LC-MS/MS Method for the Determination and Quantitation of Penicillin G and Its Metabolites in Citrus Fruits Affected by Huanglongbing

Abstract

In this study, we developed and validated a method for the extraction, identification, and quantitation of penicillin G and its metabolites (penilloic acid and penillic acid) in a variety of citrus fruits by employing sequential liquid/liquid and solid-phase extraction techniques in conjunction with UHPLC-MS/MS. Two product ion transitions per analyte were required for identification, which contributes to a high degree of selectivity. Corrected recoveries of penicillin G using an isotopically labeled internal standard were 90-100% at fortification levels of 0.1, 0.25, 1, and 10 ng/g. Absolute recoveries for penillic acid and penilloic acid were 50-75% depending on the matrix used. The limit of detection (LOD) of penicillin G and its metabolites was found to be 0.1 ng/g when 2 g of citrus was extracted. This method is useful in determining residue levels of penicillin G and its metabolites in citrus trees infected with huanglongbing bacteria after antibiotic treatment.

Dissipation and residue determination of penicillin G and its two metabolites in citrus under field conditions by DSPE/UPLC–MS/MS

“…The recoveries and LODs in this study were similar to those for other food matrices. Specially, our method has a higher LOD compared with the method for citrus in a previous study (Aldeek et al, 2015); however, the minimum recovery of the proposed method is higher than the one in that study. In addition, DSPE is simpler, economical and convenient in preparation compared with SPE, and suitable for large sample detection.…”

“…However, citrus growth is often interfered with by various diseases, such as Huanglongbing (HLB) and ulcer disease, resulting in lower quality and huge economic losses. HLB, in particular, known as the “cancer” of citrus and caused by bacteria in Candidatus Liberibacter, has become a devastating disease that threatens citrus trees worldwide (Aldeek et al, 2015). Some researches suggest that HLB can be effectively prevented and controlled by a variety of antibiotics (e.g.…”

“…Penicillin G (Benzylpenicillin) is a β ‐lactam antibiotic with a thiazolidine ring, prone to opening under the conditions of acid, base, oxidizing agents, nucleophiles, ultraviolet light, heat or polar solvents (e.g. water and alcohol), and its metabolites are mainly penillic acid and penilloic acid (Aldeek et al, 2015, 2016; Li et al, 2008). These two metabolites have no antibacterial activity, but are associated with about 3–5% of penicillin‐allergic patients (Blaha, Knevel, Kessler, Mincy, & Hem, 1976).…”

“…LC–MS/MS is the most sensitive and widely used among these methods. Aldeek et al developed and validated a method integrating solid‐phase extraction (SPE) and ultrahigh‐performance liquid chromatography (UPLC)–MS/MS for the extraction, purification and detection of penicillin G and its metabolites (penillic acid and penilloic acid) from citrus fruits and orange juice (Aldeek et al, 2015, 2017; Canzani, Hsieh, Standland, Hammack, & Aldeek, 2017).…”

Penicillin G's function, metabolites, allergy, and resistance.

“…Using the Orbitrap and triple-quadrupole based UHPLC-MS/MS platforms, we analyzed a citrus fruit sample taken from a HLB infected tree that was treated with penicillin G; the presence of the metabolites penillic acid, isopenillic acid, penilloic acid, and penicilloic acid were confirmed. Penicillin G was not found in the citrus sample, however this was anticipated, as environmental conditions on the growing field, such as heat, UV radiation, and acidic conditions of the fruit would contribute heavily to degradation after treatment [65,66].…”