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Sarcosine
Separation of potential urine biomarker from isobaric β-alanine
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printable Application Sheet
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Notes: When reversed phase columns were evaluated for their ability to separate sarcosine from beta-alanine, both compounds eluted at the
solvent front and were not separated. To achieve separation, a very intensive sample preparation has to be employed (e.g. derivatization) when
using RP methods.
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Method Conditions
| Column |
Cogent Diamond Hydride™, 4µm, 100A |
| Catalog No. |
70000-15P-2 |
| Dimensions |
2.1 x 150 mm |
| Solvents |
| A: | 50% isopropyl alcohol/ 50% DI water/ 0.1% acetic acid |
| B: | 97% acetonitrile/ 3% DI water/ 0.1% acetic acid |
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| Gradient |
| time (min.) |
%B |
time (min.) |
%B |
| 0 |
75 |
5 |
65 |
| 3 |
75 |
10 |
20 |
| 4 |
65 |
12 |
75 |
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| Post Time |
5 min |
| Injection Vol. |
1 microL |
| Flow rate |
0.6 mL/min. |
| Temperature |
50°C |
| Sample |
10 mg/L ea. of sarcosine and beta-alanine in 50:50 A:B |
| Detection |
ESI – POS - Agilent 6210 MSD TOF mass spectrometer |
Discussion
This developed LC-MS method can separate sarcosine from beta-alanine in serum and urine samples without using laborintensive sample derivatization.
Since sarcosine is considered a potential biomarker for prostate cancer risk and aggressiveness, it is essential to resolve and accurately quantify
this compound in the presence of isobaric (same m/z) beta-alanine. This objective is achieved using a Cogent Diamond Hydride™ column and a simple
gradient method presented in this application note. The developed method is sensitive, specific, quantitative, and reproducible (%RSD = 0.1). It can be
used in large scale studies with numerous samples (high throughput of the method due to simple sample preparation).
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