Morphine Hydromorphone and 6-MAM in Plasma Analyzed by LC-MS - AppNote
July 11, 2014
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Date: 11-JULY-2014 Last Updated: 28-SEPTEMBER-2026
Morphine, Hydromorphone and 6-Monoacetylmorphine (6-MAM) in Plasma
This AppNote presents a Method that could be used for a routine analysis of Plasma Samples or (whole blood samples after changing the extraction procedure) for the presence of Morphine, Hydromorphone (isobaric compounds) or 6-MAM (an indicator of Heroin use).
Peaks:
1. Morphine
286.1438 m/z [M+H]+
2. Hydromorphone
286.1438 m/z [M+H]+
3. 6-Monoacetylmorphine (6-MAM)
328.1543 m/z [M+H]+
Column: Cogent Bidentate C18 2.o™, 2.2 μm, 120 Å
Catalog No.: 40218-05P-2
Dimensions: 2.1 x 50 mm
Mobile Phase:
-- A: DI Water / 0.1% Formic Acid (v/v)
-- B: 50:50 Acetonitrile / Methanol / 0.1% Formic Acid (v/v)
Gradient:
| Time (minutes) | %B |
|---|---|
| 0 | 5 |
| 4 | 50 |
| 5 | 90 |
| 6 | 90 |
| 7 | 5 |
Post Time: 3 minutes
Injection vol.: 1 μL
Flow rate: 0.4 mL / minute
Detection: ESI – POS - Agilent 6210 MSD TOF Mass Spectrometer
Sample Preparation: To a spiked Plasma Sample (1 mL), 1 mL of an Ammonia solution (0.1% v/v) was added and vortex-mixed for 30 seconds to alkalize the Plasma. Then, the alkalized Plasma Sample was extracted with two 4 mL Ethyl Acetate aliquots by vortex for 5 minutes, and centrifuged at 4000 rpm for 8 minutes at room temperature. The supernatant was separated and evaporated to dryness under a gentle stream of Nitrogen. The residue was reconstituted with 200 µL Mobile Phase, and a 1 µL aliquot of the reconstituted solution was injected into the LC– ESI-MS for analysis.
Note : Morphine is used as an analgesic drug in the treatment of the severe pain. Hydromorphone, a semi-synthetic derivative of Morphine is used in cases of Morphine-resistant Cancer-related pain and 6-Monoacetylmorphine (6-MAM) is a specific indicator for Heroin use. All three drugs remain the selected substances in monitoring of drug abuse and addiction. Heroin use remains a serious public health issue which accounts for a substantial proportion of the health costs. Analysis of Plasma Samples for the presence of the three substances is essential in assessing short or long term exposure to the drugs.
