Chromatography was performed on a modular HPLC system from Shimadzu (Kyoto, Japan); it contained a system controller (CBM-20A), two Nexera X2 pumps, a degasser (DGU-20ASR), and a column oven (CTO-20AC). Automated extractions were carried out with a DBS-MS 500 (CAMAG, Muttenz, Switzerland). Analytes were separated on a Shim-pack GIST (4.6 × 50 mm, 5 μm STEAROYL, 227-30017-3) analytical column (Shimadzu, Kyoto, Japan). A filter frit (KrudKatcher Ultra, Phenomenex, Torrance, CA, USA) was connected upstream to the analytical column. Mobile phase A consisted of water plus 0.1% formic acid and 2 mM ammonia fluoride, while methanol supplemented with 0.1% formic acid and 2 mM ammonia fluoride was used as mobile phase B. The following stepwise gradient was applied: 40% A (0–1.0 min), 40–90% A (1.0–2.0 min), 90% A (2.0–3.0 min), and 40% A (3.01–4.0 min). The flow rate was set at 1.0 mL/min at 40 °C. The HPLC liquid stream was connected to an 8060 tandem mass spectrometer (Shimadzu, Kyoto, Japan). The mass transitions and compound specific settings were included in Table S1 .
Dbs ms 500
Manufactured by CAMAG
Sourced in Switzerland
The DBS-MS 500 is a laboratory equipment designed for the analysis of dried blood spot (DBS) samples. It provides an automated solution for the extraction and analysis of analytes from DBS samples using mass spectrometry (MS) technology. The core function of the DBS-MS 500 is to facilitate the processing and analysis of DBS samples, enabling researchers and laboratories to efficiently and accurately measure the presence and concentration of various compounds in dried blood.
Automatically generated - may contain errors
Lab products found in correlation
Modular hplc system, by Shimadzu (1 mentions)
Cto 20ac, by Shimadzu (1 mentions)
Shim pack gist, by Shimadzu (1 mentions)
Krudkatcher ultra, by Phenomenex (1 mentions)
Cbm 20a, by Shimadzu (1 mentions)
Exionlc system, by AB Sciex (1 mentions)
Triple quad 6500 mass spectrometer, by AB Sciex (1 mentions)
Neobase non derivatized msms kit, by PerkinElmer (1 mentions)
4 protocols using dbs ms 500
1
Quantitative Analysis of Compounds by HPLC-MS/MS
2
Multivitamin Analysis in Dried Blood
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The vitamin B concentrations in whole blood were gathered by the dry blood spot method (DBS). The blood samples were dropped on a special filter paper card, dried, and stored separately in a Ziploc bag with a desiccant. DBS samples were stored at −20 °C for one month before being returned to the Institute of Nutrition and Health, Wuhan University of Science and Technology. DBS samples were detected for discoloration or the absence of desiccant before analysis. Multivitamins in DBS were detected using the method of Lin Y et al. [21 ]. This method can detect multivitamins in the blood in a short time and solve the technical defects in the current detection methods of multivitamins. Dry blood spot samples were treated with a DBS-MS 500 (CAMAG, Muttenz, Switzerland). It can automatically pick up DBS cards, extract them after the identification check, and then measure them by liquid chromatography–tandem mass spectrometry. All samples were analyzed using a SCIEX ExionLC system coupled with a SCIEX Triple Quad 6500 mass spectrometer. Individuals with extreme values of vitamin B concentrations (VitB1 > 100 ng/mL, VitB2 > 70 ng/mL, and VitB6 > 90 ng/mL) were excluded.
3
DBS-MS for Tramadol and Metabolite Analysis
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A DBS-MS 500 (CAMAG, Switzerland) was connected as the front end autosampler to a Shimadzu (Kyoto, Japan) LC-MS system, see Figure 1. The extraction solvent for the DBS elution consisted of a water-methanol mixture (90/10,v/v). The elution solvent and the chromatographic conditions were optimized to achieve baseline separation and nearly symmetric peak shape for both analytes, requiring higher quantities of aqueous solution to resolve O-desmethyltramadol nicely. Each DBS card was This article is protected by copyright. All rights reserved.
photographed with a built-in camera before and after the extraction process, to document the samples, check for the presence of a blood spot, to center the extraction head position, and to verify where the extraction took place. Methanolic internal standard solution (20 μL spray volume, containing 25 ng/mL cis-tramadol-13 C, D3, and 10 ng/mL O-Desmethyl-cis-tramadol-D6) was sprayed in a homogenous layer of 1 cm 2 over the center of each spot (flow rate of 120 μL/min). Afterward, the internal standard solution was dried for 60 s at room temperature. In contrast to mixing the internal standard into the extraction solvent, this procedure enables compensating for extraction differences (recovery bias) 8, (link)9 (link) . The extraction of a 4.
photographed with a built-in camera before and after the extraction process, to document the samples, check for the presence of a blood spot, to center the extraction head position, and to verify where the extraction took place. Methanolic internal standard solution (20 μL spray volume, containing 25 ng/mL cis-tramadol-13 C, D3, and 10 ng/mL O-Desmethyl-cis-tramadol-D6) was sprayed in a homogenous layer of 1 cm 2 over the center of each spot (flow rate of 120 μL/min). Afterward, the internal standard solution was dried for 60 s at room temperature. In contrast to mixing the internal standard into the extraction solvent, this procedure enables compensating for extraction differences (recovery bias) 8, (link)9 (link) . The extraction of a 4.
4
Validation of Newborn Screening DBS Assays
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Two commercial DBS assay kits: (1) MassChrom Amino Acids and Acylcarnitines from Dried Blood/ Non-derivatised (Chromsystems Instruments
& Chemicals GmBH, Gräfelfing, Germany); and
(2) NeoBase Non-derivatized MSMS kit (with succinylacetone assay; PerkinElmer, Waltham [MA], US) were validated for use in the study. In addition to a manual puncher and an autopuncher for DBS preparation, a fully automated online extraction system (DBS-MS 500; CAMAG, Muttenz, Switzerland) was also evaluated. The precision and local reference intervals of the commercial assay kits are listed in Table 1. Our laboratory has participated in the Newborn Screening Quality Assurance Programme organised by the US Centers for Disease Control and Prevention (CDC) since 2011. The disease panel included in the study is shown in Table 2. 8, 10, 11 Step 6: Reporting Chemical pathologists were responsible for reporting of positive results to the paediatricians. The CDC cut-off for clinical decision (https://wwwn.cdc.gov/ NSQAP/Restricted/CDCCutOffs.aspx) and the Region 4 Stork Collaborative Project (https://www. clir-r4s.org/) data interpretation tools were applied during interpretation of the results.
& Chemicals GmBH, Gräfelfing, Germany); and
(2) NeoBase Non-derivatized MSMS kit (with succinylacetone assay; PerkinElmer, Waltham [MA], US) were validated for use in the study. In addition to a manual puncher and an autopuncher for DBS preparation, a fully automated online extraction system (DBS-MS 500; CAMAG, Muttenz, Switzerland) was also evaluated. The precision and local reference intervals of the commercial assay kits are listed in Table 1. Our laboratory has participated in the Newborn Screening Quality Assurance Programme organised by the US Centers for Disease Control and Prevention (CDC) since 2011. The disease panel included in the study is shown in Table 2. 8, 10, 11 Step 6: Reporting Chemical pathologists were responsible for reporting of positive results to the paediatricians. The CDC cut-off for clinical decision (https://wwwn.cdc.gov/ NSQAP/Restricted/CDCCutOffs.aspx) and the Region 4 Stork Collaborative Project (https://www. clir-r4s.org/) data interpretation tools were applied during interpretation of the results.
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