Different BBR containing micelles were administered to Apoe(−/−) mice by gavage (100 mg/kg/day of BBR). At each preset time point, five mice for each group were euthanized. The blood samples were gathered from posterior orbital venous plexus to a heparinized tube. The major organs (heart, liver, lung and adipose) were harvested and immediately immersed in liquid nitrogen and stored at −80 °C. The distribution of BBR in various formulations was also analyzed by LC‒MS/MS (Shimadzu LC-20AD-UFLC, Kyoto, Japan) described previously25 (link).
Lc 20 ad uflc
Manufactured by Shimadzu
Sourced in Japan, United States
The LC-20 AD UFLC is a high-performance liquid chromatography (HPLC) system manufactured by Shimadzu. It is designed to provide fast, efficient, and accurate liquid chromatography analysis. The system features a high-pressure pump, an autosampler, and a variety of detectors to enable a wide range of analytical applications.
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Lab products found in correlation
4 protocols using lc 20 ad uflc
1
Tissue Distribution of BBR Micelles in Mice
2
Synthesis and Characterization of Resolvin E4
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Following
a literature procedure,36 (link) a solution of
RvE4 methyl ester (2 , 5.0 μg, 14 nmol) in MeOH
was concentrated under a gentle stream of nitrogen gas, dissolved
in THF (500 μL), and cooled to −78 °C. To the resulting
solution was added 1 M LiOH (50 μL, 50 μmol) and distilled
water (one drop, ∼20 μL) and the reaction mixture was
stirred in a 4 °C cold room for 24 h. The reaction mixture was
then concentrated under a gentle stream of nitrogen gas and reconstituted
with MeOH (500 μL). The identity of the compound was verified
by UV–Vis and LC–MS/MS. The chemical yield of the RvE4
free acid1 was 69% (3.3 μg, 9.9 nmol) post saponification
(based on UV–Vis) and was determined to be >95% pure by
targeted
MRM LC–MS/MS. The physical properties of synthetic RvE4 (1 ) and biogenic RvE4 (1 ) were analyzed on a QTRAP
5500 mass spectrometer (Sciex, Framingham, MA, USA) equipped with
a LC20AD UFLC (Shimadzu, Tokyo, Japan) with a Poroshell EC-C18 column
(100 mm × 4.6 mm × 2.7 μm; Agilent Technologies, Santa
Clara, CA, USA) kept at 50 °C. RvE4 (1 ) was monitored
by targeted multiple reaction monitoring (m/z 333 > 115) and enhanced product ion mode in negative
polarity.
RvE4 (1 ) was eluted at a flow rate of 0.5 mL/min with
a gradient of LC–MS grade methanol/water/acetic acid from 50/50/0.01
v/v/v to 98/2/0.01 v/v/v. Data were acquired and analyzed with Analyst
version 1.6.2 (Sciex).8 (link)
a literature procedure,36 (link) a solution of
RvE4 methyl ester (
was concentrated under a gentle stream of nitrogen gas, dissolved
in THF (500 μL), and cooled to −78 °C. To the resulting
solution was added 1 M LiOH (50 μL, 50 μmol) and distilled
water (one drop, ∼20 μL) and the reaction mixture was
stirred in a 4 °C cold room for 24 h. The reaction mixture was
then concentrated under a gentle stream of nitrogen gas and reconstituted
with MeOH (500 μL). The identity of the compound was verified
by UV–Vis and LC–MS/MS. The chemical yield of the RvE4
free acid
(based on UV–Vis) and was determined to be >95% pure by
targeted
MRM LC–MS/MS. The physical properties of synthetic RvE4 (
5500 mass spectrometer (Sciex, Framingham, MA, USA) equipped with
a LC20AD UFLC (Shimadzu, Tokyo, Japan) with a Poroshell EC-C18 column
(100 mm × 4.6 mm × 2.7 μm; Agilent Technologies, Santa
Clara, CA, USA) kept at 50 °C. RvE4 (
by targeted multiple reaction monitoring (m/z 333 > 115) and enhanced product ion mode in negative
polarity.
RvE4 (
a gradient of LC–MS grade methanol/water/acetic acid from 50/50/0.01
v/v/v to 98/2/0.01 v/v/v. Data were acquired and analyzed with Analyst
version 1.6.2 (Sciex).8 (link)
3
Quantification of Emodin in PHELA® Extract
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The percentage content of standard phytomarker emodin in PHELA® extract was determined through RP-HPLC (Shimadzu Prominence, Kyoto, Japan) system. Built with two Shimadzu LC-20 AD UFLC reciprocating pumps, a variable Shimadzu SPD-M20A Prominence PDA detector and a Rheodyne manual injector with a loop volume of 20 μL with a C18column (Phenomenex-Luna C18, Torrance, CA, USA) (250 × 4.6 nm, 5 μm particle size) was used as the stationary phase. For analysis, the standard and test samples were dissolved in methanol to obtain a stock solution of 1 mg/mL. The standard and sample were injected at a volume of 20 μL using Hamilton Micro liters syringe (Bonaduz, Switzerland). The analysis was performed under isocratic elution using optimized mobile phase methanol and 1% acetic acid solution at a 1 mL/min flow rate. The detection of the compounds was performed at λmax254 nm. The amount of the standard present in the sample was determined through the construction of the calibration curve by plotting peak area against corresponding concentrations through linear regression using LC solution software [18 ].
4
Quantitative Analysis of Triglycerides in TFG
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Extract of TFG was standardized with respective biomarker TG by using reverse phase-high performance liquid chromatography (RP-HPLC). RP-HPLC system (Shimadzu Prominence, Kyoto, Japan) equipped with two Shimadzu LC-20 AD UFLC reciprocating pumps, a variable Shimadzu SPD-M20A Prominence PDA detector and a Rheodyne manual injector with a loop size of 20 µl was used. The peak area was calculated with LC solution software. The analysis was carried out in isocratic condition using a C18 reverse phase column having dimension of 250 mm (length) ×4.6 mm (width) with a particle size of 5 μm (Phenomenex-Luna C18, Torrance, CA, USA). Samples were filtered through Whatmman NYL 0.45 μm syringe filter and an aliquot of 20 µl of each sample was injected into injector port. Elution was carried out with methanol: Water containing 0.5% acetic acid (60:40) at a flow rate of 1 ml/min and eluate was monitored at 254 nm. TG (1 mg/ml) solution was prepared in methanol as a stock solution. Calibration curve was plotted by diluting the stock solution in the concentration range of 200–1000 μg/ml. TG present in TFG extract were identified by comparing with the retention time (Rt) in chromatographic peaks of standard with that of the extract. Percentage of TG present in TFG was determined by constructing a calibration curve.
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