ABSTRACT

This paper aims to identify whether gradual and/or total automation of

laboratories will be of general benefit to the efficiency of laboratory
procedures. First of all, automation of laboratory procedures requires funding
to procure the reagents, software and machines. Secondly, it may be difficult
for some traditional laboratories to adapt to the new automated systems. In
order to rectify whether total automation systems are worth the financial
risks and effort, surveys and experimental studies were conducted in different
laboratories

and

institutions.

The

results

indicate

that

technological

advancements are very much needed in laboratories; for example, the

software that run these automated systems are key to maintaining a larger
and more comprehensive database. Also, the Total Automation Systems of
laboratories have shown to be of key importance in providing efficient,
accurate result in less the time it takes to do it manually. It is recommended
that these techniques be also made available to undergraduate students,
which could not only provide learning opportunity for the students
themselves, but will also give use to automated systems that are otherwise
to be discarded by laboratories.

INTRODUCTION
Time brings about drastic changes to our technological advancement.
In keeping with today's fast -paced technology, automated laboratory
techniques are becoming increasingly important in modern laboratories due
to its apparent precision, accuracy and efficiency. Automated laboratory
techniques refer to laboratory procedures that involve automated machines
and/or computers that expedite the procedure. However, these techniques
are not covered in traditional undergraduate laboratories. Imposing another
hurdle to the automation of laboratory techniques is the hard transition of
traditional method to the new automated methods in laboratories which
involves the gruesome process of procurement of these machines, briefing of
staff etc. Also, seeing that these modern techniques are automated, there

remains the fact that machines also create errors. In order to address this
problem, this paper will discuss various automated techniques, it's handling
and applications vis-a-vis with traditional laboratory techniques.
MATERIALS AND METHODS
A. Automation in Clinical Biochemistry: Core, Peripheral, STAT, and Specialist
Laboratories in Clinical Chemistry
Through a self-administered survey to seniors in clinical biochemistry in NATA
GX/GYclassified laboratories in Australia, the impact of technology to
laboratory techniques was assessed. The responses were yes, no, or not
applicable and are expressed as percentages of responses. Some of the
questions sourced for descriptive answers.

B. Incorporation of Automation to Undergraduate Laboratories

REAGENTS
Distilled, deionized water and HPLC grade methanol and acetonitrile were
used without further purification.

A mixture of natural capsaicins (65 %

capsaicin, 35 % dihydrocapsaicin) was purchased from Aldrich and used to

make standards ranging from 120 ppm total capsaicins in HPLC mobile phase
(75 % methanol, 25 % water). 3-mL, 500-mg BAKERBOND speTMOctadecyl
disposable extraction columns were purchased from VWR (Cat. No. JT7020-3)
and Tabasco brand pepper sauce (McIlhenny Co., Avery Island, LA) was used
as purchased from a local grocery. Students performed manual methods for
the solid phase extraction of capsaicin and dihydrocapsaicin from pepper
sauces similar to those described previously 1,4. First, 10 mL of acetonitrile
was added to a 2-3 g sauce sample and mixed well. Then, 1 mL of this
acetonitrile extract was diluted with 9 mL water. After conditioning an SPE
cartridge with 5 mL acetonitrile followed by 5 mL water, the above solution
was passed through the cartridge, disposing of the aqueous phase. The
capsaicins were eluted with 4 mL acetonitrile, and then with 1 mL of 1 %
acetic acid in acetonitrile. High performance liquid chromatography was used
for analysis of this combined extract. Figure 1.Zymark Benchmate II
Workstation used for acetonitrile extraction, filtration and SPE cleanup of

samples prior to liquid chromatographic analysis.()

AUTOMATED EXTRACTIONS
A Zymark (Hopkinton, MA) Benchmate TMII Workstation with DOS-based
software version 3.0 was employed for the automated sample preparation
methods. The manual SPE procedure was modified in order to conduct the
extraction using the robotic workstation. Undergraduate students, as
independent study projects, developed the robotic method used for this
experiment.

Another

change

in

the

method

was

mandated

when

backpressure limitations in the system caused a leak during the SPE step.
Therefore, a filtering step was added prior to SPE to minimize particulate
matter that would otherwise be passed through the cartridge. In addition,
the order of individual steps, vortex time and speed, and solvent flow rates,
as well as other set up parameters, were all realized as important
optimization variables in the sample preparation phase of the experiment.
The method program, which was performed on ~1 gram of sauce (weighed
accurately and loaded onto the sample tray), for the laboratory experiment
reported here is shown below:
Step 1.Add 5 mL acetonitrile
Step 2.Vortex for 600 s at speed 1
Step 3.Pause for 1 minute
Step 4.Condition column with 5 mL acetonitrile
Step 5.Condition column with 5 mL water
Step 6.Prewet with 1 mL of sample and filter 0.8 mL into next tube
Step 7.Add 7.2 mL water
Step 8.Vortex for 120 s at speed 1
Step 9.Load 5 mL sample onto column
Step 10.Collect 4 mL fraction into next tube using acetonitrile
Step 11.Collect 1 mL fraction into next tube using 1 % acetic acid in
acetonitrile
Step 12.End the total time for completion of this method program is 37.4
minutes.
CHROMATOGRAPHIC ANALYSIS

All standards and capsaicin extracts were analyzed by high performance

liquid chromatography during the second three-hour laboratory period.
Capsaicin and dihydrocapsaicin were quantified by reversed-phase HPLC
using a Kratos Spectroflow 400 dual piston solvent delivery system equipped
with a 20-mL injection loop and a Kratos SF 769Z variable wavelength UV/Vis
absorption detector set at 205 nm. A ZORBAX 4.6 x 150 mm column with 5
mm C18 particles was used (Agilent Technologies, Palo Alto, CA). The mobile
phase was 75:25 methanol:water at a flow rate of 1.0 mL/min, and data was
collected using a custom program written in LabVIEW software version 4.0
(National Instruments, Austin, TX).

Extracts (as prepared above) were

injected and chromatographic peak heights were compared to calibration

curves generated from a series of capsaicin standards.()

C.

Implementing a Laboratory Automation System: Experience of a Large

Clinical Laboratory
In this research conducted by, the following topics comparing and contrasting
pre- and post-LAS have been observed and explored: turnaround time (TAT),
laboratory errors, and staff satisfaction. The benefits and limitations of
Laboratory Automation System (LAS) from the laboratory experience were
also reviewed. Question and answers regarding staff satisfaction are
descriptive while the TAT, and laboratory errors were assessed through
empirical data.

D.

Does Laboratory Automation for the Preanalytical Phase Improve Data

Quality?
In this research conducted by G. Oliveira et al., Blood from 100 volunteers
was collected into two vacuum tubes. One sample from each volunteer was
respectively

assigned

to

(G1)

traditional

processing,

starting

with

centrifugation at 1200g for 10 min, and (G2) the MODULAR PRE-ANALYTICALS

EVO-MPA system. The routine clinical chemistry tests were performed in
duplicate on the same instrument Cobas 6000 <c501> module. G1 samples

were uncapped manually and immediately placed into the instrument. G2

samples were directly fed from the MPA system to the instrument without
further staff intervention. At the end, (1) the G1 samples were stored for 6 h
at 4 C as prescribed in our accredited laboratory and (2) the G2 samples
were stored for 6 h in the MPA output buffer. Results from G1 and G2, before
and after storage, were compared.

G.

Managing the Workflow of a High-Throughput Organic Synthesis

Laboratory: A Marriage of Automation and

Information Management

Technologies
According to Dr. Burton Goodman, PhD, the procedure for this research are as
follows:
Production of libraries began with loading the appropriate linkers onto
resins, followed by complete characterization of the resin-linker intermediate.
The resins were then loaded into IRORI MiniKans and sorted using the IRORI
Autosort 10Kx sorting workstation. Chemistry was then performed using
standard laboratory glassware. Additional sorting and chemistry steps were
then performed until the compound was ready for cleavage off of the resin.
The MiniKans were then sorted into Bohdan MiniBlocks and treated with the
appropriate cleavage cocktail. Collection into 48-position racks was followed
by removal of cleavage solution through vacuum centrifugation. The
concentrate was then dissolved in a solvent mixture that allowed for standard
liquid handling automation to create 96 well plates for analysis by high
throughput flow inject NMR and LC/MS.

RESULTS AND DISCUSSION

A. Automation in Clinical Biochemistry: Core, Peripheral, STAT, and Specialist
Laboratories in Australia
According to G. Streitberg et al., Eighty-one laboratories responded, and the
locations were 63%, 33%, and 4% in capital cities, regional cities, and country

towns, respectively. Forty-two percent were public and 58% private. Clinical
biochemistry was in all core laboratories of various sizes, and most performed
up to 20 tests per sample. Thirty percent of the 121 surveyed laboratories
had plans to install an automated line. Fifty-eight percent had hematology
and biochemistry instrumentations in their peripheral laboratory, and 16%
had a STAT laboratory on the same site as the core laboratory. There were
varied instruments in specialist laboratories, and analyzers with embedded
computers were in all laboratories. Medium and large laboratories had
workstations with integrated instruments, and some large laboratories had
TLA.
B. Incorporation of Automation to Undergraduate Laboratories
Although many hot sauces have been used in this work, results reported
here are for Tabasco brand pepper sauce due to its availability and ability to
achieve a homogeneous solution compared to other oil-based sauces that
have a tendency to separate into distinct phases. Figure 2 represents
example liquid chromatograms for a capsaicin standard and for a Tabasco
extract prepared by the automated method. The baseline of the standard is
offset for clarity. Capsaicin and dihydrocapsaicin were identified in the sauce
extract

Figure

3.HPLC

calibration

plots

generated

from

capsaicin-

dihydrocapsaicin standards. The equations of the lines were y = 0.0145x +

0.0025 (r2= 0.9995) and y = 0.0111x + 0.0010 (r2= 0.9989) for capsaicin
and dihydrocapsaicin, respectively. by their chromatographic retention times
of 4.1 and 5.3 min, respectively. Other peaks in the extract chromatogram are
consistent with previous reports, such as the peak at 1.5 minutes being a
polar constituent not removed by the SPE cleanup,1and the peaks at 5.8 and
7 min being other capsaicin compounds4that were not a focus in this work.
Figure 3 presents student calibration curves generated from standards for
capsaicin and dihydrocapsaicin used to quantitate the two compounds in the
sauces. Plotting peak height as a function of capsaicin concentration yielded
straight lines with acceptable correlation coefficients (r2= 0.9995 and 0.9989
for capsaicin and dihydrocapsaicin, respectively) for quantitation. Figure 4
presents typical data obtained by students for the concentrations of capsaicin

and dihydrocapsaicin in Tabasco sauce, comparing manual and automated

sample preparation methods. The error bars on the plot represent the
standard deviation of several independent extractions (n = 7 for the manual
extractions and n = 6 for the automated extractions). The two methods
yielded statistically similar results in terms of concentrations (107 12 ppm
capsaicin and 66 12 ppm dihydrocapsaicin and 117 8 ppm capsaicin and
73 7 ppm dihydrocapsaicin for preparations done manually and by the
Benchmate, respectively), but better reproducibility was achieved with the
automated preparation method. The results obtained here are consistent with
capsaicin concentrations reported (in terms of Scoville Heat Units) for
Tabasco.5It should be noted, however, that capsaicin concentrations in this,
and other, pepper sauces can vary from bottle to bottle, so these reported
results are from the same bottle.
C. Implementing a Laboratory Automation System: Experience of a Large
Clinical Laboratory
The mean TAT for both stat and routine samples decreased post-LAS (30%
and 13.4%, respectively). In the 90th percentile TAT chart, a 29% reduction
was seen in the processing of stat samples on the LAS. However, no
significant difference in the 90th percentile TAT was observed with routine
samples. It was surprising to note that laboratory errors increased post-LAS.
Considerable effort was needed to overcome the initial difficulties associated
with adjusting to a new system, new software, and new working procedures.
D.

Does Laboratory Automation for the Preanalytical Phase Improve Data

Quality?
Results from G1 and G2, before and after storage, were compared.
Significant increases were observed in G1 compared with G2 samples as
follows:

(1)

before

storage

for

alkaline

phosphatase

(ALP),

lactate

dehydrogenase (LDH), phosphate (P), magnesium (MG), iron (FE), and

hemolysis index and (2) after storage for total cholesterol (COL), triglycerides
(TG), total protein (TP), albumin (ALB), blood urea nitrogen (BUN), creatinine

(CRE), uric acid (UA), ALP, pancreatic amylase, aspartate aminotransferase

(AST), alanine aminotransferase (ALT), g-glutamyltransferase (GGT), LDH,
creatine kinase (CK), calcium (CA), FE, sodium (NA), potassium (K), and
hemolysis index. Moreover, significant increases were observed in (3) G1after versus G1-before storage samples for COL, high-density lipoprotein
cholesterol, TG, TP, ALB, BUN, CRE, UA, AST, ALT, GGT, LDH, P, CA, MG, FE,
NA, K, and hemolysis index and (4) G2-after versus G2-before storage only for
BUN, AST, LDH, P, and CA

CONCLUSION
A. Automation in Clinical Biochemistry: Core, Peripheral, STAT, and Specialist
Laboratories in Australia
Technology evolution and rising demand for pathology services make it
imperative for laboratories to embrace such changes and reorganize the
laboratories to take into account point-of-care testing and the efficiencies of
core laboratories and TLA.
B. Incorporation of Automation to Undergraduate Laboratories
An automated sample preparation method was developed by students for the
extraction and SPE cleanup of capsaicins from commercial pepper sauce
using a robotic workstation. In addition, automation was integrated into the
undergraduate instrumental analysis laboratory by modifying an existing
experiment without utilizing additional lab time, and students were eager to
work with the workstation rather than preparing the samples by hand. The
authors hope to increase awareness of the readers of the Journal about the
important role of teaching institutions in implementing automation, and will
also realize the pedagogical benefits of
1. industrial-academic collaborations and
2. Donations of equipment that might otherwise be discarded.
C. Implementing a Laboratory Automation System: Experience of a Large
Clinical Laboratory
Although some of the known advantages and limitations of LAS have

been validated, the claimed benefits such as improvements in TAT, laboratory

errors, and staff morale were not evident in the initial months.
D.

Does Laboratory Automation for the Preanalytical Phase Improve Data

Quality?
Results show that MPA system improves the quality of laboratory testing.
Therefore, laboratory automation for the Preanalytical phase improves data
quality.
E. Automation and Expert Systems in a Core Clinical Chemistry Laboratory
Automation of the main chemistry analyzers, including immunoassay and
linking them together with preanalytical and postanalytical automation to
give total laboratory automation has given predictability to result availability.

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