Symmetry c18 analytical column

Manufactured by Waters Corporation

Sourced in Spain, Belgium, United States

The Symmetry C18 analytical column is a reversed-phase high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of organic compounds. The column features a chemically bonded C18 stationary phase that provides efficient separation and high-resolution chromatography.

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Lab products found in correlation

1260 infinity system, by Agilent Technologies (2 mentions) 6120 quadrupole mass spectrometer, by Agilent Technologies (2 mentions) Thermo ltq orbitrap mass spectrometer, by Thermo Fisher Scientific (1 mentions)

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C18 column (384 citations) Symmetry C18 column (256 citations) Symmetry C18 (133 citations) C18 analytical column (17 citations)

7 protocols using symmetry c18 analytical column

Flavonoid Extraction and Analysis from Wild-type and Transgenic Plants

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Extraction of flavonoids from shoots of wild-type plants and rol gene transgenics (4-months old) was carried out according to the reported procedure [51 (link)]. Qualitative and quantitative analysis of flavonoids was carried out using a Waters Acuity ^TM HPLC–DAD system (Waters, Spain) attached to a symmetry C-18 analytical column with dimensions of 5 μm, 3.9 × 150 mm (Waters, Spain). The wavelength was adjusted to 235–450 nm, and pressure applied was 200 psi. Separation was achieved using a mobile phase of acetonitrile with 0.5 % formic acid (A) and water with 0.5 % formic acid (B) running at a flow rate of 1 ml/min, with the following gradient (t (min), %B): (0, 95) (15, 65) (18, 10) (22, 95). The injection volume was 10 μl and retention time was 27 min. Peaks in extracts were identified by comparison with retention indices of reference standards. The analytes were detected at wavelengths specific for each metabolite with a particular retention time (Table 4).Table 4

Retention time of studied flavonoids with wavelength

S. No	Standard	Wavelength (nm)	Retention time (min)
1	Apigenin	325	20.2
2	Caffeic acid	325	8.7
3	Catechin	279	10.7
4	Isoquercetin	355	11.1
5	Quercetin	370	15.1
6	Rutin	355	10.8

Dilshad E., Ismail H., Haq I.U., Cusido R.M., Palazon J., Ramirez-Estrada K, & Mirza B. (2016). Rol genes enhance the biosynthesis of antioxidants in Artemisia carvifolia Buch. BMC Plant Biology, 16, 125.

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Profiling Bacterial Metabolites by LC-MS

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For the detection of secondary metabolites in the culture supernatants, bacteria were collected from 96-h cultures by centrifugation at 7,000 × g for 10 min and the supernatants after filtration on 0.2-μm filters were analyzed by LC-MS on an Agilent 1260 Infinity system coupled to an Agilent 6120 quadrupole mass spectrometer. The conditions for high-performance liquid chromatography (HPLC) separation were as follows: a Symmetry C₁₈ analytical column (5 μm, 4.6 mm by 150 mm; Waters Corporation) was used in positive mode, the injection volume was 100 μL, mobile phase A was 0.1% trifluoroacetic acid in water, mobile phase B was acetonitrile, and the linear gradient was 0% mobile phase B–100% mobile phase A at 0 min to 80% mobile phase B–20% mobile phase A at 40 min. UV detection was performed at 210 nm. During separation, the flow rate was 0.7 mL/min, and the column temperature was 37°C.

Lanois-Nouri A., Pantel L., Fu J., Houard J., Ogier J.C., Polikanov Y.S., Racine E., Wang H., Gaudriault S., Givaudan A, & Gualtieri M. (2022). The Odilorhabdin Antibiotic Biosynthetic Cluster and Acetyltransferase Self-Resistance Locus Are Niche and Species Specific. mBio, 13(1), e02826-21.

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LC-MS Metabolite Identification Protocol

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Sample analysis and metabolite identification were carried out on a Thermo LTQ-Orbitrap mass spectrometer (Thermo Fisher Scientific, Waltham, MA) interfaced with a 3 Ti high-performance LC pump and CTC PAL autosampler (LEAP Technologies, Carrboro, NC). The analytes were separated on a Waters Symmetry C18 analytical column (5 μm particle size, 2.1 × 150 mm; Waters, Milford, MA) with a 35-min gradient elution method. The mobile phases consisted of (A) 10 mM ammonium formate in MS-grade water and (B) MS-grade acetonitrile. The sample aliquots were eluted at a flow rate of 0.25 mL/min with 10% B over 5 min. Mobile phase (B) was gradually increased to 90% over 24 min. The column was then returned to 10% B and held for 3 min before the next injection.

Forkuo A.D., Ansah C., Pearson D., Gertsch W., Cirello A., Amaral A., Spear J., Wright C.W, & Rynn C. (2017). Identification of cryptolepine metabolites in rat and human hepatocytes and metabolism and pharmacokinetics of cryptolepine in Sprague Dawley rats. BMC Pharmacology & Toxicology, 18, 84.

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Production of Secondary Metabolites

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We investigated the production of secondary metabolites in complemented strains, by incubating 100-mL broth cultures of ΔxcnKL::odl1 (MCS5) and ΔxcnKL::odl1 (p15A-P_tet-odl-BGC-mob) strains with gentamicin and aTc at 28°C for 78 h, centrifuging the cultures, and passing the supernatants through 0.2-μm filters and concentrating them 10-fold for LC-MS analysis. Natural odilorhabdins (NOSO-95A, -B, and -C) and xenocoumacin 1 (Xcn 1) were identified in culture supernatants by LC-MS on an Agilent 1260 Infinity system coupled to an Agilent 6120 quadrupole mass spectrometer. The conditions for HPLC separation were as follows: a Symmetry C₁₈ analytical column (5 μm, 4.6 mm by 150 mm; Waters Corporation) was used in positive mode, the injection volume was 100 μL, mobile phase A was 0.1% trifluoroacetic acid in water, mobile phase B was acetonitrile, and the linear gradient was 0% mobile phase B–100% mobile phase A at 0 min to 80% mobile phase B–20% mobile phase A at 40 min. UV detection was performed at 210 nm. During separation, the flow rate was 0.7 mL/min, and the column temperature was 37°C.

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Mycotoxin Analysis by LC-MS/MS

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Liquid chromatography-tandem quadrupole mass spectrometer (LC-MS/MS) analysis was conducted, based on an ultra-high performance liquid chromatography (UHPLC) system (Waters ACQUITY, Milford, MA, USA), equipped with a symmetry guard column (3.5 μm, 10 × 2.1 mm) (Waters, Zellik, Belgium) and a symmetry C₁₈ analytical column (5 μm, 150 × 2.1 mm (Waters)). The sample was analyzed for 23 different mycotoxins, using tandem mass spectrometry (MS/MS) with a Quattro Premier™ XE tandem quadrupole mass spectrometer (Waters, Milford, MA, USA). The matrix-matched calibration curves fitted by linear regression showed a coefficient of determination (R²), ranging from 0.951 to 0.999. The average recoveries for all the mycotoxins ranged from 80% to 111%. The intraday and interday precision ranged from 3–14% and 6–20%, respectively. The LOD values were 0.5 µg·kg⁻¹ (for AFB1, AFB2, AFG1, and AFG2), 13 µg·kg⁻¹ (FB1), 17 µg·kg⁻¹ (FB2), and 53 µg·kg⁻¹ (FB3). The LOQs were 1.5 µg·kg⁻¹ for each AF, whereas the values were 26, 35, and 105 µg·kg⁻¹ for FB1, FB2, and FB3, respectively. The detailed analytical conditions were described by [31 (link),53 (link)].

Phan L.T., Tran T.M., De Boevre M., Jacxsens L., Eeckhout M, & De Saeger S. (2021). Impact of Season, Region, and Traditional Agricultural Practices on Aflatoxins and Fumonisins Contamination in the Rice Chain in the Mekong Delta, Vietnam. Toxins, 13(9), 667.

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Non-volatile Component Analysis by HPLC

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Four kinds of non-volatile components were analyzed by HPLC using an E2695 LC instrument (Waters Instruments Co., MA, USA) with an online degasser, a quaternary pump, a 2489 ultraviolet-visible detector (UVD), and an autosampler. Approximately 10 µL of the HPLC samples was analyzed with a Symmetry C-18 analytical column (250 mm × 4.6 mm, 5 µm, Waters Corp., USA) using a linear gradient with waterphosphoric acid (100:0.05, v/v) as solvent A and acetonitrile as solvent B. The gradient program was as follows: 10% B in 0-8 minutes, 10% to 30% B in 8-22 minutes, 30% to 85% B in 23-36 minutes, and 85% to 10% B in 37-66 minutes. The velocity of flow was 1.0 mL min -1 , and the column temperature was kept at 30 °C. Detection was performed at a wavelength of 220 nm in 0-25 minutes and 210 nm in 26-66 minutes.

Chang Y., Lin J., Pan S., Jing Y., Ailing G., & Deng Y. (2021). Effects of Different Drying Methods on the Contents of Nine Components and Immunomodulatory Activities of Four Components in Osmamthus fragrans Flowers. Natural Product Communications, 16(2), 1934578X2199616-1934578X2199616.

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Quantitative HPLC Analysis of Zolpidem in Nanospheres

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High-performance liquid chromatography (HPLC; Model 600 pump with UV-Vis detector; Waters, Milford, MA) was utilized for quantitative analysis of zolpidem in samples. A reverse-phase Symmetry C18 analytical column (15 x 4.6 mm, 5 µm particles; Waters) with a precolumn was used for elution; samples (20 µL) were injected and analyzed at 245 nm. The mobile phase consisted of a methanol-deionized water mixture (75:25), and a flow rate of 0.8 mL/min. A calibration curve was prepared at 245 nm using methanol as the solvent, and linearity was obtained. The zolpidem concentration contained within the nanospheres was determined by dissolving a known amount of nanospheres in 10 mL of dichloromethane, 13 (link) vortexing for 1 min, and filtering before HPLC analysis. The results obtained were expressed as encapsulation efficiency, expressed as (recovered mmol zolpidem/g nanospheres)/(loaded mmol zolpidem/g polymer) * 100.

Al-Dhubiab B.E. (2016). In vitro and in vivo evaluation of nano-based films for buccal delivery of zolpidem. Brazilian oral research, 30(1).

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