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预制的基质辅助激光解吸/电离靶板,涂有薄基质层,用于以高密度阵列形式自动进行样品沉积。

Ready-made matrix-assisted laser desorption/ionization target plates coated with thin matrix layer for automated sample deposition in high-density array format.

作者信息

Miliotis Tasso, Kjellström Sven, Nilsson Johan, Laurell Thomas, Edholm Lars-Erik, Marko-Varga György

机构信息

Department of Analytical Chemistry, Lund University, Box 124, SE-221 00 Lund, Sweden.

出版信息

Rapid Commun Mass Spectrom. 2002;16(2):117-26. doi: 10.1002/rcm.542.

Abstract

The methodology for ready-made matrix-assisted laser desorption/ionization (MALDI) target plates covered with an optimized thin layer consisting of matrix and nitrocellulose has been developed. Piezoelectric microdispensing enabled sample depositions in a high-density array format of 2000 sample depositions on a conventionally sized target plate (45 x 47 mm). The sample depositions were made reproducibly in a fully automated mode by using an in-house developed computer-controlled piezoelectric flow-through microdispenser. Additionally, the piezoelectric technique facilitated significant analyte enrichment that increased the detection sensitivity. The MS signal was obtained rapidly, generally within ten laser pulses. An airbrush device was used to generate a fine spray of matrix and nitrocellulose dissolved in acetone. The acetone evaporated instantly when reaching the target plate leaving the entire surface with a thin and uniform matrix/nitrocellulose coating consisting of very small crystals of matrix embedded in the nitrocellulose. These crystals acted as a seed-layer on subsequent analyte depositions, rendering homogeneous sample spots when using alpha-cyano-4-hydroxycinnamic acid (CHCA) as matrix. The relative standard deviation of the signal intensity between spots was (20-30)% (n = 30). The detection sensitivity was improved by restricting the sample spot diameter to 300 microm. The spot size was affected by the deposition rate and the evaporation rate of the dispensed sample volume. Mass spectra of a 25-amol peptide mixture deposition were successfully recorded.

摘要

已开发出一种用于预制基质辅助激光解吸/电离(MALDI)靶板的方法,该靶板覆盖有由基质和硝酸纤维素组成的优化薄层。压电微分配技术能够以高密度阵列形式在常规尺寸的靶板(45×47毫米)上进行2000次样品沉积。通过使用内部开发的计算机控制压电流通式微分配器,以全自动模式可重复地进行样品沉积。此外,压电技术有助于显著富集分析物,从而提高检测灵敏度。质谱信号通常在十个激光脉冲内快速获得。使用喷枪装置产生溶解在丙酮中的基质和硝酸纤维素的细喷雾。丙酮到达靶板时立即蒸发,使整个表面留下由嵌入硝酸纤维素中的非常小的基质晶体组成的薄而均匀的基质/硝酸纤维素涂层。这些晶体在随后的分析物沉积中充当种子层,当使用α-氰基-4-羟基肉桂酸(CHCA)作为基质时,可产生均匀的样品斑点。斑点之间信号强度的相对标准偏差为(20-30)%(n = 30)。通过将样品斑点直径限制为300微米提高了检测灵敏度。斑点大小受分配样品体积的沉积速率和蒸发速率影响。成功记录了25阿托摩尔肽混合物沉积的质谱。

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