Analysis of Monoethanolamine by Derivatization with Marfey's Reagent and HPLC

Abstract

A sensitive and selective method for determining the residual monoethanolamine in a developmental drug substance is developed and validated. Marfey's reagent, which is commonly used for the chiral analysis of amino acids, is reacted with the primary amine group of monoethanolamine and then analyzed by high-performance liquid chromatography-UV at 340 nm. Quantitation is performed by a standard addition method by preparing drug substance samples with added monoethanolamine ranging from 0.25-1.0 microg/mL (equivalent to 12.5-50 ppm with respect to the drug substance). The method performance is evaluated for linearity, specificity, detection and quantitation limits, accuracy, precision, and sample stability. The method is linear from 0.25-1.0 microg/mL with a coefficient of determination (r(2)) > 0.95. The accuracy and precision obtained is 105.5 +/- 4.8% (n = 3). The limits of detection and quantitation are 0.03 and 0.10 microg/mL, respectively. Instrument precision (% relative standard deviation of six injections of a derivatized 0.5 microg/mL monoethanolamine solution on two separate days) is >/= 2.0%. This method is suitable for the determination of monoethanolamine at the 25 ppm level in drug substance.

Structure determination of lipopeptides from Mycobacterium avium subspecies paratuberculosis and identification of antigenic lipopeptide probes

“…Tyr) and N -methyl amino acids showed poor reactivity against Marfey’s reagent. 24 We have developed an unprecedented chiral derivatizing agent 5 (Fig. 3) that allows for the determination of the absolute configurations of a wide range of amino acids including secondary amines by 1 H-NMR or HPLC.…”

A High‐Throughput Screening Assay for Amino Acid Decarboxylase Activity

“…In Figure 2 ( middle ) and ( right ) – at suboptimal conditions – full conversion to amine was not achieved. To provide a further independent confirmation of the extent of reaction, HPLC (Marfey’s reagent derivatization22) was conducted. However, this did not provide quantitative results for both substrate and product, rendering heavily tailing peaks.…”

MALDI-MS and HILIC ESI-MS/MS as Versatile Tools for Detection of Monoethanolamine Degradation Products in a Real Postcombustion Carbon Dioxide Capture Plant

“…In another approach, MEA can be derivatized by Marfey's reagent (1-fluoro-2,4-dinitrophenyl-5-L-alanine amide or FDAA) and analyzed by high-performance liquid chromatography (HPLC) with variable wavelength UV detection (HPLC−UV). 23 Then, the main disadvantage of these techniques is that they remain timeconsuming and labor intensive, with limited value for routine use in the screening of MEA byproducts. 24 Among new ionization techniques, matrix assisted laser desorption ionization (MALDI) coupled to a time-of-flight (TOF) analyzer has gained great interest, since an unlimited mass range can be examined with many advantages such as tolerance for buffers and contamination, simple spectra (most ions are singly charged), very high absolute sensitivity, rapid analysis, and relatively easy instrumentation.…”

“…In addition, if reaction conditions are not rigorously maintained, the incomplete derivatization could result in inaccurate estimations of the analytes. In another approach, MEA can be derivatized by Marfey’s reagent (1-fluoro-2,4-dinitrophenyl-5- l -alanine amide or FDAA) and analyzed by high-performance liquid chromatography (HPLC) with variable wavelength UV detection (HPLC–UV) . Then, the main disadvantage of these techniques is that they remain time-consuming and labor intensive, with limited value for routine use in the screening of MEA byproducts …”