Quantitative Analysis of Clenbuterol in Pig Liver by LC-MS/MS/MS

Abstract

We examined the utility of LC-MS/MS/MS in the rapid quantitative analysis of clenbuterol in pig liver. Compared with LC-MS/MS in the SRM mode, which gave a chromatogram of pig liver containing interference peaks near the target component peak, LC-MS/MS/MS generated a clear chromatogram with no interference peaks. Validation of the method yielded favorable results for both LC-MS/MS in the SRM mode and LC-MS/MS/MS, with trueness values of 95.5% and 102%, repeatability of 1.9% and 5.5%, and within-laboratory reproducibility of 2.0% and 5.1%, respectively. Moreover, the recovery of clenbuterol-d 9 used as surrogate was ≥70% in both measurement methods and thus, validation was achieved in terms of both selectivity and limit of quantification. The results suggest that LC-MS/MS/MS has highly selectivity for quantitative analysis as the influence of interference peaks is suppressed, and can be used when quantification is difficult with LC-MS/MS in the SRM mode due to matrix effects.

Determination of Clenbuterol in Various Edible Parts of Livestock Products by LC-MS/MS and LC-MS/MS/MS Methods

“…In the pig kidney (Fig. 1B), a trace peak was observed around the retention time of clenbuterol; the peak area was below the calibration range (0.1-1.5 ng/mL [23]). In the pig small intestine, no peak was observed derived from the tissue matrix (Fig.…”

“…In our previous research, the original LC-MS/MS and LC-MS/MS/MS analytical methods for clenbuterol (LOQ: 0.05 ng/g), which is equivalent to the analysis method certified by the MHLW Japan were developed. Especially, the developed LC-MS/MS/MS method is highly selective and interfering peaks derived from the food sample matrices were drastically reduced [23]. As shown in the above case reports, clenbuterol poisoning was caused by the ingestion of various livestock edible parts such as the muscle and liver.…”

“…Clenbuterol was basically determined by the previously reported LC-MS/MS and LC-MS/MS/MS methods [22,23] using a tandem mass spectrometer with multi-stage fragmentation (QTRAP 4500, Sciex, Framingham, MA, USA). The target clenbuterol was separated from other compounds by a reversed-phase C18 column (Mightysil RP-18 GP, 2.0 mm i.d.…”

“…However, in the real poisoning cases of clenbuterol, the poisoning was caused by the ingestion of various parts of livestock products such as muscle and liver (theoretically, also possible by the ingestion of other edible parts). Therefore, a higher selective analysis method applicable to various food matrices is required, and we have developed an LC-MS/MS/MS method in the previous study [23]. The LC-MS/MS/MS method was successfully applied to the pig liver samples, and wider applications to various tissues obtained from different edible animals are expected.…”

Simultaneous Determination of Seven β₂-Agonists in Livestock Products Using an LC-MS/MS System

“…However, the matrices in livestock products affected the measurements of the β 2 -agonists, and a more highly-selective analytical method should be developed. Some analytical techniques involving liquid chromatography tandem mass spectrometry (LC-MS/MS) have been mainly reported [3][4][5][6][23][24][25][31][32][33][34][35][36][37][38].…”