Journal
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
Volume 51, Issue 1-2, Pages 83-90Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.ijbiomac.2012.04.005
Keywords
Poly(gamma-glutamic acid); Spectrum properties; UV spectrophotometric method; HPLC; High-throughput; Biological samples
Funding
- National Natural Science Foundation of China [21062001, 20362001]
- National 863 Research Program of China [2006AA10Z339]
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A simple and valid ultraviolet (UV) spectrophotometric method for the determination of poly(gamma-glutamic acid) is developed. The method is based on the UV absorption spectrum of gamma-PGA in aqueous solution, which exhibits a maximum absorption wavelength at 216 nm. The results obtained were comparable to those obtained with the reported high-performance liquid chromatography (HPLC) method according to ICH guidelines. Under the proposed procedure, the calibration graph is linear over the range of 20-200 mu g/ml with regression correlation coefficient of 0.9997. Precision (%R.S.D. < 1.50) and recovery (%R. > 99.29%) are good. The limit of detection (LOD) and limit of quantitation (LOQ) are 0.39 and 1.19 mu/ml, respectively. These results agree well with those of HPLC method. Its spectrum properties studies showed that the spectrum of gamma-PGA remarkably changed with an increase in temperature due to gamma-PGA was digested into glutamate monomer. In spite of this, the determining procedure could carried out in a wide temperature range (25-50 degrees C). In addition, the method is not influenced by the molecular weight, but the measurement system need to control in pH 3.0-10.0 and ionic strength not more than 0.5 M. The proposed method is applied successfully for high-throughput quantification of poly(gamma-glutamic acid) in biological samples. The advantages of the UV method are simplicity of operation, rapidity, sensitive, low-cost and high-throughput. (c) 2012 Elsevier B.V. All rights reserved.
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