A kind of fmoc-protected amino acid purity and related substances analysis method
A technology of related substances and analysis methods, applied in the field of analysis and detection of Fmoc-protected amino acid purity related substances, can solve problems such as irreversible damage to chromatographic columns, baseline drift, system pipeline blockage, etc., and achieve the effect of avoiding sample decomposition or instrument corrosion
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Embodiment 1
[0049] A method for analyzing the purity of Fmoc-Gly-OH and related substances, comprising the following steps:
[0050] S1. Set chromatographic conditions
[0051] Instrument: high performance liquid chromatography
[0052] Column: C 18 5μm 4.6×250mm
[0053] Detector: UV, 220-260nm
[0054] Flow rate: 0.5-1.0ml / min
[0055] Column temperature: 20-30°C
[0056] Injection volume: 5-15ul
[0057] Gradient elution conditions:
[0058]
[0059] S2, prepare mobile phase
[0060] Mobile phase A: trifluoroacetic acid: water = 0.2-1: 1000; mobile phase B: trifluoroacetic acid: acetonitrile = 0.2-1: 1000;
[0061] S3, prepare Fmoc-Osu impurity control solution
[0062] Accurately weigh the relevant impurity Fmoc-Osu standard, put it in a volumetric flask, prepare a 0.5-2.0% impurity solution with acetonitrile, shake well for later use, accurately weigh the Fmoc-Gly-OH standard, and put it in a volumetric flask , dissolved in acetonitrile, and then added impurity solution...
Embodiment 2
[0089] A method for analyzing Fmoc-Phe-OH purity and related substances, comprising the following steps:
[0090] S1. Set chromatographic conditions
[0091] Instrument: high performance liquid chromatography
[0092] Column: C 18 5μm 4.6×250mm
[0093] Detector: UV, 220-260nm
[0094] Flow rate: 0.5-1.2ml / min
[0095] Column temperature: 20-30°C
[0096] Injection volume: 5-20ul
[0097] Gradient elution conditions:
[0098]
[0099] S2, prepare mobile phase
[0100] Mobile phase A: trifluoroacetic acid: water = 0.2-1: 1000; mobile phase B: trifluoroacetic acid: acetonitrile = 0.2-1: 1000;
[0101] S3, prepare Fmoc-Osu impurity control solution
[0102] Accurately weigh the relevant impurity Fmoc-Osu standard, put it in a volumetric flask, prepare a 0.5-2.0% impurity solution with acetonitrile, shake well for later use, and accurately weigh the Fmoc-Phe-OH standard, and put it in a volumetric flask , dissolved in acetonitrile, and then added impurity solution, s...
Embodiment 3
[0128] A Fmoc-Lys(Boc)-OH purity and related substance analysis method, comprising the following steps:
[0129] S1. Set chromatographic conditions
[0130] Instrument: high performance liquid chromatography
[0131] Column: C 18 5μm 4.6×250mm
[0132] Detector: UV, 220-260nm
[0133] Flow rate: 0.5-1.2ml / min
[0134] Column temperature: 20-30°C
[0135] Injection volume: 5-20ul
[0136] Gradient elution conditions:
[0137]
[0138]
[0139] S2, prepare mobile phase
[0140] Mobile phase A: trifluoroacetic acid: water = 0.2-1: 1000; mobile phase B: trifluoroacetic acid: acetonitrile = 0.2-1: 1000;
[0141] S3, prepare Fmoc-Osu impurity control solution
[0142] Accurately weigh the relevant impurity Fmoc-Osu standard product, place it in a volumetric flask, prepare a 0.5-2.0% impurity solution with acetonitrile, shake it up for later use, and accurately weigh the Fmoc-Lys(Boc)-OH standard product, place it in In the volumetric flask, dissolve with acetonitrile,...
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