One 3mm punch from a DBS using a Harris Micro-punch (Fisher Scientific) was extracted in a micro centrifuge tube with 0.5mL of a 70:30 methanol:water (v/v) lysing solution. The resulting lysing solution was then subjected to analysis as published previously [12 (link)]. Briefly, TFV-DP and FTC-TP were purified via Waters (Milford, MA, USA) QMA Solid Phase Extraction (SPE) cartridges followed by dephosphorylation to the molar equivalent parent moieties (TFV and FTC, respectively) with acid or alkaline phosphatase. The sample was then desalted and concentrated on a Phenomenex (Torrance, CA, USA) Strata-X SPE cartridge. Separation of analytes was achieved on a Phenomenex Synergi Polar RP column (2.1 × 100mm, 2.5μm) with isocratic mobile phase (0.5% acetonitrile in water with 0.1% formic acid) at 0.25mL/min flow rate. A Thermo Scientific (San Jose, CA, USA) TSQ Vantage® triple quadrupole mass spectrometer detector was used in the ESI (+) mode and the analyte [precursor/product]+ transitions (m/z) were: TFV (288.04/176.11), stable-labeled TFV-internal standard (293.04/181.11), FTC (248.10/130.00), stable-labeled FTC-internal standard (251.10/133.00). The quantifiable linear range for TFV was 2.5-2000 fmol/sample and for FTC was 0.1-200 pmol/sample [12 (link)].
Strata x spe cartridge
Manufactured by Phenomenex
Sourced in United States
The Strata-X SPE cartridge is a solid-phase extraction (SPE) product designed for sample preparation. It is used to extract and purify analytes from complex matrices. The cartridge contains a sorbent material that selectively retains target compounds while allowing interferences to be washed away.
Automatically generated - may contain errors
Lab products found in correlation
Synergi polar rp column, by Phenomenex (1 mentions)
Qma solid phase extraction cartridges, by Waters Corporation (1 mentions)
Hplc grade methanol, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Citric acid, by Thermo Fisher Scientific (1 mentions)
Doxycycline, by Merck Group (1 mentions)
Na2hpo4, by Thermo Fisher Scientific (1 mentions)
Oxalic acid, by Thermo Fisher Scientific (1 mentions)
Na2edta, by Thermo Fisher Scientific (1 mentions)
Luna c18, by Phenomenex (1 mentions)
3 protocols using strata x spe cartridge
1
Quantification of Tenofovir and Emtricitabine
2
Doxycycline Extraction and Quantification
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Doxycycline was purchased from Sigma–Aldrich Inc., St. Louis, MO, USA, and oxalic acid, Na2HPO4, citric acid, and Na2EDTA were acquired from Fisher Scientific Co. (Springfield, NJ, USA). HPLC grade methanol and acetonitrile were purchased from Merck (Darmstadt, Germany). Strata™ X SPE cartridge was acquired from Phenomenex, Torrance, CA, USA. De-ionized water was produced through Milli-Q, Germany, with 18.2 MΩ cm−1 resistivity.
3
Lipidomic Analysis of Vulvar Tissue
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Punch biopsies from the vestibule and external vulva of 11 cases and 10 controls were collected and tangentially sectioned into 3 pieces (see Fig. S2 for experimental design, available as supplemental digital content at http://links.lww.com/PAIN/B937 ). For lipidomic analysis, one piece from each site was flash frozen in an amber glass vial. Samples were then purged with argon gas and stored at −80°C until analysis. Quantitative targeted lipidomic analysis of the tissues was performed by the Wayne State University Lipidomics Core using liquid chromatography–mass spectrometry (LC-MS). Briefly, samples were prepared using C18 reverse phase cartridges (Phenomenex, StrataX SPE cartridge, 30-mg sorbent), and then subjected to reverse phase HPLC on a C18 column (Luna, C18, 3 μm, 2 mm × 150 mm, Phenomenex, CA) as published earlier.61 (link),62 (link)
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