Comparison of serum and lithium‐heparinate plasma for the accurate measurements of endogenous and exogenous morphine concentrations

Laux-Biehlmann, Alexis; Gräfe, Nadja; Mouheiche, Jinane; Stuber, Denise; Welters, Ingeborg; Delalande, François; Poisbeau, Pierrick; Garnero, Patrick; Metz‐Boutigue, Marie‐Hélène; Schneider, Francis; Goumon, Yannick

doi:10.1111/j.1365-2125.2012.04199.x

Cited by 5 publications

(9 citation statements)

References 4 publications

(6 reference statements)

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“…We discovered the EIAs to be potent ligands of the ASIC3 channel. As it was shown earlier, EIAs’ blood levels increase under pathological conditions such as infection or inflammation (Glattard et al, 2010 ; Laux-Biehlmann et al, 2012 ). Increases in EIAs’ concentration may contribute to pain symptoms in people suffering from Parkinson’s disease, as evidenced by the correlation between reduced content of EIAs and pain relief (Laux-Biehlmann et al, 2013 ).…”

Section: Perspectives For the Pain Treatmentmentioning

confidence: 71%

Endogenous Isoquinoline Alkaloids Agonists of Acid-Sensing Ion Channel Type 3

Osmakov

Koshelev

Andreev

et al. 2017

Front. Mol. Neurosci.

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Acid-sensing ion channels (ASICs) ASIC3 expressed mainly in peripheral sensory neurons play an important role in pain perception and inflammation development. In response to acidic stimuli, they can generate a unique biphasic current. At physiological pH 7.4, human ASIC3 isoform (hASIC3) is desensitized and able to generate only a sustained current. We found endogenous isoquinoline alkaloids (EIAs), which restore hASIC3 from desensitization and recover the transient component of the current. Similarly, rat ASIC3 isoform (rASIC3) can also be restored from desensitization (at pH < 7.0) by EIAs with the same potency. At physiological pH and above, EIAs at high concentrations were able to effectively activate hASIC3 and rASIC3. Thus, we found first endogenous agonists of ASIC3 channels that could both activate and prevent or reverse desensitization of the channel. The decrease of EIA levels could be suggested as a novel therapeutic strategy for treatment of pain and inflammation.

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Section: Perspectives For the Pain Treatmentmentioning

confidence: 71%

Endogenous Isoquinoline Alkaloids Agonists of Acid-Sensing Ion Channel Type 3

Osmakov

Koshelev

Andreev

et al. 2017

Front. Mol. Neurosci.

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“…In 26 selected morphine‐positive critically ill patients, eM concentrations ranged between 0.7 and 149.4 nM in plasma samples (median = 34.7 nM; 95% CI: 18.2, 50.8 nM), whereas M6G concentration ranged between 0.3 and 775.8 nM (median = 150.8 nM; 95% CI: 63.2, 238.7 nM) in the same plasma samples. As previously described by us , eM levels were highly dependent of samples preparation, lithium‐heparin plasma leading to a better eM recovery. In contrast, M6G quantification was only weakly affected by sampling procedure, as LH‐plasma and serum only differ of 8 ± 4% ( n = 18; data not shown).…”

Section: Resultsmentioning

confidence: 60%

“…Plasma samples from 39 patients were analyzed using our M6G‐specific ELISA as well as LC‐MS/MS. Plasma were collected from (i) healthy donors ( n = 8), (ii) morphine‐treated patients (morphine skin patch 12–48 H before blood collection, 20 mg/day, n = 5) and ( iii ) critically ill patients ( n = 26), naive for morphine, but positive for eM when tested with a commercial morphine‐specific ELISA .…”

Section: Resultsmentioning

confidence: 99%

“…Collected plasma were immediately stored at −80°C and were only thawed once, 15 Min before ELISA or extraction. Plasma's morphine concentrations were quantified using a morphine‐specific ELISA kit (Immunalysis) as previously described , whereas our M6G‐specific ELISA prototype was used to quantify M6G in plasma.…”

Section: Methodsmentioning

confidence: 99%

“…Collected plasma were immediately stored at À80 C and were only thawed once, 15 Min before ELISA or extraction. Plasma's morphine concentrations were quantified using a morphine-specific ELISA kit (Immunalysis) as previously described [18], whereas our M6G-specific ELISA prototype was used to quantify M6G in plasma. Different extractions cartridge have been tested for the recovery of M6G (OASIS HLB and MC-X; SOLA; Bond Elute Certify; Strata-X 33u Polymeric Reversed Phase, Strata-XL 100u Polymeric Phase, Strata C18-E, Strata SAX, Strata Screen-A, Strata X-AW 33u Polymeric Weak Anion, Strata-X-CW 33u Polymeric Weak Cation, Strata WCX, Strata-X-C 33u Polymeric Strong Cation, Strata-SL-C 100u Polymeric Strong Cation, Strata Screen-C, Strata-X-C 33u Polymeric Strong Cation; Chromabond Drug).…”

Section: Patientsmentioning

confidence: 99%

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Endogenous morphine‐6‐glucuronide (M6G) is present in the plasma of patients: Validation of a specific anti‐M6G antibody for clinical and basic research

et al. 2013

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Endogenous morphine and its derivatives (morphine-6-glucuronide [M6G]; morphine-3-glucuronide [M3G]) are formed by mammalian cells from dopamine. Changes in the concentrations of endogenous morphine have been demonstrated in several pathologies (sepsis, Parkinson's disease, etc.), and they might be relevant as pathological markers. While endogenous morphine levels are detectable using enzyme-linked immunosorbant assay (ELISA), mass spectrometry (MS) analysis was, so far, the only approach to detect and quantify M6G. This study describes the preparation of a specific anti-M6G rabbit polyclonal antibody and its validation. The specificity of this antibody was assessed against 30 morphine-related compounds. Then, a M6G-specific ELISA-assay was tested to quantify M6G in the plasma of healthy donors, morphine-treated, and critically ill patients. The antibody raised against M6G displays a strong affinity for M6G, codeine-6-glucuronide, and morphine-3-6-glucuronide, whereas only weak cross-reactivities were observed for the other compounds. Both M6G-ELISA and LC-MS/MS approaches revealed the absence of M6G in the plasma of healthy donors (controls, n = 8). In all positive donors treated with morphine-patch (n = 5), M6G was detected using both M6G-ELISA and LC-MS/MS analysis. Finally, in a study on critically ill patients with circulating endogenous morphine (n = 26), LC-MS/MS analysis revealed that 73% of the positive-patients (19 of 26), corresponding to high M6G-levels in M6G-ELISA, contained M6G. In conclusion, we show that endogenous M6G can be found at higher levels than morphine in the blood of morphine-naive patients. With respect to the interest of measuring endogenous M6G in pathologies, we provide evidences that our ELISA procedure represents a powerful tool as it can easily and specifically detect endogenous M6G levels.

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