Service Introduction

Bio-enzyme catalyzed reaction has the advantages of high specificity, mild conditions, green and environmental protection, etc., and has been more and more widely used in the production process of APIs. According to the type of reaction catalyzed by enzymes, the International Enzyme Commission (IE.C) classifies enzymes into 6 categories: transferases, hydrolases, cleavage enzymes, oxidoreductases, isomerases and ligases. The enzyme catalysts used in the preparation of AP| are process impurities and need to be controlled for residual amounts.

• Protein Residues

Protein content determination methods include Kjeldahl method, Folinol method (Lowry method), bis(acetonitrile) method, BCA method and Caulmers Brilliant Blue method (Bradford method), of which Caulmers Brilliant Blue method is favored because of the highest sensitivity and not easy to be interfered. This method can determine the amount of protein from 1 to 200 μg.

• DNA Residue

CTI adopts fluorescent staining method to detect the DNA residues of fermentation microorganisms in APIs.

• Sugar Residues

Whether it is starch, polysaccharides or oligosaccharides, the hydrolysis method can be used to convert sugars into monosaccharides, and the HPLC-CAD method can be used to detect the residues of sugars.

Service Background

Recently, CDE is concerned about the residues of enzymes, starch and DNA in the catalytic reaction during the new drug review of enzyme-catalyzed synthesized APIs. CTI Huatest has established the analytical methods for the residues of 3 components (DNA, protein and sugar).

Applicable Products

Applicable to enzyme-catalyzed, or fermentation-prepared APIs. Such as Vildagliptin, Norethindrone Hydrochloride, Atorvastatin Calcium, Rhodopsin, etc.

Service Content

Residue Detection and Methodology Validation of 3 Ingredients (DNA, Protein, Sugar) in APIs.

According to the Chinese Pharmacopoeia 2020 Edition IV “9101 Guiding Principles for Validation of Analytical Methods” for analytical methods and validation performance indicators, CTI performed methodological validation according to the requirements of impurity quantitative analytical methods. Parameters examined include specificity, linearity/range, accuracy, precision (repeatability/intermediate precision), limit of detection and limit of quantification, and solution stability.

The sensitivity of the method is such that it can fulfill most of the application scenarios.

Specialty Services

For API soluble in water, only 2~3g of sample is required for single method detection, while 20~30g of sample is required for method development and validation.