Application Note

Analysis of Physiological Amino Acids for Clinical Diagnosis on the Hitachi L-8900 Analyzer Steve Watts, Hitachi High Technologies America, Inc.

emands for analysis of amino acids that induce 30 Gly Orn Asp

D HCys P-Ser Thr

metabolic errors continue to exist. Model L-8900, the Ser 25 Lys Ala latest generation in a long line of Amino Acid Analyzers AEC Cit

20 Tau Cysthi His Hylys

from Hitachi, is a dedicated system capable of 1Mehis g-ABA Phe Glu ASA 3Mehis Val Arg simultaneously identifying and quantifying fifty (50) 15 ASA-Anhy1 Met Cys NH3

physiological amino acids for the purpose of clinical Ile a-ABA Leu Tyr 10 PEA Trp Urea Asn Sar HCit EOHNH2 Signal Intensity (mV) Intensity Signal b-Ala

diagnosis. Unparalleled performance is achieved via a Allo-Ile Gln a-AAA b-AiBA Car Pipeco 5 ALevA Ans high-resolution 3 µm ion exchange resin, a patented Cys-HCys Pro Hypro reaction column to eliminate band broadening, and a 0

unique ninhydrin delivery system with no pre-mixing 0 20 40 60 80 100 120 required. The same system configuration can also be Time (min) used to analyze amino acids related to specific metabolic Figure 1: Chromatographic separation of 50 physiological amino acids on errors in a shortened time. the Hitachi L-8900 Amino Acid Analyzer.

30 Val Discussion 25 Acetyl-Lys

A total of 50 amino acids relevant to metabolic errors can be 20 Met Allo-Ile Ile Leu analyzed in a single acquisition using current methodology on the L- 15 Phe Tyr

8900 Amino Acid Analyzer (Figure 1). Notable amino acids include Cysthi , homocitrulline, allo-, argininosuccinic acid, 10

-homocysteine mixed disulfides, homocystine, (mV) Intensity Signal argininosuccinic acid anhydride, and aminoethylcysteine (internal 5 standard). Using the same system and column configuration, key 0 amino acids related to certain metabolic errors can be targeted and 0 2 4 6 8 10 12 14 16 18 20 22 24 26 analyzed in a shortened amount of time. For example, in only 26 Time (min) minutes, it is possible to analyze eight amino acids for simultaneous screening of Maple Syrup Urine Disease (MSUD) (, allo- Figure 2: Chromatographic separation of eight key amino acids in only 26 minutes for diagnosing a variety of metabolic disorders. isoleucine, isoleucine, and ), (PKU) ( and ), () and (). Acetyl- is used as an internal standard (Figure 2). It is also common to target only tyrosine and 20 phenylalanine for the monitoring of nutrition therapy in

Phenylketonuria patients. This can be accomplished in only 10 Tyr 15

minutes (Figure 3). Phe

10

Conclusion Signal Intensity (mV) 5 Hitachi High Technologies America continually strives to provide the most technologically advanced solutions to the life science industry. 0

One example of this dedication is the Hitachi L-8900 Amino Acid 0 1 2 3 4 5 6 7 8 9 10 Analyzer’s capability for analyzing 50 physiological amino acids that Time (min) induce metabolic errors. Since all of the above analyses employ the Figure 3: Chromatographic separation of Tyrosine and Phenylalanine in same column, buffers, and reagents, the system can be only 10 minutes for monitoring nutrition therapy in PKU patients. programmed to switch automatically among them during a series of acquisitions for optimal system flexibility. Hitachi High Technologies America, Inc. Life Sciences Division 5100 Franklin Drive Pleasanton, CA 94588 Toll Free: (800) 548-9001

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