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    Home > Biochemistry News > Biotechnology News > Amino acid determination problems.

    Amino acid determination problems.

    • Last Update: 2020-10-27
    • Source: Internet
    • Author: User
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    : Please ask me now to measure
    amino acids
    (only intended to determine the three amino acids of Tianmen dongine and glutamate and alanine): is now used in the Isuzu liquid phase, LC-10AT pump and SPD-10A detector,
    chromatography
    column ods C18 column (250 X 4. 6mm), detection wavelength 254nm, column temperature 40cc, flow rate lmL.min, flow phase A:0. 1mol. L- , sodium acetate solution (pH6. 5) Acetylene (93:7), flow phase B: acetylene a water (8o:20), and the following for gradient purgation. Linear gradient escape procedure: 0. 0 min, 0%B; 13 min, 7% B; 23 min.
    23% B; 29 min, 35% B; 35 min, 40%B; 40 min, 100% B; 45 min, 100% B; 47 min, 0%B.
    the blank gradient baseline drifts, and there are large, wide peaks for more than 20 minutes, how about the same time and voltage for each gradient? I have been gradient washing all night the same ah, please ask me this method can determine amino acids?
    : AccQ derivative method, I have done a similar, for reference:
    chromatography conditions and system adaptability test with octane silane bonded silicone as a filler; After 1000 ml of water dissolved, add 2 ml of triethylamine, mix well, adjust the pH to 4.95 with phosphoric acid, filtered, that is, acetylene (90:10) as the flow phase;
    1.0ml AccQ Flour TM 2B to AccQ Flour TM 2A bottle, 50C
    heating
    5 to 10min dissolves the derivative.
    derivative amount of 10 sl for test solution,
    control product
    solution is placed in the derivative tube, plus AccQ Flour TM Borate buffer (boric acid buffer) 70ul, vortex stirring, in constant stirring add derivative 20ul, closed, heated at 55 oC 5min or room temperature 45min.
    measurement method to take this product, precision called the appropriate amount, water dissolved and diluted into a solution containing about 100ug per 1 ml, precision amount of 10 μl injection liquid chromatography, record chromatography. Another take glycine control product appropriate amount, precision weighing, the same method of determination. Based on the external standard method, the peak area is calculated, that is, the result.
    note: AccQ Flour TM 2A
    Reagent
    ; AccQ Flour TM 2B Reagent; AccQ Flour TM Borate Buffer Reagents are provided by Waters.
    : If I remember correctly, the three amino acids you want to measure do not seem to have more obvious characteristic absorption peaks at ultraviolet and visible wavelengths, you should consider using the pre-bar or post-bar derived HPLC method.
    .
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