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Group 8 mass spec automated analyser and poct the complete version
1. GROUP 8
By:
Delgado, Sharmaine Kay
Gloria, Sherina Ann
Lagos, Riza Jane
Pillora, Gin Anilou
Villaflor, Mary Queen
1
2.
3. • Mass Spectrometry
• Mass Spectrometer
• Principles
• Major Parts
• How it works
• Uses
• Types of Spectrometer
4. • An analytical technique that
measures the mass-to-charge
(m/z) ratio of charged particles.
• A technique of separating and
identifying molecules based on
its mass.
5. • A mass spectrometer is an
analytical tool used to determine
the elemental composition of an
unknown substance. It utilizes
the charged particles of
molecules to separate them.
6.
7. • A Mass Spectrometer
produces ions from the
substance under
investigation, separates them
according to their mass-to-
charged ratio (m/z) and
records the relative
abundance of each present.
9. • Different compounds can also be uniquely
identified by their mass.
Butorphanol L-dopa Ethanol
N -CH2- COOH
OH
HO -CH2CH-NH2 CH3CH2OH
HO
HO
MW = 327.1 MW = 197.2 MW = 46.1
10. • The heavier the ion, the lesser
the deflection.
• The lighter the ion, the greater
the deflection.
11.
12. • Mass spectrometers consist of four basic
parts;
• a handling system to introduce the unknown
sample into the equipment;
• an ion source, in which a beam of particles
characteristic of the sample is produced;
• an analyzer that separates the particles
according to mass; and
• a detector, in which the separated ion
components are collected and characterized.
13.
14. The sample to be analyzed enters
the instrument through the
inlet, usually as a gas, although a
solid can be analyzed if it is
sufficiently volatile to give off at
least some gaseous molecules.
15. In the ionization chamber, the sample is
ionized and fragmented. This can be
accomplished in many ways—electron
bombardment, chemical
ionization, laser ionization, electric field
ionization—and the choice is usually based on
how much the analyst wants the molecule to
fragment.
16. 3. The Mass Analyser
Here, the particles are separated into
groups by mass, and then the detector
measures the mass-to-charge ratio for each
group of fragments by electromagnetic fields.
17. 4. The Detector
Finally, a readout device—usually a
computer—records the data.
18.
19.
20.
21. • The Sample is vaporized into gas for
ionization,
• The atom is ionised by knocking one or more
electrons off to give a positive ion.
• The Ion source is maintained in a high vacuum
environment to enhance collision efficiency
and ion formation.
22.
23. The ions are accelerated so
that they all have the same
kinetic energy.
24.
25. • The ions are then deflected by a magnetic
field according to their masses. The lighter
they are, the more they are deflected.
• The amount of deflection also depends on
the number of positive charges on the ion - in
other words, on how many electrons were
knocked off in the first stage. The more the
ion is charged, the more it gets deflected.
26.
27. The beam of ions passing
through the machine is
detected electrically.
28.
29. 1. GC/MS (Gas Chromatography-Mass
Spectrometry)
• Is a method that combines
the features of Gas-Liquid
Chromatography and Mass
Spectrometry to identify the
different substances within
a sample.
30. 2. AMS (Accelerator Mass
Spectrometry)
• a ‘’tandem accelerator’’
is used to accelerate the
ions at several million
volts.
31. 3. ICP-MS (Inductively Coupled
Plasma-Mass Spectrometry)
• involves the formation of gas
containing electrons, ions and
neutral particles from Argon gas.
The sample is atomized and
ionized by this gas. In a high
vacuum mass analyzer, these
ionized atoms from gas are
passed through cones
(apertures).
32. 4. IRMS (Isotope Ratio Mass
Spectrometry)
• It is used to measure
mixture of stable isotopes. It
has two inlets that help in
repetitive measurements
with continuous supply of
sample gas.
33. 5. Tandem MS (Tandem Mass
Spectrometer)
• is a spectrometer used to
separate ions based on a
sample’s ‘’electronic’’ mass
using two or more
quadruple’s
34. 6. TIMS (Thermal Ionization-
Mass Spectrometry)
• is a mass spectrometer that
can make exact
measurements isotope ratios
of thermally ionisable
elements. This ionization can
be done by passing them
through metal ribbons under
vacuum.
35. 7. SSMS (Spark Source Mass
Spectrometry)
• can ionize the analytes in solid
samples using electric current
with two electrodes. It works
as one electrode if the sample
is metal or can be placed in a
cup-shaped electrode by
mixing with graph detected
isotopes from the sample.
36. 8. (LC/MS or LC-MS) Liquid
chromatography –mass
spectrometry
• It is used to separate
compounds chromatographically
before they are introduced to the
ion source and mass
spectrometer. LC-MS is a powerful
technique used for many
applications which has a very high
sensitivity and selectivity.
37. 9. IMS/MS or IMMS (Ion
mobility Spectrometry)
• Is a technique where ions are
first separated by drift time
through some neutral gas
under an applied electrical
potential gradient being
introduced into mass
spectrometer.
46. Automated analyzers process large volume
of tests with great precision and speed.
It permits the operator to focus on tasks that
cannot be readily automated and increased
both efficiency and capacity.
46
49. Pumped through a system of continuous
tubing. Samples are introduced in a
sequential manner, following each other
through the same network.
This analyzer is capable of analyzing
one analyte at a time.
49
50. An essential principle of the system is the
introduction of air bubbles.
Function of Air Bubbles:
The air bubbles segment each sample into
discrete packets and act as a barrier between
packets to prevent cross contamination as they
travel down the length of the tubing.
50
51. Function of Air Bubbles:
The air bubbles also assist mixing by
creating turbulent flow and provide
operators with a quick and easy check of
the flow characteristics of the liquid.
51
52. In Continuous Flow Analysis a
continuous stream of material is divided
by air bubbles into discrete segments in
which chemical reactions occur.
The continuous stream of liquid samples
and reagents are combined and
transported in tubing and mixing coils.
52
53. The tubing passes the samples from one
apparatus to the other with each apparatus
performing different functions, such as
distillation, dialysis, extraction, ion
exchange, heating, incubation, and subsequent
recording of a signal.
53
54. Continuous flow is used in some
spectrophotometric instruments in which
the chemical reaction occurs in one reaction
channel and then is rinsed out and reused
for the next sample, which may be an
entirely different chemical reaction.
54
56. Segmented Stream System
-The reaction stream is segmented with
bubbles of air or nitrogen to reduce inter-sample
dispersion.
Flow Injection Analysis
- It is low pressure and without separation.
The injected sample mixes and reacts with the
flowing stream.
56
57. It includes a peristaltic pump that continuously
aspirates sample and reagent, a variable number of
tubes constituting a manifold to circulate liquid
and a detector system.
Aspirated sample are segmented by injecting air
bubbles that should be remove before they can
reach to the detector.
57
58. At detector air bubbles are removed and each
sample is separated by washing solution, thus a
square shaped detector response is obtained, the
height of rectangle is directly proportional to
concentration of analyte.
58
60. FIA is based on the injection of a liquid
sample into a moving continuous non
segmented carrier stream of a suitable
liquid. The injected sample forms a zone
which is then transported towards a detector.
60
61. Mixing with reagent in the flowing stream
mainly occurs by diffusion-controlled process
and a chemical reaction occurs.
Detectors continuously record the physical
parameter as it changes as a result of passage of
sample material through flow cell.
61
63. Discrete analysis is the separation of each
sample and accompanying reagents in a separate
container.
Discrete analyzers have the capability of
running multiple tests on one sample at a time
or multiple samples one test at a time.
63
64. They are the most popular and
versatile analyzers and have almost
completely replaced continuous-flow
and centrifugal analyzers.
64
65. Sample reactions are kept discrete through the
use of separate reaction cuvettes, cells, slides, or
wells that are disposed of following chemical
analysis.
This keeps sample and reaction carryover to a
minimum but increases the cost per test due to
disposable products
65
66. Samples are applied to slides that are
automatically dispensed from test- specific
cartridges. Sample application is performed by
means of individual, disposable tips, thereby
eliminating the carryover problem. The sample
itself provides the liquid necessary to hydrate the
reagent layers of the slide.
66
67. The slides incubate in heated air chambers and
the color that develops is measured by
reflectance photometry from the bottom side of
the slide.
Results for each sample are collated and printed
in a report form that could be suitable for use as
the final chartable report.
67
69. Designs of Analyzer Pathway
Batch Testing- Samples are processed in concert
as a group or “batch” in the same analytical
analysis.
Sequential Testing – samples are processed
sequentially rather than in a batch.
69
70. Designs of Analyzer
Pathway
Parallel Testing- samples undergo a series of
analytical processes, usually for one analysis
at a time, often used with batch analysis.
Random access testing- a system where any
specimen can be analyze in any sequence
with regard to the initial order of the
specimens.
70
71. Patient Identification
Sampling
Sample and Specimen Transport
Dilution
Mixing
Incubation
Reaction Vessels
Analysis of Measurement
71
72. Patient identification was accomplished by
transcribing patient information onto sample
cups and print outs of test results.
With the arrival of computers, the operator could
input patient information to the laboratory
computer.
72
73. Bar code labeling systems are now
employed. The bar code was read and
would match patient data with test
results. The use of bar code labels has
served to reduce errors in matching
test results with the proper patient.
73
75. Accomplished by syringe pipette or aspirating
probe. The specimens are transferred to
sample cup, and the sample pickup device
aspirates the specimen.
In CFA, the aspirating probe is dipped into the
sample cup and the specimen is drawn up
using a peristaltic pump.
75
77. A peristaltic pump is a type of positive
displacement pump used for pumping a
variety of fluids.
77
78. Works by squeezing the tube with rollers/shoes. It
can run dry, self-prime and handle viscous or
abrasive liquids, plus, as the tube is one complete
unit, there are no seals thus making the pump leak
free and hygienic. Excellent for dosing applications.
Although this principle applies to all peristaltic
pumps the difference is in the head and the drives.
78
79. As the rollers and wiper move, a part of the
tube is pressed, causing the fluid to be pumped
onward. A restitution fluid can be sent into the
pump as the rotors and rollers moved back the
process is called 'Peristalsis„. It forms the basic
function within a Peristaltic Pump.
79
81. A piston pump (reciprocating pumps) is a
type of positive displacement pump where the
high-pressure seal reciprocates with the piston.
Piston pumps can be used to move liquids or
compress gases. Powered by an electric
motor, steam or a turbine, hydraulic drive
mechanism.
81
82. A piston pump uses the reciprocating motion of
a piston rod to move fluid along an axis through a
cylinder-shaped chamber. As the piston moves
through the cylinder, pressure builds up and forces
the fluid through the pump. The fluid flowing
through the pump pulsates due to the movement of
the piston through the cylinder.
82
84. • Reciprocating pumps will deliver fluid at
high pressure (High Delivery Head).
• They are 'Self-priming' - No need to fill
the cylinders before starting.
84
85. Discrete analyzers employ a variety of
syringe pipettes to aspirate and dispense
sample and reagents. An important
consideration for any sampling device is
specimen carry-over and therefore it should
be designed to reduce this problem.
85
86. In continuous flow analyzers, specimen
transport is accomplished using the peristaltic
pump. Air bubbles separate aliquots of the same
sample and isolate one specimen from another.
86
87. In the Dupont aca, the sample reagent pack is
transported throughout the analyzer with a chain-
driven pulley system.
Some analyzers used a motorized carousel, for
example, the Olympus Demand, to move the
reaction vessel in a circular path within the
instrument.
87
88. The Kodak Ektachem analyzers
meters the sample aliquot, by use
of a disposable sample tip
secured by an apparatus called
proboscis, onto a slide for
transport to incubation chambers
and detectors.
88
89. Sample and reagent dilutions are usually
accomplished with the syringe pipettes and
pumps. The pumps must be designed to aspirate
and deliver precise volumes of fluid.
The dilution volumes maybe adjusted by use of a
cam or programmed via a microprocessor as seen
in many discrete analyzers.
89
90. In an automated system such as
continuous analyzer mixing of a
sample and reagents is accomplished
using a glass coil inserted into the flow
path. As the sample mixture passes
through the coil, it is inverted and
mixed via gravity.
90
91. In the Beckam ASTRA systems, a
magnetically driven Teflon stirring bar
located in the bottom of the reaction
chamber is used.
The DuPont aca employs a breaker mixer
that mechanically vibrates and shakes the
pack.
91
92. Reaction mixtures that require incubation must
be conducted at constant temperatures without
significant fluctuations.
a.) heating the air around the cuvette
b.) heating metal blocks
c.) using water baths.
92
93. In CFA systems the tubing serves as reaction
vessel.
In DA, any of the following maybe used:
a.) The DuPont aca uses a sealed plastic bag that
also serves as the cuvette.
b.) The Teflon or plastic rotors in centrifugal
analyzers serves as the reaction vessels.
93
94. c.) Hitachi series and Baxters Paramax 720 ZX
use plastic cuvettes.
d.) Eastman Kodak Ektachem uses a multilayer
thin film slide. Each slide is impregnated with
reagents. Sample cup via a disposable pipette tip
onto the slide that also serves as the cuvette for
the reflectance or electrochemical measurement.
94
95. Light-emitting diodes offer direct readout of
absorbance and replace the earlier recorders with an
ink pen to trace the response of the phototube on
paper.
Computer in the laboratory instrumentation allowed
users to display results in a variety of formats and
printers provide a hard copy of patient‟s results.
95
96. Calculations, calibration curves, and
quality control are performed by the
computers, thus reducing errors and
providing more accurate results than a
non-computerized instrument.
96
97. Most automated chemistry analyzers use
photometric methods of analysis such as
spectrophotometry, fluorometry, nephelometry, an
d reflectometry.
Some analytes, for example sodium and
potassium, require the use of electrochemistry for
analysis.
Instrument manufacturer have designed
electrochemical devices based on
coulometry, amperometry, and potentiometry to
measure these and other analytes. 97
98. Automated systems based on colorimetry use
narrow-band interference filters for the isolation of
specific wavelengths. The filters are contained in a
circular disk, called a filter wheel, that rotates into
the light path. A computer controls the rotation of
the filter wheel and multiple wavelengths can be
use to analyze a specimen.
98
100. Increase the number tests performed by one
medical technologist in a given period.
Minimize the variation in results from one medical
technologist to another.
Automation eliminates the potential errors of
manual analyses as a volumetric pipetting
steps, calculation of results, and transcription of
results.
100
101. Instruments can use very small amounts of
samples and reagents.
Reduction in the variability of results and
errors of analysis through the elimination of
task that are repetitive and monotonous for
most individuals.
101
102. Faster analyses up to 120 samples per hour
Automatic data recording and preparation
Being a closed system, automation reduces
contamination
Greater accuracy and reproducibility of results as
all samples are subject to same processes
Smaller sample and reagent volumes, reduces cost
102
103. Time-consuming sample preparation steps such
as distillations, digestions, and matrix removal or
enhancement performed manually before testing
by a discrete analyzer.
Cannot perform complex chemistries such as on-
line gas
diffusion, dialysis, distillations, extractions, and
digestions
103
104.
105. • is defined as medical testing
at or near the site
of patient care outside of the
conventional laboratory. .
• brings the test conveniently
and immediately to the
patient and increases the
possibilities of the patient
receiving the test result in a
timely manner.
106. • point-of-care test systems are easy-to-use
membrane-based test strips, often enclosed by a
plastic test cassette.
• These tests require only a single drop of whole
blood, urine or saliva, and they can be performed
and interpreted by any general physician within
minutes.
•
• POCT are accomplished through the use of
transportable, portable, and handheld
instruments and test kits.
107. • Non-automated Methods- may be done by
manual rapid-testing methods using a Dipsticks
or Immunostrips.
• Instrument-Based and Automated Methods-
are automated and use a small amount of
specimen. This type of automation requires
minimal technical support and is easy to use. It
includes visual readings, display
screen, printer, infrared, wireless radio
signals, or modems.
108. • Most of the instruments utilized for POCT use
whole blood for analysis and disposable reagent
unit-dose devices.
• The most popular POCT instrument is the I-STAT
analyzer.
109.
110.
111.
112.
113.
114. • used to measure blood
gas, pH, electrolytes, and some metabolites in
whole blood specimens.
• They are also used to determine abnormal
metabolite and/or electrolyte levels in blood
and the patient’s acid-base balance and
levels of oxygen/carbon dioxide exchange.
115. • It have extensive test menus and
provide a rapid laboratory results to
expedite a patient’s diagnosis and
treatment.
• There are many compact analyzers
available for bedside testing, screening
projects, wellness centres, operating
rooms and emergency rooms.
116. • BLOOD GLUCOSE TESTING
• Blood glucose levels are measured by a meter
and use a capillary blood directly from finger
sticks.
• The blood glucose test is ordered to measure
the amount of glucose in the blood right at the
time of sample collection. It is used to monitor
glucose levels in persons with diabetes.
117. Drugs of Abuse Testing
• Drug of abuse testing are frequently
ordered on patients who exhibit symptoms
of intoxication or offer a history of drug
ingestion.
• Rapid and accurate results are critical to
manage patients effectively.
118. • Taking the sample from the wrong patient
• Taking the wrong type of sample
• Failure to follow procedure
• Incorrect result interpretation
119. • Rapid test results essential for decision-making
• A system that generates a printout of the
results
• Requires small sample volume
• Allows testing in a variety of locations
• Potential to improve patient outcome or
workflow by having results immediately
available
• Less traumatic for the patients
• Portable devices are used
120. • Potentially different reference ranges
• Costly to operate
• Minimal training of personnel to
operate the instruments
• Management of POCT is challenging
• Not all methods are appropriate for
diagnosis or monitoring treatment