WAITE LIPID ANALYSIS SERVICE

Transcription

1 WAITE LIPID ANALYSIS SERVICE INFORMATION FOR CLIENTS 1 WAITE LIPID ANALYSIS SERVICE Information for Clients

2 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 Appendix 2: Gas Chromatography Method WAITE LIPID ANALYSIS SERVICE Information for Clients

3 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 kristina.hickson@adelaide.edu.au or phone (08) (Monday-Wednesday). Staff Liu Ge Dr Liu Ge finished his PhD at the University of Adelaide in 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 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): ) 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 3 WAITE LIPID ANALYSIS SERVICE Information for Clients

4 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 medium- 4 WAITE LIPID ANALYSIS SERVICE Information for Clients

5 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 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 5 WAITE LIPID ANALYSIS SERVICE Information for Clients

6 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 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. Total Fat Percentage (Image from 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. 6 WAITE LIPID ANALYSIS SERVICE Information for Clients

7 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 FP FX Fatty Acids in Phospholipids PP PX Fatty Acids in Triglycerides GP GX Fatty Acids in Cholesterol Esters CP CX Fatty Acids in Total Lipids TP TX Total Fat Percentage AP 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. 7 WAITE LIPID ANALYSIS SERVICE Information for Clients

8 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 kristina.hickson@adelaide.edu.au. 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 kristina.hickson@adelaide.edu.au Sample Delivery Address Please notify the laboratory of expected delivery time. Mail: Waite Lipid Analysis Service School of Agriculture Food and Wine University of Adelaide PMB 1 Glen Osmond, SA, 5064 Courier: 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 if you have specific queries. 8 WAITE LIPID ANALYSIS SERVICE Information for Clients

9 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 ed to the client at the completion of the job as an Excel spread sheet to allow data manipulation and as a secure.pdf file 9 WAITE LIPID ANALYSIS SERVICE Information for Clients

10 Appendix 1: List of Fatty Acids SCIENTIFIC NAME MOLECULAR NAME COMMON NAME Saturates tetradecanoic acid 14:0 myristic acid pentadecanoic acid 15:0 pentadecanoic acid hexadecanoic acid 16:0 palmitic acid octadecanoic acid 18:0 stearic acid eicosanoic acid 20:0 arachidic acid docsosanoic acid 22:0 behenic acid tetracosanoic acid 24:0 lignoceric acid Omega 9 Family 9-tetradecenoic acid 14:1 n5 myristoleic acid 9-hexadecenoic acid 16:1 n7 palmitoleic acid 11-octadecenoic acid 18:1 n7 vaccenic acid 9-octadenoic acid 18:1 n9 oleic acid 11-eicosenoic acid 20:1 n9 eicosenoic acid 5,8,11-eicosatrienoic acid 20:3 n9 mead acid 13-docosenoic acid 22:1 n9 erucic acid 15-tetracosanoic acid 24:1 n9 nervonic acid Omega 3 family 9,12,15-octadecatrienoic acid 18:3 n3 -linolenic acid (ALA) 6,9,12,15-octadecatetraenoic acid 18:4 n3 stearidonic acid 11,14,17-eicosatrienoic acid 20:3 n3 eicosatrienoic acid (ETA) 8,11,14,17-eicsoatetraenoic acid 20:4 n3 eicsoatetraenoic acid 5,8,11,14,17-eicosapentaenoic acid 20:5 n3 eicosapentaenoic acid (EPA) 7,10,13,16,19-docosapentaenoic acid 22:5 n3 docosapentaenoic acid (DPA) 4,7,10,13,16,19-docosahexaenoic acid 22:6 n3 docosahexaenoic acid (DHA) 6,9,12,15,18,21-tetracosahexaenoic acid 24:6 n3 tetracosahexaenoic acid Omega 6 family 9,12-octadecadienoic acid 18:2 n6 linoleic acid (LA) 6,9,12-octadecatrienoic acid 18:3 n6 -linolenic acid (GLA) 11,14-eicosadienoic acid 20:2 n6 eicosadienoic acid 8,11,14-eicosatrienoic acid 20:3 n6 homo- -linolenic acid (DGLA) 5,8,11,14-eicosatetraenoic acid 20:4 n6 arachidonic acid (AA) 13,16-docosadienoic acid 22:2 n6 docosadienoic acid 7,10,13,16-docosatetraenoic acid 22:4 n6 docosatetraenoic acid 4,7,10,13,16-docosapentaenoic acid 22:5 n6 n/a Plasmogen Linked Fatty Acids 1-enyl-hexadecenoic acid n/a plasmalogen 16:0 1-enyl-octadecenoic acid n/a plasmalogen 18:0 1-enyl-1,11-octadecadienoic acid n/a plasmalogen 18:1n7 1-enyl-1,9-octadecadienoic acid n/a plasmalogen 18:1n9 Unusual Fatty Acids 9-trans-hexadecenoic acid trans 16:1n7 palmitelaidic acid 9-trans-octadecenoic acid trans 18:1n9 elaidic acid 8-eicosaenoic acid 20:1n12 n/a 5-eicosaenoic acid 20:1n15 n/a 10 WAITE LIPID ANALYSIS SERVICE Information for Clients

11 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 250 o C and the detector temperature at 300 o CFAMEs 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. 11 WAITE LIPID ANALYSIS SERVICE Information for Clients