An Overview of the Kjeldahl Method of Nitrogen Determination. Part II. Sample Preparation, Working Scale, Instrumental Finish, and Quality Control

Abstract

The Kjeldahl method was introduced in 1883 and consists of three main steps: sample digestion, distillation, and ammonia determination (titration being the primary method). The Kjeldahl method uses sulfuric acid, a variety of catalysts, and salts to convert organically bound nitrogen in samples to ammonium with its subsequent measurement (Sáez-Plaza et al., 2013). Today, this method is universally accepted and used in tens of thousands of laboratories throughout the world for nitrogen analysis in a wide variety of materials, such as foods, beverages, agricultural products, environmental samples, chemicals, biochemicals, and pharmaceuticals. However, successful analysis requires proper sampling and sample handling, which depend on the type of material. The Kjeldahl method has been validated and standardized for total (crude) protein estimation for a wide variety of food matrices, indirectly determined by their nitrogen content, and is the reference method adopted by many international organizations. The Kjeldahl procedure has several variants, based mainly on a sample size and apparatus required. A number of rapid and accurate instrumental methods have been gradually introduced that have some advantages compared to older techniques, if a large number of samples are to be run. Thus, extracted nitrogen from Kjeldahl can be determined by several other methods, i.e., spectrophotometric, potentiometric with ion selective electrode, FIA, ion chromatographic, and chemiluminescent methods. Quality control is essential for accurate and precise measurements of nitrogen by the Kjeldahl method. The importance of quality control in Kjeldahl analysis is stressed in this review. Despite some negative factors (i.e., it is hazardous, lengthy, and labor intensive), the Kjeldahl method and its variants with instrumental finish remain as accurate and reliable methods.

Keywords:

• Kjeldahl method
• sample preparation
• working scale
• nitrogen speciation
• instrumental finish
• quality control

Acknowledgments

The authors gratefully acknowledge the financial support granted by the Junta de Andalucía (Spain) through grant excellence research project P06-FQM-02029. Grateful thanks are also due to Drs. María Teresa Morales Millán and Diego Luis García González for providing us with the literature concerning JAOAC.