Spb 1 capillary column

Manufactured by Merck Group

Sourced in United States

The SPB-1 capillary column is a type of gas chromatography column manufactured by Merck Group. It is designed for the separation and analysis of a wide range of organic compounds. The column features a nonpolar stationary phase, which allows for the separation of compounds based on their boiling points and interactions with the stationary phase.

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Lab products found in correlation

18 protocols using spb 1 capillary column

PCB Analysis by GC-μECD

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Analysis of PCBs was performed in the splitless mode on an Agilent 7890 gas chromatograph equipped with a ⁶³Ni micro electron capture detector (µECD) and a SPB-1 capillary column (length, 60 m; inner diameter, 250 μm; and 0.25 µm film thickness; Supelco, St. Louis, MO, United States) (Wu et al., 2013 (link)). The following temperature program was used based on a published method (Kania-Korwel et al., 2011 (link)): hold at 80°C for 1 min, 20°C/min to 215°C, then 0.1°C/min to 219.5°C, and 20°C/min to 280°C, then hold at 280°C for 3 min. A constant helium flow rate of 1 ml/min was used for all analyses. The injector and detector temperatures were 250°C and 300°C, respectively. The ⁶³Ni-μECD used for the PCB analysis was linear up to concentrations of 1,000 ng/ml for all analytes investigated (R² > 0.999). The recovery of PCB 117 was 83 ± 7% (range: 71–97%). A detailed summary of the limits of detection, limits of quantification, and background levels of the analyzed PCBs is presented in Supplementary Table S1.

Yaghoobi B., Miller G.W., Holland E.B., Li X., Harvey D., Li S., Lehmler H.J., Pessah I.N, & Lein P.J. (2022). Ryanodine receptor-active non-dioxin-like polychlorinated biphenyls cause neurobehavioral deficits in larval zebrafish. Frontiers in Toxicology, 4, 947795.

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GC-μECD Analysis of PCBs and OH-PCBs

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Quantitative analysis of PCBs and OH-PCBs (as methylated derivatives) in sample extracts was carried out on an Agilent 7890A gas chromatography (GC) equipped with an SPB-1 capillary column (60 m length, 250 μm inner diameter, 0.25 μm film thickness; Supelco, St Louis, MO, USA) and a ^{63 (link)}Ni-micro electron capture detector (μECD) as previously reported.^{39 (link), 40 (link)} Helium was used as carrier gas with a constant flow rate of 2 mL/min. The temperature program was as follows: initial temperature 50 °C for 1 min, 30 °C/min to 200 °C, 1 °C/min to 250 °C, 10°C/min to 280 °C, and hold for 3 min. The PCBs were identified based on their retention time, with relative retention times (RRT) being within 0.5 % of the RRT of the respective PCBs standard.⁴⁵ PCBs were quantified with the internal standard using the relative response factors in both samples and the reference standard mixture. PCB levels were corrected for the recovery of the surrogate recovery standard to account for any loss of PCBs during the extraction. The hydroxylated PCB metabolites (as methylated derivatives) listed in Table S1 were not detected in any sample.

Zhang C.Y., Flor S., Ludewig G, & Lehmler H.J. (2020). Atropselective Partitioning of Polychlorinated Biphenyls in a HepG2 Cell Culture System: Experimental and Modeling Results. Environmental science & technology, 54(21), 13817-13827.

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Quantification of Hydroxylated PCB Metabolites

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The levels of hydroxylated
PCB metabolites in the concentrated sample extracts were quantified
on an Agilent 7890A gas chromatograph with a ⁶³Ni microelectron
capture detector (μECD) (Agilent, Santa Clara, California) and
an SPB-1 capillary column (60 m length, 250 μm inner diameter,
0.25 μm film thickness; Supelco, St Louis, Missouri) using the
internal standard method as described previously.^{32 (link)} OH-PCB levels, adjusted for the microsomal protein content,
are presented in Table S2.

Lehmler H.J., Uwimana E., Dean L.E., Kovalchuk N., Zhang Q.Y, & Ding X. (2022). Probing the Role of CYP2 Enzymes in the Atropselective Metabolism of Polychlorinated Biphenyls Using Liver Microsomes from Transgenic Mouse Models. Chemical Research in Toxicology, 35(12), 2310-2323.

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Fecal SCFA Quantification by GC

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Fecal short chain fatty acids (SCFAs) quantification was performed by gas chromatography. Concentrations of acetic, propionic, iso-butyric, butyric and iso-valeric acids were assessed as previously described [60 (link)].
Briefly, analyses were performed using a Varian model 3400 CX Gas chromatograph fitted with FID detector, split/splitless injector and a SPB-1 capillary column (30 m × 0.32 mm ID, 0.25 μm film thickness; Supelco, Bellefonte, PA, USA). Quantification of the SCFAs was obtained through calibration curves of acetic, propionic, iso-butyric, butyric, and iso-valeric acid in concentrations between 0.25 and 10 mM (10 mM 2-ethylbutyric acid as internal standard). Results are expressed as mg/g of wet weight of feces.

Ceccarani C., Bassanini G., Montanari C., Casiraghi M.C., Ottaviano E., Morace G., Biasucci G., Paci S., Borghi E, & Verduci E. (2020). Proteobacteria Overgrowth and Butyrate-Producing Taxa Depletion in the Gut Microbiota of Glycogen Storage Disease Type 1 Patients. Metabolites, 10(4), 133.

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Cholesterol Quantification by GC-FID

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This analysis was performed using a Varian (Palo Alto, CA) model 3300 gas chromatograph fitted with a flame ionization detector, split/splitless injector, and an SPB-1 capillary column (30 m × 0.32 mm i.d., 0.25 μm film thickness; Supelco, Bellefonte, PA) and helium as the carrier gas (2 mL/min). Injector and detector temperatures were 250 and 300°C, respectively. The initial oven temperature was 50°C and was increased by 20°C/min to 230°C, then by 4°C/min, and then held at 250°C for 25 min. Data processing was performed using AZUR V5.0 software (Analytical Technology, Brugherio, Italy). The concentration of cholesterol was calculated using the internal standard method. All assays were repeated in triplicate.

Albano C., Morandi S., Silvetti T., Casiraghi M.C., Manini F, & Brasca M. (2018). Lactic acid bacteria with cholesterol-lowering properties for dairy applications: In vitro and in situ activity. Journal of dairy science, 101(12).

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Quantitative Analysis of PCB-153 and 3-OH-PCB-153

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Quantitative analyses of PCB-153 and 3-OH-PCB 153 in sample extracts were carried out on a 7890A gas chromatography (GC) system (Agilent) equipped with a SPB®-1 capillary column (

60 m \times 250 μ m \times 0.25 μ m

film thickness; Supelco) and a ⁶³Ni-micro electron capture detector (Agilent) as previously reported (Wu et al. 2011 (link)), with minor modifications and described below. Helium was used as carrier gas with a constant flow rate of

2 mL / min

. The injector and detector temperatures were

240 ° C

and

300 ° C

, respectively. The column temperature program was initially set as

50 ° C

, held for 1 min, then gradually increased to

200 ° C

30 ° C / min

, increased to

250 ° C

1 ° C / min

, increased to a final temperature of

280 ° C

10 ° C / min

, and held there for 3 min. The PCB-153 and 3-OH-PCB-153 were identified by the retention time of their authentic standards. The relative retention time (RRT) of all analytes was within 0.5% of the RRT of the respective standard. PCB 153 and 3-OH-PCB 153 were quantified with the internal standard method as described (Kania-Korwel et al. 2007 (link)). Levels were corrected for recoveries below 100%.

Egusquiza R.J., Ambrosio M.E., Wang S.G., Kay K.M., Zhang C., Lehmler H.J, & Blumberg B. (2020). Evaluating the Role of the Steroid and Xenobiotic Receptor (SXR/PXR) in PCB-153 Metabolism and Protection against Associated Adverse Effects during Perinatal and Chronic Exposure in Mice. Environmental Health Perspectives, 128(4), 047011.

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Quantification of Hydroxylated PCB Metabolites

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The levels of hydroxylated PCB metabolites in the concentrated sample extracts were quantified on an Agilent 7890A gas chromatograph with a ⁶³Ni-micro electron capture detector (μECD) (Agilent, Santa Clara, California, USA) and an SPB-1 capillary column (60 m length, 250 μm inner diameter, 0.25 μm film thickness; Supelco, St Louis, MO, USA) using the internal standard method, as described.^{32 (link)} OH-PCB levels, adjusted for the microsomal protein content, are presented in Table S2.

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Quantification of Hydroxylated PCB 91 Metabolites

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To determine the levels of hydroxylated PCB 91 metabolites, sample extracts were analyzed on an Agilent 7890A gas chromatograph with a ⁶³Ni-micro electron capture detector (μECD) and a SPB-1 capillary column (60 m length, 250 μm inner diameter, 0.25 μm film thickness; Supelco, St. Louis, MO, USA) as reported earlier.^{43 (link)} PCB 204 was added as internal standard (volume corrector) prior to GC analysis. PCB 91 metabolites, as the corresponding methylated derivatives, were quantified based on their respective relative response factors as described previously.^{40 (link), 44 (link)} The average RRTs of the metabolites, calculated relative to PCB 204, were within 0.5% of the RRT for the respective standard.⁴² Formation rates of OH-PCBs are presented adjusted for nmol P450 or microsomal protein content (Tables S3–S6).

Uwimana E., Li X, & Lehmler H.J. (2018). HUMAN LIVER MICROSOMES ATROPSELECTIVELY METABOLIZE 2,2′,3,4′,6-PENTACHLOROBIPHENYL (PCB 91) TO A 1,2-SHIFT PRODUCT AS THE MAJOR METABOLITE. Environmental science & technology, 52(10), 6000-6008.

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Quantification of Fecal SCFAs and Calprotectin

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We performed short chain fatty acids (SCFAs) and calprotectin quantification from stool samples.
Concentrations of acetic, propionic, iso-butyric, butyric, and iso-valeric acids were assessed by gas liquid chromatography in accordance with the method proposed by Weaver et al. (1989 (link)) with slight modifications described in Borgo et al. (2017 (link)). Analyses were performed using a Varian model 3400 CX Gas-chromatograph fitted with FID detector, split/splitless injector and a SPB-1 capillary column (30 m × 0.32 mm ID, 0.25 μm film thickness; Supelco, Bellefonte, PA, USA). Results are expressed as mg/g of wet weight of feces. Quantification of the SCFAs was obtained through calibration curves of acetic, propionic, iso-butyric, butyric, and iso-valeric acid in concentrations between 0.25 and 10 mM (10 mM 2-ethylbutyric acid as internal standard). SCFA data on the same cohort have been previously described in Verduci et al. (2018 (link)).
Fecal calprotectin concentrations were measured by a commercial ELISA kit (Calprotectin ELISA Kit, Immundiagnostik, Bensheim, Germany), according to manufacturer instructions.

Bassanini G., Ceccarani C., Borgo F., Severgnini M., Rovelli V., Morace G., Verduci E, & Borghi E. (2019). Phenylketonuria Diet Promotes Shifts in Firmicutes Populations. Frontiers in Cellular and Infection Microbiology, 9, 101.

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GC-μECD Analysis of Metabolite Levels

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Sample
extracts were analyzed on an Agilent 7890A gas chromatograph with
a ⁶³Ni-micro electron capture detector (GC-μECD)
and a SPB-1 capillary column (60 m length, 250 μm inner diameter,
0.25 μm film thickness; Supelco, St Louis, MO) as reported previously.^{28 (link)} Both the inlet and detector temperatures were
set to 250 °C. The initial oven temperature was 50 °C, held
for 1 min, and then increased by 30 °C/min until it reached 200
°C. The temperature increased by 1 °C/min until 250 °C,
then by 10 °C/min to a final temperature of 280 °C. The
injector was operated in the splitless mode. Since authentic standards
of the metabolites were not available, relative metabolite levels
are presented as area of the major metabolite relative to the area
of the internal standard (PCB 204). The RRTs of all metabolites, calculated
relative to PCB 204, were within 0.5% of the average RRT for the respective
metabolite.³³

Uwimana E., Maiers A., Li X, & Lehmler H.J. (2016). Microsomal Metabolism of Prochiral Polychlorinated Biphenyls Results in the Enantioselective Formation of Chiral Metabolites. Environmental Science & Technology, 51(3), 1820-1829.

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