The plasma samples were extracted using HPLC grade methanol (CROMASOLV, Sigma-Aldrich). Subsequently, nitrite and nitrate levels were measured by a sensitive and selective measurement HPLC system (ENO-20 Eicom Japan), which uses reverse phase chromatography to separate nitrite from nitrate. Thereafter, nitrate was reduced to nitrite through a reaction with cadmium and reduced copper inside a reduction column. Reduced nitrite was then derivatized with Griess reagent and the level of diazo compounds were analyzed by detection at 540 nm as previously described in detail.10 (link)
Cromasolv
Manufactured by Merck Group
Sourced in United States
CROMASOLV is a line of high-purity solvents and reagents designed for use in analytical and preparative chromatography applications. The products in this line are manufactured to meet stringent quality standards and provide consistent performance.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using cromasolv
1
Plasma Nitrite/Nitrate Measurement by HPLC
2
Plasma Nitrite/Nitrate Measurement by HPLC
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The plasma samples were extracted using HPLC grade methanol (CROMASOLV, Sigma-Aldrich). Subsequently, nitrite and nitrate levels were measured by a sensitive and selective measurement HPLC system (ENO-20 Eicom Japan), which uses reverse phase chromatography to separate nitrite from nitrate. Thereafter, nitrate was reduced to nitrite through a reaction with cadmium and reduced copper inside a reduction column. Reduced nitrite was then derivatized with Griess reagent and the level of diazo compounds were analyzed by detection at 540 nm as previously described in detail.10 (link)
3
Measuring Nitrite and Nitrate Levels
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The plasma samples were extracted using HPLC grade methanol (CROMASOLV, Sigma-Aldrich, Burlington, MA, USA), and the saliva samples were centrifuged at 10,000× g for 10 min at 4 °C before dilution with carrier solution containing 10% HPLC grade methanol. Dialysate samples were analyzed undiluted. Subsequently, nitrite and nitrate levels were measured HPLC (ENO-20 Eicom, Kyoto, Japan), which uses reverse phase chromatography to separate nitrite from nitrate. Thereafter, nitrate was reduced to nitrite through a reaction with cadmium and reduced copper inside a reduction column. Reduced nitrite was then derivatized with Griess reagent, and the level of diazo compounds was analyzed by detection at 540 nm as previously described in detail [25 (link)].
4
Biological Sample Preparation for Nitrite/Nitrate Analysis
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Blood samples were immediately centrifuged at +4 °C (4700×g, 5 min) and plasma was stored at −80 °C until analyses. The plasma samples were extracted using HPLC grade methanol (CROMASOLV, Sigma-Aldrich) and the methanol was pre chilled in the freezer using a glass bottle to avoid nitrite/nitrate contamination from different plastic materials. In brief, 100 µL methanol was added to 100 µL plasma in 1.5 mL Eppendorf tubes tested for nitrite/nitrate contamination. The tubes were vortexed and then centrifuged for 10 min 4 °C 10,000g.
The saliva samples were centrifuged for 10 min at 4 °C and 10,000g before dilution with carrier solution containing 10% HPLC grade methanol.
The salivary glands removed from the animals were weighted, cut in smaller pieces and immediately placed in ice-cold methanol 1 mL/g tissue. The tissue was then homogenized using a bullet blender with Zirccox beads. After homogenization, the samples were centrifuged for 10 min at 4 °C 10,000g. The supernatant was then transferred to another tube and stored overnight at −20 °C.
The saliva samples were centrifuged for 10 min at 4 °C and 10,000g before dilution with carrier solution containing 10% HPLC grade methanol.
The salivary glands removed from the animals were weighted, cut in smaller pieces and immediately placed in ice-cold methanol 1 mL/g tissue. The tissue was then homogenized using a bullet blender with Zirccox beads. After homogenization, the samples were centrifuged for 10 min at 4 °C 10,000g. The supernatant was then transferred to another tube and stored overnight at −20 °C.
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