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Файл:Ординатура / Хирургия / Библиотека им академика М.И. Перельмана / Книга_5733_Библиотеки_им_академика_М_И_Перельмана
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157
Selenious acid in concentrated sulfuric acid
Procedure:
Place a small sample of the substance on a white ceramic plate or glass
spot plate.
Add one or two drops of the Mecke reagent.
Observation:
Hydrocodone usually yields a green color that may turn to dark blue upon
reaction with the Mecke reagent.
3. Froehde Reagent Test:
Reagent Composition:
Molybdenum trioxide in concentrated sulfuric acid
Procedure:
Place a small sample of the substance on a white ceramic plate or glass
spot plate.
Add one or two drops of the Froehde reagent.
Observation:
Hydrocodone typically results in a purple color change when reacting with
the Froehde reagent.
4. Liebermann Reagent Test:
Reagent Composition:
Sodium nitrite in concentrated sulfuric acid
Procedure:
Place a small sample of the substance on a white ceramic plate or glass
spot plate.
Add one or two drops of the Liebermann reagent.
Observation:
Hydrocodone generally turns a deep blue or black color with the
Liebermann reagent.
General Procedure for Spot Tests
Preparation:
Ensure that the reagents are fresh and properly labeled.
Use clean spot plates or surfaces for testing.
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Sample Application:
Use a small amount of the sample for testing to prevent wastage and
ensure safety.
Reagent Addition:
Add the reagent dropwise to the sample to observe any color change.
Observation and Interpretation:
Note the immediate color change and any subsequent changes.
Compare the observed color with standard reference charts for
interpretation.
Safety Precautions:
Wear gloves and eye protection while handling reagents, as many are
corrosive.
Perform tests in a well-ventilated area or under a fume hood.
Limitations
Specificity: Color tests can be nonspecific and may produce similar color
changes for different substances.
Quantification: These tests do not provide quantitative information about
the concentration of hydrocodone.
Interferences: Other substances present in the sample may interfere with
the test results.
Applications
Field Testing: Useful for on-the-spot drug screening by law enforcement or
at events.
Preliminary Identification: Helps in quickly identifying the presence of
hydrocodone before more specific and sensitive instrumental analysis.
Chemical Methods:
1. High-Performance Liquid Chromatography (HPLC)
Procedure:
Sample Preparation: Extract hydrocodone from the biological matrix (e.g.,
blood, urine) using solid-phase or liquid-liquid extraction.
Mobile Phase: Use a mixture of water with an acid (like formic acid or
phosphoric acid) and an organic solvent (like acetonitrile or methanol).
Column: Employ a C18 reversed-phase column.
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Detection: Use UV detection at a wavelength specific for hydrocodone (e.g.,
around 280 nm).
Advantages:
High sensitivity and specificity.
Suitable for complex biological matrices.
2. Gas Chromatography-Mass Spectrometry (GC-MS)
Procedure:
Derivatization: Convert hydrocodone into a more volatile derivative using
reagents like N-Methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA).
Injection: Introduce the sample into the GC-MS system.
Column: Use a capillary column with a non-polar stationary phase.
Detection: Identify hydrocodone based on its mass-to-charge ratio (m/z).
Advantages:
High sensitivity and specificity.
Can differentiate between hydrocodone and its metabolites.
3. Liquid Chromatography-Mass Spectrometry (LC-MS/MS)
Procedure:
Sample Preparation: Similar to HPLC, extract hydrocodone from the
biological matrix.
Mobile Phase: Use a gradient of water with formic acid and acetonitrile.
Column: Employ a C18 or other suitable column.
Detection: Use tandem mass spectrometry for high sensitivity and
specificity.
Advantages:
Extremely sensitive and specific.
Suitable for low concentration detection in complex matrices.
4. Ultraviolet-Visible (UV-Vis) Spectroscopy
Procedure:
Sample Preparation: Dissolve the sample in a suitable solvent (e.g.,
methanol).
Wavelength Selection: Measure absorbance at a specific wavelength where
hydrocodone shows maximum absorbance (e.g., around 280 nm).
Advantages:
Simple and quick.
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Suitable for initial screening.
5. Infrared (IR) Spectroscopy
Procedure:
Sample Preparation: Prepare a thin film of the sample or mix with
potassium bromide (KBr) to form a pellet.
Measurement: Record the IR spectrum and identify characteristic
absorption bands of hydrocodone.
Advantages:
Provides structural information.
Non-destructive.
6. Enzyme-Linked Immunosorbent Assay (ELISA)
Procedure:
Sample Preparation: Dilute the biological sample appropriately.
Antigen-Antibody Reaction: Add the sample to wells coated with
antibodies specific for hydrocodone.
Detection: Add an enzyme-linked secondary antibody and a substrate that
produces a color change.
Advantages:
Rapid and suitable for high-throughput screening.
High sensitivity.
7. Radioimmunoassay (RIA)
Procedure:
Sample Preparation: Similar to ELISA, prepare the biological sample.
Antigen-Antibody Reaction: Mix the sample with radioactively labeled
hydrocodone and antibodies.
Separation: Separate bound from free antigen using a solid-phase or
liquid-phase separation method.
Measurement: Measure radioactivity using a scintillation counter.
Advantages:
High sensitivity and specificity.
Suitable for low concentration detection.
Summary of Steps for HPLC and LC-MS/MS
High-Performance Liquid Chromatography (HPLC)
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Preparation of Mobile Phase: Mix water with an acid (e.g., formic acid) and
acetonitrile.
Column Equilibration: Equilibrate the C18 column with the mobile phase.
Sample Injection: Inject the extracted sample into the HPLC system.
Detection and Quantification: Detect hydrocodone using UV detection and
quantify using a calibration curve.
Liquid Chromatography-Mass Spectrometry (LC-MS/MS)
Preparation of Mobile Phase: Prepare a gradient of water with formic acid
and acetonitrile.
Column Equilibration: Equilibrate the C18 column with the mobile phase.
Sample Injection: Inject the extracted sample into the LC-MS/MS system.
Detection and Quantification: Detect and quantify hydrocodone using
tandem mass spectrometry.
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HYDROMORPHINOL
(α-Oxymorphol)
Chemical name: (5α,6α)-4,5-epoxy-17-methyl-morphinan-3,6,14-triol
Molecular structure:
Physical properties:
Appearance: solid
Colour: white to light brown solid
Melting point: >231°C (dec.)
Solubility: Water soluble (20.6 mg/mL)
Molecular framework: Aromatic heteropolycyclic compounds
Molecular weight: 303.35 g/mol
Chemical Properties:
Chemical formula: C17H21NO4
Chemical classification: Opium Derivative
Functional groups present in Hydromorphinol:
- Phenol: The -OH group attached to an aromatic benzene ring.
- Alcohol: A hydroxyl (-OH) group attached to a saturated carbon atom.
- Ether: An oxygen atom connected to two alkyl or aryl groups (an
oxygen bridge within the molecule).
- Tertiary Amine: A nitrogen atom attached to three alkyl or aryl
groups.
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Ph of Hydromorphinol: The pH of a solution of hydromorphinol depends on
several factors, including its concentration and the solvent used. Hydromorphinol
itself is a solid compound, and its pH is typically considered in the context of its
aqueous solutions. Since hydromorphinol is a weak base (due to the presence of
the tertiary amine group), its solutions are likely to be slightly basic. However,
precise pH values would depend on the concentration of the solution and whether
any salts (such as hydromorphinol hydrochloride) are used.
For a precise pH value of a hydromorphinol solution, specific experimental
measurement is required. If you have a specific concentration or formulation in
mind, you could determine the pH by using a pH meter or referring to empirical
data provided in pharmacological or chemical literature.
Methods of Analysis
Physical Methods:
1. Colorimetric Reagents: Some chemical reagents react with opioids to produce
characteristic colors. While specific tests for hydromorphinol may not be readily
available, you can use general opioid reagents.
Froehde Reagent: This reagent is used for testing alkaloids, which includes
opioids.
Procedure: Add a few drops of Froehde reagent (a mixture of molybdic
acid or ammonium molybdate in sulfuric acid) to the sample.
Expected Result: Observe the color change. Opioids typically produce a
purple to blue color.
2. Spot Tests: Spot tests can provide a quick indication of the presence of an opioid.
Liebermann’s Reagent: This is a mixture of potassium nitrite and sulfuric
acid.
Procedure: Place a small sample of the substance on a white ceramic plate.
Add a drop of Liebermann’s reagent.
Expected Result: The presence of opioids often results in a color change,
such as a deep red or purple color.
3. Microcrystalline Tests: These tests involve the formation of characteristic
crystals when the substance reacts with specific reagents.
Procedure: Place a small amount of the sample on a microscope slide. Add
a drop of a suitable reagent, such as gold chloride or platinic chloride.
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Expected Result: Characteristic crystals form, which can be observed
under a microscope. Compare the crystal shapes with reference images to
identify the substance.
4. Solubility Tests: Solubility tests can help in preliminary identification by
observing how the substance dissolves in different solvents.
Procedure: Test the solubility of the sample in water, ethanol, chloroform,
and other solvents.
Expected Result: Record the solubility in each solvent. Hydromorphinol
may have specific solubility characteristics that can help distinguish it
from other substances.
5. Melting Point Determination: While not a color test, determining the melting
point can provide valuable information.
Procedure: Use a melting point apparatus to measure the melting point of
the sample.
Expected Result: Compare the measured melting point with the known
melting point of hydromorphinol.
Limitations and Considerations
Sensitivity: These tests are less sensitive and specific compared to
chromatographic or spectroscopic methods.
Safety: Handle all reagents and samples with care, following appropriate
safety protocols.
Reference Standards: It's important to have reference standards for
comparison to interpret the results accurately.
Chemical methods:
1. High-Performance Liquid Chromatography (HPLC)
HPLC is widely used for the separation, identification, and quantification
of components in a mixture. For hydromorphinol:
Procedure: Samples are dissolved in a suitable solvent and injected into
the HPLC system. The mobile phase and stationary phase are selected to
optimize the separation of hydromorphinol from other components.
Detection: UV or fluorescence detectors are commonly used.
2. Gas Chromatography-Mass Spectrometry (GC-MS)
GC-MS combines gas chromatography and mass spectrometry to identify
different substances within a test sample.
Procedure: The sample is vaporized and then separated in a gas
chromatograph. The separated substances are identified and quantified by
mass spectrometry.
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Detection: Provides highly sensitive and specific analysis of
hydromorphinol.
3. Liquid Chromatography-Mass Spectrometry (LC-MS)
LC-MS combines liquid chromatography and mass spectrometry.
Procedure: Similar to HPLC, but with mass spectrometry as the detector,
providing higher sensitivity and specificity.
Detection: Often used for detailed structural analysis and quantification.
4. Fourier Transform Infrared Spectroscopy (FTIR)
FTIR measures the infrared spectra of absorption or emission of a solid,
liquid, or gas.
Procedure: The sample is prepared (often as a pellet or thin film) and
exposed to infrared radiation. The resulting spectrum provides
information about the molecular structure.
Detection: Useful for identifying functional groups and molecular
structures.
5. Nuclear Magnetic Resonance (NMR) Spectroscopy
NMR spectroscopy exploits the magnetic properties of certain atomic
nuclei.
Procedure: The sample is dissolved in a suitable solvent and placed in a
magnetic field. Radiofrequency radiation is used to perturb the magnetic
alignment of the nuclei, and the resulting signals are analyzed.
Detection: Provides detailed information about the molecular structure
and dynamics.
6. Ultraviolet-Visible Spectroscopy (UV-Vis)
UV-Vis spectroscopy measures the absorbance of UV or visible light by a
substance in solution.
Procedure: The sample is dissolved in a solvent, and its absorbance is
measured at different wavelengths.
Detection: Useful for determining the concentration and purity of
hydromorphinol.
7. Capillary Electrophoresis (CE)
CE separates ions based on their electrophoretic mobility with the use of
an applied voltage.
Procedure: The sample is introduced into a capillary filled with an
electrolyte solution. An electric field is applied, causing ions to migrate at
different rates.
Detection: Often coupled with UV or MS detection for enhanced analysis.
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8. Thin-Layer Chromatography (TLC)
TLC is a simple, quick, and inexpensive method to analyze mixtures.
Procedure: The sample is spotted on a TLC plate and developed in a
solvent system. The separated components are visualized using UV light or
chemical reagents.
Detection: Provides a qualitative analysis of hydromorphinol.
Sample Preparation
Extraction: Using solvents to isolate hydromorphinol from biological
matrices or formulations.
Derivatization: Chemical modification to improve detectability or stability
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