Nicotinamide in Veterinary Practice

What is Nicotinamide?

Nicotinamide (also known as Niacinamide) is a form of vitamin B3, which is necessary for the body’s processing of lipids, amino acids, and glucose. A deficiency can have an impact on appetite as well as red blood cell production. Vitamin B3 is an essential vitamin for cats, because they cannot produce it in their bodies and must consume it).

Nicotinamide is given as a nutritional supplement to cats to control phosphorus levels in their blood. This is important for cats with chronic kidney disease as their kidneys cannot excrete phosphorus and this accumulates in their blood, leading to hyperphosphatemia.

Additionally, nicotinamide can also be used in conjunction with other medications to treat inflammatory skin conditions in dogs, such as lupoid or pemphigoid (1).

How is it given?

Nicotinamide is administered orally as a tablet or capsule. It can also be combined with water to form a liquid or administered with or without food (2).

Side effects 

The most common side effects are loss of appetite, vomiting, diarrhoea, and sleepiness. Seizures and an increase in liver enzymes are two serious side effects. However, because this supplement is used in conjunction with tetracycline, side effects from niacinamide may not be solely attributable to it (1).

Our work

We offer analysis of nicotinamide in feline plasma. Contact us here for more information

References

  1. VCA Animal Hospital. Niacinamide. (Internet) (Cited on 2021 Nov 28) Available from: https://vcahospitals.com/know-your-pet/niacinamide

Pesticide Residues in Food & on Plants

The use of pesticides 

The agriculture industry faces pressure to protect yields and crops to ensure they can keep up with the high demand and produce large amounts of food. To do this they need to use pesticides (fig.1).

pesticide residues in plant material

Fig 1. Application of pesticides in plants

Pesticides are used to protect agricultural production and planting from insects, weeds, fungi and other pests. Hence, increasing yields and food production. However, pesticide residues can be toxic to humans and cause acute and chronic health problems.

To prevent these health effects, a few countries signed the 2001 Stockholm Convention, an international treaty aimed at eliminating or restricting the production and use of persistent organic pollutants, such as dichlorodiphenyltrichloroethane (DDT) and lindane, which can persist in soil and water for years (1).

A pesticide’s toxicity is determined by its function and other factors, such as the dose to which the person has been exposed and the route by which the exposure occurred (inhalation, swallowing, etc). Today, none of the pesticides authorized for use on food crops are genotoxic, however if exposed to levels above a certain limit, it can cause serious long-term health effects such as cancer (1).

Who’s at risk?

People who work in agriculture are most at risk, as they are in direct contact with pesticides. The general population have less exposure through water and food.

Our work

It is important to be aware that pesticides can have serious consequences if used in quantities above the safe limit set by WHO. We offer analysis of pesticide residues in plant material. To send your samples for analysis contact us here. 

References

  1. World Health Organization. Pesticides Residues in Food. (Internet) (Updated 2018 Feb; cited on 2021 Nov) Available from: https://www.who.int/news-room/fact-sheets/detail/pesticide-residues-in-food

General Lab Skills Course – A review from our interns

Our general lab skills course is designed to give you the basic skills needed to work in a GLP laboratory environment. You will receive in-house certification once the training is completed. To enroll on this course no previous experience is needed, however depending on your experience the training will be tailored to your needs.

What will you learn?

  • Pipetting
  • Preparation of serial dilutions
  • Preparation of buffers
  • Proper use of balances
  • Use of a pH meter
  • Centrifuge use
  • Use of PLT (pipette leak testing)
  • General lab safety

Who can enroll?

To enroll on this course no previous experience is needed, however depending on your
experience the training will be tailored to your needs.

Why enroll on this course?

This courses is beneficial for graduates or students that may be entering a laboratory environment for the first time or for scientists returning from a career break. This training will improve your laboratory skills which are fundamental to ensure your results are reliable and accurate. Moreover, this training is given on a 1-to-1 basis which provides the trainee a more practical experience and time to develop and improve their skills with laboratory equipment.

Review from our interns

“This internship has been an amazing experience. Before I was not confident with my laboratory skills and this internship has given me the confidence and all the basic skills needed to work in laboratory and get accurate results. I am very grateful that I got this opportunity.” – Gomes

” I am very grateful that I had my first “real world” laboratory experience here at Bionanalysis. The support from Cali has been exceptional. Being a member of this analytical team made learning new, complex skills more enjoyable. This experience has opened a new possible career path, which I had not fully considered pursuing.”- Eloisa

 

General Lab Skills Course

Eloisa changing a buffer on the HPLC

How to book?

Click here to book your training 

 

Alfaxalone in Veterinary Practice

What’s Alfaxalone and its function

Alfaxalone (Fig.1) is a neuroactive steroid that has anesthetic properties by acting on the gamma-aminobutyric acid subtype A (GABAA) receptors. It can act as a GABA agonist, when administered in high concentrations or modulate the chloride ion transport across the membrane of neuronal cells when given at low concentrations. Alfaxalone has different mechanisms of action, depending on the dose administered.

Alfaxalone in Veterinary Practice

Fig.1 Chemical Structure of Alfaxalone

This drug is currently being used in veterinary practice as an injectable anesthetic, induction agent or sedative for animals (1).

Advantages

There is little change in cardiac output or blood pressure, when clinically relevant doses of Alfaxalone are administered to healthy patients (2). Moreover, Alfaxalone has a high therapeutic index, a short duration of action, and is noncumulative (2). Because of these properties, this drug is ideal for use as an induction agent or for providing injectable anesthesia (2).

Furthermore, Alfaxalone can also be administered intramuscularly, facilitating the administration on agitated animals (2).

Side effects

Respiratory depression is the most common side effect: Alfaxalone can lower respiratory rate, minute volume, and blood oxygen saturation and cause apnea (1). Hence, this drug should be administered slowly over a period of at least 60 seconds or until anesthesia is produced, as apnea can occur if given too quickly (1). 

Furthermore, Alfaxalone can lead to depressive effects on the central nervous system such as reduced blood flow to the brain, intracranial pressure, and body temperature (1).

Animals recovering from anesthesia by Alfaxalone, should be taken to a quiet and dark area as they can be very agitated and react poorly to external stimuli such as noise and light (1).

Method

Using our LC-MS, we have analyzed Alfaxalone in feline plasma and estimated the values of LLOQ and ULOQ to be 8 ng/ml and 8000 ng/ml, respectively.

For more information about this analysis contact us by email. 

References

  1. Wikipedia. Alfaxalone. (Internet) (Updated on 2020 Oct; cited on 2021 Jul) Available from: https://en.wikipedia.org/wiki/Alfaxalone
  2. Martin J, Kennedy, DVM, Diplomate ACVAA. Alfaxalone for anesthesia in dogs and cats: what you should know. (Internet) Medvets. (Updated 2017 Sept 20; cited on 2021 Oct 15th) Available from: https://www.medvetforpets.com/alfaxalone-anesthesia-dogs-cats-know/

Mass Spectrometry Training

One of the most used pieces of equipment for analysing a sample is the mass spectrometer. It works by separating ions based on their mass/charge ratio and sending them through a magnetic or electrical field. The theory mass spectrometry (MS) was put forward in the late 1890s, but it wasn’t until the early 20th Centrury that an instrument was built and mass spectrometry continues to evolve into the twenty-first century. Nowadays, MS allows scientists to observe qualitative and quantitative changes in different compounds  and can be used to explore details about different biomolecules.

We offer a 5-day mass spectrometry training course that will ensure you learn all the skills needed to master the LC-MS. Our training services are normally offered on a one to one basis so that you can take the most out of this course. If you are currently working on a research project that requires the use of LC-MS to obtain data, you can let our lab manager know and she will tailor the course to your needs.

mass spectrometry training

Fig. 1 Shimadzu LCMS-8040

Training goals:

  • Learn how to set up and run HPLC analysis
  • Understand method development and which parameters need to be considered (mobile and stationary phase, temperature, pH, etc)
  • Sample preparation methods
  • LC-MS basics and method development
  • How to interpret HPLC and LC-MS results

During the course, as it is one to one, you have the opportunity to ask any questions you might have about MS and HPLC.

Structure of the course:

Day 1

  • Health and Safety
  • Standard laboratory protocol training

Day 2 

  • How to prepare samples
  • Basics of HPLC use

Day 3

  • HPLC method development 
  • Data analysis
  • Q&A

Day 4 

  • LC-MS basics
  • LC-MS method development

Day 5

  • LC-MS data analysis
  • Quiz
  • Q&A

The training course is taught by our lab manager, Dr. Carolyn Hyde, who has over 30 years of experience. If you are looking to master the HPLC and LC-MS instruments, contact us to book your training.

Click here to contact us about our mass spectrometry training course.

Analysis of Mannitol in Human Plasma

Mannitol is an osmotic diuretic that is physiologically inactive in humans and is found naturally in fruits and vegetables as a sugar or sugar alcohol. Mannitol raises the osmolality of blood plasma, allowing more water to move from tissues including the brain and cerebrospinal fluid into interstitial fluid and plasma (1).

analysis of mannitol in human plasma

Fig.1 3D Model of Mannitol

Mannitol can be used to:

  • Reduce Inter-cranial Pressure

Mannitol is administered intravenously and becomes a new solute in the plasma. Thus, increasing the tonicity of the plasma, which will pull water into the intravascular space from the brain parenchyma. The water will then be excreted through the kidneys.

  • Reduce Intra-ocular Pressure

Due to an increase in the plasma's tonicity by mannitol, water from the vitreous humous of the eye is drawn out to the intravascular space.  Finally, the water and mannitol will be excreted by the kidneys. The intraocular pressure is reduced as the vitreous humor’s water content decreases.

  • Promote Diuresis in Acute Renal Failure

Intravenous mannitol is sometimes given to those who have acute renal failure. Even in acute renal failure, the kidneys are responsible for a large portion of mannitol excretion. As mannitol is expelled, it attracts water, boosting the patient’s water output and assisting in the prevention or treatment of fluid overload caused by oliguria in acute renal failure.

  • Excrete Toxic Materials

Mannitol can be used to promote the excretion of harmful chemicals, substances, and medications, similar to how it is used to treat oliguria in acute renal failure.

  • Prevent Intradyalytic Hypotension

Hemodialysis patients frequently experience intradialytic hypotension and dialysis disequilibrium symptoms. The reason for this is a decrease in intradialytic osmolality. By increasing serum osmolality, mannitol can help prevent intradialytic hypotension.

Administration

Mannitol is administered intravenously when used for medical purposes (2). Mannitol
is available in a variety of quantities, ranging from 5% mannitol (5 gm mannitol
dissolved in 100 mL solution) to 25% mannitol (25 gm of mannitol dissolved in 100 mL
of fluid) (2). The amount of mannitol administered depends on the type of condition
that is being treated (2).

  • Increased Intercranial Pressure – dosages range from 0.25 g/kg to 2 g/kg
  • Increased Intraocular Pressure – dosages range from 0.25 g/kg to 2g/kg
  • Oliguria treatment – 0.2 g/kg
  • Excretion of toxic materials – 0.25 g/kg to 2 g/kg

Side effects

  • Heart failure
  • The movement of free water into the intravascular region might cause or worsen electrolyte imbalances. Hyponatremia, hypokalemia, and hypocalcemia are some of the electrolyte disorders that might occur.
  • Worsen cerebral edema. Frequent mannitol doses can cause mannitol to penetrate the blood-brain barrier, worsening cerebral edema because the mannitol that has leeched into the brain attracts water into the brain rather than out of it.
  • Dehydration due to diuresis

Method

We have analyzed mannitol in human plasma. Using our HPLC instruments (Fig. 2), we have determined our LLOQ and ULOQ to be 7.5 and 1500 µg/ml, respectively.

HPLC mannitol analysis

Fig. 2 HPLC

To book our instruments please contact us here.

References

1. National Library of Medicine. Mannitol. (Internet) (Cited on Aug 2021) Available from: https://pubchem.ncbi.nlm.nih.gov/compound/Mannitol

2. StatPearls. Mannitol. (Internet) (Cited on Aug 2021) Available from: https://www.ncbi.nlm.nih.gov/books/NBK470392/

Our HPLC Training Courses

Looking for A HPLC training COURSE?

High Performance Liquid Chromatography is a powerful analytical tool. We offer HPLC training courses which will give you all the skills needed to master the HPLC instrument.

Who is this course for?

HPLC training

Fig.1 Our HPLC system

HPLC training is for any person (no previous experience needed) who is looking to improve their laboratory skills or learn how to master the HPLC instrument. Depending on your previous experience we will prepare a tailored training to cover everything you may need to know.

Both HPLC training courses are taught on a 1 to 1 basis (maximum 2 people) so that you can take the most out of this training and have plenty of time to adapt to the instrument. We focus on giving you a complete and practical experience, hence by the end of the training, you will feel confident to run by yourself your own set of samples on the instrument.

Course Options

We offer two HPLC courses:

  • 3 Day HPLC Training
  • 1 Day HPLC Refresher Course

The 3 day course covers all the HPLC basics, standard laboratory training, sample preparation, method development and data analysis. It is the perfect choice for someone that may want to learn how to set and run HPLC to improve their analytical skills. This is a complete course that aims to teach you all the fundamentals you need to run your own samples.

The 1 day course covers HPLC basics and method development, however it is subject to the trainee’s previous experience. This course is advised to someone that already has some experience with HPLC and wishes to remind themselves about HPLC basics/method development or is looking for advice on method development for their own research project.

Structure of the 3 Day Course

  • Day 1 – Essential reading & standard laboratory training
  • Day 2 – Sample preparations & method development
  • Dat 3 – Method development (continuation) & data analysis

At the end of the course, there is a Q&A with our manager, you can clarify any questions you might have.

How to book

Fill out the form in the contact us page, or if you prefer you can email us or call us directly. Please click here to book your HPLC training course!

 

 

 

 

Amlodipine in Veterinary Practice

Amlodipine has been used to treat high blood pressure in humans, but it has now been approved for veterinary practice. It is given to cats and dogs to treat hypertension and other associated diseases.

About Amlodipine

This drug is classified as a calcium channel blocker. The heart and arteries contract more forcefully when calcium is present, thus calcium channel blockers relax and widen blood arteries by inhibiting calcium. This effect will lower the blood pressure which prevents heart diseases, heart attacks and strokes.

amlodipine in veterinary practice

Fig.1 Chemical Structure of Amlodipine

Side effects

Like all medicines, amlodipine can have side effects. In animals, common side effects include vomiting, lack of appetite, diarrhea and sleepiness (1). Serious side effects such as severe weakness, changes in urination, abnormal bleeding or rapid heart rate are rare, but can also occur (1).

Hypertension in Cats

Hypertension in cats is a moderately frequent and possibly fatal condition. The most prevalent kind of hypertension in cats is secondary hypertension, which is caused by an underlying systemic condition that impairs the body’s regulatory mechanisms responsible for maintaining normal blood pressure (2). Among these conditions, chronic kidney disease and hyperthyroidism are the most common (2).

Injury to organs can happen due to hypertension. The most prevalent target organ injury found in cats with hypertension is ocular disorders, which can range from retinal detachment resulting in blindness to more subtle indications like hemorrhage or swelling in the back of the eye (2).

Hence, not only can amlodipine treat hypertension, but it can also be given to cats to prevent damage to kidneys, heart, brain, and eyes.

Analysis of Amlodipine in Feline Plasma

Using our HPLC system and LC-MS, we have examined amlodipine in feline plasma. The LLOQ and ULOQ were set for the method as 0.5 ng/ml and 100ng/ml respectively for Amlodipine. The Limit of Detection was calculated as -0.001 ng/ml in feline plasma.

References

  1. Rania Gollakner. Amlodipine. (Internet) VCA. (cited on 2021 Jul) Available from: https://vcahospitals.com/know-your-pet/amlodipine-besylate
  2. Cornell Feline Health Center. Hypertension: Feline hypertension. (Internet) College of Veterinary Medicine. (updated 2021; cited on 2021 Jul) Available from: https://www.vet.cornell.edu/departments-centers-and-institutes/cornell-feline-health-center/health-information/feline-health-topics/hypertension

Developing Good Pipetting Techniques

hOW IMPORTANT IS IT TO DEVELOP GOOD Pipetting Techniques?

In order to achieve optimal and reliable results for our diagnostic services we ensure all staff are
trained to a high degree in basic laboratory skills, such as liquid handling.

Pipettes are used daily in our lab for liquid handling. Two factors contribute to good laboratory
pipetting: tools and techniques. At the Bio-analysis centre we use mainly single channel manual
‘transferpette’ pipettes (Figure 1) which are known to be highly accurate at various volumes; however, the
most important factor is the expertise of the operator.

developing good pipetting techniques

Fig. 1 Single Manual Channel Transferpette

There are two main pipetting techniques: forward pipetting and reverse pipetting (/back pipetting).
Forward pipetting aspirates from the first stop and ejects until the second, whereas reverse
pipetting aspirates from the second stop and ejects until the first.

Other factors include:

– Immersion angle and depth
– Pre-rinsing
– Speed
– Temperature of environment and samples
– Performing pipette leak testing daily

Prolonged poor ergonomics while using pipettes can result in repetitive strain injuries (RSI) and
carpel tunnel syndrome (CTS) (El-Helaly et al., 2017). Studies have shown how higher work duration
increases the risk of developing RSI and CTS (El-Helaly et al., 2017, Bjorksten et al., 1994) . During our
training courses we recognise this issue and work with you to improve your pipetting technique
giving you the confidence to work comfortably in a laboratory environment.

Good laboratory pipetting is fundamental in our work producing accurate and precise data from the
HPLC and MS-LC. We are experienced in pipetting difficult solutions such as plasma and serum,
which are notoriously bubbly and therefore difficult to pipette!

Click here to find out more about our 1 day lab skills course.

References:
BJORKSTEN, M. G., ALMBY, B. & JANSSON, E. S. 1994. Hand and shoulder ailments among laboratory
technicians using modern plunger-operated pipettes. Appl Ergon, 25, 88-94.

EL-HELALY, M., BALKHY, H. H. & VALLENIUS, L. 2017. Carpal tunnel syndrome among laboratory
technicians in relation to personal and ergonomic factors at work. J Occup Health, 59, 513-
520.

Liquid Chromatography Method Development

Why is Method Development important? 

In the pharmaceutical sector, liquid chromatography procedures are used to assay compounds and quantify impurities present within medicinal products. Method development allows the creation of the best method to test for impurities and compounds which help ensure the quality, safety and efficacy of new products and drugs.

Liquid Chromatography Method Development 

Method development is the process by which the optimal conditions are found for a particular compound to separate it from any contaminants. To design the best method for a particular sample, the following parameters and conditions must be optimized:

  • Selection of chromatographic mode (Reverse Phase, Normal Phase,
    Size exclusion, HILIC, etc.)
  • Selection of stationary phase (C18, C8, Phenyl-hexyl, Biphenyl,etc.)
  • Selection of detector (UV, RID, MS, ELSD)
  • Selection of mobile phase (which buffers; pH of buffers)
  • Flow rate
  • Preparation of samples (protein precipitation, Solid Phase Extraction,
    etc)
  • Injection volume
  • Concentration
  • Calibration Range

HPLC Method Development

Our Nexera XR HPLC system (Fig.1) is ideal for developing procedures for a variety of material, ranging from complicated combinations of tiny molecules or proteins to single chemical purification.

HPLC method development

Fig. 2 Nexera XR HPLC System

Mass Spectrometry Method Development

We have the expertise to design methodologies to quantify most drug-like substances using our Shimadzu LCMS-8040 (Fig.2).

Mass spectrometry method development

Fig.2 Shimadzu LCMS-8040

Our manager Dr. Carolyn Hyde has over 20 years of experience developing methods on HPLC and LC-MS. Click here to contact us for more information on our services. 

References

Avoomeen. Why Pharma Companies Must Invest in Method Development & Validation (Internet). (Cited on 16th July) Available from: https://www.avomeen.com/pharma-companies-invest-method-development/