Advance Instrumentation

and Techniques
 Qualitative analysis vs Quantitative analysis

 Most analyses today are carried out with specially designed electronic
instruments controlled by computers

 These instruments make use of the interaction of electromagnetic radiation and

matter, or of some physical property of matter, to characterize the sample
being analyzed.

 Often these instruments have automated sample introduction, automated

data processing, and even automated sample preparation.

 To understand how the instrumentation operates and what information it can

provide requires knowledge of chemistry, physics, mathematics, and
engineering.
 The approach used to solve problems may include the following
steps:
1. Defining the problem and designing the analytical method
2. Sampling and sample storage
3. Sample preparation
4. Performing the measurement
5. Assessing the data
6. Method validation

7. Documentation
LABORATORY INFORMATION SYSTEMS (LIS)

LIS is a system of computer software designed to handle laboratory data. An LIS

achieves its function via a central computer, a number of input/output
devices, and the computer software
The functions of an LIS include:
(1) Database of patient information
(2) Compilation of specimen test results
(3) Production of patient reports
(4) Production of ancillary reports
(5) Data storage
The major Pros of an LIS
1. Diminishes time-consuming paper work
2. Stores patient data
3. Stores QC data
4. Offers immediate access to results
5. Reduces the chance of an assay being ran on the wrong patient when barcode is used
6. Managerial reports (tests per month, workload units, TATs)
INSTRUMENT SELECTION PROCESS
A. Needs assessment
1. Characteristics of the individual laboratory
2. Test volume (Random access vs. batch testing)
3. Throughput (TAT for STATs)
4. Connectivity (LIS)
5. Test menu for present and future needs
6. FDA approved
7. Sample dilution (manual vs. automatic) Cost
8. Ease of troubleshooting 1. Instrument itself
9. Special electrical or plumbing requirements 2. Reagents
10. Waste management 3. Contracts (service, labor, travel)
11. Amount of heat generated 4. Interface costs (LIS)
12. Safety features
B. Minimum standards for quality
1. Linearity 2. Sensitivity 3. Specificity 4.Accuracy
5. Precision 6. Stability of hardware/software (bar coding and ID features)
7. Stability and availability of reagents
 STAT Tests - Test results that are urgently needed for the diagnosis or treatment of the
patient. The delay can be life threatening.

 ASAP (As Soon As Possible) – Test results that are needed as soon possible for the
diagnosis or treatment of the patient.

 Routine – Tests that are collected and batched for efficiency and cost effectiveness. These
results are not needed on an immediate basis for diagnosis or treatment

 TAT (Turn Around Time) – The interval of time between when a sample is received by the
laboratory and the results are reported
PROBLEM SOLVING
A. Define the problem
B. Create scenarios
C. Implement solution
D. Examine solution
E. Reflect (on problem/solution)

1. Quality assurance (QA) is a comprehensive set of policies, procedures, and practices

that are followed to ensure that a laboratory’s results are reliable. QA evaluates the quality of
the services provided. QA provides a way to prevent problems as well as deal with problems
that occur

2. Quality control (QC) ensures that a particular test method is working properly and that
results of that test are reliable

QA includes record keeping, maintenance and calibration of equipment,

proficiency testing, quality control, and training of personnel.
 Writing a Laboratory Notebook
The following information is essential:-
• Date
• Team
• Objective(s): Statement of the problem or task
• Bibliographical references
• Experimental plan
• Safety comments (special hazards)
• Environmental protection, waste disposal
• Chemicals used, approach (g, ml, moles), purity
• Very detailed information on the apparatus used
• Procedures
• All manipulations and observations
The observations made and the data recorded will lead to the acceptance
or rejection of the stated hypothesis, and will decide what future
experiments may be done.
 They need to be:
• Recorded honestly.
• Recorded as you go along, in the notebook, in ink, immediately.
• Do not trust to memory, even for a minute or so – someone
talks to you, and that data are forgotten.
• Do not use odd scraps of paper or the edge of your lab coat to
record data.
• The raw data are precious. Information on where raw data are
stored is essential.
• use good penmanship
• Interpretation of experimental data
• Discussion and conclusion
• relate your results to your hypothesis – do they support or
refute it? Comparisons must be as quantitative as possible. Of
course a simple analysis practical will only produce a result
• record any ideas you have, however brief – if you don ’ t write
them down, you will forget them
Laboratory logbook. A page number, B date, C title/statement of purpose, D team member, E
objective of experiment, F background, G description of procedure, H important details, I
signature
Laboratory logbook.
J error crossed off with
explanation,
K computation of data
and answer to the question
posed by the experiment,
L important information
 Accuracy and Precision
 Accuracy is a measure of how close a measured analytical result to the true
answer.
 We often work with an “accepted” true value or “accepted reference value”
 Accuracy is evaluated by analyzing known, standard samples.
Such as standard reference materials; certified reference materials
 Another way of assessing accuracy is to spike a sample with a known amount of
the pure analyte The sample is analyzed and the amount of added analyte
recovered is reported.
- Such as A spike recovery of 100% would indicate that all of the added
analyte was measured by the analytical method
 The accuracy of a single measured quantity is the difference between the true
value and the measured value divided by the true value and expressed as a
percent.
 • Accuracy is the nearness of a test result to the true value.

 Precision is a measure of how close replicate results on the same sample are
to each other.
Imprecise and inaccurate Accurate but imprecise

Precise but inaccurate Precise and accurate

 Replicate Determinations of Analyte in a Samplea
% Analyte
Analyst 1b Analyst 2C Analyst 3d
10.0 8.1 13.0
10.2 8.0 10.2
10.0 8.3 10.3
10.2 8.2 11.1
10.1 8.0 13.1
10.1 8.0 9.3
Average (%) 10.1 8.1 11.2
Absolute error e 0.0 2.0 1.1
Standard deviation 0.089 0.13 1.57
a Accepted true answer is 10.1+0.2% (obtained independently). dResults are imprecise and inaccurate
b Results are precise and accurate eAbsolute error = true value - measured value
C Results are precise but inaccurate
 Different analytical techniques have different requirements, these are the most
common :
(i) Selectivity – The ability to detect only the analyte and not any other component that
may be present in the sample or in the sample solution.
(ii) Precision – The tightness and lack of scatter of results.
(iii) Linearity – The direct relationship between the amount or concentration of an
analyte and the detector signal produced. The linearity plot should normally have a small
y intercept.
(iv) Recovery (accuracy) – The ability to detect all of the analyte and have the detector
response of the analyte be the same in both the sample and the standard.
(v) Sensitivity – The ability to accurately detect and quantitate low levels of analyte
when this is required.
(vi) Robustness – The insensitivity of the method’s ability to generate accurate results
when small changes occur in test conditions and parameters.
Practicability – ability of a method to be easily repeated
Reliability – ability of a method to maintain its accuracy and precision for an
extended period of time under different variables
Sensitivity
The sensitivity of a test in the ability of the test to identify
correctly affected individuals
Proportion of persons testing positive among affected individuals

Affected persons
(Positive by gold standard)

Persons testing positive Persons testing negative

(True positives) (False negatives)

Sensitivity = True positives / Affected persons

Estimate the 95% confidence interval
Specificity
The specificity of a test in the ability of the test to identify
correctly non-affected individuals
Proportion of person testing negative among non affected individuals

Non-affected persons
(Negative by gold standard)

Persons testing negative Persons testing positive

(True negatives) (False positives)

Specificity = True negatives / Non-affected persons

Estimate the 95% confidence interval
 Types of Errors

 There are two principal types of error in analysis: determinate or systematic error and
indeterminate or random error
1. Determinate Error

 Caused by faults in the analytical procedure or the instruments used in the analysis.
 Implies that the cause of this type of error may be found out and then either avoided or
corrected
 A particular determinate error may cause the analytical results produced by the
method to be always too high; another determinate error may render all results too
low.
 Can arise from uncalibrated balances, improperly calibrated volumetric flasks or
pipettes, malfunctioning instrumentation, impure chemicals, incorrect analytical
procedures or techniques, and analyst error
 Analyst error
 The person performing the analysis causes these errors.
 They may be the result of inexperience, insufficient training, or being “in a hurry”.
 An analyst may use the instrument incorrectly, perhaps by placing the sample in the
instrument incorrectly each time or setting the instrument to the wrong conditions for
analysis
Some other analyst-related errors are
(1) carelessness

(2) transcription errors

(3) calculation errors

 Reagents and instrumentation

 Analytical method
 2. Indeterminate Error

 Include the limitations of reading balances, scales such as rulers or dials, and
electrical “noise” in instruments
 Indeterminate errors arise from sources that cannot be corrected, avoided, or even
identified, in some cases

 All analytical procedures are subject to indeterminate error

Generalized instrumentation system
 Assignment 1
Q1: What are the main causes that contribute to poor accuracy and
Precision?

Q2: How we can avoid the systemic errors?

Q3: What are the main sources of the errors that you find in your work?

Q4: Mentions different types of centrifuge that present in your work and
state the maintenances that you do ?