Complete automation will lead to human prohibition in pharma industries. Analytical techniques in drug discovery and development stage generate large amounts of data that is not possible for humans to statistically analyze.

1. Introduction to Automated
Methods of Analysis
Mr. Sanket P. Shinde
Assistant Professor
Pune-Maharashtra.

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IUPAC define automation as ‘the use of combinations of mechanical and
instrumental devices to replace, refine, extend or supplement human effort
and facilities in the performance of a given process, in which at least one
major operation is controlled without human intervention, by a feedback
mechanism’.
Automated devices function without human intervention and are conceived
to make decisions with the aid of feedback system.
The highest level automation is when the device is completely automatic.
Completely devices are self-monitoring and self-adjusting, have greater
independence than other automatic devices.
Introduction

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Human Prohibition Avoid Contamination Cost Cutting
Accuracy & Precision Large Number of Sample

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Need for Automation
▪ There is a growing trend for minimal human participation in laboratory
process.
▪ Large number of analytes is to be determined.
▪ Growing demand for cost reduction in personnel and material
expenditure.
▪ All types of errors arising from human factor need to be eliminated.
▪ Hospitals, industries and laboratories urgently require data for process
controls.
▪ Some chemicals, especially reagents, are expensive and unstable.
▪ Analytical techniques in drug discovery and development stage generate
large amounts of data that is not possible for humans to statistically
analyze.

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▪ Process large number of samples.
▪ Process Control.
▪ Determination of multiple components from the same sample.
▪ Facilitate analytical techniques.
▪ Reduce human intervention to cut errors and cut cost.
▪ Lower consumption of expensive reagents.
▪ Increase sample throughput.
▪ Handle large number of samples.
▪ Process samples with low concentration of analytes.
Objectives of Automation

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▪ Faster analysis.
▪ Automatic data recording.
▪ Closed system hence free from contamination.
▪ Human intervention minimal hence free from human error arising out
of boredom, fatigue and carelessness,
▪ Greater accuracy and reproducibility as all samples are subject to
same process.
▪ Reduced costs on account of smaller samples and expensive reagents.
▪ Automatic sample preparation and range changing.
Benefits of Automation

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There are two types of automatic analysis techniques
1. Discrete Technique : In this technique, each sample undergoes analysis
in a separate chamber. The samples are analyzed either parallel or
sequentially. Therefore, the discrete technique is also known as batch
analysis.
Following are the salient features of this technique-
▪ Each individual sample is treated as a separate entity and analyzed
accordingly.
▪ Both the sample and the reagent are metered into discrete reaction
vessels.
▪ In some cases the reagent according to the analyte is already present in
the reaction vessel and thus the sample only needs to be introduced into
the reaction vessel.
▪ In the reaction vessel the reagent is mixed with the sample and then
color measurement occurs.
Types of Automatic Analysis Technique

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2. Continuous Technique : In this technique, the samples flow
continuously and sequentially in the tube each sample separated by an
air bubble. Each sample is mixed sequentially with reagent in the tube
and then passed to the detector.
There are two types of continuous techniques;
a. Segmented Flow Technique
▪ In this technique a peristaltic pump pumps sample and reagent in to the
chamber through tubes.
▪ The mixture is then passed into the detector for analysis.
▪ The aspirated samples are separated by air bubbles that should be
removed before the mixture reaches the detector.
▪ Once, the air bubble is removed each sample is separated by washing
solution.
b. Flow Injection Analysis
▪ FIA is based on the injection of a liquid sample into a moving
continuous no segmented carrier stream of a suitable liquid. The injected
sample forms a zone which is then transported towards a detector.

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1. Sequential Automation Process : The sequential automation process is
also known as serial automation process. In this process the analysis is
carried out sequentially and as such the second one begins only after the
completion of the first. It is also possible to configure the sequential
automation process with online collection of sample and determination
from a flowing stream.
2. Parallel Automation Process : In this process more than one automated
chemical operations is performed simultaneously. Thus some or all of the
below mentioned chemical operations can be performed simultaneously.
1) Obtaining sample 2) Sample preparation
3) Component separation 4) Analytes detection
5) Report generation
3. Hybrid Automation Process : As the name itself suggests this process is a
combination of the above two processes. Thus, some processes are
conducted sequentially while some are conducted in parallel.

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The basic automatic analysis system comprises of the following -
▪ Sampling unit
▪ Proportioning pump unit
▪ Manifolds unit
▪ Dialyzers unit
▪ Constant bath temperature unit
▪ Detector unit
▪ Recorder unit
The steps in automatic analysis system are -
1. Sample collection
2. Sample pre-treatment and measurement
3. Control instrument parameters
4. Calculation of results
5. Check results for conformance to set parameters
6. Generate and present report

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1. Sampling Unit
▪ The primary function of the sampling unit is to introduce unmeasured
sample into the system.
▪ The sampling unit consists of a circular turntable carrying cups and
rotating at a predetermined speed.
▪ The movement of the turntable is synchronized with the movements of
the sampling crook.
▪ The sampling crook is a thin tube that aspirates the sample into the
cups.
▪ The movement of the turntable and the sample mixers attached enable
the sample to mixed before and during aspiration. The cups are washed
between samples.

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2. Proportioning Pump Unit
▪ The function of the proportioning pump is to continuously push the
sample, reagents, air and gasses through the analytical chain.
▪ The proportioning pump comprises of roller head assembly and multiple
flexible plastic tubes, one coming from sampler and other from reagent
bottles and one simply drawing air.
▪ These tubes are placed length wise along the platen platform.
▪ The roller head assembly is driven by a constant speed gear motor
which when switched on compresses the tube against the platen driving
the liquid before them,

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3. Manifolds Unit
▪ A manifold comprises of mixing and delay coils, pump tubes,
transmission tubes, platter and fittings.
▪ The mixing coil is used to mix the sample and delay a specimen for a
completion of a chemical reaction before reaching the colorimeter.
▪ Air is introduced into the reagents tubes through the tubes in the
manifold thereby producing a series of bubbles at regular intervals in
the stream known as bubble pattern.
▪ Fittings are employed in manifolds to join stream of liquids, split a
stream or introduce air to the stream.

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4. Dialyzer Unit
The function of dialyzer is to remove protein cells to ensure an
interference free analysis. The dialyzer comprises of a pair of mirror
grooved Perspex plates clamped with a semi permeable cellophane
membrane which goes into the centre and returns outside all on its own.
5. Constant Temperature Bath Unit
The primary function of constant temperature thermostatically
controlled bath is to ensure constant temperature of the stream. Once,
the stream leaves the dialyzer it is combined with additional reagents.
6. Detector Unit: Detectors such as colorimeter and flourimeter are used.
7. Recorder Unit
The function of the recorder is to keep a record of the analysis and
present when required. The most common recorder is de voltage null-
balance potentiometer recorder.

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The Good Automated Laboratory Practices (GALP) provides important
guidelines for the management of automated laboratory, and have ensured that
work can be carried out efficiently and effectively while saving time and
minimizing human intervention.
The basic principles of GALP are as follows:
▪ The system should be able to assure the integrity of all entered data.
▪ The formulae and algorithm employed by the system should be reliable,
accurate and appropriate.
▪ The system should provide an audit trail that should track data entry and
modification to the person responsible for the same.
▪ The change control procedure must be capable of tracking the system
operation and application software.
▪ Appropriate user procedure must be followed.
▪ The system should possess alternative plan for system failure, disaster
recovery and unauthorized access.

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▪ Flow injection analysis (FIA) is characterized by an automated,
continuous flow approach to perform chemical analysis.
▪ It is based on injecting a small, well-defined volume of sample into a
continuously flowing carrier stream whereby a concentration gradient of
the sample is created.
▪ The sample zone is gradually dispersed into the carrier by axial and radial
diffusion as it is moved through the channel under laminar flow
conditions.
▪ Reagents may be added at various confluence points and mix with the
sample zone under the influence of radial dispersion to produce reactive
or detectable species.
▪ The obtained peak height or area can be used to quantify the analyte. The
whole procedure, including sample injection, transport, reaction, and
detection, can be performed in a relatively short time.
Flow Injection Analysis

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Flow Injection Analysis

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Sample and Reagent Transporter
The solution in a flow-injection analysis is moved through the system by a
peristaltic pump. The peristaltic pump is a device in which the fluid is
squeezed through plastic tubes.
Peristaltic Pump

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Sample Injectors and Detectors
▪ The injectors and detectors employed in flow-injection analysis are similar
to those employed in HPLC in terms of design and performance.
▪ The sample solution should be infected rapidly as a pulse without
disturbing the flow of the carrier stream.
▪ Detection in flow-injection procedures has been carried out by automatic
absorption and emission instruments, fluorometers, electrochemical
systems, refractometers, spectrophotometers and photometer.
Separations in flow-injection analysis
▪ Separation in flow injection analysis is carried out by dialysis, liquid/liquid
extraction, and by gaseous diffusion.
▪ Dialysis is used to separate inorganic ions like chloride, sodium, or small
organic molecules such as glucose, from high molecular weight species
such as proteins.
▪ Extraction is another separation technique that is used in flow injection
analysis.

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There are mainly five types of flow injection analysis
▪ Syringe based injection system
▪ Injection with rotary valve
▪ Proportional injection
▪ Merged injection
▪ Hydrodynamic injection.
Types of Flow Injection Analysis

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