Method for detecting purity of 5-aminolevulinic acid hydrochloride (5-ALA)

Abstract

The invention discloses a method for detecting the purity of 5-aminolevulinic acid hydrochloride (5-ALA). The method comprises the following information and related steps that a mobile phase A (MPA) is a 0.2% heptafluorobutyric acid aqueous solution, and a mobile phase B (MPB) is a 0.2% heptafluorobutyric acid acetonitrile solution; a chromatographic column adopts octadecylsilane chemically bonded silica as a filler for separation; preparing of a sample solvent is carried out, wherein water/acetonitrile in a ratio of 95/5 as the sample solvent, wherein the solvent is also used as a blank solution; preparation of a system applicability solution is carried out, wherein 100 mg of a 5-ALA system applicability reference substance and 5 ml of a sample solvent are uniformly mixed, and the sample is diluted with the sample solvent to a scale; preparing of a sample solution is carried out, namely preparing is performed by using a sample instead of a 5-ALA system applicability reference substance under the same conditions, subsequently the solutions are injected into a high performance liquid chromatograph according to a sample injection sequence, and a chromatogram of the solution under the wavelength of 265 nm is measured. Through the mode, compared with an existing USP method, the method has the advantages of being high in principal component retention, good in chromatographic peak shape and higher in detection accuracy, and is more suitable for purity analysis of the compound.

Description

technical field [0001] The invention relates to the technical field of chromatographic analysis, in particular to a detection method for the purity of 5-aminolevulinic acid hydrochloride (5-ALA). Background technique [0002] The small molecular compound 5-aminolevulinic acid hydrochloride (5-ALA) is a prefix compound of tetrahydropyrrole, which is an essential substance for organisms to synthesize chlorophyll, heme, vitamin B12, etc., and is widely used in the field of agriculture. It has important applications in medicine, feed industry, cosmetics industry and organic synthesis intermediates. For the above-mentioned small molecule amino acid purity analysis, the conventional analysis methods in the prior art are thin layer chromatography, pre-column / post-column Derivatization and, in some specific cases, direct use of HPLC. [0003] Thin-layer chromatography (TLC) method is easy to operate, easy to develop color, and has a fast development speed, but the specificity of TL...

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Problems solved by technology

[0007] This kind of direct use of high performance liquid chromatography to determine the related substances of small molecule amino acids will have a big hidden danger. Since amino acids are zwitterionic compounds, within the pH range that conventional chromatographic columns can tolerate, there are often two This configuration is likely to be reflected in the chromatogram as two peaks that can be transformed into each other, which brings difficulty to the analysis; on the other hand, having a suitable retention factor K (usually 1-20) is a basic requirement for the development of analytical methods. Requirements, for 5-ALA, this USP method has almost no retention of the compound, the retention factor is only 1.0, so the specificity of the detection method cannot be guaranteed

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