Bilateral striata were isolated from frozen brains, weighed and homogenized in 0.2 M perchloric acid. Homogenates were centrifuged at 6000×g for 20 min at 4 °C. The supernatants were resolved by high-performance liquid chromatography with a Hypersil BDS C18 reversed phase column (3 µm particle size, 130 Å pore size, 100 × 4.6 mm; Phenomenex) at a flow rate of 1 ml/min as previously reported [12 (link)]. The mobile phase comprised of citric acid (31.9 g/l), sodium acetate (2 g/l), octanesulfonic acid (460 mg/l), EDTA (30 mg/l) and methanol (15%), pH 3.6. DA, and its metabolite, 3,4-dihydroxyphenylacetic acid (DOPAC) were detected by redox oxidation using an ESA 5014 analytical cell and ESA Coulochem II electrochemical detector (Thermo Fisher Scientific) with reducing (E1) and oxidizing (E2) electrodes set at − 200 mV and + 250 mV, respectively. The chromatograms were analyzed using the Chromeleon Chromatography Data System (V 6.2; Dionex). DA and DOPAC levels were normalized to the initial wet tissue weight and expressed as pmol/mg.
Chromeleon chromatography data system
Manufactured by Thermo Fisher Scientific
Sourced in United States
Chromeleon Chromatography Data System is a software solution designed for data acquisition, processing, and management in chromatographic analysis. It provides a comprehensive platform for the control and monitoring of chromatographic instruments, as well as the analysis and reporting of chromatographic data.
Automatically generated - may contain errors
Lab products found in correlation
Dad 3000rs, by Thermo Fisher Scientific (2 mentions)
Flexible fused silica capillary tubing for trap columns, by Molex (2 mentions)
Polymeric reverse phase plrp s, by Agilent Technologies (2 mentions)
Fs360 50 8 n 20, by New Objective (2 mentions)
Esa coulochem 2, by Thermo Fisher Scientific (1 mentions)
Dionex ultimate 3000 hplc system, by Thermo Fisher Scientific (1 mentions)
Acc 3000, by Thermo Fisher Scientific (1 mentions)
Tcc 3000sd, by Thermo Fisher Scientific (1 mentions)
Eclipse xdb c18 column, by Agilent Technologies (1 mentions)
Tcc 100, by Thermo Fisher Scientific (1 mentions)
Asi 100, by Thermo Fisher Scientific (1 mentions)
Uvd340u detector, by Thermo Fisher Scientific (1 mentions)
Capcell pak c18 column, by Shiseido (1 mentions)
C phycocyanin, by Merck Group (1 mentions)
Lpg 3 00 pump, by Thermo Fisher Scientific (1 mentions)
Dionex ultimate 3000 uhplc, by Thermo Fisher Scientific (1 mentions)
Ultimate 3000, by Thermo Fisher Scientific (1 mentions)
Simca p version 13, by Sartorius (1 mentions)
Ultimate 3000 rs autosampler, by Thermo Fisher Scientific (1 mentions)
Dionex ultimate 3000 uhplc system, by Thermo Fisher Scientific (1 mentions)
13 protocols using chromeleon chromatography data system
1
Quantification of Dopamine and Metabolite
2
HPLC Quantification of Chlorogenic Acid and Hyperoside
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HPLC was performed on Dionex Ultimate 3000 HPLC system (Dionex, Germany) equipped with a pump (LPG 3×00), an auto sampler (ACC-3000), a column oven (TCC-3000SD), and diode array UV/VIS detector (DAD-3000(RS)). HPLC chromatogram data were processed using Dionex Chromeleon™ Chromatography Data System.
Chromatographic separation was conducted on Agilent eclipse XDB-C18 column (150 mm × 4.6 mm, 5 μm) at column temperature 30°C The mobile phase consisted of aqueous with 0.1% trifluoroacetic acid solution (A) acetonitrile (B) using gradient elution system of 5% (B) at 0–5 min and 5–50% (B) at 5-40 min and flow rate was 1.0 mL/min. The ultraviolet (UV) wavelength was selected and monitored at 254 nm according to the wavelength of chlorogenic acid and hyperoside.
Chromatographic separation was conducted on Agilent eclipse XDB-C18 column (150 mm × 4.6 mm, 5 μm) at column temperature 30°C The mobile phase consisted of aqueous with 0.1% trifluoroacetic acid solution (A) acetonitrile (B) using gradient elution system of 5% (B) at 0–5 min and 5–50% (B) at 5-40 min and flow rate was 1.0 mL/min. The ultraviolet (UV) wavelength was selected and monitored at 254 nm according to the wavelength of chlorogenic acid and hyperoside.
3
HPLC Quantification of Salvianolic Acid B in Herbal Extracts
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The quantitation of major compound, salvianolic acid B, in SM extract was performed by HPLC analysis. The HPLC-DAD system consisting of a chromatographic pump (P680, Dionex, Idstein, Germany), an automated sample injector (ASI-100, Dionex), and a thermostatted column compartment (TCC-100, Dionex) equipped with an UVD 340U detector (Dionex). The chromatogram of HPLC system was recorded by using Dionex’s Chromeleon™ Chromatography Data System. The column used in this work was a Capcell Pak C-18 column (150 mm × 4.6 mm, I.D., 5 μm, Shiseido Co., Ltd., Japan.). The column temperature was set to 30 °C. The mobile phase consisted of (A) 0.1 % formic acid in water and (B) 100 % acetonitrile at a flow rate of 1.0 mL/min. The gradient condition was as follows: 0–5 min, 10 %–30 % B; 5–25 min, 30 %–40 % B; 25–30 min, 40 %–100 % B.
4
Capillary-Based Liquid Chromatography Setup
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Flexible fused silica capillary tubing for trap columns (O.D.: 360 μm, I.D.: 150 μm) and analytical columns (O.D.: 360 μm, I.D.: 50 μm and 75 μm) were purchased from Polymicro Technologies (Phoenix, AZ). Polymeric reverse-phase (PLRP-S, dP = 5 μm, pore size = 1000 Å, Agilent Technologies; Santa Clara, CA) [34 (link)] was packed in fritted capillary tubing to make trap columns (packing length: 2 cm) and analytical columns (packing length: 15 cm) using a home-made pressure cell. Spray emitters (360 μm O.D., 50 μm I. D., 8 μm tip, Cat. No: FS360-50-8-N-20) were purchased from New Objective (Woburn, MA) and micro tee assemblies (Cat. No: P-888) for vented tees were purchased from Upchurch Scientific (Oak Harbor, WA). Conventional LC plumbing was installed as shown in Fig.1A . Briefly, the trap column, vented tee, analytical column, high voltage tee and spray emitter were assembled in order from the switching valve. The novel LPOx plumbing was assembled as shown in Fig. 1B . The stator for the 10-port switching valve (SV2) (Cat. No: 6041.0012) was purchased from Dionex (Sunnyvale, CA). The dimensions of all capillaries which have not been mentioned are: O.D. 360 μm and I.D 30 μm. Valve switching of SV1 and SV2 were programmed into the LC method using the Chromeleon chromatography data system (Dionex, version 6.8)
5
Identification of Phycocyanin in Spirulina maxima
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To identify the phycocyanin from water extracted S. maxima sample, extracted S. maxima and purchased C-phycocyanin were analyzed by HPLC-DAD. C-phycocyanin from Spirulina sp. was purchased from Sigma–Aldrich (St. Louis, MO, USA, Cat # P2172). The high-performance liquid chromatography (HPLC) (Dionex) equipped with an LPG 3 × 00 pump, an ACC-3000 autosampler, a DAD-3000 (RS) diode array ultraviolet (UV)/visible detector, and a column oven. Both samples were separated on a Jupiter C5 column (5 μm, 300 Å, 4.6 mm × 250 mm) at 25°C. The mobile phase consisted of 20% (v/v) aqueous acetonitrile (ACN) solution containing 0.1% (v/v) trifluoroacetic acid (TFA) and all reagents were purchased from J. T. Baker as HPLC grade. The flow rate was 1.0 mL/min and the concentration of S. maxima sample was 100 ppm and phycocyanin was 1000 ppm. The injection volume of each sample was 20 μL. The output signal of the detector was recorded using a Dionex Chromeleon™ Chromatography Data System. The UV wavelength was 580 nm and 640 nm, respectively, and the chromatograms were acquired at 580 nm.
6
Capillary-Based Liquid Chromatography Setup
7
Monosaccharide Analysis via GC-MS
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LAM samples were hydrolyzed with 2 M trifluoroacetic acid. The resulting monosaccharides were converted to alditol acetates and analyzed on a TSQ 8000 Evo triple-quad GC-MS (Thermo Scientific). The mass spectrum was scanned from m/z 50 to 500, data analysis was performed using Chromeleon Chromatography data system (Thermo Scientific). Quantitation was based on 3-O-methyl -glucose, used as an internal standard (45 ).
8
Chromatographic Analysis of Acorus Species
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Peaks above the S/N ratio of the chromatogram were labeled and manually integrated using version 7.2 of the Chromeleon Chromatography Data System software (Thermo Fisher Scientific) to distinguish between the different species of Acorus. Principal component analysis (PCA) and Hierarchical clustering analysis (HCA) of the relative peak areas were performed using the SPSS for Windows 16.0 software (SPSS Corporation, Armonk, NY, USA) to differentiate between the different peaks. Summary data were expressed as the mean ± standard deviation (SD) for n = 4.
9
UHPLC Analysis of Furanocoumarin Biotransformation
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A Dionex UltiMate 3000
UHPLC (Thermo Fisher Scientific) with a PDA detector, equipped with
a C18 Kinetex column (2.1 × 100 nm, 1.7 μm, Thermo Scientific,
Waltham, MA) was used for HPLC analysis. Program setup, data collection,
and analysis were conducted using Chromeleon Chromatography Data System
(CDS) software version 6.80 (Thermo Fisher Scientific). The detector
was set to record at 250 nm for furanocoumarin biotransformation products
analysis, simultaneously with UV spectrum monitoring in the range
of 190–380 nm. The injection volume was 1 μL, the flow
rate was 0.2 mL/min, and the column temperature was 25 °C. HPLC
eluents consist of 0.1% acetic acid (v/v) in water (A) and MeCN (B),
and a multistep gradient program was employed.
For the analysis
of the A. dahurica extract and biotransformation
products of furanocoumarins, solution B was started at 10%, increased
to 30% for 1 min, to 40% for 11 min, to 50% for 15 min, to 60% for
3 min, to 80% for 2 min, to 90% for 1 min, and held for 7 min.
UHPLC (Thermo Fisher Scientific) with a PDA detector, equipped with
a C18 Kinetex column (2.1 × 100 nm, 1.7 μm, Thermo Scientific,
Waltham, MA) was used for HPLC analysis. Program setup, data collection,
and analysis were conducted using Chromeleon Chromatography Data System
(CDS) software version 6.80 (Thermo Fisher Scientific). The detector
was set to record at 250 nm for furanocoumarin biotransformation products
analysis, simultaneously with UV spectrum monitoring in the range
of 190–380 nm. The injection volume was 1 μL, the flow
rate was 0.2 mL/min, and the column temperature was 25 °C. HPLC
eluents consist of 0.1% acetic acid (v/v) in water (A) and MeCN (B),
and a multistep gradient program was employed.
For the analysis
of the A. dahurica extract and biotransformation
products of furanocoumarins, solution B was started at 10%, increased
to 30% for 1 min, to 40% for 11 min, to 50% for 15 min, to 60% for
3 min, to 80% for 2 min, to 90% for 1 min, and held for 7 min.
10
Size-Exclusion Chromatography for Variant Analysis
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The separation of size
variants was performed with an UltiMate 3000 HPLC system (Thermo Fisher
Scientific) using a TSKgel G3000SWXL size-exclusion chromatograph
column (Tosoh Bioscience, cat no. 08541, 5 μm, 7.8 × 300
mm). Data were recorded by UV detection at 280 nm (UV280), and data analysis was done using the Chromeleon chromatography
data system (CDS) software (Thermo Fisher Scientific). The mobile
phase buffer composed of 0.2 M potassium phosphate and 0.25 M KCl
at pH 6.2 was used. A molecular weight standard demonstrated the system
suitability. Prior to loading, the samples were diluted to 30 mg/mL
with formulation buffer, and 5 μL (150 μg) of sample was
injected per run. The following analytical conditions were applied:
0.5 mL/min flow rate, 25 ± 2 °C column temperature, 5 ±
4 °C auto-sampler temperature, and 30 min under isocratic conditions.
Relative areas under the curve (AUCs) were used to calculate the percentage
areas from the total area of all peaks observed.
variants was performed with an UltiMate 3000 HPLC system (Thermo Fisher
Scientific) using a TSKgel G3000SWXL size-exclusion chromatograph
column (Tosoh Bioscience, cat no. 08541, 5 μm, 7.8 × 300
mm). Data were recorded by UV detection at 280 nm (UV280), and data analysis was done using the Chromeleon chromatography
data system (CDS) software (Thermo Fisher Scientific). The mobile
phase buffer composed of 0.2 M potassium phosphate and 0.25 M KCl
at pH 6.2 was used. A molecular weight standard demonstrated the system
suitability. Prior to loading, the samples were diluted to 30 mg/mL
with formulation buffer, and 5 μL (150 μg) of sample was
injected per run. The following analytical conditions were applied:
0.5 mL/min flow rate, 25 ± 2 °C column temperature, 5 ±
4 °C auto-sampler temperature, and 30 min under isocratic conditions.
Relative areas under the curve (AUCs) were used to calculate the percentage
areas from the total area of all peaks observed.
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