SAE, HMPP, and drugs were mixed well and exposed to a spot light source (xenon lamp) for photopolymerization within 1.0 minute. The amount of each formulation containing SAE, HMPP, and drugs had a weight ratio of 30.0/2.0/5.0 (30.0/2.0/1.0 for PAMBA). The formed membrane was extracted by double-distilled water at 30°C without stirring. A small amount of solution was extracted at regular intervals, and then measured by high-performance liquid chromatography (HPLC) to detect the drug concentration using an LC-20A system with an Inertsil ODS-3 column (both from Shimadzu, Kyoto, Japan) at a flow rate of 1.0 mL⋅min−1 and column temperature of 30°C. Measurements were carried out with an injection volume of 20.0 µL and a detection wavelength of 208 nm for EACA and AMCHA, 220 nm for PAMBA, and 254 nm for EDDA. The mobile phase was a mixture of acetonitrile with 1.0% phosphate buffer (10/90) containing 0.055% of sodium hexanesulfonate (prepared by addition of 0.055% sodium hexanesulfonate and phosphoric acid at pH 2.2) for EACA and AMCHA; methanol/phosphate buffer (pH 2.5) (40/60) for PAMBA; acetonitrile with 0.06% tetrabutylammonium hydroxide (pH 6.5 with phosphoric acid) solution (20/80) for EDDA.
Inertsil ods 3 column
Manufactured by Shimadzu
Sourced in Japan
The Inertsil ODS-3 column is a reversed-phase high-performance liquid chromatography (HPLC) column. It is designed for the separation and analysis of a wide range of organic compounds. The column features a spherical silica gel with a chemically bonded octadecyl (C18) stationary phase, providing high efficiency and reproducible separations.
Automatically generated - may contain errors
Lab products found in correlation
Hplc system, by Shimadzu (2 mentions)
0.22 μm membrane filter, by Merck Group (1 mentions)
Prominence lc solution system, by Shimadzu (1 mentions)
Spd m20a, by Shimadzu (1 mentions)
Rid 10a, by Shimadzu (1 mentions)
Lc 20a, by Shimadzu (1 mentions)
Lc 10atvp pump, by Shimadzu (1 mentions)
Marfey s reagent, by Merck Group (1 mentions)
Automatic amino acid analyzer, by Hitachi (1 mentions)
8 protocols using inertsil ods 3 column
1
Photopolymerized Drug-Loaded Membranes
2
HPLC Analysis of Sugars and Organic Acids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sugar and organic acid contents were determined by high-performance liquid chromatography (HPLC), according to the method of Toshima et al. [23 (link)]. Each fraction (0.02 g) was dissolved in 5 mL of ultrapure water and passed through a 0.22-μm membrane filter (Millipore, Bedford, MA, USA) prior to analysis. For sugar content analysis, the extracts were subjected to HPLC using a UF-Amino station system (Shimazu, Kyoto, Japan) equipped with a refractive index detector (RID-10A, Shimadzu) and Asahipak NH2P-50 4E column (Showadenko, Tokyo, Japan). The chromatographic conditions were as follows: solvent, 75% (v/v) acetonitrile (CH3CN); column temperature, 40 °C; flow rate, 1.0 mL/min. Retention times and spectra were compared with those of pure standards of glucose, fructose, and sucrose. For organic acid content determination, each fraction was analyzed by reverse-phase HPLC using a Prominence LC solution system (Shimadzu) equipped with a photodiode array detector (SPD-M20A, Shimadzu) and Inertsil ODS3 column (Shimadzu). The chromatographic conditions were as follows: solvent, 75% (v/v) CH3CN; column temperature 40 °C; flow rate, 1.2 mL/min. Retention times and spectra were compared with those of pure standards of quinic acid, citric acid, and malic acid. Results are expressed as milligrams per gram dry weight of each fraction. Sample extracts were analyzed three times.
3
Quantitative Analysis of Bioactive Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The appropriate amount of harpagide, harpagoside, angloside C and cinnamic acid was accurately measured, dissolved by methanol, placed in a vial (volume 5 ml) and shaken to make a control product. The concentration was 1 g/L, 0.1 g/L, 1 g/L, 0.1 g/L to make a control solution. The water extract of SR was gradient elution at 1 ml/min, the detection wavelength was 210/280 nm, and acetonitrile-0.03 % phosphoric acid aqueous solution was used as mobile phase. Sample 10ul each time for 6 consecutive times. The precision, stability and repeatability of the reaction system were measured 24 h after the preparation of the control products. The control product was mixed with the sample to make the test solution, and the recovery rate and sample content (concentration) were measured.
Shimadzu Inertsil ODS-3 column was used in this study. The mobile phase A used in this study is 0.03 % phosphoric acid solution, and the mobile phase B is acetonitrile. The gradient elution procedure is shown in the following table. The flow rate used in this study was 1.0 ml/min, the detection wavelength was 210 nm and 280 nm, and the sample size was 10ul. The mobile phase gradient elution process is as follows.
Shimadzu Inertsil ODS-3 column was used in this study. The mobile phase A used in this study is 0.03 % phosphoric acid solution, and the mobile phase B is acetonitrile. The gradient elution procedure is shown in the following table. The flow rate used in this study was 1.0 ml/min, the detection wavelength was 210 nm and 280 nm, and the sample size was 10ul. The mobile phase gradient elution process is as follows.
| Time | mobile phase B (acetonitrile) | mobile phase A (0.03 % phosphoric acid solution) |
|---|---|---|
| 0∼10min | 10 %∼25 % | 90 %∼75 % |
| 10∼20min | 25 %∼30 % | 75 %∼70 % |
| 20∼30min | 30 %∼40 % | 70 %∼60 % |
| 30∼40min | 40 %∼50 % | 60 %∼50 % |
4
Quantifying Cryptotanshinone and Tanshinone IIA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CMC-OHA/TNCs of 1.00 g was placed into 100 mL of methanol and sonicated for 10 min to cause the cryptotanshinone and tanshinone IIA to completely dissolve in methanol. The samples were filtered through a 0.2 μm membrane filter, and the content of cryptotanshinone and tanshinone IIA were determined by HPLC (LC-20A, Shimadzu, Tokyo, Japan). Analyses were performed on inertsil ODS-3 column (250 × 4.6 mm, 5 μm, Shimadzu, Tokyo, Japan) with column temperature maintained at 30 °C. The mobile phase consisted of methanol and 0.1% phosphoric acid (85:15, v/v). The flow rate was 1 mL·min−1 and the detection wavelength was 254 nm. Tests of each sample were carried out three times and the results were recorded as an average.
5
Monosaccharide Composition Analysis of IOP-A2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Two milligrams of IOP‐A2 were dissolved in 1 mL of 2 M HCl/methanol and hydrolyzed at 80°C for 8 h. The sample was further hydrolyzed with 1 mL of 2 M trifluoroacetic acid (TFA) at 120°C for 1 h. The hydrolysate was derivatized with 1‐phenyl‐3‐methyl‐5‐pyrazolone (PMP) prior to the application of a DIKMA Inertsil ODS‐3 column (4.6 mm × 150 mm) and analyzed with a Shimadzu HPLC system (LC‐10ATvp pump and SPD‐10AVD UV–VIS detector). The monosaccharide composition was determined and quantified by taking the retention time and peak area of a standard monosaccharide as reference. (Needs & Selvendran, 2010 ).
6
Quantification of S-Adenosyl Methionine in Microbial Cultures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For measurement of SAM produced by M. purpureus, the fermentation culture was centrifuged at 8,000 × g for 10 min to collect mycelia and medium supernatant, respectively. The medium supernatant was filtered with a 0.22 μm pore membrane and used for SAM determination. The wet mycelia were dried at −80°C by vacuum freezing and incubated in the solution containing 4 ml of 1.5 M perchloric acid and 4 ml of 1.5 M ammonia water at 4°C for overnight. The mixture was centrifuged at 10,000 × g for 15 min to collect the supernatant for SAM determination. SAM was determined by high-performance liquid chromatography (HPLC) (SHIMADZU, Shanghai, China) as previously described with modifications (Wang et al., 2012 (link)). The HPLC analysis conditions were as follows: Inertsil ODS-3 column (4.6 mm × 250 mm, 5 μm, SHIMADZU, Shanghai, China); injection volume 20 μl; oven temperature 30°C; flow rate 1 ml/min; the eluate was monitored at 260 nm. Mobile phase A (10 mM ammonium formate buffer, pH 3.5) and phase B (methanol) were run on an isocratic elution program (95% phase A buffer and 5% phase B buffer). The standard chemicals SAM was purchased from Macklin Biochemical Co., Ltd (Shanghai, China).
7
Amino Acid Analysis of Pseudoalterin Hydrolysate
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PG (OD600 ≈ 1.0) was digested with 20 μg ml−1 pseudoalterin in 20 mM Tris-HCl (pH 9.0) at 25 °C for 24 h. The released amino acids in the hydrolysate were analyzed by an Automatic Amino Acid Analyzer (Hitachi L8900, Japan). The d /l -amino acids and peptides in the hydrolysate were also analyzed by HPLC. The d /l -amino acids in the hydrolysate were derivatized with FDAA (Nα-(2,4-dinitro-5-fluorophenyl)-l -alaninamide, Marfey’s reagent; Sigma) as described by Hess et al.48 (link). Derivatized amino acids were separated with a linear gradient of formic acid (30 mM, pH 2.6)/acetonitrile on an HPLC with an Inertsil ODS-3 column (250 × 4.6 mm; 5 μm particle size) (Shimadzu, Japan) at a flow rate of 1.5 ml min−1 and detected at 340 nm. Glycine, d -alanine, l -alanine, l -lysine, d -glutamate, AG, AGG, AGGG, AGGGG, AGGGGG, GG, GGG, GGGG, GGGGG, Ae, AeK, AeKa, AeKaA, eK, eKa, eKaA, Ka, KaA, and aA at concentrations of 0.5–4.0 mg ml−1 served as standard solutions.
Peptides AaKAGGGGGA and lactic acid-AeKAGG were synthesized by ChinaPeptides Co., Ltd (China). Each of these peptides (50 μg) was hydrolyzed using 2.5 μg pseudoalterin in 25 μl of 10 mM Tris-HCl (pH 9.0) at 20 °C for 25 h. The hydrolysates were subjected to LC-MS analysis to determine the molecular masses of the released peptides. The sequences of the released peptides were determined by using ExPASy tools.
Peptides AaKAGGGGGA and lactic acid-AeKAGG were synthesized by ChinaPeptides Co., Ltd (China). Each of these peptides (50 μg) was hydrolyzed using 2.5 μg pseudoalterin in 25 μl of 10 mM Tris-HCl (pH 9.0) at 20 °C for 25 h. The hydrolysates were subjected to LC-MS analysis to determine the molecular masses of the released peptides. The sequences of the released peptides were determined by using ExPASy tools.
8
HPLC-based Ascorbic Acid Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AA content was determined using highperformance liquid chromatography (HPLC) following the protocols of Lui et al. (2013) with modifications. Briefly, MFC was dissolved in 2.5% w/v metaphosphoric acid. Clear extracts (20 μL) were injected into an Inertsil® ODS-3 column (4.6 x 250 mm) of a Shimadzu HPLC system (Shimadzu Co., Kyoto, Japan). The mobile phase was pure deionized water adjusted with metaphosphoric acid to pH 2.2. Flow rate and column temperature were 0.8 mL/min and 40 , A UV-vis photodiode array detector at 254 nm was applied to quantify the AA. Results were calculated using L-ascorbic acid as an external standard and expressed as mg/ 100 g sample.
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!