HPLC Method for Analysis of Hydroxyproline, Creatine, Creatinine on Newcrom AH by SIELC Technologies
High Performance Liquid Chromatography (HPLC) Method for Analysis of Hydroxyproline, Creatine, Creatinine
Hydroxyproline is a non-essential amino acid primarily found in collagen, the main structural protein in connective tissues such as skin, bones, and cartilage. It plays a crucial role in maintaining collagen stability and is often used as a biomarker for collagen turnover and connective tissue disorders. Creatine is a naturally occurring compound found in muscles and is used to produce energy during high-intensity exercise. It is synthesized in the body from amino acids and stored in muscles for quick energy release. Creatinine, on the other hand, is a waste product produced from the breakdown of creatine. It is filtered by the kidneys and excreted in urine. The measurement of creatinine levels in the blood and urine is commonly used to assess kidney function, as high levels may indicate impaired kidney function or other health issues.
Hydroxyproline, Creatine, and Creatinine can be analyzed and separated using a Newcrom AH mixed-mode stationary phase column. The analysis utilizes an isocratic method with a simple mobile phase consisting of water, acetonitrile (MeCN), and formic acid as a buffer. Mass spectrometric detection was carried out in ESI-SIM mode, monitoring the ions [M+H]⁺ at m/z 132.00, 132.05, 114.00.
| Column | Newcrom AH, 3.2 x 100 mm, 5 µm, 100 A |
| Mobile Phase | MeCN/H2O – 10/90% |
| Buffer | Formic Acid – 0.5% |
| Flow Rate | 0.5 ml/min |
| Detection | ESI-SIM: [M+H]⁺ 132.00, 132.05, 114.00 |
| Sample | 1. 0.6 mg/mL, 2. 0.15 mg/mL, 3. 0.06 mg/mL |
| Injection volume | 1 µl |
| LOD* | 1. 3.8 ppb, 2. 4.0 ppb, 3. 2.1 ppb |
| Class of Compounds | Amino Acid-Derived compounds |
| Analyzing Compounds | Hydroxyproline, Creatine, Creatinine |
Application Column
Newcrom AH
Column Diameter: 3.2 mm
Column Length: 100 mm
Particle Size: 5 µm
Pore Size: 100 A
Creatinine
Hydroxyproline
