CLINICAL CHEMISTRY - PAGE 1 TO 23 (IN HOUSE REVIEW)

International Organization for Standards

Meaning of ISO

International Union of Pure and Applied Chemistry

Meaning of IUPAC

Clinical and Laboratory Standards Institute

Meaning of CLSI

Substance Abuse and Mental Health Services Administration

This administration certifies laboratories to conduct forensic drug testing for federal agencies

SI units

Recommended for the following:

1. Because compounds react on a molar basis

2. Recommended report of analytes

Liter

SI units uses this type of unit to reference volume.

Celsius

Predominant practice for temperature measurement

Kelvin

SI designation for temperature

400 to 700 nm

Range for visible spectrum (nm)

Infrared

700 nm is what type of light?

Ultraviolet

<400 nm is what type of light?

Spectrphotometry

Process of measuring light transmitted by a solution.

1. Tungsten light bulb

2. Hollow cathode lamp

3. Deuterium discharge lamp and Mercury arc lamp

3 types of light/radiant source of POLYchromatic Light.

Tungsten light bulb / Incandescent tungsten / Tungsten iodide lamp

Most common light/radiant source in spectrophotometry.

1. Entrance Slit

2. Unwanted stray light

In spectrophotometry this part minimizes the unwanted _____ light

Stray Light

In spectrophotometry this is the most common cause of loss of linearity.

1. Deteriorated optics

2. Light dispersed

3. Extraneous room light

Stray light can be caused by 3 factors.

Absorbance Error

This is the major effect of stray light is the error of what.

Sharp cutoff filters

Type of filter best for measuring stray light.

Neutral Density Filter + Didymium Filter

These two filters combined verify the absorbance accuracy and linearity

650 nm

A linearity study is performed on a visible spectrophotometer at _______ nm.

Absorbance:

1. 0.20

2. 0.41

3. 0.62

4. 0.79

5. 0.92

Concentration of Standard in a linearity study will have absorbance reading of what:

1. 10.0 mg/dL

2. 20.0 mg/dL

3. 30.0 mg/dL

4. 40.0 mg/dL

5. 50.0 mg/dL

Monochromator

In spectrometry this isolates specific wavelength of light.

Grating

This is the most used monochromator in a spectrophotometry.

Colored Glass Filter

This a least expensive, not very precise, simple, inexpensive and useful monochromator in a spectrophotometry.

Prism

This type of monochromator in a spectrophotometry refracts short wavelengths more than long wavelengths.

1. Wavelength Accuracy

2. Narrow Natural Bandpass

3. Highest Absorbance

A quality assurance procedure that verifies a spectrophotometer is reading the correct wavelength by scanning a substance with a ___________ (sharp absorbance peak) and confirming that the ________ absorbance occurs at the substance's known wavelength

Exit Slit

In spectrophotometry this controls the bandwidth of light the leaves the monochromator and reaches the sample

1. Narrower the bandpass

2. Greater the photometric resolution

3. Width of the exit slit

The ________ the bandpass, the ________ the photometric resolution. Bandpass can be made smaller by reducing the ________ of the exit slit.

Cuvet

In spectrophotometry this is also called the absorption cell/analytical cell/sample cell. It holds the solution.S

1. Square cuvette

2. Round cuvette

In spectrophotometry, (1) ________ cuvette has advantage over (2) ________ cuvette

Alumina silica cuvette

In spectrophotometry this material is the most commonly used for cuvettes.

Glass cuvettes

In spectrophotometry this material is used for visible light range but absorbs UV light.

Quartz cuvette

In spectrophotometry this material is used for UV radiation for cuvet.

1. Beer’s Law

2. Directly Proportional

3. Inversely Proportional

This mathematical formula explains that the absorbance of an unknown substance is ________ proportional to the amount of light absorbed and _______ proportional to the amount of light transmitted.

A = 2.0 - log %T (%T means transmittance it can be 1.0 or something)

Formula of Beer’s Law

1. Cu = Au/As x Cs

2. Absorbance of unknown (Au)

3. Absorbance of standard (As)

4. Concentration of known (Cs)

Example:

A cholesterol standard is 200 mg/dL with an absorbance (As) of 0.30. The patient's sample has an absorbance (Au) of 0.45. Calculate the patient's cholesterol concentration.

Solution:

Cu = (Au / As) × Cs

Cu = (0.45 / 0.30) × 200

Cu = 1.5 × 200

Cu = 300 mg/dL

Calculating the Concentration of Unknown:

1. It’s formula

2. Meaning of Au

3. Meaning of As

4. Meaning of Cs

Lambert Law

This explains that the absorbance increases exponentially with an increase in the light path.

Photodetector

In spectrophotometry this detects and converts transmitted light.

1. Photomultiplier tube

2. Sensitive

3. Room Light

In spectrophotometry this is the most common type of photodetector and the most _______. This should not be exposed to __________ because it will burn out.

1. Barrier layer cell or Photocell or Photovoltaic cell

2. External Power Source

In spectrophotometry this photodetector is the simplest detector and requires no external __________ source.

Photodiode

In spectrophotometry this photodetector respond to a wide range of wavelength from UV to Visible.

1. Phototube

2. Photocell

3. Outside Voltage

In spectrophotometry this photodetector is the same as ______ but it differs in the _______ voltage required for operation.

Meter or Read out device

In spectrophotometry this displays the output of the process.

1. Blanking Technique

2. Color of the Reagent

3. Zero Absorbance

4. Use Sample Blank

5. Ultracentrifugation

(1) This is a method to correct unwanted absorbance not caused by the analyte being measured. This is how to do it.

i. Contains the same reagents used for the test

ii. Reagent blank corrects for absorbance caused by the color of (2) ____________

iii. Adjust the spectrophotometer to (3) ____________

iv. (4) ___________ correction for interfering chromogens or lipemia

v. (5) ___________ is used to clear the serum or plasma of chylomicrons

1. Sample Blank

2. Sample

(1) This is used to subtract the intrinsic absorbance of the sample usually caused by hemolysis, icterus, turbidity, or drug interference. It is performed by substituting the (2) _______ for the reagent.

Single beam spectrophotometer

This is the most simplest type of spectrophotometer. It does one measurement at a time.

1. Double Beam Spectrophotometer

2. Two photodetectors

(1) This type of spectrophotometer permits automatic correction of sample and reference absorbance. (2) This use how many photodetectors?

1. Reflectometry or Reflectance Photometry

2. Nonlinear

3. Bilirubinometer

4. Automated reagent strip reader

(1) This method uses diffused light illuminates a reaction mixture in a carrier and REFLECTED light is measured. The intensity of light reflected is (2) _______ in relation to the concentration of the analyte. Examples of these are (3) _________ and Automated (4) _________ reader

1. Flame Emission Photometry

2. Blue flame

(1) This measures the light emitted by a single atom burned in a flame. This flame color (2) ______ is the hottest. Light intensity of atoms that are emitting energy = (3) ___________.

Sodium = Golden Yellow

Potassium = Violet

Lithium/Rubidium = Crimson red or Red

Magnesium/Copper = Green to Blue-green

Flame Emission Photometry uses these substances and each of them have a corresponding color.

• Sodium =

• Potassium =

• Lithium/Rubidium =

• Magnesium/Copper =

Atomic Absorption Spectrophotometry (AAS)

This method measures light absorbed by atoms dissociated by heat.

1. Atomizers

2. Nebulizer

3. Flame or a graphite furnace

AAS uses this instrument to convert ions to atoms. It uses two types of this instrument.

1. Calcium

2. Copper

3. Magnesium

4. Lead

5. Aluminum

6. Lithium

7. Zinc

AAS uses these 7 unexcited trace metals.

1. Flameless Atomic Absorption Spectrophotometry

2. Graphite Cylinder / Furnace

(1) AAS that uses electricity to break the chemical bonds instead of flames. It uses this (2) ________ cylinder / furnace to hold the sample where electricity passes through to evaporate and atomize the sample.

1. Fluorometry or Molecular Luminescence Spectrophotometry

2. Excitation Monochromator

3. Emission monochromator

4. Quenching Effect

(1) Method of measuring the amount of light intensity emitted by a molecule after excitation by electromagnetic radiation. It requires a (2) ________ monochromator and an (3) __________ monochromator. It’s more sensitive than spectrophotometer. It has a known disadvantage called the (4) ___________ effect.

1. pH

2. Temperature

3. Chemicals

In fluorometry quenching effect has 3 factors.

1. Chemiluminescence

2. Fluorometry

3. Enzyme (Bioluminescence)

(1) A method of using chemical or electrochemical reaction. It does not use excitation radiation and monochromator. It is more sensitive than (2) __________. This (3) catalyst = __________ can enhance the chemiluminescence.

1. Luminol

2. Acridinium esters

3. Dioxetane

4. Ruthenium

Chemiluminescence uses oxidation reaction of these 4 substances.

1. Turbidimetry

2. Concentration

3. Size

(1) Measures light transmitted (or attenuated) by a particle in a solution. This is used for abundant large particles. It is dependent on two factors (2) _______ and (3) ________

1. Nephelometry

2. Wavelength

3. Particle size

(1) Determines the amount of scattered light. The detector of this method is directly proportional to concentration. It is also for measuring Ag-Ab complexes. This method depends on these factors (2) ___________ and (3) Particle size

Ampholyte

In electrophoresis this molecule, such as a protein, whose net charge can be either positive or negative.

1. Isoelectric point

2. Migrate

(1) In electrophoresis this pH point is where protein has no net charge. Particle with no net charge will not (2) ________ in electrophoresis.

1. Anode (positive electrode)

2. Cathode (negative electrode)

In electrophoresis if the:

1. Particles migrate towards ________ if negatively charged

2. Particles migrate towards __________ if positively charged

1. 8.6 pH

2. Albumin

3. a1 globulin

4. a2 globulin

5. Beta globulin

6. Gamma globulin

At pH (1) _____ proteins migrate and divide into: (2) _________, (3) ________ globulin, (4) ________ globulin, (5) _______ globulin, and (6) ________ globulin.

Iontophoresis

Type of electrophoresis that is described as the migration of small ions

Zone electrophoresis

Type of electrophoresis that is described as the migration of charged macromolecules (protein)

1. Capillary electrophoresis

2. In narrow-bore fused silica capillaries

3. Separated by Electro-Osmotic Flow (EOF)

4. Positive charge moves faster

5. Negative charge move slower

(1) Type of electrophoresis where separating is performed in a narrow-bore fused (2) __________. This is where molecules are separated by (3) ___________ flow. The positive charge moves (4) ___________ and Negative charge moves (5) __________

Isoelectric Focusing

This method in electrophoresis is ideal for separating proteins with identical size (Molecular weight and mass) but with different net charge.

1. High resolution protein electrophoresis

2. 12 discrete zones

(1) This method in electrophoresis separates proteins into as many as (2) _____ discrete zones to be visualized in high resolution.

1. Net electric charge

2. Size and Shape

3. Strength

4. Chemical and physical properties

5. Temperature

Factors Affecting the Velocity of Migration

a. Net (1) __________ of the molecule
b. (2) _____ and _______ of the molecule
c. (3) ________ of the electric field
d. (4) __________ and _________ properties of the supporting medium
e. Electrophoretic (5) _____________

1. Chromatography

2. Mobile phase

3. Stationary phase

(1) This is a method / process of separating the mixture components in a solution. It has two basic components (2) _____ phase (Gas or Liquid) and _____ phase (Solid or Liquid).

Retention factor value

In chromatography this is the value for the distance leading edge to front of components moves / total distance of solvent.

1. Paper chromatography (Whatman paper) - simple and inexpensive

2. Thin later chromatography (silica gel, alumina, cellulose, or crosslinked dextran) - Most faster and sensitive

In chromatography the stationary phase can use this materials:

1. ______ chromatography ( 1 example)

2. ____ _____ chromatography (4 example)

1. Gas chromatography

2. Slower Elution

3. Faster Elution

In chromatography this method is used for mobile phase it’s principle is:

• Uses volatile compounds elute in order of increasing the boiling point

• Lower boiling point means _____ elution

• High boiling point means ______ elution

1. Gas Chromatography - Mass Spectrometry (GS-MS)

2. 15 days

3. Discarded

(1) Combination of processes for confirmatory test for drug abuse

(2) A positive result in this must be challenge within _____ days

(3) If not challenged within the timeline, the samples should be ___________

1. Liquid Chromatography

2. High Performance Liquid Chromatography

3. Reverse-phase HPLC

This chromatography is a separation technique where the mobile phase is a liquid and the stationary is a solid packed inside a column. This has two types:

1. The stationary phase of this type is polar and the mobile phase is nonpolar. It uses high pressure to force solvent through a packed column for rapid, efficient separation.

2. This one the stationary phase is nonpolar and mobile phase is more polar than the stationary phase. This is most widely used mode in clinical labs for drug monitoring, hormones, and proteins.

HPLC (Normal Phase)

This type of liquid chromatography used in rapid HbA1c measurement.

1. MS/MS (Tandem Mass Spectrometry)

2. Vacuum

This method of mass spectrometry is used to detect inborn errors of metabolism. The ___________ is needed to prevent collision between fragments.

1. MALDI-TOF

2. Nonlinear function

3. More time

(1) This method for testing is used for peptides and protein analysis. The time required for an ion with a given mass to reach the detector is a (2) ______ function of the mass, with larger irons requiring more (3) ______ than smaller ions

1. Volumetric Method

2. Schales and Schales method (Chloride Test)

(1) This method’s principle is unknown sample + known sample with an indicators. It uses the (2) _________ and _______ method (_____ test)

1. Osmometry

2. Osmolality

3. Colligative

4. Glucose, Urea nitrogen, and Sodium

Two formulas of Osmolality:

A. 2(Na) + glucose/20 + BUN/3 = mOsm/kg

B. 1.86 Na + glucose/18 + BUN/2.8 + 9

1. This is the measurement of osmolality (Serum, plasma, or urine).

2. Defined as the number of osmoles of solute per kilogram of solvent

3. Measuring changes in __________ properties of solution (osmotic pressure, boiling point, freezing point, and vapor pressure)

4. Osmotic particles includes this three substances.

1. Freezing point depression osmometry

2. Volatile substances

3. Lowered by 1.86°C

4. Lowered by 0.3 mmHg

5. Raised by 0.52°C

(1) Osmometry method that is commonly used for measuring the changes in colligative properties in a solution. It is sensitive to (2) __________ substances while vapor pressure osmometer is not.

• In this method, the temperature is lowered by (3) 1.____°C

• In vapor pressure it is lowered by (4) 0.__°C mmHg or torr

• In boiling point it is raised by (5) 0.__°C

1. Lipids

2. Protein

This two substances is not a contributor to osmolality

1. Osmolal gap

2. Osmotically Active

(Osmolal gap = Measured osmolality - calculated osmolality)

(1) The difference between the measured osmolality and the calculated osmolality. This gap indirectly indicates the presence of (2) ______ active substances.

Electrochemistry Techniques

This method measures the current voltage. by the activity of a specific ion

Ion-Selective Electrode

This electrochemistry technique contains selective membrane that response primarily to a specific ion; when the electrode is placed in a solution, a potential voltage develops across the membrane.

1. Glass aluminum silicate - Sodium

2. Antibiotic Valinomycin gel - Potassium

3. Dioctylphenyl phosphonate - Calcium

4. Nonactin and Monactin - NHA4+ analysis

Selective membrane for each of this substances in ISE:

1. Sodium

2. Potassium

3. Calcium

4. NH4+

1. Amperometry

2. Clarke or Clark electrode

1. This electrochemistry technique measures the amount of current produced through the oxidation or reduction of the substance to be measured at an electrode held at a fixed potential.

2. This type of electrode is used for pO2

1. Potentiometry

2. Calomel

3. Silver or Silver chloride

4. Uses pH and pCO2

5. Severinghaus electrode

1. This electrochemistry technique measures the potential difference between an indicator electrode and a reference electrode.

2. Reference electrode of this technique is (2) _________, (3) ________ and _________

3. The uses of this technique is for ______ and ______

4. This electrode is used for this technique

1. Coulometry

2. Faraday’s law

3. Chloride coulometer

1. This electrochemistry technique is based on _____ law

2. Number of equivalent weights of a reactant oxidized or reduced is directly proportional to the quantity of ELECTRICITY (CURRENT/TIME) used in the reaction.

3. This coulometer must have a current across the generator electrodes for this substance to be measured.

Polarography

This electrochemistry technique uses a voltage at which sudden or abrupt rise in current occurs is characteristic of the electrochemical reaction is involved.

Anodic Stripping Voltammetry

This is based on the electrochemistry technique polarography. The trace metal ions in the solution are reduced and plated onto the anodic (working) electrode. The metal is stripped off the anode > measured as current.

1. Analyte

2. Reagent

3. Catalyst

4. Calibration

5. Control

1. A chemical substance that is the subject of chemical

2. A compound or a mixture added to a system to cause a chemical reaction or test if a reaction occurs.

3. Anything that speeds up the rate of chemical reaction but is not produced or consumed in the reaction.

4. Process of testing and adjusting an anaylzer’s readout to establish correlation between measured and actual concentrations.

5. A sample that is chemically and physically similar to the unknown specimen and is tested in exactly the same manner.

1. Open reagent system

2. Closed reagent system

1. This type of reagent are from many vendors that may be adapted to an analyzer

2. This type of reagent is the proprietary reagents from the instrument manufacturer are suitable for use on the analyzer

Standard/Calibrator

A material known concentration, used for calibrating an instrument.

Type I reagent water

Type of reagent with maximum water purity for ultramicrochemical analysis, tissue and/or cell culture methods, trace metals.

Type II reagent water

Type of reagent with acceptable in chemistry, hematology, microbiology, immunology, and other clinical tests. Acceptable for routine analytic requirements.

Type III reagent water

This reagent water is for most general laboratory tests.

1. Continuous Flow Analyzer

2. Centrifugal Analyzer

3. Discrete Analyzer

1. Samples flow through a common reaction vessel or pathway

2. Analyzer that uses centrifugation to transfer specimen and reagents

3. Analyzer that is capable of running multiple tests — one sample at a time