Fractions B1 and B3 were subjected to further separations using a Shimadzu LC-20AR HPLC system (Shimadzu, Kyoto, Japan) equipped with a reverse phase column (Shim-pack PREP-ODS, 250 mm × 20 mm, 5 μm, Shimadzu, Kyoto, Japan), a binary pump, and a UV-Visible detector (SPD-20AV, Shimadzu, Kyoto, Japan). Fractions were redissolved in 0.1% formic acid in water and injected into the column. Compounds were eluted with a linear gradient with 0.1% formic acid in water (A) and 0.1% formic acid in CH3CN (B) as the mobile phase at a flow rate of 10 mL/min. Eluent B was increased from 5–45% in 3 min, then from 45 to 53% in 20 min, then held at 100% for 6 min, and finally set back to 5% for 4 min. The detection wavelength was set at 214 nm. Purified compounds Qb1 (2 mg, retention time at 13.5 min) and S13 (1 mg, retention time 20.5 min) were obtained by manual collection from B3 and B1, respectively.
Shim pack prep ods
Manufactured by Shimadzu
Sourced in Japan
The Shim-pack PREP-ODS is a high-performance liquid chromatography (HPLC) column designed for preparative-scale separations. It features an octadecylsilane (ODS) stationary phase for the separation of a variety of organic compounds. The column is suitable for use in preparative HPLC applications.
Automatically generated - may contain errors
4 protocols using shim pack prep ods
1
Isolation and Purification of Bioactive Compounds
2
HPLC Analysis of Rice Bran Oil Extracts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The rice bran oil extracts were dissolved in methanol, were further filtered with a 0.45 μm filter and were analyzed by HPLC. δ-Tocotrienol standard sample was purchased from Sigma (Sigma Co., Ltd., St. Louis, MO, USA). HPLC analysis was performed on a Shimadzu Prominence series apparatus with a fluorescence detector (Prominence RF-20A/Axs, Shimadzu Co., Ltd., Kyoto, Japan). The excitation wavelength was 296 nm and the emission wavelength was 325 nm, which were operated on fluorescence detector. The Hypersil Gold PFP column (250 mm × 4.6 mm i.d., 5 μm, ThermoFisher Scientific, Waltham, MA, USA) was used as an analytical column. The eluents were methanol/H2O (85:15, vol/vol) at a flow rate of 0.8 mL/min. All the results were recorded, and the peaks were integrated by the chromatography software Labsolution LC (Shimadzu, Kyoto, Japan). δ-Tocotrienol was purified by Shim-pack PREP-ODS (20.0 mm × 250 mm, 15 µm, Shimadzu, Kyoto, Japan). The eluents include acetonitrile, tetrahydrofuran, methanol, 1% ammonium acetate (684:220:68:28) and the flow rate was 8 mL/min. The δ-tocotrienol standard that was purchased from Chromadex, Inc. (Irvine, CA, USA) was used to determine the absorption peak of δ-tocotrienol.
3
Semipreparative HPLC Purification of DTPA–DIG
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DTPA–DIG was isolated by semipreparative RP-HPLC using a Shimadzu
equipment (Shimadzu Corp., Japan) composed of wo LC6AD pumps and a
manual injection valve (Rheodyne 7125, Rheodyne, CO). The injection
was performed using a 1.000 μL sample loop, and detection was
carried out at 200 and 220 nm, employing a dual-wavelength detector
(SPD M10A) controlled by LCsolution software v.1.25 (Shimadzu Corp.).
Separation was performed on an ODS column (Shim-pack PREP-ODS; 250
× 4.6 mm2 i.d., 5 μm), using a linear gradient
of 10–100% ACN with 0.1% formic acid (v/v) for elution in 20
min, at a flow rate of 7 mL/min. The obtained peak fractions were
pooled, and the acetonitrile was removed in a rotary evaporator at
30 °C, followed by lyophilization to eliminate the water.
equipment (Shimadzu Corp., Japan) composed of wo LC6AD pumps and a
manual injection valve (Rheodyne 7125, Rheodyne, CO). The injection
was performed using a 1.000 μL sample loop, and detection was
carried out at 200 and 220 nm, employing a dual-wavelength detector
(SPD M10A) controlled by LCsolution software v.1.25 (Shimadzu Corp.).
Separation was performed on an ODS column (Shim-pack PREP-ODS; 250
× 4.6 mm2 i.d., 5 μm), using a linear gradient
of 10–100% ACN with 0.1% formic acid (v/v) for elution in 20
min, at a flow rate of 7 mL/min. The obtained peak fractions were
pooled, and the acetonitrile was removed in a rotary evaporator at
30 °C, followed by lyophilization to eliminate the water.
4
Isolation and Characterization of 8-oxo-erythraline
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Mn (salen) oxidation reaction was submitted to semipreparative LC using the following conditions: C18 column (Shim-pack Prep-ODS, 5 μm, 20 mm × 25 cm, Shimadzu), flow rate 9 mL/min and acetonitrile (B) and H2O (A) both with TFA 0.02% (v/v) as solvents. The elution profile was 0–25 min – 15-65% B, 25–30 min – 25-80% B, 30–32 min – 80-100% B and 32–35 min – 100% B. The compound 8-oxo-erythraline (2) was isolated from this procedure and characterized by MS and NMR.
8-oxo-erythraline (2): brown amorphous solid;1H NMR (CDCl3, 400 MHz) δH 6.79 (dd, J = 2.3, 10.2 Hz, H-1), 6.65 (s, H-14), 6.64 (s, H-17), 6.26 (dbr, J = 10.2 Hz, H-2), 6.01 (sbr, H-7), 5.86 (d, J = 1.0 Hz, O-CH2-O), 5.83 (d, J = 1.0 Hz, O-CH2-O), 3.81 (m, H-3), 3.69 (m, H-10), 3.58 (ddd, J = 3.7, 6.9, 12.4 Hz, H-10), 3.27 (s, –OCH3), 3.07 (ddd, J = 7.5, 9.5, 15.8 Hz, H-11), 2.91 (ddd, J = 3.7, 6.9, 15.8 Hz, H-11), 2.73 (dd, J = 4.9, 11.1 Hz, H-4), 1.62 (t, J = 11.1 Hz, H-4); 13C NMR (CDCl3, 100 MHz) δC 171.6 (C-8), 157.7 (C-6), 147.2 (C-15), 146.1 (C-16), 136.8 (C-2), 129.5 (C-13), 127.7 (C-12), 123.8 (C-1), 119.7 (C-7), 109.5 (C-17), 105.0 (C-14), 101.2 (O-CH2-O), 74.8 (C-3), 67.3 (C-5), 56.5 (OCH3), 41.1 (C-10),38.0 (C-4), 27.3 (C-11). ESI MS (pos. ion mode) m/z 312.1232 [M + H]+ (calcd. for C18H18NO4+ 312.12303).
8-oxo-erythraline (2): brown amorphous solid;1H NMR (CDCl3, 400 MHz) δH 6.79 (dd, J = 2.3, 10.2 Hz, H-1), 6.65 (s, H-14), 6.64 (s, H-17), 6.26 (dbr, J = 10.2 Hz, H-2), 6.01 (sbr, H-7), 5.86 (d, J = 1.0 Hz, O-CH2-O), 5.83 (d, J = 1.0 Hz, O-CH2-O), 3.81 (m, H-3), 3.69 (m, H-10), 3.58 (ddd, J = 3.7, 6.9, 12.4 Hz, H-10), 3.27 (s, –OCH3), 3.07 (ddd, J = 7.5, 9.5, 15.8 Hz, H-11), 2.91 (ddd, J = 3.7, 6.9, 15.8 Hz, H-11), 2.73 (dd, J = 4.9, 11.1 Hz, H-4), 1.62 (t, J = 11.1 Hz, H-4); 13C NMR (CDCl3, 100 MHz) δC 171.6 (C-8), 157.7 (C-6), 147.2 (C-15), 146.1 (C-16), 136.8 (C-2), 129.5 (C-13), 127.7 (C-12), 123.8 (C-1), 119.7 (C-7), 109.5 (C-17), 105.0 (C-14), 101.2 (O-CH2-O), 74.8 (C-3), 67.3 (C-5), 56.5 (OCH3), 41.1 (C-10),38.0 (C-4), 27.3 (C-11). ESI MS (pos. ion mode) m/z 312.1232 [M + H]+ (calcd. for C18H18NO4+ 312.12303).
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!