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Phenylalanine, Tyrosine & Tryptophan HPLC Assay Kit

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$570.00
SKU:
PNL31-H100

 Product Description

Package insert view pdf

 

 

Phenylalanine, Tyrosine & Tryptophan HPLC Assay kit:
For Research Use Only
Size:  100 Tests
Incubation Time:  15 minutes
Sample Type:  Plasma, Whole Blood, Filter Spots   
Sample Size: 50 µL

Product Developed and Manufactured in the Germany
Product Support in the USA

Intended Use

The Eagle Biosciences Phenylalanine, Tyrosine & Tryptophan HPLC Assay kit is intended for the quantitative determination of phenylalanine, tyrosine and tryptophan in whole blood, plasma and filter spots. This Phenylalanine, Tyrosine & Tryptophan HPLC Assay Kit is for research use only and is not for use in diagnostic procedures.

Assay Principle

For the determination of Phenylalanine, Tyrosine & Tryptophan a sample preparation has to be performed. Therefore a precipitation solution, containing an internal standard, is added. After centrifugation the supernatant is injected into the HPLC system. The isocratic separation via HPLC at 30°C lasts 10 minutes. The chromatograms are recorded by a UV-detector. The quantification is performed with the delivered plasma calibrator; the concentration is calculated via the “internal standard method” by integration of the peak heights respectively areas.

Whole blood and plasma

  1. Pipette 50 µl sample, CAL or CTRL into the sample vials.  The sample vials contain the internal standard dissolved in precipitation solution.
  2. Mix briefly (5 sec.) on a vortex mixer.
  3. Leave the tubes for 15 minutes at 2-8°C and centrifuge afterwards at 10.000g for 5 minutes.
  4. Inject 20 µl of the supernatant for chromatography into the HPLC-system.

Filter spots

  1. Put the filter spot (with dried patient blood) in a sample vial.  The sample vials contain the internal standard dissolved in precipitation solution.
  2. Mix briefly (5 sec.) on a vortex mixer.
  3. Incubate the tubes for 30 minutes at 37°C, leave them for 10 min in an ultrasonic bath and centrifuge afterwards at 10.000g for 5 minutes.
  4. Inject 20 µl of the supernatant for chromatography into the HPLC-system

Typical Chromatogram

phenylalanine-tyrosine-tryptophan.png

Assay Background
Phenylketonuria is the most abundant genetic disorder of the amino acid metabolism. It is characterized by a lack of phenylalanine hydroxylase which causes an increase of phenylalanine in cells and body fluids.  Due to the lack of enzyme activity, phenylalanine, normally metabolised to tyrosine, is converted to phenylpyruvic acid.  Left untreated, this can cause brain damage and progressive mental retardation. Additionally high concentrations of phenylalanine inhibit various enzymes which are involved in the synthesis of neurotransmitters and melanin. The incidence of occurrence is about 1 in 15000, varying widely in different populations.

Untreated children with phenylketonuria are normal at birth, but rather early they don´t attain early developmental milestones, show mental retardation, hyperactivity, convulsions, low pigmentation of the hair and alterations of the skin.  An early diagnosis within the first days after birth is very important. Nevertheless the blood level of phenylalanine has to be controlled regularly because reduced levels can lead to proteolysis with disturbance of growth.

The Eagle Biosciences Phenylalanine, Tyrosine & Tryptophan HPLC Assay kit makes it possible to determine phenylalanine, tyrosine and tryptophan in an easy, fast and precise method. The Phenylalanine, Tyrosine & Tryptophan HPLC Assay kit includes all reagents for preparation and separation of the samples with exception of the columns (IC4000rp) and the controls (IC4000ko). Both can be supplied by Eagle Biosciences.  Besides the complete test kits it is possible to order all components separately.  Please request our single component price list.

Conversion to molarity:

Analyte

Molecular weight

Conversion

Phenylalanine

165.2

mg/dl x 60.5 = µmol/l

Tyrosine

181.2

mg/dl x 55.2 = µmol/l

Tryptophan

204.2

mg/dl x 49.0 = µmol/l

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Eagle Biosciences, Inc.
20A NW Blvd., Suite 112
Nashua, NH 03063
Email: info@eaglebio.com
Toll Free: 866-411-8023
International: +617-419-2019

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