RSpak DE-413, a polymer-based reversed phase chromatography column, can be used with aqueous eluents with no organic solvent. In this application, simultaneous analysis of C1 compounds including formaldehyde, methanol, and formic acid was performed using an aqueous solution of phosphoric acid as an eluent.
Column:Shodex RSpak DE-413 (4.6 mm I.D. x 150 mm) x 2
Eluent:10 mM H3PO4 aq.
Flow rate:1.0 mL/min
Detector:RI, UV (190 nm)
Column temp.:50 ℃
Wine contains furfural and guaiacol. Furfural has butterscotch and caramel notes, and guaiacol has a charcoal, smoky note, both contribute to wine flavoring. In this application, furfural and guaiacol were analyzed using RSpak DE-413, a polymer-based reversed phase chromatography column.
Column:Shodex RSpak DE-413 (4.6 mm I.D. x 150 mm)
Eluent:2 mM HClO4 aq./CH3CN=50/50
Flow rate:1.0 mL/min
Detector:UV (210 nm)
Column temp.:40 ℃
ε-caprolactam, a residual monomer in nylon 6, was analyzed using Asahipak ODP-50 4D, a reversed-phase chromatography column. Analysis of ε-caprolactam can be performed without an influence from extract contaminants.
Sample pretreatment
- Add 10 mL of pure water to 0.5 g of nylon 6 pellets. Seal and heat at 100 ℃ for 2 hours.
- Once the mixture is cooled to room temperature, filter through a 0.5μm filter. Use the filtrate as a test sample.
Column:Shodex Asahipak ODP-50 4D (4.6 mm I.D x 150 mm)
Eluent:H2O/CH3CN=80/20
Flow rate:0.7 mL/min
Detector:UV (210 nm)
Column temp.:40 ℃
3,4-Dimethylpyrazole phosphate (DMPP) is a nitration inhibitor added to solid fertilizers. The Japanese regulation for “Testing Methods for Fertilizers (2022*)” mentions to use an octadecylsilylated silica gel column for DMPP analysis. A calibration curve for DMPP obtained using Silica C18M 4D showed a high linearity with a coefficient of determination (R2) 0.9999 in the concentration range 0.5 to 50 μg/mL.
Urea fertilizer, one of the target fertilizers, was analyzed. The extract was prepared in accordance with the Testing Methods for Fertilizers. DMPP was not detected in the extract of the fertilizer analyzed. As a next step, addition recovery test was performed using the same sample (spiked DMPP concentration 0.185 mg/g). A good result was obtained with an average recovery rate of 102 % (n=3).
*The version at the time of the application acquisition.
Sample preparation
- (1)Measure 1.00 g of urea fertilizer and put it in a 200-mL erlenmeyer flask. (For the spike recovery test DMPP was added to the sample at this point.)
- (2)Add 100 mL of water to (1) and stir with a magnetic stirrer for 10 minutes.
- (3)After allowing it to stand, transfer a portion of the supernatant to a 1.5-mL glass-stoppered centrifugal sedimentation tube.
- (4)Centrifuge (8000 ~ 10000 x g for about 5 minutes) and use the supernatant as an injection sample.
Column:Shodex Silica C18M 4D (4.6 mm I.D x 150 mm)
Eluent:10 mM NaH2PO4 aq./CH3CN=1000/175
Flow rate:0.7 mL/min
Detector:UV (224 nm)
Column temp.:40 ℃
Ceftazidime is a cephem antibiotic used to treat bacterial infections. Some formulations of ceftazidime contain arginine. KW402.5-4F, an aqueous SEC (GFC) column, was used to separate ceftazidime and arginine, and a good separation was obtained.
Column:Shodex KW402.5-4F (4.6 mm I.D. x 300 mm)
Eluent:1.15 g/L NH4H2PO4 (pH 2.0 adjusted with H3PO4)/CH3CN=25/75
Flow rate:0.5 mL/min
Detector:UV (206 nm)
Column temp.:30 ℃
Carboplatin, a type of anticancer drug, is called a platinum agent because it contains platinum in its structure. A silica-based amino column is used in Assay of Carboplatin listed in the United States Pharmacopeia and the National Formulary (USPNF 2022 Issue 1*). With reference to this USP monograph, carboplatin was analyzed using Asahiak NH2P-50 4E, a polymer-based amino column. We confirmed that NH2P-50 4E was also suitable for the analysis.
*The version at the time of the application acquisition.
Column:Shodex Asahipak NH2P-50 4E (4.6 mm I.D. x 250 mm)
Eluent:CH3CN/H2O=83/17
Flow rate:1.5 mL/min
Detector:UV (230 nm)
Column temp.:25 ℃
USP-NF GENERAL CHAPTERS lists <121.1> Physicochemical Analytical Procedures for Insulins. The target analytes included in this method are insulin analogues, animal-derived insulin, and human insulin. The analysis of high molecular weight proteins should be carried out with a column filled with L20 packing material and meets following requirements.
System suitability requirements
Retention times:
Polymeric insulin complexes: 13 – 17 min
Covalent insulin dimer: about 17.5 min
Insulin monomer: 18 – 22 min
Peak-to-valley ratio*: ≥ 2.0
*The ratio of the height of the covalent insulin dimer peak to the height of the valley between the covalent insulin dimer peak and the insulin monomer peak.
Below lists the monographs using this method (at 2023 June).
Insulin |
Insulin Injection |
Insulin Human |
Insulin Human Injection |
Insulin Aspart |
Insulin Aspart Injection |
Insulin Glargine |
Insulin Glargine Injection |
Insulin Lispro |
Insulin Lispro Injection |
Insulin Zinc Suspension |
Extended Insulin Zinc Suspension |
Prompt Insulin Zinc Suspension |
Isophane Insulin Suspension |
Isophane Insulin Human Suspension |
Human Insulin Isophane Suspension
and Human Insulin Injection |
The PROTEIN KW-802.5 confirmed the requirements were met for the analysis of insulin beef.
- Column:Shodex PROTEIN KW-802.5 (8.0 mm I.D. x 300 mm)
- Eluent:0.1 wt% L-Arginine aq./CH3CN/CH3COOH=13/4/3
- Flow rate:0.5 mL/min
- Detector:UV (276 nm)
- Column temp.:25 ℃
Proteoglycan, a type of glycoprotein, is found in cartilages of humans and animals. They are glycosaminoglycans bound to proteins. Proteoglycan promotes skin cell growth and the production of hyaluronic acid and collagen. Proteoglycan itself also has a high-water retention capability. Using those advantages, proteoglycans are used in functional foods and cosmetics, expecting the effects such as anti-aging of the skin, alleviation of arthritis symptoms, and inflammation suppression.
In this application, proteoglycan in a commercial supplement was analyzed using OHpak SB-806M HQ, an aqueous SEC column. A good linear calibration was achieved. Analysis of a commercial supplement showed a close to a 100 % recovery rate, compared to the manufacturer’s information provided.
Sample preparation
- Grind a supplement in a mortar to make it into powders.
- Measure 270 mg of the powder and add 15 mL of 4-M guanidine hydrochloride solution.
- Sonicate for 15 minutes and then centrifuge at 1800 x g for 10 minutes.
- Collect the supernatant. Add another 15 mL of 4-M guanidine hydrochloride to the residue and repeat step (3). Repeat this process one more time (i.e., a total of 3 times adding the 4-M guanidine hydrochloride).
- Ultrafiltrate (MW: 50,000-100,000) all collected supernatant from step (4) at 2400 x g for 30 minutes.
- Add 5 mL of water to the “proteoglycan fraction” which did not pass through the ultrafiltration membrane. Centrifuge at 1800 x g for 10 minutes. Repeat this step five times to wash the proteoglycan fraction.
- Add eluent to the proteoglycan fraction to make up the volume of 20 mL.
- Filtrate with 0.45μm membrane and use it as an injection sample.
Columns : Shodex OHpak SB-806M HQ (8.0 mm I.D. x 300 mm) + SB-G 6B (6.0 mm I.D. x 100 mm)
Eluent : 25 mM KH2PO4 + 25 mM Na2HPO4 aq.
Flow rate : 0.5 mL/min
Detector : RI
Column temp. :40 ℃