WAITE LIPID ANALYSIS SERVICE
INFORMATION FOR CLIENTS
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WAITE LIPID ANALYSIS SERVICE – Information for Clients
Contents
Waite Lipid Analysis Service .............................................................................................................................................. 3
Contact .............................................................................................................................................................................. 3
Staff ................................................................................................................................................................................... 3
Liu Ge ............................................................................................................................................................................ 3
Kristina Hickson ............................................................................................................................................................. 3
Services offered at Waite Lipid Analysis Service ............................................................................................................... 3
What sort of analysis do you need?.................................................................................................................................. 4
Fatty Acids in Total Lipids.............................................................................................................................................. 4
Phospholipids ................................................................................................................................................................ 4
Triglycerides .................................................................................................................................................................. 5
Free Fatty Acids ............................................................................................................................................................. 5
Cholesterol Esters ......................................................................................................................................................... 6
Total Fat Percentage ..................................................................................................................................................... 6
Waite Lipid Analysis Service Routine Methods ................................................................................................................. 7
Price of Analysis ................................................................................................................................................................ 7
Quotes ............................................................................................................................................................................... 7
Billing ................................................................................................................................................................................. 7
University of Adelaide clients ....................................................................................................................................... 8
Non University of Adelaide clients ................................................................................................................................ 8
Booking-in Samples for Analysis ....................................................................................................................................... 8
Sample Delivery Address................................................................................................................................................... 8
Sample Presentation ......................................................................................................................................................... 8
Biological Material ............................................................................................................................................................ 9
Sample Containers ............................................................................................................................................................ 9
Sample Numbering ........................................................................................................................................................... 9
Report Format ................................................................................................................................................................... 9
Appendix 1: List of Fatty Acids ........................................................................................................................................ 10
Appendix 2: Gas Chromatography Method .................................................................................................................... 11
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WAITE LIPID ANALYSIS SERVICE – Information for Clients
Waite Lipid Analysis Service
Professor Robert Gibson’s Fatty Acid Laboratory, situated on the Waite Campus of the University of Adelaide has
underpinned his world class research for many years. Waite Lipid Analysis Service (WLAS) has been set up as a
business group under Adelaide Research and Innovation (ARI), the University’s commercial arm, to provide internal
and external clients with the same high quality service for all your fatty acid analysis needs.
Contact
For all enquiries please contact Kristina Hickson via email [email protected] or phone (08) 8313 6162
(Monday-Wednesday).
Staff
Liu Ge
Dr Liu Ge finished his PhD at the University of Adelaide in 2013. He has considerable experience in fatty acid research
using sophisticated techniques such as gas and liquid chromatography (GC, HPLC), and mass spectrometry
(LCMSMS). His research at the University of Adelaide led to the development of a dried blood spot technique for
determining the omega-3 fatty acid status from capillary blood which has been patented. He has also worked for
large pharmaceutical companies developing and optimising manufacturing processes for various pharmaceutical
products.
Kristina Hickson
Kristina has a Bachelor of Applied Science Degree with Honours. Additionally she has over eight years’ experience
working in a NATA accredited environmental commercial laboratory, specialising in instrumentation including gas
and liquid chromatography (GC, HPLC) and liquid chromatography mass spectrometry (LCMS and LCMSMS).
Services offered at Waite Lipid Analysis Service
WLAS prides itself in providing accurate and reproducible analysis of fat content and fatty acid composition in a wide
range of sample types including human biological fluids such as whole blood plasma, red blood cells and breast milk,
animal and plant tissue, oils and spreads, foodstuffs, animal feeds, bacteria, fungi and yeast. Our methods can
generally be adapted to extract lipid from most sample types so, if your type of sample is not listed, please contact
us to see if it can be done.
A new and exciting development at WLAS is the PUFAcoat dried blood spot (DBS). The PUFAcoat paper, developed
by WLAS researcher Dr Liu Ge and Prof Gibson (Liu, G., Mühlhäusler, B. S., Gibson, R.A. 2014. A method for long term
stabilisation of long chain polyunsaturated fatty acids in dried blood spots and its clinical application. Prostaglandins,
Leukotrienes and Essential Fatty Acids. 91(6): 251-260) is a special filter paper which stabilises fatty acids in biological
samples at room temperature for periods of several weeks at room temperature. We have been using the PUFAcoat
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WAITE LIPID ANALYSIS SERVICE – Information for Clients
cards in our clinical trials routinely since 2013 with huge success. From 2015, we are offering WLAS clients the option
to purchase PUFAcoat cards for their samples – which offers significant convenience and savings in the costs of
sample storage, transport and fatty acid analysis compared to traditional methods. If you are interested in finding
out more, please contact us.
WLAS offers a number of options for sample analysis. We can accurately determine the total amount of lipid (fat) in
a sample as well as the amount of the separate lipid classes – phospholipids, free fatty acids, triglycerides and
cholesterol esters. We can then assess the fatty acid composition of the total sample and/or one or more of these
lipid classes. Results can be reported either as a percentage of the total lipid or in absolute amounts.
Appendix 1 provides a list of the specific fatty acids we generally include in our reports. If you are interested in a
fatty acid that does not appear on the list, please let us know – since we have the capacity to assess many others.
What sort of analysis do you need?
Different lipid fractions are analysed depending on the sample type, the convention in that particular field (what do
other researchers report in their publications?) and what questions you want to answer. To assist you in deciding
which analysis you require, we have provided some basic information on the different lipid classes below. However,
we are able to work with you to determine the most appropriate analysis for the sample type and your biological
question:
Fatty Acids in Total Lipids
The analysis of the fatty acid composition of the total lipids is routine for samples that are meant to be eaten (by
animals or humans), since it provides information on the total amount of different fatty acids in that material, and
therefore, the amount of each fatty acid that the person (or animal) would obtain from consumption. Common
sample types that we process for total lipids include human food (e.g. meat, seafood, eggs, processed foods, grains,
pulses and nuts) and beverages, prey items (e.g. insects, snails, frogs, fish), animal feeds (e.g. formulated diets for
aquaculture, pets, production animals) and edible pasture plants (e.g. grass, feed grains and lucerne). Total lipid
analysis can also be used with complex samples such as whole blood or dried blood spots as an ‘overview’ to see
how variable the fatty acid composition can be between individuals.
In our new PUFAcoat DBS method, we also assess total fatty acids in a drop of whole blood, plasma or breast milk
and have shown that the results are highly correlated with the values obtained by measuring fatty acid status of the
phospholipids. This is the simplest and therefore cheapest type of analysis, since we don’t need to separate the
sample into its different lipid fractions. A total fat percentage analysis is also available on request.
Phospholipids
Phospholipids are a class of lipids that are the major component of cell membranes. They are composed of a polar
head group and two fatty acids that may, or may not, be the same as each other (in the Figure below they are the
same). Since it takes time for fatty acids consumed in the diet to be incorporated into the cell membranes, assessing
the fatty acid composition of the phospholipid fraction of blood or plasma provides a good indication of medium4
WAITE LIPID ANALYSIS SERVICE – Information for Clients
long term lipid status. For this reason, the fatty acid composition of phospholipids is generally used for the
assessment of patient fatty acid status clinically and in clinical trials involving fatty acid interventions. We routinely
measure the phospholipid fatty acid composition of plasma, red blood cells and tissues from animal trials.
(Image from http://www.uic.edu/classes/bios/bios100/lectf03am/phospholipid.jpg)
Triglycerides
Triglycerides are made up of a glycerol backbone attached to 3 fatty acids (that may or may not be the same as each
other). Triglycerides are the main form of lipid in animal fats and vegetable oils and are a common type of fat that
accounts for about 95 per cent of all dietary lipids.
The triglyceride component in blood plasma is generally thought to be more reflective of recent fat consumption (i.e.
hours-days before a sample is collected) and therefore isn’t recommended if you are interested in knowing about
long-term status. For example, if a patient who normally has very low levels of omega-3 fats in the diet eats salmon a
few hours before a sample is collected, the level of omega-3 fats in the triglyceride component would be quite high
(and therefore not give you an accurate indication of their ‘basal’ status). Once digested, triglycerides circulate in the
bloodstream to be used as an energy source, with excess being stored in fat depots in the body.
(Image from nuchiro.wordpress.com)
Free Fatty Acids
Free Fatty acids are carboxylic acids with unbranched carbon tails that, when bound, form phospholipids and
triglycerides. They are usually found in low concentrations/proportions in tissues because they are more reactive
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WAITE LIPID ANALYSIS SERVICE – Information for Clients
than phospholipids and triglycerides. Higher amounts of free fatty acids are found in fats and oils that are rancid,
oxidised or otherwise degraded.
While free fatty acids make up a relatively small percentage of the total lipid fraction in blood and most tissues,
recent studies have shown that this lipid fraction may play a more important role in the biological activity of fatty
acids – and we are currently working on new methods to improve the efficiency of isolation.
(Image from https://hashtagbcbg.files.wordpress.com/2014/04/fatty-acid-structure1.png)
Cholesterol Esters
Cholesterol esters are a dietary lipid, and derived from cholesterol and although both are considered sterols (a
subclass of fat) cholesterol esters are formed when the body produces enzymes to produce a metabolic reaction
with the cholesterol itself. The ester bond is formed between the carboxylate group of a fatty acid and the hydroxyl
group of cholesterol. Cholesterol esters have a lower solubility in water due to their increased hydrophobicity.
(Image from http://lipidlibrary.aocs.org/Primer/content.cfm?ItemNumber=39303)
Total Fat Percentage
We routinely measure the total amount of fat in samples and report it as the fat percentage (or g of fat per 100g of
sample). This information is provided to all clients who have fatty acids assessed, but it is also possible to have as a
stand-alone analysis. The total fat percentage of a sample can be useful if you are interested in the total amount of
lipid in a particular sample, but not in what fatty acids are present.
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WAITE LIPID ANALYSIS SERVICE – Information for Clients
Waite Lipid Analysis Service Routine Methods
Please refer to the table below for our most routine testing conducted. We provide analysis as either a qualitative
measurement as a percentage (%) result, or quantitatively as mg/100g.
Type of Analysis
Qualitative Code
Quantitative Code
Free Fatty Acids
Fatty Acids in Phospholipids
Fatty Acids in Triglycerides
Fatty Acids in Cholesterol Esters
Fatty Acids in Total Lipids
Total Fat Percentage
FP
PP
GP
CP
TP
AP
FX
PX
GX
CX
TX
AX
Speak with the laboratory manager if you have fatty acids requirements other than those above. We may be able
to help you.
Price of Analysis
There is a range of prices depending on the category of client and the type of samples, so contact the laboratory for
the necessary information. Sometimes research samples require extra preparation before analysis; the cost will be
passed on to the client, and will be specified in the quote.
Quotes
Obligation-free quotes are provided to all clients prior to undertaking analyses, once we have discussed your
requirements and agreed on the type of analyses that you would like. Quotes are beneficial for the clients as they
provide an upfront cost for the analysis. Prices are fixed for a length of time so ensure potential prices increases are
factored in for long-term projects. Quotes also allow you to raise purchase orders to ensure funding is reserved
within the client organisation.
Billing
For long term projects, invoices are generally generated quarterly for jobs completed in the previous three months,
and are issued in the first week of March, June, September and December. This enables bills to be settled before the
end of the financial year or before grants terminate at the end of December.
Shorter term projects will be billed on completion of the analyses and submission of the report.
If you need to have a different billing schedule, please talk to us.
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WAITE LIPID ANALYSIS SERVICE – Information for Clients
University of Adelaide clients
If debit codes commence “UNIAD”, the transaction will be a simple Journal Entry. You will receive a quarterly log of
batch details. All you have to do is nominate a debit code on the attached authority, sign it and return it to the
laboratory.
Non University of Adelaide clients
External clients will be required to sign the ‘Waite Lipid Analysis Service Agreement’ from ARI; for more information
please contact Kristina Hickson at [email protected]. External clients will receive a quarterly log of
batch details with an invoice from ARI attached. Payment is by cheque or direct debit to ARI. Direct debit
instructions are included on the Tax Invoice. Please provide the mailing address and ABN of your institution when
booking in samples. To ensure funds are available to pay invoices, raise a purchase order prior to sending samples.
Booking-in Samples for Analysis
The Waite Lipid Analysis Service booking form acts as the contract between Waite Lipid Analysis Service and the
client. There are four sheets within the Excel based form SAMPLES LIST, CLIENT DETAILS & CHECKLIST, BATCH
DETAILS and WLAS TERMS & CONDITIONS.
Internal clients are required to read the T&C’s outlined in the document and sign the form with sample submission.
External Clients are required to not only fill out the booking form but sign into an agreement with ARI for invoicing
purposes. For more information please contact Kristina Hickson at [email protected]
Sample Delivery Address
Please notify the laboratory of expected delivery time.
Mail:
Courier:
Waite Lipid Analysis Service
School of Agriculture Food and Wine
University of Adelaide
PMB 1
Glen Osmond, SA, 5064
Waite Lipid Analysis Service
University of Adelaide
c/o Level 1, Receiving Bay
Wine Innovation Central Building
Cnr Paratoo Rd & Hartley Grove,
Urrbrae, SA, 5064
Hand Deliver:
Waite Analytical Services
Room GS20B
Main Waite Building
School of Agriculture Food and Wine
University of Adelaide
Entrance 1
Waite Road, Urrbrae, SA, 5064
Sample Presentation
No analysis can be better than the sample you provide, so care must be taken in sample collection, identification,
labelling and handling. Please email if you have specific queries.
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WAITE LIPID ANALYSIS SERVICE – Information for Clients
Biological Material
Hazardous material must be identified on the booking form.
Blood and tissue samples etc. must be kept cool from sampling to delivery, apart from the PUFAcoat cards which can
be at room temperature.
Sample Containers
Samples must be presented in clean sealed containers. Containers small enough to fit into tube holders to keep them
in order for extraction is preferred but containers in a sealable bag are okay for larger samples as long as they are
sequentially numbered.
Sample Numbering
A unique number that identifies both client and sample e.g. your initials or trial code + number (preferably sequential)
must be clearly written on the outside of the container. These ID’s must match the sample list provided in the booking
forms. Samples must be presented in the order of the list.
Report Format
Reports provided will list all of the recorded fatty acids in either percentage or in mg/g. There is a standard report
format that lists all of the main fatty acids along with a table summarising the data. If you require your data in a
specific format we can accommodate this but if a new template or macro needs to be setup extra charges may be
incurred. Results will be emailed to the client at the completion of the job as an Excel spread sheet to allow data
manipulation and as a secure .pdf file
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WAITE LIPID ANALYSIS SERVICE – Information for Clients
Appendix 1: List of Fatty Acids
SCIENTIFIC NAME
MOLECULAR NAME
COMMON NAME
Saturates
tetradecanoic acid
pentadecanoic acid
hexadecanoic acid
octadecanoic acid
eicosanoic acid
docsosanoic acid
tetracosanoic acid
14:0
15:0
16:0
18:0
20:0
22:0
24:0
myristic acid
pentadecanoic acid
palmitic acid
stearic acid
arachidic acid
behenic acid
lignoceric acid
14:1 n5
16:1 n7
18:1 n7
18:1 n9
20:1 n9
20:3 n9
22:1 n9
24:1 n9
myristoleic acid
palmitoleic acid
vaccenic acid
oleic acid
eicosenoic acid
mead acid
erucic acid
nervonic acid
18:3 n3
18:4 n3
20:3 n3
20:4 n3
20:5 n3
22:5 n3
22:6 n3
24:6 n3
-linolenic acid (ALA)
9,12-octadecadienoic acid
6,9,12-octadecatrienoic acid
11,14-eicosadienoic acid
18:2 n6
18:3 n6
20:2 n6
linoleic acid (LA)
-linolenic acid (GLA)
eicosadienoic acid
8,11,14-eicosatrienoic acid
5,8,11,14-eicosatetraenoic acid
13,16-docosadienoic acid
7,10,13,16-docosatetraenoic acid
4,7,10,13,16-docosapentaenoic acid
20:3 n6
20:4 n6
22:2 n6
22:4 n6
22:5 n6
homo--linolenic acid (DGLA)
arachidonic acid (AA)
docosadienoic acid
docosatetraenoic acid
n/a
n/a
n/a
n/a
n/a
plasmalogen 16:0
plasmalogen 18:0
plasmalogen 18:1n7
plasmalogen 18:1n9
trans 16:1n7
trans 18:1n9
20:1n12
20:1n15
palmitelaidic acid
elaidic acid
n/a
n/a
Omega 9 Family
9-tetradecenoic acid
9-hexadecenoic acid
11-octadecenoic acid
9-octadenoic acid
11-eicosenoic acid
5,8,11-eicosatrienoic acid
13-docosenoic acid
15-tetracosanoic acid
Omega 3 family
9,12,15-octadecatrienoic acid
6,9,12,15-octadecatetraenoic acid
11,14,17-eicosatrienoic acid
8,11,14,17-eicsoatetraenoic acid
5,8,11,14,17-eicosapentaenoic acid
7,10,13,16,19-docosapentaenoic acid
4,7,10,13,16,19-docosahexaenoic acid
6,9,12,15,18,21-tetracosahexaenoic acid
stearidonic acid
eicosatrienoic acid (ETA)
eicsoatetraenoic acid
eicosapentaenoic acid (EPA)
docosapentaenoic acid (DPA)
docosahexaenoic acid (DHA)
tetracosahexaenoic acid
Omega 6 family
Plasmogen Linked Fatty Acids
1-enyl-hexadecenoic acid
1-enyl-octadecenoic acid
1-enyl-1,11-octadecadienoic acid
1-enyl-1,9-octadecadienoic acid
Unusual Fatty Acids
9-trans-hexadecenoic acid
9-trans-octadecenoic acid
8-eicosaenoic acid
5-eicosaenoic acid
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WAITE LIPID ANALYSIS SERVICE – Information for Clients
Appendix 2: Gas Chromatography Method
FAMEs are separated and measured on a Hewlett-Packard 6890 gas chromatograph (CG) equipped with a
50m capillary column (0.32mm internal diameter SGE, Victoria) coated with 70% cyanopropyl
polysilphenylene-siloxane (BPX-70) (0.25µm film thickness) which is fitted with a flame ionization detector.
We use helium as the carrier gas and the split-ratio is set at 20:1. The injector temperature is usually set at
250oC and the detector temperature at 300oCFAMEs are identified based on the retention time of standards
using the Chemstation software and a normalised percentage calculated based on response factors. For
samples with an internal standard a further calculation based on how much standard was added was
performed to convert the data to a concentration value.
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WAITE LIPID ANALYSIS SERVICE – Information for Clients