维持机体电荷平衡和生理功能中起着极其重要的作用。因此,开发一种快速、灵敏检测L-Arg的方法,在生命科学和营养健康领域具有十分重要的意义。
目前,检测精氨酸的方法主要有比浊法、分光光度法、液相色谱法、毛细管电泳法、质谱分析法、表面等离子共振法、荧光分析法等,其中Ding等制备了基于二芳基-罗丹明衍生物的荧光传感器,发现其与Cu2+的复合物显示出对L-Arg的灵敏识别能力,检测下限为2.2μmol/L。然而,这些仪器分析的方法存在设备笨重、价格昂贵、操作繁琐等缺点,而电化学传感器具有简便、快速、灵敏等优点,已引起了广泛关注。其中,基于双酶(精氨酸酶I和脲酶)和电活性聚苯胺修饰的铂电极(PANi/Nafion/Pt),可应用于葡萄酒和果汁样品中L-Arg的检测,检测下限为38μmol/L。随后,Stasyuk等发展了基于过饱和人肝脏精氨酸酶I重组酵母细胞的L-精氨酸生物传感器,该传感器具有响应快速(60s)、检测限低(0.085mmol/L)等优点。但是,这几种检测精氨酸的电化学传感器都需要酶的参与,使其应用受到局限。
近年来,由于多肽能模拟蛋白质的多种结构和功能特征,且多肽易于合成与修饰、成本低、易于保存、有良好的稳定性,基于多肽的电化学传感器逐渐应用于蛋白质、抗原等生物分子的检测。Zhao等使用二茂铁(FC)功能化螺旋肽测定酶活性前列腺特异性抗原(PSA),其电化学测量原理是在PSA存在下电极表面发生溶蛋白性裂解,导致电流信号下降;该电极制备简单,具有良好的选择性,检测下限为0.2ng/mL。Yang等筛选出对双酚A(BPA)表现出特异性的七肽,自组装于金电极表面,随着捕获的BPA分子的增多而响应峰电流下降,线性响应范围为1~5000nM,检测下限为0.7nM。HWang等将Noro-1肽固定在金电极表面上,可应用于人类诺如病毒的检测,重现性与稳定性好,检测下限为99.8nM。然而迄今为止,基于多肽复合膜用于检测L-精氨酸的无酶电化学传感器尚未见报道。
发明内容
本发明旨在克服现有技术的不足,提供一种L-精氨酸的检测方法及传感器。
为了达到上述目的,本发明提供的技术方案为:
所述L-精氨酸的检测方法包括如下步骤:
(1)合成二茂铁功能化十六肽二硫代环戊烷(FC-P16 Peptide);所述二茂铁功能化十六肽二硫代环戊烷结构式如式(I)所示:
(2)制备FC-P16 Peptide/AuE多肽复合膜修饰电极:将金电极于Piranha溶液中浸泡后清洗,然后抛光,洗净并用N2吹干;将吹干后的金电极浸入二茂铁功能化十六肽二硫代环戊烷浓度为30~80μmol/L的二茂铁功能化十六肽二硫代环戊烷溶液和三(2-羧乙基)膦浓度为10~80μmol/L的磷酸盐缓冲溶液(PBS,10mmol/L,pH=7.4)中20~30h;再将金电极于6-巯基-1-己醇浓度为0.5~2.0mmol/L的6-巯基-1-己醇溶液中浸泡2~30min后,用PBS沿着金电极表面冲洗以除去非特异性吸附的其它物质,用N2吹干即得FC-P16 Peptide/AuE多肽复合膜修饰电极;
(3)以FC-P16 Peptide/AuE多肽复合膜修饰电极为工作电极,以银/氯化银电极作为参比电极,以铂丝电极作为对电极,构成三电极体系;然后采用循环伏安法和示差脉冲伏安法考察不同修饰电极的电化学行为,并采用示差脉冲伏安法对不同浓度的L-精氨酸进行测试,绘制工作标准曲线,再采用标准加入法对待测样品中的L-精氨酸进行检测。
优选地,步骤(2)中对金电极表面进行抛光是分别用1.0μm、0.3μm和0.05μm的氧化铝粉对金电极表面进行抛光。优选地,步骤(2)中所述金电极直径为3mm。
优选地,步骤(3)中是在10mmol/L磷酸盐缓冲溶液(PBS,pH=7.4)、2.0mmol/L[Fe(CN)6]4-/3--10mmol/L PBS溶液、含有1.0×10-5mol/L精氨酸的PBS(10mmol/L,pH=7.4)溶液中采用循环伏安法和示差脉冲伏安法考察不同修饰电极的电化学行为,采用示差脉冲伏安法对不同浓度的L-精氨酸进行电流响应与浓度关系测试;示差脉冲伏安法参数为:振幅0.05V,脉冲周期0.5s,抽样宽度0.02,脉冲宽度0.2s,循环伏安法参数为:采样间隔0.001V,扫速100mV/s。
所述检测L-精氨酸的传感器包括作为工作电极的多肽复合膜修饰电极;所述多肽复合膜修饰电极包括金基质(5),所述金基质(5)表面修饰有多肽复合膜层(6),所述多肽复合膜层(6)中含有多肽分子(7);所述多肽分子(7)为二茂铁功能化十六肽二硫代环戊烷分子,所述二茂铁功能化十六肽二硫代环戊烷分子的氨基酸序列为GGGGFGHIHEGYGGGG(SEQID NO.2),两端的-GGGG-为连接序列(Linker)。
优选地,所述多肽复合膜层(6)中还含有6-巯基-1-己醇分子(8)。
优选地,所述金基质(5)的厚度为1.0~5.0mm,所述多肽复合膜层(6)的厚度为2~20nm。
优选地,所述传感器对L-精氨酸的浓度存在良好的线性关系,检测的线性范围为1.0×10-13~1.0×10-7mol/L,检出限为1.0×10-13mol/L。
本发明首先筛选出了L-精氨酸(L-Arg)特异性肽序列FGHIHEGY(SEQ ID NO.1),基于此序列,以两端的-GGGG-为连接序列(Linker),合成了两端分别具有二茂铁与1,2-二硫代环戊烷-3-正丁基残基侧链的二茂铁功能化多肽,即,二茂铁功能化十六肽二硫代环戊烷(FC-P16 Peptide),其中含有碳链骨架桥—(CH2)4—,其氨基酸序列如SEQ ID NO.2所示。该肽末端具有二硫基团,还原剂三(2-羧乙基)膦(简称:TCEP)的加入打断了二硫键而形成两个硫醇基团,硫醇基团能在金电极表面通过Au-S键结合,从而避免使用任何连接剂。本发明通过将二茂铁功能化多肽修饰到金电极的表面上,并以6-巯基-1-己醇(简称:MCH)为封闭剂,制得新型的多肽复合膜修饰电极,既FC-P16 Peptide/AuE。通过循环伏安法(CV)与示差脉冲伏安法(DPV)证实在电极表面形成了L-Arg特异性肽分子的自组装单层(SAM)。实验结果证明,多肽复合膜修饰电极对L-Arg具有良好的电化学响应信号,说明多肽复合膜修饰电极在生物传感监测方面具有潜在的应用价值。
总之,本发明设计合成了一种含有FGHIHEGY氨基酸序列的二茂铁功能化十六肽二硫代环戊烷(FC-P16 Peptide),即二茂铁功能化十六肽-3-正丁基-1,2-二硫代环戊烷(FC-P16-C4-DTCP),通过三(2-羧乙基)膦(TCEP)还原二硫键将该多肽分子自组装于金电极(AuE)表面,并用6-巯基己醇(MCH)封闭金表面,获得二茂铁功能化多肽修饰金电极(FC-P16Peptide/AuE)。通过循环伏安法(CV)与示差脉冲伏安法(DPV)探讨L-Arg在不同修饰电极上的电化学行为,发现FC-P16 Peptide/AuE对L-Arg表现出良好的电化学响应特性。在10mmol/L磷酸盐缓冲溶液(PBS,pH=7.4)中,该修饰电极的DPV响应峰电流与L-Arg的浓度在1.0×10-13~1.0×10-7mol/L范围内存在良好的线性关系,检测限为1.0×10-13mol/L。该多肽复合膜修饰电极具有良好的重现性、重复性与选择性,在生命科学与营养健康方面有潜在的应用价值。
附图说明
图1为基于多肽复合膜修饰电极的L-精氨酸检测传感器的工作结构示意图;
图2为二茂铁功能化十六肽二硫代环戊烷(FC-P16-C4-DTCP,FC-P16 Peptide)的序列;
图3为不同修饰电极在2.0mmol/L[Fe(CN)6]3-/[Fe(CN)6]4-(浓度比1:1)-PBS(10mmol/L,pH=7.4)溶液中的循环伏安图:(a)AuE,(b)FC-P16 Peptide/AuE;
图4为AuE(曲线a)和FC-P16 Peptide/AuE(曲线b)电极在PBS溶液(10mmol/L,pH=7.4)中的循环伏安曲线图:(a)AuE,(b)FC-P16 Peptide/AuE;
图5为AuE(曲线a)和FC-P16 Peptide/AuE(曲线b)电极在含有1.0×10-5mol/L的精氨酸的PBS溶液(A)和不含精氨酸的PBS溶液(B)中的示差脉冲伏安曲线图:(a)AuE,(b)FC-P16 Peptide/AuE;
图6为FC-P16 Peptide/AuE对不同浓度L-Arg上的示差脉冲伏安曲线图(A)及峰电流与浓度对数的响应关系曲线图(B)、分段线性关系曲线图(C,D)。
图1中:1、银/氯化银电极;2、铂丝电极;3、表面修饰的金电极;4、待测溶液;5、金基质;6、多肽复合膜层;7、多肽分子;8、6-巯基-1-己醇分子;9、L-Arg。
具体实施方式
实施例中以超纯水制备多肽母液(1.0mmol/L),并在使用时用磷酸盐缓冲溶液(PBS,10mmol/L,pH=7.4)稀释至所需浓度,0.20mol/L的NaOH和0.20mol/L的HCl用来调节磷酸盐缓冲溶液的pH值;所用试剂均为分析纯(AR),实验用水均为超纯水(电阻率≥18.3MΩ·cm)。以下描述中,氨基酸的描述均采用英文缩写。
一、实验过程
1、制备FC-P16 Peptide/AuE多肽复合膜修饰电极
将金电极(直径3mm)浸泡在Piranha溶液中5min,用超纯水清洗。依次用1.0μm、0.3μm、0.05μm的Al2O3粉打磨抛光成镜面,分别在超纯水、无水乙醇、超纯水中超声5min,清洗干净并用N2吹干。室温下,吹干后的金电极浸入含有40μmol/L的二茂铁功能化十六肽二硫代环戊烷(FC-P16-C4-DTCP,FC-P16 Peptide)和50μmol/L的三(2-羧乙基)膦(TCEP)的磷酸盐缓冲溶液(PBS,10mmol/L,pH=7.4)中24小时。为了阻断金表面上余下的活性位点并使多肽能垂直于表面,将修饰电极在1.0mmol/L的6-巯基-1-己醇(MCH)溶液中浸泡5min,用PBS沿着金表面冲洗以除去非特异性吸附的其它物质,用N2吹干,得FC-P16 Peptide/AuE多肽复合膜修饰电极,于4℃下保存备用。
2、L-Arg电化学检测
参见图1,以表面修饰的金电极作为工作电极,银/氯化银(饱和氯化钾)电极1作为参比电极,铂丝电极2作为对电极的三电极体系,使用电化学工作站对L-Arg 9进行电化学检测,用10mmol/L的PBS为缓冲溶液。采用循环伏安法(CV)与示差脉冲伏安法(DPV)考察不同修饰电极在10mmol/L磷酸盐缓冲溶液(PBS,pH=7.4)、2.0mmol/L[Fe(CN)6]4-/3--10mmol/L PBS溶液、含有1.0×10-5mol/L精氨酸的PBS(10mmol/L,pH=7.4)溶液中的电化学行为,并采用示差脉冲伏安法对不同浓度的L-Arg进行电流响应与浓度关系测试,绘制标准工作曲线;DPV的参数设置如下:振幅0.05V,脉冲周期0.5s,抽样宽度0.02,脉冲宽度0.2s,CV的参数设置如下:采样间隔0.001V,扫速100mV/s。
其中,所述表面修饰的金电极3即为本发明所述检测L-精氨酸的传感器中的多肽复合膜修饰电极;所述多肽复合膜修饰电极包括金基质5,所述金基质5表面修饰有多肽复合膜层6,所述多肽复合膜层6中含有多肽分子7和6-巯基-1-己醇分子8,所述多肽分子7为二茂铁功能化十六肽二硫代环戊烷分子(FC-P16 Peptide),即二茂铁功能化十六肽-3-正丁基-1,2-二硫代环戊烷(FC-P16-C4-DTCP),所述二茂铁功能化十六肽二硫代环戊烷分子的氨基酸序列为GGGGFGHIHEGYGGGG,两端的-GGGG-为连接序列(Linker)。所述金基质5的厚度为1.0~5.0mm,所述多肽复合膜层6的厚度为2~20nm。
3、样品处理与测定
采用标准加入法对猪血清样品(待测溶液4)中L-Arg进行检测。猪血清样品(来源于5头活体三元杂小猪,体重均为7~15kg)由中国科学院亚热带农业生态研究生(长沙)提供。分别将5种不同的猪血清样品50.00μL加入pH=7.4的PBS缓冲溶液(4.950mL)中稀释100倍,再向猪血清溶液中加入不同浓度的L-Arg,采用DPV进行测定。
二、实验结果与分析
1、L-Arg在电极表面的电化学行为
通过循环伏安法和交流阻抗法测量以观察多肽复合膜修饰电极的组装情况,从图3可以发现在2.0mmol/L[Fe(CN)6]3-/[Fe(CN)6]4-(1:1)-10mmol/L PBS溶液中,裸金电极(AuE,a曲线)处观察到了一对十分明显的氧化还原峰,说明在裸金电极上存在快速的电子转移;与裸金电极(AuE)相比,FC-P16 Peptide/AuE电极(b曲线)处不易观察到明显的氧化还原峰。显然,当二茂铁功能化十六肽二硫代环戊烷(FC-P16-C4-DTCP,FC-P16 Peptide)组装到金电极上之后,形成的多肽复合膜阻挡了电极表面与溶液的电子传递,因此其电流值下降很大。
同时,考察了AuE、FC-P16 Peptide/AuE电极在10mmol/L,pH=7.4的PBS溶液中的循环伏安(CV)行为(图4)。如图4所示,AuE(曲线a)未有明显的氧化还原峰,FC-P16Peptide/AuE(曲线b)电极在0.45—0.55V附近有较明显的氧化还原峰,这说明多肽首端连接的二茂铁在界面上发生了氧化还原反应。
考察AuE、FC-P16 Peptide/AuE电极在含有1.0×10-5mol/L的精氨酸的10mmol/LPBS溶液(图5A)和不含精氨酸的PBS溶液(图5B)中的示差脉冲伏安(DPV)行为。由图5A可知,L-Arg在AuE电极上的还原峰不明显,说明L-Arg在AuE表面较难被还原(曲线a);而相对于AuE来说,FC-P16 Peptide/AuE电极对L-Arg的响应增强(曲线b),响应峰电流增大了,峰电位为Ep=0.32V处,说明FC-P16 Peptide/AuE电极上的多肽分子能够很好地结合L-Arg。作为对比如图5B所示,AuE在不含精氨酸的PBS溶液中未有明显的还原峰(曲线a),而FC-P16Peptide/AuE电极在0.32V附近有较明显的还原峰(曲线b),这说明多肽首端连接的二茂铁在界面上发生了还原反应。因此,可以证明二茂铁基团连接在多肽分子上。
2、线性范围和检出限
在优化的实验条件下,FC-P16 Peptide/AuE电极通过示差脉冲伏安法对不同浓度的L-Arg进行检测(图6A),从图中我们发现,当L-Arg的浓度从1.0×10-13mol/L升到1.0×10-4mol/L的过程中,响应峰电流在稳定地减小(图6B),这说明当越来越多的精氨酸结合到多肽上后,增大了金电极表面与二茂铁之间的空间位阻,电子传递效率降低。从图6C中可知,L-Arg的还原峰电流与其浓度在1.0×10-13mol/L~1.0×10-7mol/L范围内呈现出良好的线性关系,线性方程为I=0.006537lgc-0.4054,相关系数R=0.9967,检测下限为1.0×10-13mol/L;而且从图6D中可知,L-Arg的还原峰电流与其浓度在1.0×10-7mol/L~1.0×10- 4mol/L范围内也有一定的线性相关性,线性方程为I=0.02408lgc-0.2841,相关系数R=0.9656。将本发明制备的FC-P16 Peptide/AuE电极与其它文献报道的精氨酸电极进行比较(见表1),可以发现本发明所制备电极具有更加优异的性能,尤其是与无酶电极相比。
Table 1 Comparison of performance with different modified electrodes
Note:ADI:arginine deiminase;PANi:Polyaniline;SPE:screen-printedelectrode;PtE:Pt electrode;U/A:urease and arginase I;EA#14.3aptamer:96unitthiolated G-quadruples DNA;ISFET:ion-selective field effect transistor.
参考文献:
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[2]Stasyuk N,Smutok O,Gayda G,Vus B,Koval’chuk Y,Gonchar M.Bi-enzymel-arginine-selective amperometric biosensor based on ammonium-sensingpolyaniline-modified electrode[J].Biosensors&Bioelectronics,2012,37(1):46-52.
[3]Stasyuk N Y,Gayda G Z,Gonchar M V.L-Arginine-selective microbialamperometric sensor based on recombinant yeast cells over-producing humanliver arginase I[J].Sensors and Actuators B:Chemical,2014,204:515-521.
[4]Carter Z A,Kataky R.A G-quadruplex aptamer based impedimetricsensor for free lysine and arginine[J].Sensors and Actuators B:Chemical,2017,243:904-909.
[5]Sheliakina M,Arkhypova V,Soldatkin O,Saiapina O,Akata B,DzyadevychS.Urease-based ISFET biosensor for arginine determination[J].Talanta,2014,121:18-23.
4、电极的重现性、重复性
将同一批次相同条件下制备的6支多肽复合膜修饰电极检测1.0×10-8mol/L的L-Arg,得到的相对标准偏差为2.1%,说明该多肽修饰电极具有良好的重现性。采用同一支电极对1.0×10-8mol/L的L-Arg连续测定3次,得到的相对标准偏差为0.56%,说明该电极具有良好的重复性。
5、抗干扰测试
在pH=7.4的PBS(10mmol/L)为底液的三电极体系中,考察了常见氨基酸物质对FC-P16 Peptide/AuE修饰电极检测L-Arg的影响,在L-Arg(1.0×10-8mol/L)存在下加入50倍浓度的干扰组分,结果表明加入甲硫氨酸(Met)、酪氨酸(Tyr)、异亮氨酸(lle)、天冬氨酸(Asp)、谷氨酰胺(Gln)、亮氨酸(Leu)、缬氨酸(Val)、苏氨酸(Thr)、丙氨酸(Ala)、苯丙氨酸(Phe)、脯氨酸(Pro)、组氨酸(His)、谷氨酸(Glu)、甘氨酸(Gly)、赖氨酸(Lys)、色氨酸(Trp)后,修饰电极的峰电流几乎未发生明显变化,说明FC-P16 Peptide/AuE修饰电极对L-Arg具有良好的选择性。
本发明设计了对L-精氨酸(L-Arg)具有特异性结合作用的氨基酸序列,引入二茂铁探针,进而构建了基于二茂铁功能化十六肽复合膜的修饰电极(FC-P16 Peptide/AuE),该多肽复合膜修饰电极对L-Arg具有良好的选择性、重复性、重现性与低的检出限,可应用于猪血清样品中L-Arg的测定,在生命科学与营养健康方面具有重要的应用前景。
序列表
<110> 长沙理工大学
<120> 一种L-精氨酸的检测方法及传感器
<141> 2019-07-05
<160> 2
<170> SIPOSequenceListing 1.0
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Phe Gly His Ile His Glu Gly Tyr
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<211> 16
<212> PRT
<213> 未知(null)
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Gly Gly Gly Gly Phe Gly His Ile His Glu Gly Tyr Gly Gly Gly Gly
1 5 10 15
Claims (8)
1.一种L-精氨酸的检测方法,其特征在于,所述方法包括如下步骤:
(1)合成二茂铁功能化十六肽二硫代环戊烷;所述二茂铁功能化十六肽二硫代环戊烷结构式如式(I)所示:
(2)制备二茂铁功能化多肽修饰金电极:将金电极于Piranha溶液中浸泡后清洗,然后抛光,洗净并用N2吹干;将吹干后的金电极浸入二茂铁功能化十六肽二硫代环戊烷浓度为30~80μmol/L的二茂铁功能化十六肽二硫代环戊烷溶液和三(2-羧乙基)膦浓度为10~80μmol/L的磷酸盐缓冲溶液中20~30h;再将金电极于6-巯基-1-己醇浓度为0.5~2.0mmol/L的6-巯基-1-己醇溶液中浸泡2~30min后,用PBS沿着金电极表面冲洗以除去非特异性吸附的其它物质,用N2吹干即得二茂铁功能化多肽修饰金电极;
(3)以二茂铁功能化多肽修饰金电极为工作电极,以银/氯化银电极作为参比电极,以铂丝电极作为对电极,构成三电极体系;然后采用循环伏安法和示差脉冲伏安法考察不同修饰电极的电化学行为,并采用示差脉冲伏安法对不同浓度的L-精氨酸进行测试,绘制工作标准曲线,再采用标准加入法对待测样品中的L-精氨酸进行检测。
2.如权利要求1所述的方法,其特征在于,步骤(2)中对金电极表面进行抛光是分别用1.0μm、0.3μm和0.05μm的氧化铝粉对金电极表面进行抛光。
3.如权利要求1所述的方法,其特征在于,步骤(2)中所述金电极直径为3mm。
4.如权利要求1所述的方法,其特征在于,步骤(3)中是在10mmol/L磷酸盐缓冲溶液、2.0mmol/L[Fe(CN)6]4-/3--10mmol/L PBS溶液、含有1.0×10-5mol/L精氨酸的10mmol/L磷酸盐缓冲溶液中采用循环伏安法和示差脉冲伏安法考察不同修饰电极的电化学行为,采用示差脉冲伏安法对不同浓度的L-精氨酸进行电流响应与浓度关系测试;示差脉冲伏安法参数为:振幅0.05V,脉冲周期0.5s,抽样宽度0.02,脉冲宽度0.2s,循环伏安法参数为:采样间隔0.001V,扫速100mV/s。
5.一种检测L-精氨酸的传感器,特征在于,所述传感器包括作为工作电极的多肽复合膜修饰电极;所述多肽复合膜修饰电极包括金基质(5),所述金基质(5)表面修饰有多肽复合膜层(6),所述多肽复合膜层(6)中含有多肽分子(7);所述多肽分子(7)的结构式如权利要求1中的式(I)所示。
6.如权利要求5所述的传感器,其特征在于,所述多肽复合膜层(6)中还含有6-巯基-1-己醇分子(8)。
7.如权利要求5所述的传感器,其特征在于,所述金基质(5)的厚度为1.0~5.0mm,所述多肽复合膜层(6)的厚度为2~20nm。
8.如权利要求5至7任一项所述的传感器,其特征在于,所述传感器对L-精氨酸的浓度存在良好的线性关系,检测的线性范围为1.0×10-13~1.0×10-7mol/L,检出限为1.0×10- 13mol/L。
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