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 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 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 close to 100 % recovery rate, compared to the manufacturer's information provided.
- (1)Grind a supplement in a mortar to make it into powders.
- (2)Measure 270 mg of the powder and add 15 mL of 4-M guanidine hydrochloride solution.
- (3)Sonicate for 15 minutes and then centrifuge at 1800 x g for 10 minutes.
- (4)Collect the supernatant. Add another 15 mL of 4-M guanidine hydrochloride to the residue and repeat the step (3). Repeat this process one more time (i.e., total of 3 times adding the 4-M guanidine hydrochloride).
- (5)Ultrafiltrate (MW: 50,000-100,000) all collected supernatant from step (4) at 2400 x g for 30 minutes.
- (6)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.
- (7)Add eluent to the proteoglycan fraction to make up the volume to 20 mL.
- (8)Filtrate with 0.45μm membrane and use it as an injection sample.
Sample: 50 µL
Proteoglycan standard solution (0.1 mg/mL)
Pretreated supplement extract solution
- 1.Proteoglycan
- Column
- :Shodex OHpak SB-G 6B (6.0 mm I.D. x 100 mm) + SB-806M HQ (8.0 mm I.D. x 300 mm)
- Eluent
- :25 mM KH2PO4 + 25 mM Na2HPO4 aq.
- Flow rate
- :0.5 mL/min
- Detector
- :RI
- Column temp.
- :40 ℃
Sample Name Index
Operation Manual / Certificate of Analysis
Operation Manuals and Certificate of Analysis / Inspection Certificate for the following products can be downloaded here.
Product Name Index
Applications
- Sample Load (KW-803)
- Effects of Flow Rate (KW-802.5, KW-803)
- Standard Proteins (1) (KW-802.5)
- Standard Proteins (2) (KW-802.5 and KW-803)
- Standard Proteins (14) (KW-802.5)
- Standard Proteins (15) (KW-803)
- Standard Proteins (16) (KW-804)
- Standard Proteins (18) (KW-803)
- Standard Proteins (19) (KW-804)
- Standard Proteins (23) (GF-510 HQ)
- Standard Proteins (24) (KW402.5-4F)
- Standard Proteins (25) (KW403-4F)
- Standard Proteins (26) (KW404-4F)
- Standard Proteins (27) (KW405-4F)
- Recovery of Proteins (GF-310 HQ and GS-320 HQ)
- Recovery of Proteins (KW-802.5 and KW-803)
- Recovery of Proteins (KW-803)
- Proteins in Human Serum (3) (KW-803)
- Dimer and Trimer of BSA
- Crude Albumin (Chicken Egg) (1) (KW-803)
- Crude Lipoxidase
- Crude Catalase
- Crude Hexokinase
- Human Serum (1) (SB-802 HQ)
- Tryptic Digest of Human IgG
- Comparison of Chromatograms of Protein mixtures
- Comparison between KW402.5-4F and KW-802.5
- Recovery of Proteins (KW402.5-4F, KW403-4F)
- Human Control Serum
- Analysis of Whey in Yogurt (KW403 4F + KW402.5 4F)
- Lectin Protein
- Peptide Analysis (SEC)
- PEG-Chymotrypsin (KW-802.5)
- SEC Analysis of PEG-Asparaginase (SB-806 HQ)
- Human IgM (2)
- Poly-Lysine
- Avidin (2)
- α1 Acid Glycoprotein
- Analysis of Gelatin (SB-806M HQ)
- Analysis of Insulin Glargine According to JP Method (KW-802.5)
- Analysis of Human Insulin According to JP Method (KW-802.5)
- Analysis of High Molecular Impurities in Insulin Aspart According to Pharmacopeia Method (KW-802.5)
- Analysis of High Molecular Weight Protein According to USP-NF GENERAL CHAPTERS Physicochemical Analytical Procedures for Insulins (KW-802.5)
- Comparison of SEC Separation of Standard Proteins
- Comparison of SEC Separation of Polyclonal Antibody
- Comparison of SEC Separation of Monoclonal Antibody
- Comparison of Separation of BSA Aggregates (LW-803, KW-803)
- Monitoring Papain Digestion of Humanized Monoclonal IgG (LW-803)
- Analysis of Lipoproteins in Serum (KW-804)
- Comparison of SEC Separation of Standard Proteins between LW-403 4D and LW-803
- Durability Test by Number of Sample Injections (LW-403 4D)
- Analysis of Epoetin According to USP-NF Method (LW-803)
- Comparison of Separation of Proteins with Conventional Device and Semi-micro Type Device (LW-403 4D)
- Comparison of IgG Separation with Conventional Device and Semi-micro Type Device (LW-403 4D)
- Particle Size Distribution Determination of Hepatitis B Surface Antigen Virus-like Particle (HBsAg VLP) (KW405-4F)
- Analysis of Hemocyanin (KW405-4F)
- Analysis of Somatropin According to EP Method (LW-803)
- SEC Analysis of Rituximab (LW-803)
- SEC/MALS Analysis of Norovirus VLP (SB-805 HQ)
- Analysis of Thymosin α1 (KW-802.5)
- SEC/MALS Analysis of Exosome (SB-806 HQ)
- Molecular Weight Measurements of Low Molecular Weight Heparins According to USP-NF GENERAL CHAPTERS <209> (KW-803 + KW-802.5)
Applications (Related Information)
- Calibration Curves for KW-800 Series (Pullulan)
- Calibration Curves for KW-800 Series (Proteins)
- Calibration Curves for KW-800 Series (PEG, PEO)
- Calibration Curves of KW400-4F (1) (Pullulan)
- Calibration Curves of KW400-4F (2) (Proteins)
- Calibration Curves of KW400-4F (3) (PEG, PEO)
- Calibration Curves for GF-HQ Series (Aqueous Eluent: Proteins)
- Calibration Curve for LW-803 (Proteins)
- Calibration Curve for LW-403 4D (Proteins)