Reconstituting Lyophilised Controls

Reconstituting Lyophilised Controls

What is Lyophilisation?

Lyophilisation or ‘freeze drying’ is the process by which water is removed from a product after it is frozen and placed under a vacuum, allowing the ice to change directly from solid to vapor without passing through a liquid phase. The process consists of three separate processes:

  1. Freezing
  2. Primary Drying (Sublimation)
  3. Secondary Drying (Desorption)

There are many benefits to using a lyophilised control including; improved product shelf-life and enhanced stability of volatile analytes. For example, many lyophilised controls have a shelf life of up to four years from the date of manufacture resulting in a reduction of costly new lot validation studies. Furthermore, lyophilised controls can be aliquoted and refrozen to extend the working stability of the product.

Reconstituting Lyophilised QC Material

The process of reconstitution involves adding a specified volume of distilled water to lyophilised QC material. The water should completely dissolve the lyophilised contents, giving a liquid solution, which is ready for analysis.

Reconstitution is a straightforward process, but requires a high level of precision. Small errors can have serious implications to the reconstituted material:

  • If too much water is pipetted during reconstitution, the material will be heavily diluted and results will be lower than expected
  • If too little water is pipetted during reconstitution, the material will not be sufficiently diluted, and results will be higher than expected
  • If the correct volume of water is pipetted, but a small amount of water gets stuck in the pipette tip due to poor pipetting technique, results will be higher than expected

If a lyophilised control has been reconstituted incorrectly the contents of the vial will be wasted. It is therefore vitally important that controls are reconstituted with care.

Materials and Methods Required

The list of requirements for an accurate and consistent reconstitution technique is not extensive, but each requirement is vital. Labs should have:

  • Calibrated volumetric pipettes
  • Sterile, appropriately sized pipette tips
  • Distilled water, or other reconstitution fluid as specified
  • Technician with good pipetting technique
  • Lyophilised QC stored according to manufacturer’s specifications

How to Reconstitute Lyophilised QC Material

Each different lyophilised control may require slightly different preparation, always refer to the instructions for use before reconstituting control material. The below guide provides a general overview of the reconstitution process, using the Randox Human Assayed Chemistry Premium Plus control (HN1530) as an example

  1. Place the vial of lyophilised QC on a flat surface, carefully remove the lid and the rubber stopper making sure not to spill any material
  2. Using a calibrated pipette and sterilised pipette tip, add exactly 5ml of distilled water directly into the QC vial, ensuring no water is left in the pipette tip, or on the rim/side of the vial
  3. Place the rubber stopper and lid firmly back onto the QC vial, and leave to stand for 30 minutes
  4. After 30 minutes, gently invert the QC vial 10-15 times to ensure the contents is completely dissolved, making sure to avoid the formation of foam. It is important that you DO NOT SHAKE the vial. Alternatively place the vial on a roller for 30 minutes to ensure the contents is thoroughly mixed
  5. Once satisfied all material has been completely dissolved, proceed to use the QC product in accordance with the ‘Control’ section of the individual analyser application
  6. Once finished, refrigerate any unused material. It is good practice to label the vial with the date of reconstitution to prevent the use of material outside of the recommended stability period
  7. Prior to reusing lyophilised material, mix the contents thoroughly by gentle inversion, as highlighted in Step 4

Additional Considerations

It is important to remember that there may be slightly different reconstitution requirements for different QC material. For this reason, it is vital that the instructions provided on the QC Kit Inserts are closely followed.

Reconstituting lyophilised QC can be time-consuming. Therefore, Randox Acusera offer convenient 5ml distilled water serum diluent to assist laboratories with reconstitution of lyophilised controls. These user-friendly pour over vials streamline the reconstitution process and eliminate the risk of pipetting errors.

If you have any further questions regarding lyophilised controls or would like to contact us, please do so by emailing us at acusera@randox.com or use the contact us button provided.


Liver health: do you know your limits?

Are you taking part in Dry January? Giving up alcohol can do your insides a lot of good, and it’s great news for your liver in particular. It may be that after the festive period our liver needs a little bit of rest!

Did you know that alcohol consumption across the UK increases by a staggering 41 percent more than the annual monthly average in December? That’s more than anywhere else in the world.

The effects of alcohol on your health really depend on how much you drink and how often, but as the statistics show, more of us increase our uptake of alcohol over the festive period.

So how does this impact our body?

The results of over indulging vary from a hangover, a poor night’s sleep, to causing an irregular heartbeat, and in some cases, excessive alcohol intake can lead to liver damage. This can be a very serious condition, given the liver’s vital role in the body.

The liver plays a central role in all metabolic processes. In fat metabolism, it breaks down fats and produces energy. When we intake alcohol or drugs, the liver metabolizes the drug and detoxifies chemicals.  And it also makes proteins important for blood clotting and other functions.

Following these processes, the liver also secretes bile that ends up back in the intestines and helps the digestion of fats and oils, otherwise known as lipids.

As Randox Health Expert Dr. Gary Smyth explains:

“The liver is one of the most complex organs in the body and also one of the most important. Although it is very resilient, each time it has to filter alcohol some of its cells die.  The liver can develop new cells, but abuse over a prolonged period reduces its ability to regenerate, causing serious damage.”

It is not just heavy drinking over years that can cause liver disease – binge drinking is also a culprit and can lead to your liver becoming fatty and inflamed. The best advice is to drink in moderation. Simple tips like taking a glass of water in-between alcoholic drinks are key to staying hydrated.

Know your units;

  • According to drinkaware.co.uk, unit guidelines are now the same for men and women.
  • Both are advised not to regularly drink more than 14 units a week
  • This equates to 6 pints of 4% beer / 6 glasses of 13% wine / 14 glasses (25ml) of 40% spirits
  • But don’t save up your 14 units, it’s best to spread evenly across the week.
  • If you want to cut down the amount you’re drinking, a good way is to have several drink-free days each week.
  • If you’ve had a heavy drinking session, avoid alcohol for 48 hours.

What does one unit of alcohol look like?

One unit of alcohol is the amount of alcohol an average adult can process within one hour so that so that there’s no alcohol left in their bloodstream.

One unit of alcohol equates to:

  • 218ml of standard 4.5% cider
  • 76ml of standard 13% wine
  • 25ml of standard 40% whiskey
  • 250ml of standard 4% beer
  • 250ml of a standard 4% alcopop

How many units are in my drink?

  • Small glass white / rosé / red wine (125ml 12%) = 1.5 units
  • Standard glass white / rosé / red wine (175 ml 12%) = 2.1 units
  • Large glass white / red / rosé wine (250ml 12%) = 3 units
  • Pint of lager / beer / cider (5.2%) = 3 units
  • Bottle of lager / beer / cider (330ml 5%) = 1.7 units
  • Single small shot of spirits (25ml 40%) = 1 unit*

*taken from NHS Live Well Guidelines

Having your liver health checked after Christmas is a great way of tracking any changes that you may need to make to your lifestyle, for better or for worse – essential for helping you prevent liver disease and allowing you to take early action if it is diagnosed.

At Randox we offer a comprehensive menu of liver function tests to determine the health of your liver.  Provided by Randox to a wealth of hospitals, laboratories and research facilities across the globe, these tests are also directly available to you, the consumer, via our Randox Health clinics.

They include:

  • Alanine Aminotransferase (ALT) – an enzyme mainly found in the liver. Liver injury or disease will release ALT into the bloodstream, thus elevating serum ALT levels.  Moderately high or mildly elevated ALT levels can be associated with chronic liver disease, such as cirrhosis, which is scarring of the liver.

 

  • Aspartate Aminotransferase (AST) – an enzyme found predominantly in the heart, liver and skeletal muscles. Cell injury or disease will release AST into the bloodstream, thus elevating blood AST levels. Increased AST levels may be associated with hepatitis (inflammation of the liver), cirrhosis (scarring of the liver), or drug-induced liver injury.

 

  • Gamma-Glutamyltransferase (GGT) – an enzyme found mainly in the liver. Increased levels of GGT in the blood may indicate bile duct injury, hepatitis (inflammation of the liver), cirrhosis (scarring of the liver), liver necrosis (death of liver tissue), liver tumours or the use of drugs that are toxic to the liver.  A high GGT level is frequently associated with increased alcohol consumption, as this liver enzyme is involved in the breakdown and removal of alcohol from the body.

 

  • Glutamate Dehydrogenase (GLDH) – an enzyme located within the mitochondria (energy-producing machinery) of cells, particularly within liver tissue. Significant liver cell damage may cause release of GLDH into the bloodstream.  Toxic liver damage, liver cell necrosis (cell death) or hypoxic liver disease (where liver cells are deprived of oxygen) may cause an increase in GLDH. Measurement of GLDH in combination with other liver markers may help distinguish between different causes of liver dysfunction.

 

  • Bilirubin – a yellowish-brown pigment found in bile (a fluid produced in the liver that facilitates digestion in the intestine). Increased levels may be associated with liver or bile duct blockage (eg due to gallstones), hepatitis (inflammation of the liver), cirrhosis (scarring of the liver), trauma to the liver, a drug reaction, long-term alcohol abuse or rare inherited disorders (eg Dubin-Johnson syndrome which is characterised by mild jaundice).

 

  • Albumin – produced by the liver, albumin is the most abundant protein in the blood. Albumin plays in important role in maintaining blood pressure and transporting a wide variety of small molecules, such as hormones, vitamins and drugs, throughout the body. Various conditions are associated with decreased albumin levels, including kidney and liver diseases.

 

  • Copper – an essential mineral that plays a part in many enzyme systems within the body. Excess or deficiency of copper is very rare, however raised copper levels may be caused by chronic liver disease or acute hepatitis (inflammation of the liver).

And when used in conjunction with the wide variety of other tests available within the world’s most comprehensive and personalised health testing menu, you can obtain an understanding of your full body health like never before.

That’s why we don’t test in isolation, which can give a patchy representation of your health and can fail to pick up on related symptoms elsewhere in the body. We test up to 350 tests across 25 areas of your health – giving you the power to take your health into your own hands.

Contact the Randox Health team today to determine the health of your liver, and of your body.

Call 0800 2545 130 or click here.

 

 


Protecting Pets from the Threat of Mycotoxins

Pet Food companies worldwide are working towards constantly improving and maximising the quality of their product. The problematic topic of mycotoxin contamination in pet feed is quickly becoming a major cause for concern. This is due to the risk they pose for animal health and with the  increasing prevalence of mycotoxins globally the focus is on pet food companies to meet EU and FDA regulations and maximise the quality of their product.

What are Mycotoxins?

Mycotoxins are naturally occurring metabolites that are produced by certain moulds and with the ability to develop and grow on a variety of crops they can affect large amounts of feed and increasingly, pet food. If a sample tests positive even for low levels of contamination the toxins are still strong enough to cause illness in animals, and if low levels are consumed over a long period of time this can result in chronic illnesses including; cancer, organ damage and neurological disorders.

The main mycotoxins of concern in pet food are;

  • Deoxynivalenol (DON)
  • Fumonisins (FUM)
  • Zearalenone (ZEN)
  • Aflatoxins
  • Ochratoxin
  • T-2 Toxin

Contamination can occur in any country around the world and at any stage of production. Herein lies the issue of how to prevent mycotoxin pollution, to tackle the issue head on and work towards a mycotoxin free product is the joint responsibility of feed producers, supply chain partners and quality control laboratories ensuring the complete safety of the product.

How can you tell if an animal has ingested pet food contaminated with mycotoxins?

In terms of animal health, mycotoxins can cause a variety of problems. Severity and symptoms can vary from animal to animal but general symptoms include; hyperactivity, vomiting, high temperature and loss of coordination. If you suspect your pet has been affected by mycotoxins you must bring them to the vet for immediate treatment.

The European Union currently regulate all the mycotoxins listed above and are subject to maximum or recommended residue limits. In the US, FDA regulations are limited to aflatoxins, DON and fumonisins, see table below for FDA regulations. If mycotoxin levels in feed fail to meet FDA standards, mass amounts of feed may need to be destroyed as grain producers are prohibited from mixing contaminated feed with clean feed to reduce the mycotoxin levels.

PetsMycotoxinCommodityLevel
Immature AnimalsAflatoxinsCorn/ peanut/ other ingredients20 ppb
Adult PetsAflatoxinsCorn/ peanut/ cottonseed meal/ other ingredients20 ppb
DONGrain/ grain byproducts, not to exceed 40% of diet5 ppm
FumonisinsCorn/ corn byproducts, not to exceed 50% of the diet10 ppm

How do we tackle the problem?

Safe, reliable screening solutions for different variations of mycotoxins are available that can ensure only mycotoxin free feed is produced. Randox Food Diagnostics have created mycotoxin screening platforms as a response to increased levels of mycotoxins being found in feed globally.

The platforms use patented Biochip Array Technology (BAT) so pet food producers can test for multiple toxins from a single sample. Randox Food Diagnostics have a range of mycotoxin Biochip Arrays available with customised arrays available to suit the specific screening needs of certain producers. Each Biochip format uses a straightforward extraction process with a 50µl sample of feed, available tests include; Fumonisins, Ochratoxin A, Aflatoxin G1/G2, Aflatoxin B1, Paxiline, Ergot Alkaloids, Diacetoxyscirpenol, Deoxynivalenol, T2 Toxin and Zearalenone.

For more information on mycotoxin screening with Randox Food Diagnostics contact info@randoxfooddiagnostics.com

 

 

 


Obesity: the disease, the problems, and the power of prevention

Earlier this year the World Obesity Federation made the stark statement that: “The early diagnosis and treatment of childhood obesity could be considered similar to vaccination.”

Essentially, they want to see this condition treated in the same way as chicken pox, measles and mumps: tackled – in the hope of eradication – by a strategic approach founded on proactive policies and early prevention.

Obesity in children and adolescents has risen tenfold in the last 40 years, according to a recent study by The Lancet. In Britain, one in ten young people aged between 5 and 19 is obese. Worryingly, the prevalence of obesity is actually higher in younger children than older ones.

The WHO first called for obesity to be understood as a disease in 1948, but back then it wasn’t even considered a risk factor for cardiovascular disease. In 1997 the WHO held a special conference on obesity and stated that: “the global epidemic projections for the next decade are so serious that public health action is urgently required.”

Then it was alarmed that the prevalence of men with a BMI greater than 30 was 15% and 16.5% in women. To think that it has now risen dramatically to 67% for men and 57% for women, highlights just how serious a problem obesity poses to society.

The calls for more countries to officially recognise it as a disease is based on the position that obesity meets the definition of a chronic, relapsing, progressive disease that causes organ damage.

Women and men who are obese are 12.5 and 5.2 times (respectively) more likely to develop diabetes than people who are a healthy weight. 90% of people with Type 2 diabetes are obese.

People with diabetes are then at a greater risk of a range of chronic health conditions including cardiovascular disease, blindness, amputation, kidney disease and depression than people without diabetes. Diabetes leads to a two-fold excess risk for cardiovascular disease, and diabetic retinopathy is the leading cause of preventable sight loss among people of working age in England and Wales.  About one in twenty people have diabetes, yet people with diabetes account for one quarter to one third of hospital admissions for cardiovascular disease.

According to Government figures released this year, people who have Type 2 diabetes are 28.4% more likely to die early than their peers.

Getting in front of this wave of diabetes will not only bring down the numbers of people affected but also see a positive impact on the numbers of obese people. As with all conditions – the earlier they are identified, the better. To do this, new methods of diagnosis are being developed.

A radical new test for a protein found in our blood called adiponectin can identify pre-diabetes. This is a game-changing diagnostic tool that empowers people with the knowledge that they are at risk, but may be able to avoid it through relatively simple lifestyle changes.

The adiponectin test is available from Randox – both for clinical use and also through our Randox Health clinics.  We have developed the most comprehensive health checks available on the market. These are so sensitive that in a range of conditions including diabetes we are able to identify signs of pre-illness.  This enables clients to make often simple changes to stay healthy.

We know that prevention works. The NHS carried out a study in 2016 which revealed an average 26% reduction in new cases of Type 2 diabetes in those participating in a diabetes prevention programme, compared with usual care.

 

To find out more, click here.

For further information please email: randoxpr@randox.com


Diagnosing diabetes with the RX series

Diabetes is a lifelong condition that causes a person’s blood sugar level to become too high.

If you have diabetes, your body is unable to break down glucose into energy. This is because there’s either not enough insulin to move the glucose, or the insulin produced doesn’t work properly [1] which can lead to serious health complications.

The RX series range of analysers have one of the largest test menus available on the market which includes an extensive diabetes testing panel. Tests within the RX series diabetes panel allow for Diagnosis, Monitoring and Risk Assessment of Diabetes.

Adiponectin

An adiponectin test system is a device intended for the quantitative in vitro determination of adiponectin concentration in human serum or plasma.

Adiponectin is a protein hormone, produced and secreted by fat cells (adipocytes), which is normally found in reasonably high concentrations within the blood. Adiponectin regulates the metabolism of lipids and glucose and influences the body’s response to insulin and inflammation.

Adiponectin levels are inversely correlated with abdominal visceral fat (AVF) levels, which have proven to be a strong predictor of several pathologies including metabolic syndrome, type 2 diabetes mellitus (T2DM), cancers and cardiovascular disease (CVD). It is widely recognised that people who are overweight are at a higher risk of developing T2DM, however measure waist circumference and Body Mass Index (BMI) are not enough. As such adiponectin levels are a much more reliable indicator of at-risk patients.

A number of key publications have advocated the testing of adiponectin in clinical settings and concluded that higher adiponectin levels are associated with a lower risk of T2DM across diverse populations.[2]

Fructosamine

A fructosamine test system is a device intended for the quantitative in vitro determination of glycated protein (fructosamine) concentration in human serum or plasma.

Fructosamine is a mid-term indicator of diabetic control as it can provide information on a person’s averge blood glucose levels over the preceding 14-21 days.

 Due to the shorter time span of fructosamine, it is often used to evaluate the effectiveness of medication changes and to monitor the treatment of gestational diabetes.

HbA1c

A Haemoglobin A1c test system is a device intended for the quantitative in vitro determination of Haemoglobin A1c concentration in whole blood.

 In a diabetic patient, where blood glucose levels are abnormally elevated, the level of HbA1c also increases proportionally to the level of glucose in the blood and has been widely accepted as an indicator of the mean daily blood glucose concentration over the preceding 6-8 weeks. It is therefore, a long term indicator of diabetic control.

Read our poster on Randox’s development of a new latex enhanced immunoturbidimetric assay for the rapid direct measurement of glycated haemoglobin (HbA1c) applicable to RX series analysers by clicking here. 

 Diagnosing diabetes with the RX series

The RX series range of clinical chemistry analysers have many benefits when testing patients for diabetes. With analysers ranging from the RX misano semi-automated analyser to the RX modena which can perform up to 1200 tests per hour the RX series analysers offer a suitable platform for your laboratory, ensuring results are received in a time efficient manner. Windows based software and easily recognisable icons ensure that the RX series analysers are easy to use and allows for an enhanced laboratory productivity. Laboratory cost savings can also be achieved with a low water consumption available on each RX series analyser.

Other RX series analyser features include:

Diabetes Test Menu:

Consolidate your testing with a comprehensive diabetes testing panel available on the RX series analysers. A large number of tests can be carried out on one platform, including direct HbA1c testing, providing consolidation opportunities and real cost savings.

Accurate Testing:

High quality results are achieved first time, every time. This saves operator time and avoids unnecessary additional costs of repeat testing and reduces the possibility of patient misdiagnosis.

Unrivalled performance:

Built in inventory management system automatically calculates remaining reagent volume and the number of tests available. Superior performance means minimal downtime and swift reporting of results.

If you would like more information in relation to the RX series testing capabilities please contact us by emailing: theRXseries@randox.com


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