The HPLC system consisted of a pump (PU-4180; Jasco, Tokyo, Japan), an ultraviolet detector (UV-4075; Jasco), and an autosampler (AS-4550; Jasco). The HPLC column was a Capcell Pak C18 MG II (Osaka Soda Co., Ltd., Tokyo, Japan) reversed-phase column (250 nm × 4.6 nm i.d.) and Capcell Pak C18 MG II guard column (10 mm × 4.0 mm; Osaka Soda) at a temperature of 20 °C. The mobile phase was 0.5% KH₂PO₄ (pH 4.5) and acetonitrile (43:57, v/v), and the flow rate was 1.0 mL/min. Sample detection was carried out at 250 nm.
Uv 4075
Manufactured by Jasco
Sourced in Japan, United States
The UV-4075 is a dual-beam ultraviolet-visible (UV-Vis) spectrophotometer designed for accurate and reliable measurements. It is capable of analyzing the absorption characteristics of liquid and solid samples across a wide wavelength range.
Automatically generated - may contain errors
Lab products found in correlation
Pu 4180, by Jasco (7 mentions)
Co 4061, by Jasco (2 mentions)
As 4150, by Jasco (2 mentions)
Reax 2, by Heidolph (2 mentions)
Md 4010, by Jasco (2 mentions)
Extrema, by Jasco (2 mentions)
As 4550, by Jasco (1 mentions)
Rezex roa organic acid h column, by Phenomenex (1 mentions)
Chomnav software, by Jasco (1 mentions)
Lc 4000, by Jasco (1 mentions)
Ri 4030, by Jasco (1 mentions)
As 4050, by Jasco (1 mentions)
Dismic 13hp045an, by Advantec (1 mentions)
Agilent 1100 series, by Agilent Technologies (1 mentions)
Chemstation, by Agilent Technologies (1 mentions)
Adventurer pro as214, by Ohaus (1 mentions)
Tskgel ods 100z, by Tosoh (1 mentions)
Pu 1580, by Jasco (1 mentions)
Lg 1580 02, by Jasco (1 mentions)
Lc net 2 adc, by Jasco (1 mentions)
14 protocols using uv 4075
1
HPLC Analysis of Compounds
2
Monitoring Microbial Growth and Metabolites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell growth during cultivation was monitored by measuring OD600 on a Genesys 20 spectrophotometer (Thermo Fisher).
The concentration of glucose and organic acids in cultivation broth was analyzed by a Jasco (Tokyo, Japan) LC-4000 high-performance liquid chromatography system equipped with an autosampler (AS-4150), a pump (PU-4180), a column oven (CO-4061), a UV detector (RI-4030), and an RI detector (UV-4075). The mobile phase consisted of 5 mM H2SO4 and was pumped at constant flow rate of 0.8 mL/min. A Rezex ROA-organic acid H + column (Phenomenex, Torrance, CA, USA) maintained at 80 °C was used to separate the metabolites in the cultivation broth. The concentration of each compound was determined by Jasco ChomNAV software (version 2.03.03).
The concentration of glucose and organic acids in cultivation broth was analyzed by a Jasco (Tokyo, Japan) LC-4000 high-performance liquid chromatography system equipped with an autosampler (AS-4150), a pump (PU-4180), a column oven (CO-4061), a UV detector (RI-4030), and an RI detector (UV-4075). The mobile phase consisted of 5 mM H2SO4 and was pumped at constant flow rate of 0.8 mL/min. A Rezex ROA-organic acid H + column (Phenomenex, Torrance, CA, USA) maintained at 80 °C was used to separate the metabolites in the cultivation broth. The concentration of each compound was determined by Jasco ChomNAV software (version 2.03.03).
3
Synthesis of Kapakahine F Fluorescent Probes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Kapakahine F (2 , 0.5 mg, 0.7 μmol) in 33.3 μL DMF was added to 66.6 μL of the solution of reagent 3a in DMF (20 mg/mL) and was stirred at room temperature in the dark for 20 h. In this experiment, we used synthetic kapakahine F (provided by Prof. Phil Baran). After quenching with MeOH, the fraction was further purified by reversed-phase HPLC [COSMOSIL 5C18-AR II (φ10 × 250 mm), flow rate; 2 mL/min, isocratic elution with 50% acetonitrile 0.05% TFA, detection; 220 nm JASCO UV4075] yielding Kap F-5-FL (6a , 370 µg) which gave an [M − H]− ion peak at m/z 1171.4579 (calcd. C67H63N8O12 1171.4571).
Kap F-5,6-FL(6a/b ) was also obtained following the same procedure but with 3a/b instead of 3a.
Kap F-5,6-FL(
4
Synthesis of Kap A-5-FL and Kap A-5,6-FL
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A portion (100 μL) of 3a in N,N-dimethylformamide (DMF) (20 mg/mL, 6.8 μmol) was added dropwise to kapakahine A (1 , 0.5mg, 0.5 μmol) dissolved in 50 μL of DMF. In this experiment, we used kapakahine A isolated from natural sources. The reaction mixture was stirred at room temperature in the dark for 20 h. After quenching with MeOH, the reaction mixture was subjected to ODS flash column chromatography eluting with 100% MeOH. The 100% MeOH eluting fraction was further purified by reversed-phase HPLC [COSMOSIL 5C18-ARⅡ (φ10 × 250 mm), flow rate; 2 mL/min, isocratic elution with 55% acetonitrile 0.05% TFA, detection; 220 nm, JASCO UV4075] to yield Kap A-5-FL (5a : 340 µg). In the ESIMS analysis, 5a gave an [M − H]− ion peak at m/z 1522.6737 (calcd. C85H92N11O16 1522.6729).
Kap A-5,6-FL (5a/b ) was also prepared following the same procedure but with 3a/b instead of 3a.
Kap A-5,6-FL (
5
Isolation and Purification of Carotenoid Isomers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All-trans β-apo-8′-carotenal (purity ≥ 96.0%) was purchased from SIGMA and used as it received. Light from W-halogen lamp (50 W) was irradiated for 3 h to n-hexane solution of the all-trans isomer to generate the cis isomers using iodine as a photocatalyst [31 (link)]. The cis isomers were isolated and purified by the use of a set of HPLC (JASCO PU-4180 and UV-4075). For isolation of the cis isomers, three conditions were adopted. Condition 1 (for the isolation of 13-cis, 9,13′-cis, 9-cis, and 7-cis isomers): Column, Inertsil SIL-100A 7.6 × 250 mm, 5 µm; Eluent, diethyl ether/n-hexane 4/96 v/v; Flow rate, 2.5 mL/min; Detection wavelength, 450 nm. Condition 2 (for the isolation of the 13′-cis and 15-cis isomers): Column, LiChrosorb Si 60 7.6 × 300 mm, 5 µm; Eluent, diethyl ether/n-hexane 3/97 v/v; Flow rate, 1.5 mL/min; Detection wavelength, 450 nm. Condition 3 (for the isolation of the 13,13′-cis and 9,13-cis isomers): Column, Daisopak SP 60 6.0 × 250 mm, 5 µm; Eluent, diethyl ether/n-hexane 4/96 v/v; Flow rate, 1.0 mL/min; Detection wavelength, 450 nm.
6
Synthesis of Fluorescent Ester Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
6-(Fluorescein-5-carboxamido) hexanoic acid succinimidyl ester (5-SFX: 3a , 1.3 mg, 2.2 µmol, Thermo Fisher Scientific Inc., Waltham, MA, USA) were mixed with 1 µL of triethylamine (TEA, 7.2 µmol) in 645.6 µL of MeOH. The reaction mixture was stirred at room temperature in the dark for 20 h. The reaction was quenched with excess MeOH and the dried reaction mixture was subjected to reversed-phase HPLC [COSMOSIL 5C18-ARⅡ (φ10 × 250 mm, Nacalai Tesque Inc., Kyoto, Japan), flow rate; 2 mL/min, isocratic elution with 35% acetonitrile 0.05% TFA, detection; 220 nm, JASCO UV4075] yielding 5-FL-COOMe (4a , 550 µg). Compound 4a gave an [M − H]− ion peak at m/z 502.1511 (calcd. C28H24NO8 502.1507) in the ESIMS.
Methyl esters4a and 4b were also obtained by HPLC separation of the reaction mixture obtained following the same procedure but with a 1:1 mixture of 5- and 6-SFX (3a/b ) instead of 3a with adjustment of the amounts of triethylamine.
Methyl esters
7
Quantifying Phytate in Meat Substitutes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phytate was analyzed as inositol hexaphosphate (InsP6) by high-performance ion chromatography (HPIC) according to Carlsson et al. [29 (link)]. A sample of freeze-dried and ground meat substitute (0.5 g) was extracted in duplicate with 10 mL of 0.5 mol/L HCl for three hours using a laboratory shaker (Heidolph Reax 2; Heidolph Instruments, GmbH, Schwabach, Germany). Then, 1 mL was removed, centrifuged and filtered to remove fat before transferring the extract to an HPLC vial. The chromatography setup consisted of an HPLC pump (model PU-400oi; Jasco Inc., Easton, MD, USA) for the eluent and an RHPLC pump (model PU-4180; Jasco, Oklahoma City, OK, USA) equipped with a PA-100 guard column and a CarboPac PA-100 column. InsP6 was eluted with an isocratic eluent of 80% HCl (1 mol/L) and 20% MilliQ water at 0.8 mL/min, subjected to a post-column reaction with ferrous nitrate, and detected at 290 nm in a UV–visible HPLC detector (UV-4075; Jasco, Oklahoma City, OK, USA). Columns were run under ambient temperature. Each sample had a run time of 7 min, and the InsP6 concentration was calculated using external standards covering the range of 0.1–0.6 μmol/mL.
8
HPLC Quantification of Taro Oxalates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Oxalic acid was quantified using high‐performance liquid chromatography (HPLC).23 To extract soluble and total oxalates, taro powder was mixed with distilled water and 0.5 mol L–1 hydrochloride solution (20 mg mL−1). Samples were vortexed (Se‐08, Taitec, Saitama, Japan) and sonicated for 30 min each and centrifuged at 1600×g for 10 min (Model 3500, Kubota, Tokyo, Japan). The supernatant was collected and filtered through a 0.45 μm filter (DISMIC®‐13HP045AN, Advantec) and used as the sample for analysis. The HPLC unit comprised a pump (PU‐4180, Jasco, Japan), a detector (UV‐4075, Jasco, Tokyo, Japan), an autosampler (AS‐4050, Jasco, Tokyo, Japan), and a column oven (SLC‐25A, MEE, Japan). The HPLC analysis was performed using a C18 reversed‐phase column (Cosmosil 5C18‐PAQ, 4.6 mm I.D. × 250 mm, Nacalai Tesque, Kyoto, Japan) maintained at 40 °C. The eluent used was 20 mM phosphate buffer (pH 2.5) with a flow rate of 1.0 mL min−1, a detection wavelength at 254 nm, and an injection volume of 10 μL. The calibration curve was obtained using oxalic acid anhydride (Wako, Japan) as standards (0, 31.3, 62.5, 125, 250, 500, and 1000 μg mL−1, n = 3). Insoluble oxalate was calculated as the difference between the total and soluble oxalates.
9
RP-HPLC Method Validation for Pharmaceutical Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RP-HPLC analysis was performed on a Jasco Extrema (JASCO, Inc., 28600 Mary’s court, Easton, Maryland 21601, USA) connected with an in built autosampler (AS-4150), quaternary pump (PU-4180) with an on-line degasser, column oven (CO-4061), UV/Vis detector (UV-4075) and an interface box LC-NetII/ADC. Chromatographic data was collected and analysed using ChromNAV Ver.2 software. A second HPLC system (Agilent 1100 series) equipped with variable wavelength detector (G1314A VWD), autosampler (G1313A) and isocratic pump system (G1310A). This system automated with a Chemstation (Agilent technologies) software and which was used for intermediate precision (repeatability) testing and data collection. Chromatographic separations were achieved by using SOLAS 100 Å C18 150 mm × 4.6 mm 5 µm column (Glantreo limited, Ireland). Different HPLC columns such as SOLAS ODS C18 150 mm × 4.6 mm 5 µm, SOLAS ODS C18 150 mm × 4.6 mm 3 µm, SOLAS BDS C18 150 mm × 4.6 mm 5 µm, SOLAS BDS C18 150 mm × 4.6 mm 3 µm and EIROSHELL C18 150 mm × 4.6 mm 2.6 µm (Glantreo limited) were used for this method development purpose. Each Chemical and test samples were accurately weighed using an analytical balance (Adventurer Pro AS214, Ohaus corporation, Pine Brook, NJ USA).
10
HPLC Analysis of Tea Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After filtration with a 0.45‐μm filter, tea infusion and tea leaf extract samples were analyzed using HPLC (Extrema, Jasco). HPLC analysis conditions were as follows: C18 reverse‐phase column (TSKgel ODS‐100Z, 5 μm, 4.6 mm I.D. × 150 mm; Tosoh) and guard column (TSKgel Guardgel ODS‐100Z, 5 μm, 3.2 mm I.D. × 15 mm; Tosoh) were maintained at 30°C, and detection was performed at 280 nm. The mobile phase was 0.05% phosphoric acid (A) and methanol:acetonitrile (3:2) binary solution system (B) with the following conditions: 0–8 min with B solution 25% constant and a gradient of 8–18 min with a direct increase in B solution up to 75%. Individual compounds were identified with highly selective spectral data in combination with their retention times and an ultraviolet photodiode array detector (UV‐4075 and MD4010, Jasco).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!