Separation analysis of macrolide antibiotics with good performance on a positively charged C18HCE column

Abstract

The separation of basic macrolide antibiotics suffers from peak tailing and poor efficiency on traditional silica-based reversed-phase liquid chromatography columns. In this work, a C18HCE column with positively charged surface was applied to the separation of macrolides. Compared with an Acquity BEH C18 column, the C18HCE column exhibited superior performance in the aspect of peak shape and separation efficiency. The screening of mobile phase additives including formic acid, acetic acid and ammonium formate indicated that formic acid was preferable for providing symmetrical peak shapes. Moreover, the influence of formic acid content was investigated. Analysis speed and mass spectrometry compatibility were also taken into account when optimizing the separation conditions for liquid chromatography coupled with tandem mass spectrometry. The developed method was successfully utilized for the determination of macrolide residues in a honey sample. Azithromycin was chosen as the internal standard for the quantitation of spiramycin and tilmicosin, while roxithromycin was used for erythromycin, tylosin, clarithromycin, josamycin and acetylisovaleryltylosin. Good correlation coefficients (r(2) > 0.9938) for all macrolides were obtained. The intra-day and inter-day recoveries were 73.7-134.7% and 80.7-119.7% with relative standard deviations of 2.5-8.0% and 3.9-16.1%, respectively. Outstanding sensitivity with limits of quantitation (S/N ≥ 10) of 0.02-1 μg/kg and limits of detection (S/N ≥ 3) of 0.01-0.5 μg/kg were achieved.

Development and Application of an MSALL-Based Approach for the Quantitative Analysis of Linear Polyethylene Glycols in Rat Plasma by Liquid Chromatography Triple-Quadrupole/Time-of-Flight Mass Spectrometry

“…Both are suitable for hydrophilic compounds, provide higher pH stability and give better separation of PEGs and good peak shape with minimum carryover. 44 Their large pore size (300 Å) helps to avoid blockade of the pores by large molecules thereby improving column efficiency and extending column life. Low MW PEGs.…”

“…Separation of PEGs was performed on either a BEH 300 C18 column (2.1 × 50 mm 300 Å, Waters) or a Venusil ASB C8 column (2.1 × 50 mm 300 Å, Agela Technologies). Both are suitable for hydrophilic compounds, provide higher pH stability and give better separation of PEGs and good peak shape with minimum carryover . Their large pore size (300 Å) helps to avoid blockade of the pores by large molecules thereby improving column efficiency and extending column life.…”

Residue Determination in Honey

“…In addition, Canada authorities fixed an MRL in honey equal to 200 μg/kg as sum of tylosin A and B ( Table 1). The most significantly published procedures are summarized in Table 4 [31][32][33][34][35][36][37]. Lincomycin belongs to the group of lincosamides, and its activity against Paenibacillus larvae strains has been reported.…”

Synthesis of a stationary phase based on silica modified with branched octadecyl groups by Michael addition and photoinduced thiol-yne click chemistry for the separation of basic compounds

“…These extracts mainly consist of protoberberine alkaloids [32], which are always positively charged. As the a polar-copolymerized stationary phase C18HCE with positively charged surface was beneficial for the separation of alkaloids and basic macrolides [33,34], TFA was added into the mobile phase for the protoberberine alkaloids separation. 4 under the optimal chromatographic conditions, respectively.…”