Lungs were removed and immersed immediately in liquid nitrogen prior to storage at −80 °C. Lung tissue was ground to a fine powder under liquid nitrogen. Ground lung tissue was mixed in 1 ml distilled water on ice in Pyrex® tubes and incubated in 125 μl 50 % TCA (Sigma Aldrich) at 4 °C for 20 min. Samples were centrifuged at 528 × g for 10 min at 4 °C. The lungs were then hydrolyzed in 1 ml of 12 N HCL overnight at 110 °C prior to reconstitution in 2 ml of distilled water. 200 μl of hydrozylate, or hydroxyproline standard (Sigma Aldrich), were oxidized in 500 μl chloramine T (Sigma Aldrich) for 20 min at room temperature (RT) prior to the addition of 500 μl of Ehrlich’s solution (Sigma Aldrich) and incubation at 65 °C for 15 min. Samples were then incubated at RT for 2 hours prior to colorimetric analysis at an absorbance of 550 nm. Hydroxyproline per mg of lung tissue was calculated against a standard curve.
Pyrex tube
Manufactured by Merck Group
Pyrex® tubes are laboratory glassware designed for a variety of scientific and industrial applications. They are made of borosilicate glass, which is known for its thermal resistance and chemical durability. Pyrex® tubes are available in various sizes and shapes to accommodate different experimental requirements.
Automatically generated - may contain errors
Lab products found in correlation
Hydroxyproline standard, by Merck Group (2 mentions)
Chloramine t, by Merck Group (2 mentions)
Ehrlich s solution, by Merck Group (2 mentions)
Hydrozylate, by Merck Group (2 mentions)
Salicylic acid, by Merck Group (1 mentions)
Hplc system, by Waters Corporation (1 mentions)
10.6 m hcl, by Merck Group (1 mentions)
Discovery dsc 18, by Merck Group (1 mentions)
Acetic acid, by Merck Group (1 mentions)
5 protocols using pyrex tube
1
Quantifying Lung Collagen Content
2
Quantifying Lung Collagen Content
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Lungs were removed and immersed immediately in liquid nitrogen prior to storage at −80 °C. Lung tissue was ground to a fine powder under liquid nitrogen. Ground lung tissue was mixed in 1 ml distilled water on ice in Pyrex® tubes and incubated in 125 μl 50 % TCA (Sigma Aldrich) at 4 °C for 20 min. Samples were centrifuged at 528 × g for 10 min at 4 °C. The lungs were then hydrolyzed in 1 ml of 12 N HCL overnight at 110 °C prior to reconstitution in 2 ml of distilled water. 200 μl of hydrozylate, or hydroxyproline standard (Sigma Aldrich), were oxidized in 500 μl chloramine T (Sigma Aldrich) for 20 min at room temperature (RT) prior to the addition of 500 μl of Ehrlich’s solution (Sigma Aldrich) and incubation at 65 °C for 15 min. Samples were then incubated at RT for 2 hours prior to colorimetric analysis at an absorbance of 550 nm. Hydroxyproline per mg of lung tissue was calculated against a standard curve.
3
Extraction and Quantification of Analytes
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Authentic samples and in-house QCs were thawed and vortexed. RfB materials were reconstituted according to manufacturer's specifications. Three-hundreds microliters of each calibrator, sample and QCs were pipetted into 13x100 mm Pyrex ® tubes (Sigma Aldrich), spiked with 30 µL IS and vortexed for 2 min. Two mL of Nhexane:ethyl-acetate mixture (9:1) were added before tubes were vigorously vortexed for 5 min and centrifuged for 15 min (2000 g, RT). The lower aqueous layer was frozen in ice bath, while the upper organic layer was decanted in 12x75 mm glass tubes (Laboindustria, Arzergrande, Italy) and dried under nitrogen flow. Samples were dissolved in 100 µL 50% methanol and transferred into autosampler glass vials (Agilent Technology, Santa Clara, CA). Each batch included authentic samples, a set of calibrators and external QCs, and three replicates of in house QCs placed at batch beginning, middle and end.
4
Characterization of Bismuth Subsalicylate
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Commercially available Pepto-Bismol suspension manufactured by P&G was purchased from the supermarket Foodland Pukalani, HI, USA. Prior to performing 3DED and scanning transmission electron microscopy (STEM) measurements, 5 mL of the suspension was washed with 50 mL of water and centrifuged at 6000 × g for 5 min whereafter it was left to dry overnight under ambient conditions (this washed powder is referred to as BSS-PB).
Commercially available bismuth subsalicylate (BSS, CAS: 14882-18-9) was purchased from Sigma-Aldrich (BSS-SA). The untreated powder was used for structure determination.
Laboratory-made bismuth subsalicylate was prepared by adding 72 mg bismuth(iii ) oxide (Aldrich, 99.999% trace metal basis) and 128 mg salicylic acid (Sigma-Aldrich, ACS reagent, ≥99%) to a 5 mL Pyrex tube filled with 3 mL of deionized water and sealed with a PTFE cap. The tube was heated to 140 °C while stirring at 800 rpm for 3 h. The solid was then filtered off, washed with 50 mL of hot deionized water (>95 °C), and left to dry in ambient conditions overnight.
Commercially available bismuth subsalicylate (BSS, CAS: 14882-18-9) was purchased from Sigma-Aldrich (BSS-SA). The untreated powder was used for structure determination.
Laboratory-made bismuth subsalicylate was prepared by adding 72 mg bismuth(
5
Furosine Quantification in NFDM
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Furosine (FUR) was measured in NFDM as described by Resmini et al. (1990) with minor modifications. Between 110 and 140 mg of NFDM was accurately weighed into a screw-cap Pyrex tube with a PTFE-lined septa along with 8 mL of 10.6 M HCl (Sigma Aldrich). Nitrogen gas was bubbled through the samples for 2 min and then capped and heated to 110°C for 23 h. After cooling to room temperature, the hydrolysate was filtered and 0.5 mL of filtrate was loaded onto a solid-phase extraction column (500 mg Discovery DSC-18, Supelco) previously conditioned with 5 mL of methanol followed by 10 mL of water. The FUR was eluted with 3 mL of 3 M HCl. Detection was performed with a Waters HPLC system (Milford, MA). A FUR dedicated column was used (Altima C8, 250 × 4.6 mm, 5μm, Alltech-Grace, Columbia, MD) with isocratic conditions. The mobile phase consisted of 0.4% acetic acid (Sigma Aldrich) and the column temperature was 35°C. Ultraviolet detection was performed at 280 nm. A 5-point external standard curve ranging from 50 to 400 ng was constructed for quantification.
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