Using the Shapiro–Wilk test, all data were assessed for normality. One-way ANOVA (for multiple groups) and student's t-test (for two groups) were employed for comparisons (SPSS 26.0). P < 0.05 was considered statistically significant. Pearson's test was employed for correlation analyses. Graphs were generated using GraphPad Prism 8.0. Fatty acid profile data were processed using Quant-My-Way (Agilent Technologies). All data were expressed as the mean ± standard error of the mean (SEM).
Quant my way
Manufactured by Agilent Technologies
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Quant-My-Way is a versatile laboratory instrument that enables precise quantification of a wide range of analytes. It utilizes advanced techniques to provide accurate and reliable measurement capabilities.
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4 protocols using quant my way
1
Fatty Acid Analysis Protocol
2
Fatty Acid Profiling of Broiler Samples
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In total, 0.1 g of the samples was extracted in chloroform and dichloromethane mixture. Fatty acid profiles were analyzed using the Agilent gas chromatography system (Agilent 7820; Agilent Technologies, USA) with the CP-Sil 88 gas chromatographic column (100 m × 0.25 mm × 0.25 μm, Agilent, USA) selected based on the properties of different compounds, 1 μL injection volume, 10:1 split ratio, and high-purity helium as the carrier gas at a flow rate of 1.0 mL/min. Initially, the temperature of the column was maintained at 100 °C for 5.0 min. Then, it was gradually increased to 240 °C at 4 °C/min for 15 min. Subsequently, a mass spectrometry system (Agilent 5977; Agilent Technologies, USA) was used to identify and quantify the fatty acids. Data were acquired using MassHunter GC/MS Acquisition (Agilent Technologies) and analyzed using Quant-My-Way (Agilent Technologies). The detection of fatty acid composition in broilers was completed by Sanshu Biotechnology Co., Ltd.
3
Oxylipin Profiling by UPLC-MS/MS
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Oxylipin profiling was performed using UPLC with an Agilent Ultivo QQQ MS system coupled to an Agilent 1290 Infinity II UPLC system (Agilent, Santa Clara, CA, USA). Chromatographic separation of the oxylipins was achieved using a gradient of water, methanol, and acetonitrile, all with 0.1% acetic acid (v/v). The acquisition parameters were as previously described [32 (link)] with minor modifications, and the MS data were used for quantification. Surrogate analytes and internal and external standards were used to monitor extraction efficiency and ensure accurate quantitation with standard curves. The acquired data were quantified using Quant-My-Way (Agilent, Santa Clara, CA, USA) using 9 isotope-labeled internal standards. Here we report the data for oxylipins with >80% of values above the limit of detection (43 of 62 oxylipins) and for 4 PUFAs: arachidonic acid (ARA), linoleic acid (LA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) (Cayman Chemical, Ann Arbor, MI, USA). UPLC was performed in 2 separate batches, ensuring that repeat measures across time for the same participant were all included in the same batch.
4
Quantifying Oxylipins by LC-MS/MS
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The PGE2, 5-HETE, 12-HETE, and 20-HETE quantification was performed on an Agilent Ultivo QQQ MS system coupled to an Agilent 1290 Infinity II UPLC system (Agilent, Santa Clara, CA, USA). Chromatographic separation of oxylipins was achieved using a gradient of water, methanol, and acetonitrile all with 0.1% acetic acid (v/v). Acquisition parameters were as previously described with minor modifications [20 (link)]. The acquired data were quantified by Quant-My-Way (Agilent, Santa Clara, CA, USA) using calibration curves.
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