Sciex triple quad 5500

LC-MS/MS analyses were performed on a SCIEX Triple Quad 5500 mass spectrometer (SCIEX, Concord, ON, Canada) equipped with Turbo Ion Spray interface. The HPLC systems consisted of an CTC PAL autosampler (LEAP Technologies, Morrisville, NC, USA) equipped with a model 1290 binary pump (Agilent, Santa Clara, CA, USA). All instruments were controlled and synchronized by SCIEX Analyst software (version 1.6.3). Mobile phases compositions were 0.1% formic acid in water (mobile phase A) and 0.1% formic acid in acetonitrile (mobile phase B). The gradient for PFAS was maintained at 10% B for 0.8 min, followed by a 0.6-min linear increase to 95% B, and kept at 95% B for 4 min before a linear decrease to 10% in 0.1 min. The column was equilibrated at 5% B for 0.5 min before injecting samples. The total run time for each injection was 6 min. The chromatographic separation was conducted on a Phenomenex Kinetex C18 100Å 30 × 2.1 mm column (Torrance, CA, USA) with a flow rate of 0.5 mL/min. The injection volume for each sample was 5 µL. Quadrupoles Q1 and Q3 were set on unit resolution and mass over charge (m/z) of the analytes and are shown in Table S2. Multiple-reaction-monitoring (MRM) mode using specific precursor/product ion transitions was used for quantification. Data processing was performed using SCIEX Analyst software (version 1.6.3).

ME was analyzed using liquid chromatography–electrospray ionization-tandem mass spectrometry (LC−ESI−MS/MS) to recognize the phenolic and flavonoid compounds in the plant extract. The separation system was carried out using Exion LC AC and SCIEX Triple Quad 5500 + MS/MS, and the ionization was performed using an electrospray ionization (ESI).

Analytes were measured using UPLC-MS/MS, which was equipped with an LC-30A ultra-performance liquid chromatography (Shimadzu, Kyoto, Japan) and Sciex Triple Quad 5,500 tandem triple quadrupole mass spectrometer (AB Sciex, Framingham, MA, United States). Chromatographic separation was performed using an XSelect HSS T3 column (2.1 mm × 100 mm, 2.5 µm, Waters, Milford, MA, United States) at 40°C by gradient elution. The mobile phase consisted of water (A) and acetonitrile (0.1% formic acid, B). The elution procedure was as follows: 0–1.0 min, 50% B; 1.0–1.5 min, 50%→98% B; 1.5–3.0 min, 98% B; 3.0–3.1 min, 98%→50% B; 3.1–4.1 min, 50% B. The flow rate was 0.35 mL min⁻¹, and the injection volume was 6 µL.
The positive ion mode with multiple reaction detection was used. The monitored ion pairs were m/z 526.5→72.2 for almonertinib, 468.3→255.3 for sorafenib-d₃, 437.3→145.0 for rivaroxaban, 460.3→443.4 for apixaban and 440.4→145.0 for rivaroxaban-d₄ (Figure 2). The mass spectrometer conditions, including delustering potential (DP) and collision energy (CE) of the compounds, are shown in Table 1. Other parameters of the mass spectrometer were as follows: ion source gas 1, 60.0 psi; ion source gas 2, 50.0 psi; curtain gas, 25.0 psi; source temperature, 500°C; ion spray voltage, 5,500 V.

An Agilent 1290 UPLC system was coupled with an Agilent 6530C Q-TOF-MS/MS (Agilent Technologies, USA); A Waters ACQUITYTM I-Class UPLC (Waters Technologies, USA) was coupled with a SCIEX Triple Quad 5,500 (AB SCIEX, USA); an XS105 analytical balance was purchased from METTLER TOLEDO (Shanghai, China); a KQ-800VSM Bench ultrasonic cleaner was purchased from Kunshan Ultrasonic Instrument Co., Ltd. (Jiangsu, China); LC-MS-grade acetonitrile was purchased from Merck KGaA (Darmstadt, Germany); HPLC-grade formic acid was purchased from Kermel (Tianjin, China); ultra-pure water was purchased from Watsons (Guangzhou, China).
A total of 31 reference standards (adenosine, guanosine, chlorogenic acid, loganin, caffeic acid, schaffetaside, rutin, forsythoside A, ferulic acid, sibemine, fritillin B, isochlorogenic acid A/B/C, quercitrin, rosmarinic acid, salvianolic acid B, liquiritin, lindenaza, baicalin, genistein, forsythin, liquiritigenin, mullein isoflavones, bergamot esters, baicalein, formononetin, glycyrrhizic acid, glycyrrhetinic acid, wogonin, and ligustilide) were purchased from the National Institutes for Food and Drug Control, Jiangsu Yongjian Pharmaceutical Technology, Ltd., and Shandong WoDeSen Bioscience Technology, Ltd. The purities of all the reference standards were over 96.0%. Samples of Jb. L were produced by Shandong Hongjitang Pharmaceutical Group, Ltd.

hiPSC-Heps were incubated in William’s E Medium supplemented with the Hepatocyte Maintenance Supplement Pack (CM4000; Gibco; Durham, NC, USA) containing a cocktail of CYP probe substrates (phenacetin 50 µM (for CYP1A2), diclofenac 5 µM (for CYP2C9), mephenytoin 50 µM (for CYP2C19), bufuralol 10 µM (for CYP2D6), midazolam 5 µM (for CYP3A)) at 37 °C. After 60 min incubation, the incubation media were collected and samples were kept at − 80 °C until performing LC–MS/MS analyses. The formed metabolites of CYP probe cocktail (acetaminophen, 4′-hydroxydiclofenac, 4′-hydroxy-S-mephenytoin, 1′-hydroxybufuralol and 1′-hydroxymidazolam, respectively) were quantified with the LC–MS/MS (Prominence Ultra-Fast Liquid Chromatography (Shimadzu, Kyoto, Japan)/SCIEX Triple Quad 5500 + (AB Sciex LLC, Framingham, MA)).

Cells were incubated in William’s E Medium supplemented with the Hepatocyte Maintenance Supplement Pack (CM4000; Gibco; Durham, NC, USA) containing a cocktail of CYP probe substrates (phenacetin 50 µM (for CYP1A2), diclofenac 5 µM (for CYP2C9), mephenytoin 100 µM (for CYP2C19), bufuralol 10 µM (for CYP2D6), midazolam 5 µM (for CYP3A)) at 37 °C. After 60 min incubation, the incubation media were collected and samples were kept at −80 °C until performing liquid chromatog-raphy-tandem mass spectrometry (LC-MS/MS) analyses. The formed metabolites of CYP probe cocktail (acetaminophen, 4′-hydroxydiclofenac, 4′-hydroxy-S-mephenytoin, 1′-hydroxybufuralol, and 1′-hydroxymidazolam, respectively) were quantified with the LC-MS/MS (LC-20A (Shimadzu, Kyoto, Japan)/SCIEX Triple Quad 5500+ (AB Sciex LLC, Framingham, MA, USA)).

DRV concentrations in formulation, cell lysates, mouse plasma, and tissue samples were measured using the established LC-MS/MS method we have previously detailed [28 (link),29 (link)]. Prior to collection for in vitro study, differentiated U1 cells, also known as U1 macrophages, underwent a washing process with PBS and were then harvested using RIPA buffer. Quantification of DRV and the internal standard RTV (at a concentration of 50 ng/mL) was conducted using an AB SCIEX Triple Quad 5500 tandem mass spectrometer, which featured an electron spray ionization source operating in positive mode. The separation of these compounds was achieved through a liquid chromatography system, specifically the LC-20AD XR model from Shimadzu, MD. Data acquisition and analysis were carried out in multiple-reaction monitoring (MRM) mode using MultiQuant^® software (Version 3.0.2) by AB Sciex (Foster City, CA, USA). To mitigate matrix effects, calibration curves were constructed using blank control samples, such as plasma or tissue homogenates, tailored to the types of samples being analyzed. These controls showed no significant interference. All samples underwent centrifugation at 10,000 rpm using a Centrifuge SORVALL LEGEND X1R by Thermo Scientific, USA, after which the supernatant was further processed for LC-MS/MS analysis.