The potency of HPLC-DAD and LC-MS/MS combined with ion chromatography for detection/purification of levulinic acid and bio-compounds from acid hydrolysis of OPEFB

• Published in: RSC advances Vol. 12; no. 44; pp. 28638 - 28646
• Main Authors: Saengsen, Chatcha, Sookbampen, Orawan, Wu, Shuke, Seetasang, Sasikarn, Rongwong, Wichitpan, Chuaboon, Litavadee
• Format: Journal Article
• Language: English
• Published: Cambridge Royal Society of Chemistry 07-10-2022; The Royal Society of Chemistry
• Subjects: Acids; Biomass; Carbohydrates; Cellulase; Cellulose; Chemistry; Chromatography; Column chromatography; Glucose; Glucosidase; High performance liquid chromatography; Hydrolysis; Hydroxymethylfurfural; Inorganic acids; Ion exchange; Levulinic acid; Ligands; Mass spectrometry; Optimization; Purification; Qualitative analysis; Stability analysis
• ISSN: 2046-2069; 2046-2069
• DOI: 10.1039/d2ra03563d

This work reports a new strategy for the detection and purification of levulinic acid (LA) and bio-compounds from the acid hydrolysis and enzymatic treatment of oil palm empty fruit bunch (OPEFB) through high-performance liquid chromatography (HPLC) techniques combined with ion/ligand chromatography. The detections of LA, biomass-saccharides, hydroxymethylfurfural (HMF), and furfural were successfully elucidated by optimizing the multiple reaction monitoring mode (MRM) and liquid chromatography conditions using a Pb
2+
ligand exchange column in the liquid chromatography with tandem mass spectrometry (LC-MS/MS) approach. High-performance liquid chromatography with diode-array detection (HPLC-DAD) combined with an H
+
ion exchange column also showed potency for detecting chromophoric compounds such as LA, HMF, furfural, and acid (by-products) but not biomass-saccharides. Both techniques showed acceptable validation in terms of linearity, limit of detection (LOD), limit of quantitation (LOQ), accuracy, precision, and stability in both quantitative and qualitative analysis. However, the LC-MS/MS approach showed higher sensitivity for detecting LA and HMF compared with HPLC-DAD. Samples comprised of cellobiose, glucose, HMF, and LA from the acid hydrolysis of cellulose to LA with a mineral acid, and the biocatalysis of cellulase and β-glucosidase catalyzed cellulose (from OPEFB) to glucose were successfully monitored through the LC-MS/MS approach. In addition, using the optimal HPLC conditions obtained from LC-MS/MS, the purification of LA from other substances obtained from the hydrolysis reaction of cellulose (5 g) was successfully demonstrated by HPLC-DAD equipped with a fraction collector combined with an H
+
ion exchange column at gram-scale of 1 g LA with a purification rate of 0.63 g ml
−1
min
−1
.
The analytical approach for detection and purification levulinic acid from and bio-compound in hydrolysis biomass.