So welcome to the session three
of the Waters Webinar Week focusing on innovations
and applications for the clinical workflow. I'm Godo Bosch, and
I will be your host. Before we get into
the presentation, I'd like to cover a
few practical details. Some of you may be already
familiar with this setup, but I will still cover it. At the bottom of your screen,
we have attached some material related to the talk today. Which can be downloaded
or consulted. Some are documents and
some others are web pag
es. On the top left is a
very important feature. It's the Q&A box. You can type in your
questions as they come during the
presentation, and we'll do our best efforts
to answer all of them at the end of the presentation. And on the right is another
very important thing-- a very short survey for--
or no, three questions that we'd like you
to answer and give us some feedback so that
we can improve further for future events like these. So for today, we
have two presenters. I will start with
Isha Cho
wdhary, who is a Senior Global Product
Manager in the Clinical Business Unit, which has been
created 18 months ago by Waters Corporation. She joined Waters
three years ago and has recently supported
the development and launch of the MassTrak
immunosuppressant sets as well as the mass
track steroid serum sets. Starting the presentation
will be Dominic Foley, who is a Senior Scientist
working at Waters' Mass Spec Headquarters in
Wilmslow in the UK. And he's part as well of
the Clinical Business Un
it. And he joined Waters 10
years ago and has specialized in the development of LC-MS/MS
clinical research applications with a particular focus on
steroid hormone analysis as well as biomolecules. Their presentation title
is, "Enabling Harmonization of Steroid Hormone LC-MS/MS
Methods Through Use of MassTrak Steroid Serum Sets." And without further ado, I'll
pass it on to Dom and Isha. Thank you, Godo. So thanks for joining today. This is the outline of what
we plan to discuss today. I will firs
tly
cover the importance of high-quality
analytical methods in steroid measurements, and how
the perception of these methods have changed over
the last 20 years. This will include emphasis
on the value of LC-MS/MS and the confidence in the
results it can provide. Isha will then
discuss the Waters workflow and the MassTrak
sets and how these can benefit your laboratories. I will then follow this
up with an LC-MS method for steroid hormone analysis
using the Waters workflow. And that includes the
use of
the MassTrak track sets RUO. Finally, Isha will
then finish up the presentation with further
resources and collateral. So the importance of
high-quality analytical methods in steroid measurement. High-quality analytical methods
provide a number of benefits, including aiding the consistency
of data interpretation, which can improve the identification
of abnormal results. This can help minimize
additional testing related to poor performance of the
methods, saving time and money. This consis
tency can result
in greater harmonization of data across
different laboratories, improving the overall quality
of cross-collaborative research. For steroids that exhibit
diurnal and age-related changes in addition to plasma
protein binding, an analytical method that meets
the measurement performance specifications can be critical. And as you can see on the
right-hand side of the slide, a steroid such as
testosterone falls into this category based
on biological variation, and in this case taken
f
rom the EFL database. The desirable bias and
precision specifications are around 6%,
indicating the need for a high-quality
analytical method associated with this analysis. So there are challenges in
steroid hormone measurements, and I thought I'd use
testosterone to shine a spotlight upon
these challenges. These challenges have been
fully recognized in the last 20 years, and in 2003 there was an
editorial in Clinical Chemistry that asked whether
widely-adopted immunoassay techniques for
testost
erone analysis are better than a
guess and this was based on method comparisons
between immunoassay platforms and mass spectrometry and
later corroborated by others. It demonstrated that
testosterone immunoassay results from females
were inaccurate and sometimes missed the
target values by 200% to 500%. And these studies resulted in
the generation of an Endocrine Society position statement on
testosterone, stating that-- stating that direct assays for
testosterone should be avoided, and methods
using extraction
and chromatography...chromatography...[AUDIO OUT] Similar issues were
highlighted across different steroid
hormone methods, which led to the Endocrine Society
and Neurology Society guidelines highlighting issues of using
immunoassays for steroids and indicating MS methods are
the preferable measurement. Then in 2013, a
positioning statement was released by the Journal
of Clinical Endocrinology and Metabolism, somewhat
controversial at the time, which recognized that for
high-imp
act steroid research, the era of immunoassays
was coming to a close and the acceptance
guidelines on manuscripts was going to reflect
this from January 2015. However, this
recommendation was later suspended pending
further review because of the wide-ranging
impacts this would have on the clinical research. However, based on the growing
use of LC-MS/MS in clinical laboratories in 2014, CLSI
released an approved guideline on the use of and implementation
of LC-MS/MS methods in clinical labs. And w
ithin that, they state that
LC-MS/MS assays are superior with regard to selectivity
due to combination of the LC separation and mass selection. And the power and value of
LC-MS/MS is demonstrated in this slide where we have
a series of different steroid isomers that could interfere
with each other during immunoassay measurements, and
even just mass spectrometry alone where we have
11-deoxycortisol, 21-deoxycortisol,
corticosterone, all isomers of each other. 17-hydroxyprogesterone,
11-deoxycorti
costerone are isomers, as
well as testosterone and epitestosterone. And what we see here is that
we can separate them out and be detected using LC-MS.
So the orthogonality of the chromatography with
the MRM mass spectrometry is a particular powerful tool
that enhances the method, precision, and accuracy
for these steroids, providing greater confidence
in the results we obtain. So one key difference in
the use of LC-MS methods for steroid analysis
is that they are often based on
laboratory-develo
ped tests, and these are validated
to regulatory guidelines such as CLSI, FDA, and EMEA
method validation guidelines. And this means that method
harmonization and the quality of laboratory materials
used becomes more important, and as part of lab
quality management systems are increasing demands on
demonstrating the quality of clinical methods using LDTs. One aspect of this
is the growing need for metrological, traceable
calibration materials to replace in-house prepared
materials to aid complia
nce with ISO 15189. And these are based
on requirements for quality and competence
in medical laboratories. This metrological
traceability is a process that related measurement
values to a reference standard and is a tool to ensure
more accurate results. So with that in mind,
this is just something that we thought we'd ask
in terms of a quick poll. So if possible, in the
next 20 or 30 seconds, I just want to get
some quick feedback from the audience on
a question around, is metrological traceabi
lity
for your CALs and QCs important to you? And it's either yes or
no in this instance. So we just want
to a quick idea of is it a high priority
in your laboratory in regards to the production of
calibration and QC materials? So I'll just give you another
five or ten seconds on that, and then hopefully we
do get some poll results from the attendees. And then what the
intention is then is to pass over to
Isha, who will take you through some of the
benefits of the new MassTrak sets for steroid an
alysis. And we have 100%, so
yes, it's important. So over to you, Isha. Thanks, Dom. And thank you so
much for our audience for participating in that poll. So to meet your
laboratory's need to develop these high-quality
analytical methods for steroid measurements,
Waters offers a suite of products for the
end-to-end endocrinology workflow. The products included in
this workflow help support a wide range of endocrinology
applications such as steroids, proteins and peptides,
amines, and vitamins.
So if you look at the overall
workflow starting from sample prep, we offer Andrew+ robots
that execute sample prep steps with a high level of
reproducibility and full experimental traceability. Additionally for sample prep,
we offer precision consumables and standards that are produced
with low lot-to-lot variability and tested rigorously in-house. Our MassTrak standards,
such as calibrators and QCs, are metrologically traceable
to both reference material as well as reference
measurement procedu
res, and help with steroid
method standardization. For LC-MS/MS, our ACQUITY
I-Class PLUS LC systems coupled with either the XEVO TQ-S micro
or the XEVO TQ-S Absolute mass spectrometers help support
robust quantification of a broad range of
endocrinology analytes, especially those that are
found at the very low end of the concentration
range in the human body. And then finally on the back
end, our data acquisition and reporting softwares such
as MassLynx and TargetLynx offer a range of data
proc
essing functionalities with the ability
to bidirectionally communicate with labs'
LIMS systems as well. So I wanted to take a deep
dive into the MassTrak steroid serum sets which form a critical
component in the steroid LC-MS workflow. So here's an overview of the
MassTrak Steroid Serum Sets family. A set will contain at
least a Cal and QC, and in some instances
contains additional products like the internal
standards and optimization mixes that are critical to
method development as well. There
are three different
steroid sets that we offer. Set 1 includes CALs, QCs,
IS, and optimization mix, and covers a panel of 12
different steroid hormones indicated on the bottom
left-hand side of the slide. This panel was designed
keeping in mind the medium to low
throughput labs that typically get requests to
test multiple steroids in one sample and have less pressure
on meeting short turnaround time expectations. Sets 2 and 3 contain
CALs and QCs. Set two includes a panel
of four steroid hormone
s. Set 3 includes a panel of five. And these smaller
panels are more suitable to your high
throughput labs that typically get requests to test fewer
number of steroid analytes per sample and have
higher expectations to meet short turnaround times. All of the Cal, QC, IS,
and optimization sets can be ordered
individually, and some of the components like
the IS and OptiMaxes are compatible across all
three pairs of Cal and QC sets. All three sets are
available in RUO format currently, with the Cal
and
QC set one also available in the IVDD format in Europe. We've launched these RUO sets
1 2 and 3 in June this year, and we're very pleased
to announce the upcoming launch of the
IVDR-approved sets 2 and 3 that should become orderable in
the next two to three months. Set 1 IVDR will then soon
follow up with a launch in Q2 next year. Here are some high-level
features of the sets. Concentration ranges for
all of the analytes included in the three calibrator
sets are included on the table on the
left. And as you hover down the
table, right off the bat you can see that there are wide
concentration ranges in place for all analytes
included in these CALs to support various analytical
and workflow needs. In terms of configuration
for the CALs, we offer at least six different
levels including the blank. The QCs are offered in
either a three-level or a four-level format. The CALs, QCs, optimization
mix, and the IS-- all of them contain lyophilized
steroid compounds with a target recon volume
of 2 mils for the CALs, QCs, and optimization mix, while the
IS offered with a target recon volume of 25 mils. Additionally, these products
have long in-use stability. So we report up to three
months at -ten degrees Celsius for all of the four components
and a stability at one month at 2 to 8c for CALs and QCs. And finally, all
of the CALs and QCs are prepared from
different CRM sources and the values are
assigned and traceable to either certified reference
material or reference measurement pro
cedures
like Ghent or RFB. So all of the features
that I just spoke about in the earlier slide were meant
to confer four main benefits to your lab. So with using these
ready-to-use steroid serum sets, you can save time
and resources that would have otherwise been spent
on producing these in-house. And with 3X longer
in-use stability compared to some of
the more competitor products on the market,
we allow the consumption of this product for a long
period of time as well. Secondly, you can
comply
with ISO 15189, as well as CLIA C-62A,
some of the guidelines that Dom talked about
earlier in the slide, because these CALs and QCs are
metrologically traceable to not just CRM, but to reference
measurement procedures for certain analytes. And we take a lot of
effort in making sure that our CALs and QCs are
sourced from two different CRM sources. Thirdly, the wide concentration
ranges for our CALs and QCs have really helped
support a wide range of analytical and workflow
needs that you may have
. And then lastly, you can
observe high performance in pure lab comparisons
like EQA schemes that you may participate
in by using these sets that are manufactured with high
lot-to-lot consistency and tested in-house using those
select EQA materials as well. So I wanted to double-click
on that final benefit. Apologies. Let me just go back. Not quite sure why the
photo is showing, but-- so we go through
in-house testing-- very, very rigorous
testing schedules for all of the calibrator and
QC sets
that we make in -house. The testing is performed using
a combination of EQA materials as well as in-house
QC panels and the mean lot-to-lot
deviations for all of the analytes
included in a set are then examined
against EQA ALTM. If the plot on the
right showed fine, you would have seen some
sample data for steroid set 1 calibrators with those mean
deviations plotted for all 12 analytes. And for all 12 analytes for
those three different lots prepared, we've shown
deviations of less than 9.3% agai
nst EQA ALTM. So using the
MassTrak steroid sets enables your lab to maintain
those harmonization performance over time using different
manufactured lots of CALs and QCs. I'll hand it over to Dom. OK. Thank you, Isha. So Isha has described some of
the benefits of the MassTrak sets RUO, so I thought
what I'll do now is just take you through the
application of those sets with an in-house developed LC-MS
method for some corticosteroids and androgens. So this is, again-- is a
Waters workflow slide f
or LC-MS of steroid hormone analysis. And what I thought
I'll do is just pull out some of those products,
consumables, and informatics platforms to demonstrate
what we're actually using with this analysis. So we're using Oasis
SPE consumables. We've got the MassTrak sets, the
columns, the ACQUITY UPLC HSS column, the ACQUITY UPLC
I-Class PLUS system linked with the Xevo TQ-S
macro, and we're using the MassLynx and
TargetLynx informatics platforms and before I proceed
to the methodology we applie
d as a Waters solution, I thought
I'd just ask a quick question around sample preparation. So on my travels, I know
all different customers use different sample
preparation techniques for steroid analysis,
so I thought I just want to ask the audience,
what is the predominant sample preparation technique you
use for steroid analysis? Do you use solid
phase extraction, supported liquid extraction,
liquid-liquid extraction, or protein precipitation as
the main technique in your lab? There's four, s
o I'll give
you another five or 10 seconds to answer that before I
move on to the next slide. OK. So I'll move on to
the next slide now, just to see the results. And we have a
combination of two. So solid phase extraction, 50%
from people who've answered. And then 50% from the attendees
for protein precipitation. So that's interesting,
really, in regards to our workflow, which
we'll see on the next slide. So this is the Waters
system solution. As you can see in the
sample prep consumables, we ar
e using the Oasis SPE
PRiME HLB micro Elution plate. So we are using solid
phase extraction for this Waters solution. And that is in combination
with the MassTrak sets for that analysis. And we're able to automate this
on the Hamilton style liquid handling robot. And there are other automation
platforms available to do this, but our focus was on
Hamilton in this instance. And we were able to adopt that
sample preparation to perform the separation on the ACQUITY
UPLC I-Class+ FL system with separ
ation on the HSST three
column with T3 Vanguard and detection on the TQ-S
macro mass spectrometer. The runtime for this is 5.5
minutes injection to injection. Just some more details around
the actual sample preparation for this method. So we thaw and mix the samples
in centrifuge prior to use. We use 100 microliters of
sample for the analysis. We add internal
standard, and then we precipitate briefly with
addition of methanol to disrupt the protein
binding associated with steroids in serum. We d
ilute that with water. This helps with the binding of
the analytes to the solid phase extraction material. And we centrifuge that
to create a pellet at the bottom of the plates
or microcentrifuge tube. We take that supernatant
and add that to the plate after we've equilibrated
and conditioned the plate. So we load that
supernatant slowly onto the plate to ensure that
it binds well to the chemistry. We wash the wells with
high pH and low pH-- so 0.1% ammonia in 35% methanol,
followed by 0.1% form
ic acid in 35% methanol. So this is just
to help remove any of those potential
unknown interferences you may get from
sample to sample. And then we elute the analytes
with acetonitrile and methanol from the plates. So we only use 30 microliters. And then we
subsequently dilute that with 70 microliters of water. So this allows us to inject
the elute effectively directly from the extraction
onto the LC-MS system. And some more details around the
ACQUITY UPLC I-Class+ FL system set up. So we're usi
ng ammonium
acetate, 0.1% formic acid in water and methanol
for this method. So just before I
move on actually, this application
note is available. So some of the resources will
be highlighted at the end. And also within
the webinar itself, there are also links to
some of these resources. So we have a weak
washer, 40% methanol, strong wash in methanol, and
it's a fairly basic gradient setup from one to 3.5 minutes
and an increase of 20% B over that time frame. And as I said it's, 5.5
minutes inj
ection to injection. And some details around
the TQ-S micro setup. So we're looking at positive
ion mode and also actually negative ion mode. So that's actually not
necessarily fully correct there. So DHA sulfate is in
negative ion mode. Capillary voltage of 1 kilovolt,
150 degrees source temperature. 600 degrees desolvation
temperature. And just moving to
the bottom, unit mass resolution on the MS1. And also we just narrow the
resolution slightly in MS2. Sometimes we do this
just to help improv
e the selectivity of the
assay to a certain degree at the lower end of the
concentration range. And this can sometimes help
reduce some of the background noise associated with
this type of analysis, and then can improve
your signal to noise. And the MRM conditions
for this analysis-- we have obviously the
sets 2 and sets 3. There are different
analytes within that sets, but also the sets do share some
analytes like androstenedione and 17-OHP. I'm not going to talk all the
way through these trans
itions, but just to highlight,
we are looking at as I said previously,
a negative ion compound in DHEA sulfate. And the rest are in
positive ion mode. And each has an
internal standard either deuterated or
isotopically labeled 13C3 internal standard. And here we have some
chromatographic separation. I don't think the picture
there is at full resolution, but you can make out the
separation between some of those critical
steroid isomers that I previously discussed
on the HSS T3 column. So we have
the 11-
and 21-deoxycortisol and corticosterone towards
the top of the chromatogram, the 17-OHP and 21-OHP towards
the bottom of the chromatogram, and also testosterone
and epitestosterone. And also, there's some
high-level performance characteristics around
analytical sensitivity, and linearity. So, what I've done here,
is just highlighted what we achieved for
analytical sensitivity of the C1 calibrator
in MassTrak sets 2 and 3, that provide a signal
to noise greater than 10 to 1. As you can se
e, signal to
noise across both sets using this method on a
continuous macro is more than enough, in
terms of performance. I've also highlighted one of
the calibration lines for-- in this case, testosterone. And this spans all the three
orders of dynamic range. So it's demonstrating
that we do have linearity across that range. And that's the steroid within
both sets that has the widest dynamic range across the sets. Some performance characteristics
associated with precision. So, this was determin
ed by
extracting and quantifying replicates of a
four-level QC material over a four-day period. We actually had 48
replicates across each set. Repeatability was assessed
by analyzing four replicates at each QC level. What this demonstrates,
is actually, we've got really good
precision for total precision and repeatability in both
sets using this method. So, set 2, we're seeing
less than or equal 6.9%. In set 3, is slightly
more elevated at 9.9%, but still within
what you would expect in terms of
a good method. There are slightly
higher deviations for the 21-deoxycortisol. This is probably one of the more
challenging analytes associated with this analysis, but as I
said, it's still within 10%. Moving on to some
performance characteristics. So we looked at agreement
to some EQA material and also in house panels. And so there was only
material available for testosterone,
androstenedione 17-OHP, DHEAS, and cortisol. Unfortunately, at this moment
in time, as far as I'm aware, there's no EQA
for
11-deoxycortisol and 21-deoxycortisol. So we use some in house panels,
which were independently gravimetrically prepared
to assess the accuracy of these compounds. The data is obtained
over a four-day period, and compared to the
mass spectrometry mean for each sample. In the case of the
ones that we did have, EQA S4 and the mean percent
difference from the target was tabulated. The mean bias for these samples
was within 5% across the range. It demonstrates
that we've actually got good metro
logical
traceability, and a good enough routine
measurement procedure in place using this in-house
developed method. And Isha did cover some of the
benefits of the MassTrak sets workflow. But what I thought, is
just highlight the benefits of the overall workflow
we're employing for this RUO analysis. So we're looking at savings
of up to three months of skilled labor time spent
on in-house production, using the MassTrak sets. And these sets can be
used in up to 20 sample batches with each set. Th
ey can be used for-- depending on your
throughput, you could end up using one
set for every one or two months of analysis. The overall cost per sample for
the calibrator is around $0.30, or 20 pounds depending on
where you're coming from around the world. It's up to three hours
to first sample result. So, that's taking
into account you've already prepared the sets,
you perform the analysis of a typical batch between
48 to 96 samples, which takes about two hours. Then, the LC-MS/MS analysis
for t
he calibrators, followed by the first sample,
takes about three hours overall when you incorporate both
the sample preparation and the analysis time. There is one month
in-use stability, which Isha has already
highlighted with these CALs and QCs. We've demonstrated
that, when we do analyze the accuracy compared
to EQAs, it's within 10%. We have the benefits
of the MassLynx LIMS interface as well. The back end, where
we've got exchange of data flow between the
MassLynx and any LIMS systems you ha
ve in place. So that improves your
efficiency in the lab, and also you have the
injection-to-injection time as well. Which gives us 5.5 minutes
over the course of separation of seven steroid hormones. Very briefly, I
just want to cover some competitive comparisons. So this is based on
obtaining some materials from different vendors, these
are the calibration materials. We compared EQA
LC-MS method bias across the vendors using this
in-house clinical research method. And as you can see, both
vend
or A and Vendor B do show quite large
increases in variation from that target concentration. What this shows, is the
MassTrak serum sets, steroid serum sets
was shown to have the lowest level of deviation
from those target values. So in summary, just
like to highlight that the MassTrak steroid
serum calibrator and QC sets contain metrological,
traceable calibrators. That does aid in
compliance to 15189. Significant time
savings in resource can be made by using
these pre-made materials. And it's
demonstrated that these
materials are reproducible, and accurate across
manufacturing lots, and do cover the relevant
ranges for each steroid hormone that you may be interested in. Using an in-house
method we've developed, demonstrated excellent
performance of the MassTrak steroid serum sets, using the
Waters LC-MS workflow solution. So before we finish
up today, I'd just like to pass
back to Isha just to discuss some
additional resources. Thanks, Dom. So I wanted to leave the
audience with some
tools and resources that you
can leverage to find more information about the product. I've hyperlinked
the landing page for the MassTrak steroid
RUO sets family over here. Please feel free. Can easily be found on the
Waters website as well. This landing page serves
as your one stop shop solution to find out everything
you need about the product. So, on the product
web page itself, we have links to the app notes
the instructions for use. We also have the functionality
to place the order online d
irectly for these
products, and then we also have an
ordering guide as well, to help you sort through
all of the different SKUs that we offer. And then finally, I wanted
to make you all aware that we have a global
end of promotion on going for these sets. This is for web orders only. The discount code is CON15
and the different rates of discounts applied dependent
on the units that you purchase. All of the sets are included
in this promotion, so the steroids, and
the immuno sets. This promotion
is ongoing
till the end of Jan. So please feel free
to utilize this to stock up on your
sets, supplies, before the end of
the year (2023). And with that, I'd like to
finish off our presentation. Thank you so much to all of
our scientific calibrators who have helped
provide their feedback all throughout the development
process for this product. That has really helped
in designing the product as best as possible. Thank you so much
for our audience for listening in today. All of the information
is
hyperlinked into the presentation. And if you have
any more questions, please feel free to
visit our website, or you can get in touch
with your local Waters representative. Thank you. Thank you for the presentation. Now it's Q&A time. So let's see what came in. So we had a few questions. Probably the first one, I
guess, would be more for Isha. Are there any plans for
estrone and estradiol to be added in the sets? There are short-term
plans to add the estrogens and the
aldosterone into our sets.
But, we have yet to put down a
firm launch date on the ground. We will reach back
out to collaborators, such as yourself,
Laura, to help make sure that we've designed a
concentration ranges that are suitable for your lab. And we'll send out
routine notifications as we get closer to the
launch of those products. But, what we anticipate, is sort
of augmenting our existing sets with those additional analytes. Very good. So we have another
question, probably for Dom, or maybe Isha. How did you asses
s the accuracy? Did you spike the
analytes in the matrix at different concentrations? Did you purify
the plasma samples where you have
spiked the analytes through your purification
method presented here? So, two questions
in one, or three. I can perhaps try
and answer that one. A lot of the
accuracy assessments are based on the external
quality assurance programs. So we have the different
steroid hormone programs that we choose the
UK NEQAS scheme. But there are others available. Looking at, eff
ectively,
agreements to the mass spectrometry methods which
are seen as the gold standard in this instance. So effectively, we get
hold of those samples, and we measure them using the
in-house developed method. We extract the samples. The serum samples,
we extract those alongside our calibrators,
and QC materials, which are meteorologically
traceable. And that, in theory,
will provide us with a good assessment of
how our method is performing in respect to those mass
spectrometry measured samples
on the EQS scheme. So we take that mean
measurement from the EQA scheme, and compare to what the
calculated concentration is using our methodology. I believe that is the correct. Maybe the correct answer
in regards to the question. But if you do want to follow
up that, then feel free. Yeah, definitely. I mean, if there are
further questions. Well, first of
all, you can still type them in into
the Q&A box, or even ask us after we close
this session if you have additional questions coming in. We
have another one here. It's about, well, how we
manage the different orders of magnitude of
the concentrations between the steroids
analyzed in a single panel. Example-- DHEAS is a
hundred-fold higher than testosterone. So, question is--
yeah, how do we do without swamping
the detector or losing some analytes in the analysis? Yeah, so I can answer
that one as well, Godo. So, DHEA sulfate is-- perhaps, it's
easy, and difficult to analyze at the same time. It's that abundant
that you can detect it
, but then, because
it's so abundant, you can sometimes get issues
around detector saturation, peak broadening,
things like that. And so, we are able to
balance that in the analysis. Typically, DHEA sulfate
doesn't recover as well through the solid
phase extraction, compared to other steroids. You do get some loss in
recovery as a result of that, particularly with our method. So that does actually help in
regards to that overall problem that you're describing. So, we are able to
offset some of t
hat as well, with manipulation
of the detector conditions we have in the mass spectrometer. So we can almost reduce
the sensitivity we have, or even almost flatten
the sensitivity, we get from the mass spec to
reduce that saturation effect, if need be. So there are things
you can do in order to address that
particular problem. Typically, you want to reduce
the sensitivity of the DHEA sulfate, and obviously improve
the sensitivity for the others, such as testosterone. OK Thank you. We have two mo
re. What automation is the steroid
panel method compatible with? Sorry, what was that? Godo, can you repeat that? Yeah. Well, how can we automate,
or what automation is available for
the steroid panel? Yeah, yeah. So, as I said in
the presentation, we have this automated
on the Hamilton STAR, but effectively,
this is transferable across different
automation platforms. We previously had this
automated on a Tecan Freedom EVO 100 system. But since then, we've
gradually moved over to the STAR to per
form
our automation methods. Theoretically, Andrew+
Pipetting Robot could do this. Any sort of platform
could do the method is fairly compatible with
different systems, I'd say. It's a fairly routine
precipitation and SPE method. So, I don't envisage people
having too many problems if they chose to adopt it on
their automation platform. OK, Yeah I mean fair. And last one. So, what mass
spectrometers are most suited for the analysis
of the MassTrak set, so for the steroids
that you described? Her
e, you have the TQ-S micro. Well, what else
would you consider? Yeah, so we typically
advise the TQ-S micro for most of our steroid
hormone methods. But it would also work
on an TQ-XS, Xevo TQ-XS, or with Xevo TQ Absolute. So they have the wider
dynamic range detector, which can help compensate for
the increase in sensitivity we do observe on those
systems, because you get that extra order
of dynamic range when you do move to those. So, I'd say those
are the systems you'd be looking at to run th
ese. I wouldn't necessarily
advise the use of a Xevo TQD for these,
because some of them are a bit more challenging
and wouldn't necessarily be compatible, particularly
with the MassTrak sets or EVO for this analysis. So certainly Xevo
TQ-S micro is maybe a go-to, and then Xevo TQ-XS or
an Absolute as the next option. OK. Thank you. Thanks very much. Yeah, I'm not seeing any
new questions coming in. So, well, thanks again
for the presentations. It was very insightful. I thank as well, of course,
the attendants for being here. We will have another
session at the hour, but for that, we'll have
to log out of this one and log in to the next one. So hopefully, we'll see all of
you in the next presentation. Thank you. Buh-bye. Thank you. Thank you. Bye.
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