An efficient immunoaffinity chromatographic method for extraction and purification of papaverine from samples of pericarpium papaveris and food products

Abstract

A highly selective immunoaffinity column was obtained by coupling anti-papaverine polyclonal antibodies to CNBr-activated Sepharose 4B. It was found that the coupling efficiency and performance of the immunoaffinity column were greatly improved by prolonging the coupling reaction time from 3 h at 20 degrees C with shaking to incubation overnight at 4 degrees C after the 3 h shaking reaction. The pH and ionic strength were observed to be the most important factors that influence the binding of papaverine to the immunoaffinity column. Using 0.01 mol/L phosphate buffered saline (PBS, pH 8.3) and methanol-water (80:20, v/v) as the loading and eluting solutions, respectively, papaverine was first retained on the column and then quantitatively eluted out with a mean recovery of 86% at a loading concentration of 1 microg/mL. When applied to real samples of pericarpium papaveris and food products, the established immunoaffinity column showed high efficiency in removing the matrix interferences in the samples and satisfactory recovery results were obtained. The method was useful for extraction and purification of papaverine from related samples.

Novel immunoassay and rapid immunoaffinity chromatography method for the detection and selective extraction of naringin in Citrus aurantium

“…Purification or extraction of natural bioactive compounds based on IAC is a more concise, cheaper, environmentally friendly, and time‐saving strategy. Recently, we developed a potential method for the separation of glycyrrhizic acid and 20 ( R / S ) gisenoside epimers . In this study, the water extract of Citrus aurantium (CA‐EXT) was directly loaded onto an IAC after filtering, and then the column was washed with DW and eluted with 0.1 M glycine‐HCl buffer (pH 2.7) containing 0.5 M NaCl.…”

“…Furthermore, it is environmentally friendly because no organic reagents are used in the extraction process. According to current studies, IAC has been applied to the isolation of proteins and small molecules, e.g., the purification of a‐momorcharin from bitter melon seeds and extraction of papaverine from samples of pericarpium papaveris . We have also resolved 20( S )‐ginsenoside Rg2 (GsRg2) and 20( R )‐GsRg2 using IAC based on anti‐GsRg2 mAbs , and we have separated glycyrrhizic acid (GA) from Glycyrrhiza uralensis .…”

Amantadine‐functionalized magnetic microspheres and stable isotope labeled internal standards for reducing matrix effect in determination of five opium alkaloids by liquid chromatography‐quadrupole linear ion trap mass spectrometry

“…The proposed methodology was compared with other representative techniques reported in recent years for the analysis of opium alkaloids in food samples. As can be seen in Table , the reported techniques mainly are the couples of LLE, SPE, QuEChERS, and MSPE with GC, GC‐MS, HPLC, and LC‐MS/MS . Though some HPLC methods without sample pretreatment gave good recovery, the chromatographic column and instrument were easily polluted because the extraction solution contained a number of impurities .…”

“…In 2011, the National Health and Family Planning Commission of the People’s Republic of China (RPC) announced a list of foods that may be illegally added with nonedible substances or abuse food additives and took illegal addition of poppy husk as a key monitoring project . Opium alkaloids are generally detected by gas chromatography (GC), gas chromatography coupled to mass spectrometry (GC‐MS), high‐performance liquid chromatography (HPLC) using ultraviolet or fluorescence detectors, and liquid chromatography tandem mass spectrometry (LC‐MS/MS) . Of these, (LC‐MS/MS) with multiple reaction monitoring (MRM) provides more reliable, quantitative data of the analytes at very low concentrations than other detection technologies, and thus it has become the preferred technique for the analysis of opium alkaloids.…”

“…In order to reduce the influence of the ME, several effective methods have been proposed, including sample pretreatment, matrix‐matched standards, stable isotope‐labeled internal standards, and optimum chromatographic conditions . Until now, pretreatment of opium alkaloids in samples has been carried out mainly by liquid–liquid extraction (LLE), quick, easy, cheap, effective, rugged, safe (QuEChERS), solid‐phase extraction (SPE) using an immunoaffinity column as well as mixed‐mode cation exchange sorbents, and magnetic solid‐phase extraction (MSPE) . Among these, LLE is complex to operate and uses large quantities of highly toxic organic reagents (about 200 mL) .…”