Simultaneous detection of 15 antibiotic growth promoters in bovine muscle, blood and urine by UPLC-MS/MS

<p>An analytical method was established for the rapid detection of antibiotic growth promoters (AGPs) in bovine muscle, and bovine blood and bovine urine, using ultra high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). After the addition of an aqueous solution of EDTA-Na₂, the pH of bovine urine samples was directly adjusted to 5.2 by acetic acid-ammonium acetate and purified by HLB solid-phase extraction cartridge; bovine muscle and bovine blood samples processing were extracted with acetonitrile (ACN) and ACNwater (90:10; v/v) without any purification step. The samples were then centrifuged, concentrated and analysed by UPLC-MS/MS on an ACQUITY UPLC® BEH C18 column using gradient elution. The developed method was validated and mean recovery percentages at three spiked levels were 74–119%, 76–115% and 76–119%, respectively, in bovine muscle, bovine blood, and bovine urine. The relative standard deviation (RSD) ranged from 1.0% to 14.7% in spiked bovine muscle, bovine blood and bovine urine. The limits of detection (LOD) of all analytes were in the ranges 0.11–3.82 µg kg⁻¹, 0.10–2.49 µg kg⁻¹ and 0.06–4.53 µg kg⁻¹ in bovine muscle, bovine blood, and bovine urine, respectively. The method was sensitive, accurate and was applied to monitor real samples. To the best of our knowledge, this is first method available for simultaneous determination of several classes of APGs in bovine muscle, and bovine blood and bovine urine.</p

The results of 55 patients detected by MLPA.

<p>A total of 43 cases (78.2%) showed 22q11.2 heterozygous deletion, of whom 40 (93.0%) exhibited typical 3-Mb deletion, while 3 (7.0%) showed proximal 1.5-Mb deletion; no case was found having atypical deletion on 22q11.2. Only 1 case (1.8%) had 3-Mb duplication. None of the chromosomal abnormalities in the MLPA kit were found in the other 11 patients.</p

Data of MLPA analysis with P250-B1 DiGeorge kit.

<p>The four graphs represent four patients’ data analyzed by MLPA. In each graph, the spots represent MLPA probes, the upper green line indicates a peak ratio of 1.3 and any probes above this line represent a duplication, the lower green line indicates a peak ratio of 0.75 and any probes below this line represents a deletion, and the probes between the two lines are considered as normal two copies. (<b>A</b>): A patient with 22q11.2 deletion spanning 3Mb TDR (red spots). (<b>B</b>) A patient with 22q11.2 deletion spanning proximal 1.5Mb (red spots) within TDR. (<b>C</b>): A patient with 22q11.2 duplication (red spots) mapping to 3Mb TDR. (<b>D</b>) A patient with normal copy probes.</p

Characteristic face of VCFS.

<p>These Chinese VCFS patients all presented with a characteristic face, consisting of vertically long face, narrow palpebral fissures, fleshy nose with a broad nasal root, flattened malar region, retrognathia, and sometimes overfolded helix (E) or cup-shaped ear (F).</p

Summary of patient data.

<p>M, male; F, female; CHD, congenital heart disease; +, presence of symptoms; −, absence of symptoms or no CVNs in MLPA detection; del, deletion; dup, duplication.</p

Number of patients presenting with major clinical features of VCFS.

<p>All cases with characteristic faces exhibited 22q11.2 heterozygous deletions.</p

Surgical features stratified by group.

<p>OT = operative time; WIT = Warm ischemia time; LOS =  length of stay; EBL =  estimated blood loss.</p

Multivariate analyses of selected outcomes.

<p>Multivariate analyses of selected outcomes.</p

Demographic and clinical characteristics of patients.

<p>VS  =  versus; MI-OPN =  mini-incision open partial nephrectomy; OPN =  open partial nephrectomy; LPN =  laparoscopic partial nephrectomy; BMI =  body mass index; eGFR  =  estimated glomerular filtration rate; SD =  standard deviation.</p

The supra-12th rib mini-flank approach for managing renal lesions.

<p>Renal clear cell carcinoma in the upper pole of the right side (A, D, G): The length of incision after MI-OPN (A); The length of the lesion after MI-OPN (D); Coronal reconstruction demonstrates a renal mass in the upper pole of the right kidney (G). Renal angiomyolipoma in the left side (B, E, H): The length of incision after MI-OPN (B); The length of the lesion after MI-OPN (E); Axial computed tomography (CT) scan shows a mass in the left kidney with low-density dark areas (H). Duplex kidney in the right side (C, F, I): C The length of incision after MI-OPN (C); The length of the lesion after MI-OPN (F); Coronal reconstruction demonstrates the right duplex kidney (I).</p