Sarcosine Analyzed with LCMS - AppNote
May 1, 2012
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Separation of Potential Urine Biomarker from Isobaric ß-Alanine
This developed LCMS method can separate Sarcosine from Beta- Alanine in serum and urine samples without using labor intensive 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.
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).
Column: Cogent Diamond Hydride™, 4μm, 100Å
Catalog No.: 70000-15P-2
Dimensions: 2.1 x 15mm
Mobile Phase:
--A: 50% Isopropyl Alcohol / 50% DI Water / 0.1% Acetic Acid
--B: 97% Acetonitrile / 3% DI Water / 0.1% Acetic Acid
Gradient:
Temperature: 50˚C
Post Time: 5 minutes
Flow rate: 0.6mL / minute
Detection: ESI – POS - Agilent 6210 MSD TOF Mass Spectrometer
Injection vol.: 1μL
Sample Preparation: 10mg / L each of Sarcosine and Beta-Alanine in 50:50 A:B
Note: 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.
Attachment
No 129 Sarcosine Analyzed with LCMS pdf 0.2 Mb Download File
This developed LCMS method can separate Sarcosine from Beta- Alanine in serum and urine samples without using labor intensive 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.
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).
Peaks:
1. Sarcosine
2. ß-Alanine
Column: Cogent Diamond Hydride™, 4μm, 100Å
Catalog No.: 70000-15P-2
Dimensions: 2.1 x 15mm
Mobile Phase:
--A: 50% Isopropyl Alcohol / 50% DI Water / 0.1% Acetic Acid
--B: 97% Acetonitrile / 3% DI Water / 0.1% Acetic Acid
Gradient:
| Time (minutes) | %B |
| 0 | 75 |
| 3 | 75 |
| 4 | 65 |
| 5 | 65 |
| 10 | 20 |
| 12 | 75 |
Post Time: 5 minutes
Flow rate: 0.6mL / minute
Detection: ESI – POS - Agilent 6210 MSD TOF Mass Spectrometer
Injection vol.: 1μL
Sample Preparation: 10mg / L each of Sarcosine and Beta-Alanine in 50:50 A:B
Note: 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.
Attachment
No 129 Sarcosine Analyzed with LCMS pdf 0.2 Mb Download File