Phenylalanine

Manufactured by Cambridge Isotopes

Phenylalanine is an essential amino acid used as a laboratory reagent. It is a white crystalline powder that is soluble in water and other polar solvents. Phenylalanine is commonly used in biochemical and cellular research applications.

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3 protocols using phenylalanine

Stable-Isotope Internal Standards Protocol

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The following stable-isotope internal standards were used for this study: amino acid standard mix NSK-A containing methionine, arginine, ornithine, isoleucine, and phenylalanine (Cambridge Isotope Laboratories), d3-creatine (Sigma Aldrich), and d3-5’-methylthioadenosine (13C Molecular, Fayetteville, NC).

Tang X., Keenan M.M., Wu J., Lin C.A., Dubois L., Thompson J.W., Freedland S.J., Murphy S.K, & Chi J.T. (2015). Comprehensive Profiling of Amino Acid Response Uncovers Unique Methionine-Deprived Response Dependent on Intact Creatine Biosynthesis. PLoS Genetics, 11(4), e1005158.

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Phenylalanine Kinetics in ICU Patients

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In ICU patients, arterial lines and nasogastric feeding tubes/jejunostomy catheters that were already in situ for routine clinical care, were used for infusion and sampling; no additional invasive procedures were performed. Positioning of feeding tubes was confirmed by reviewing recent radiographs, or by air insufflation and epigastric auscultation. After baseline blood samples, tracer infusion was added to ongoing enteral feeding by a separate infusion pump connected to the feeding tube by a three-way stopcock. An infusion of L-1-13 C phenylalanine (Cambridge Isotope Laboratories Inc, Tewksbury, MA; isotopic enrichment 99%, tested for chemical and isotopic purity, prepared as 10 mg/mL in aqueous solution) was started as a primed continuous infusion at a dose calculated to yield 30% enrichment of total (i.e. nutrition þ tracer) dietary phenylalanine content, with a priming bolus equivalent to a 1-h dose. Blood samples were taken every 30 min for the following 12 h.

Liebau F., Király E., Olsson D., Wernerman J, & Rooyackers O. (2021). Uptake of dietary amino acids into arterial blood during continuous enteral feeding in critically ill patients and healthy subjects. Clinical nutrition (Edinburgh, Scotland), 40(3).

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Amino Acid Standards for Metabolomic Analysis

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Amino acid standards (alanine, arginine, aspartic acid, cysteine,
glutamic acid, glycine, histidine, iso leucine, leucine, lysine, methionine,
phenyl alanine, proline, serine, threonine, tyrosine, valine, glutamine,
asparagine, GABA, citrulline, ornithine, taurine, tryptophan, 5-HTP,
norvaline, and kynurenine) were purchased from Sigma (St. Louis, MO).
Isotopically labeled amino acid standards (alanine (¹³C₃,¹⁵N), arginine (¹³C₆, ¹⁵N₄), aspartic acid (¹³C₄, ¹⁵N), cystine (¹³C₆, ¹⁵N₂), glutamic acid (¹³C₅, ¹⁵N), glycine (¹³C₂, ¹⁵N), histidine
(¹³C₆, ¹⁵N₃), iso leucine
(¹³C₆, ¹⁵N), leucine (¹³C₆, ¹⁵N), lysine (¹³C₆, ¹⁵N₂), methionine (¹³C₅, ¹⁵N), phenyl alanine (¹³C₉, ¹⁵N), proline (¹³C₅, ¹⁵N),
serine (¹³C₃, ¹⁵N), threonine (¹³C₄, ¹⁵N), tyrosine (¹³C₉, ¹⁵N), valine (¹³C₅, ¹⁵N), citrulline (d4), GABA (¹³C₄), glutamine
(¹³C₅), asparagine(¹³C₄), ornithine (d6), tryptophan (d8), kynurenine (d4)) were obtained
from Cambridge Isotope Laboratories (Andover, MA). Stock solutions
of each compound were prepared at a concentration of 500 ng/μL
and stored at −80 °C. A 10-point calibration curve (0.01–100
pmol/μL) was prepared by serial dilutions and spiked with internal
standards at a final concentration of 5 pmol/μL. Mass spectrometry-grade
formic acid was purchased from Sigma-Aldrich (St Louis, MO), and HPLC-grade
acetonitrile was purchased from Fisher Scientific (Fair Lawn, NJ).

Spain O, & Funk C. (2022). Detailed Characterization of the Cell Wall Structure and Composition of Nordic Green Microalgae. Journal of Agricultural and Food Chemistry, 70(31), 9711-9721.

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