Method for separating and detecting D-p-hydroxyphenylglycine and enantiomer thereof

Abstract

The invention discloses a method for separating and detecting D-p-hydroxyphenylglycine and an enantiomer thereof. The method comprises: step 1, carrying out a derivatization reaction through adoptionof a derivatization reagent and D-p-hydroxyphenylglycine at a certain reaction temperature in a reaction solvent to prepare a derivatized product; and step 2, analyzing the derivatized product by using normal-phase high performance liquid chromatography, and separating and detecting derivatized D-p-hydroxyphenylglycine and an enantiomer thereof. The derivatization reagent is one of di-tert-butyl dicarbonate, 9-fluorenylmethyl chloroformate, and benzyl chloroformate, and the high performance liquid chromatography takes a normal-phase chromatographic column as a separation column. According to the method, D-p-hydroxyphenylglycine and an enantiomer derivative thereof can be efficiently separated, the baseline separation is achieved, and the separation degree is more than 1.5. The baseline issmooth and steady, and the peak pattern is good. The method is great in the specialization and high in detection sensitivity and is beneficial for fast accurately detecting the content of the enantiomer in D-p-hydroxyphenylglycine.

Description

technical field [0001] The invention belongs to the technical field of drug analysis, and in particular relates to a method for separating and detecting D-p-hydroxyphenylglycine and its enantiomers. Background technique [0002] D-Hydroxyphenylglycine can be used in the production of β-lactam semi-synthetic antibiotics, and the main drugs produced with it include cephalexin, cephradine, cefaclor, amoxicillin (amoxicillin), cefoperazone, etc. These The drug has a wide range of uses, and it has killing effects on Gram-positive bacteria, Gram-negative bacteria, Toxoplasma, spirochetes, etc. Its antibacterial ability has surpassed penicillin, and it is more and more widely used in clinical practice. D-p-Hydroxyphenylglycine is also used in the photosensitive field and as an analytical reagent for iron, phosphorus, silicon, etc. Therefore, D-p-hydroxyphenylglycine is an important chemical, chemical and pharmaceutical intermediate, and also an important chiral source for the synt...

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Problems solved by technology

Ha Jing et al. used cyclodextrin and hydroxypropyl β-cyclodextrin as chiral mobile phase additives to study the resolution of D,L-p-hydroxyphenylglycine in a reversed-phase high-performance liquid chromatography system, and established a β-cyclodextrin -Cyclodextrin is a chiral mobile phase additive method for separating p-hydroxyphenylglycine enantiomers, using ODS column (250mm×4.6mm), with V (methanol-β cyclodextrin): V (pH 4.5 phosphate Buffer)=30:70 is the mobile phase, the flow rate is 0.2mL / min, the column temperature is 25°C, and the detection wavelength is 230nm; but this method still has some disadvantages, adding chiral additives to the mobile phase, the consumption of additives is large, and the analytical method High cost and long system equilibration time

Shi Junqing et al studied the resolution of 8 commercial chiral chromatographic columns for D,L-p-hydroxyphenylglycine, the results showed that OD, AD, IA, AS, IB, (S,S)-whelk-O1 and QN-AX 7 D, L-p-Hydroxyphenylglycine could not be resolved successfully by any chiral column, and racemic p-Hydroxyphenylglycine could only be separated on the Teicoplanin stationary phase Chirobitotic T column, but this column is not an ordinary experiment Commonly used chromatographic columns in laboratories, and the chromatographic columns are expensive and rarely used, which limits the wide application of this method

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