Metanephrines in Urine - HPLC

Order No.: 2020-B, for 100 tests
Parameters:
3-Methoxytyramine, Metanephrine, Normetanephrine

No pH adjustment after acidic hydrolysis
Only 1 sample clean up column required
Suitable for automated sample preparation

Assay validated according to IVDR (=> Declaration of Conformity)

3-Methoxytyramine

Metanephrine

Normetanephrine

Clinical relevance
This assay allows the simultaneous quantitative determination of total (sum of free and conjugated metabolites) metanephrine, normetanephrine and 3-methoxytyramine in human urine samples by HPLC.

It is intended to be used for patients in whom the urinary levels of total normetanephrine, total metanephrine and total 3-methoxytyramine are of clinical importance, primarily as an aid to diagnosis and monitoring of suspected pheochromocytoma and paraganglioma.

Furthermore, it is intended for patients in whom the urinary levels of total normetanephrine and total 3-methoxytyramine are of clinical importance, as an aid to diagnosis and monitoring of suspected neuroblastoma.

 

Assay characteristics
Sample preparation is carried out manually or semi-automated on ASPEC® (simple liquid handling system), and analytic separation is done via high performance liquid chromatography with electrochemical detection (HPLC-ECD). The use of specific internal standards ensures high precision and reliability in quantitation of the analytes.

The assays 2020-C and 6000-C are available as alternative methods that allow the combined determination of catecholamines and metanephrine with one column.

Detailed performance evaluation data can be found in Appendices II and III of the instruction manual.

More Information
Method of Analysis HPLC
Number of Tests 100
Please note The information listed here, including the sample preparation, is not sufficient for using the product. Please read the information provided in the instruction manual, which includes detailed information on limitations associated with the use of the product in line with its intended purpose. Detailed performance evaluation data can be found in Appendices II and III of the instruction manual.
Lower and Upper Limit of Quantitation

Normetanephrine: 4.5 µg/L – 3000 µg/L
Metanephrine: 7.3 µg/L – 5900 µg/L
3-Methoxytyramine: 9.9 µg/L – 5900 µg/L

Specimen 24-hour urine. Alternatively spot urine may also be used. In this case, results must be related to creatinine.
Sample Preparation

The information on the manual sample preparation presented here is not sufficient for use in the laboratory. For a detailed step by step description, please refer to the instruction manual.

Acidic hydrolysis
• Pipette 100 µL Internal Standard into a sealable hydrolysis tube.
• Add 1 mL sample/calibrator/control.
• Adjust pH using 2 N HCl.
• Seal the tube and incubate at 95°C for 30 min, then cool down.

Neutralisation
• Add 6 mL Neutralisation Buffer to the hydrolysed urine.

Extraction
• Agitate the Sample Clean Up Column.
• Take off the column cap and remove the sealing nipple.
• Allow the equilibration buffer to flow through completely, discard effluent.
• Apply the entire urine dilution onto the Sample Clean Up Column, allow to flow through, discard effluent.
• Wash with 10 mL high-purity water, discard effluent.
• Wash with 5 mL Wash Buffer, discard effluent.
• Add 5 mL Elution Buffer to the Sample Clean Up Column and collect the eluate into a suitable vial.

Continued sample preparation prior to injection
• Add 150 µL glacial acetic acid to the eluate, mix briefly.
• Inject 20 µL of the eluate into the HPLC system.

 

For the semi-automated sample preparation with the Gilson® ASPEC®, please refer to the instruction manual.

Run Time 15 min (Internal Standard 2024) --- 20 min (Internal Standard High Resolution 2044/HR)
Injection Volume 20 µL (manual sample preparation)
Flow Rate 1.0 mL/min
Column Temperature +20 to +25 °C
Potential approx. +780 mV to +840 mV
Additional Info For the Chromsystems HPLC analysis of metanephrines in urine any isocratic HPLC system with electrochemical detector is suitable.
Parameters 3-Methoxytyramine, Metanephrine, Normetanephrine
The following components are included in the kit:
The following products are not included in the kit but are required for the application of the method:

3-Methoxytyramine

Metanephrine

Normetanephrine

Clinical relevance
This assay allows the simultaneous quantitative determination of total (sum of free and conjugated metabolites) metanephrine, normetanephrine and 3-methoxytyramine in human urine samples by HPLC.

It is intended to be used for patients in whom the urinary levels of total normetanephrine, total metanephrine and total 3-methoxytyramine are of clinical importance, primarily as an aid to diagnosis and monitoring of suspected pheochromocytoma and paraganglioma.

Furthermore, it is intended for patients in whom the urinary levels of total normetanephrine and total 3-methoxytyramine are of clinical importance, as an aid to diagnosis and monitoring of suspected neuroblastoma.

 

Assay characteristics
Sample preparation is carried out manually or semi-automated on ASPEC® (simple liquid handling system), and analytic separation is done via high performance liquid chromatography with electrochemical detection (HPLC-ECD). The use of specific internal standards ensures high precision and reliability in quantitation of the analytes.

The assays 2020-C and 6000-C are available as alternative methods that allow the combined determination of catecholamines and metanephrine with one column.

Detailed performance evaluation data can be found in Appendices II and III of the instruction manual.

More Information
Method of Analysis HPLC
Number of Tests 100
Please note The information listed here, including the sample preparation, is not sufficient for using the product. Please read the information provided in the instruction manual, which includes detailed information on limitations associated with the use of the product in line with its intended purpose. Detailed performance evaluation data can be found in Appendices II and III of the instruction manual.
Lower and Upper Limit of Quantitation

Normetanephrine: 4.5 µg/L – 3000 µg/L
Metanephrine: 7.3 µg/L – 5900 µg/L
3-Methoxytyramine: 9.9 µg/L – 5900 µg/L

Specimen 24-hour urine. Alternatively spot urine may also be used. In this case, results must be related to creatinine.
Sample Preparation

The information on the manual sample preparation presented here is not sufficient for use in the laboratory. For a detailed step by step description, please refer to the instruction manual.

Acidic hydrolysis
• Pipette 100 µL Internal Standard into a sealable hydrolysis tube.
• Add 1 mL sample/calibrator/control.
• Adjust pH using 2 N HCl.
• Seal the tube and incubate at 95°C for 30 min, then cool down.

Neutralisation
• Add 6 mL Neutralisation Buffer to the hydrolysed urine.

Extraction
• Agitate the Sample Clean Up Column.
• Take off the column cap and remove the sealing nipple.
• Allow the equilibration buffer to flow through completely, discard effluent.
• Apply the entire urine dilution onto the Sample Clean Up Column, allow to flow through, discard effluent.
• Wash with 10 mL high-purity water, discard effluent.
• Wash with 5 mL Wash Buffer, discard effluent.
• Add 5 mL Elution Buffer to the Sample Clean Up Column and collect the eluate into a suitable vial.

Continued sample preparation prior to injection
• Add 150 µL glacial acetic acid to the eluate, mix briefly.
• Inject 20 µL of the eluate into the HPLC system.

 

For the semi-automated sample preparation with the Gilson® ASPEC®, please refer to the instruction manual.

Run Time 15 min (Internal Standard 2024) --- 20 min (Internal Standard High Resolution 2044/HR)
Injection Volume 20 µL (manual sample preparation)
Flow Rate 1.0 mL/min
Column Temperature +20 to +25 °C
Potential approx. +780 mV to +840 mV
Additional Info For the Chromsystems HPLC analysis of metanephrines in urine any isocratic HPLC system with electrochemical detector is suitable.
Parameters 3-Methoxytyramine, Metanephrine, Normetanephrine
The following components are included in the kit:
The following products are not included in the kit but are required for the application of the method: