How to read an HPLC report
To ensure you are receiving the highest quality products from RRV Team our peptides are all HPLC tested prior to delivery. We guarantee a 99% or greater purity for all of our peptides. We can also include a copy of the HPLC with your order so you can confirm the quality.
Below is an example based on our TB 500 HPLC report describing the process and explaining the results.
TB 500 HPLC Report Explained
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY – HPLC
High performance liquid chromatography is a powerful tool in analysis. This page looks at how it is carried out and shows how it uses the same principles as in thin layer chromatography.
High performance liquid chromatography is basically a highly improved form of column chromatography. Instead of a solvent being allowed to drip through a column under gravity, it is forced through under high pressures of up to 400 atmospheres. That makes it much faster.
It also allows you to use a much smaller particle size for the column packing material which gives a much greater surface area for interactions between the stationary phase and the molecules flowing past it. This allows a much better separation of the components of the mixture.
The other major improvement over column chromatography concerns the detection methods which can be used. These methods are highly automated and extremely sensitive.
The column and the solvent
Confusingly, there are two variants in use in HPLC depending on the relative polarity of the solvent and the stationary phase.
Normal phase HPLC
This is essentially just the same as you will already have read about in thin layer chromatography or column chromatography. Although it is described as “normal”, it isn’t the most commonly used form of HPLC.
The column is filled with tiny silica particles, and the solvent is non-polar – hexane, for example. A typical column has an internal diameter of 4.6 mm (and may be less than that), and a length of 150 to 250 mm.
Polar compounds in the mixture being passed through the column will stick longer to the polar silica than non-polar compounds will. The non-polar ones will therefore pass more
Reversed phase HPLC
In this case, the column size is the same, but the silica is modified to make it non-polar by attaching long hydrocarbon chains to its surface – typically with either 8 or 18 carbon atoms in them. A polar solvent is used – for example, a mixture of water and an alcohol such as methanol.
In this case, there will be a strong attraction between the polar solvent and polar molecules in the mixture being passed through the column. There won’t be as much attraction between the hydrocarbon chains attached to the silica (the stationary phase) and the polar molecules in the solution. Polar molecules in the mixture will therefore spend most of their time moving with the solvent.
Non-polar compounds in the mixture will tend to form attractions with the hydrocarbon groups because of van der Waals dispersion forces. They will also be less soluble in the solvent because of the need to break hydrogen bonds as they squeeze in between the water or methanol molecules, for example. They therefore spend less time in solution in the solvent and this will slow them down on their way through the column.
That means that now it is the polar molecules that will travel through the column more quickly.
Reversed phase HPLC is the most commonly used form of HPLC.
Looking at the whole process
A flow scheme for HPLC
Injection of the sample
Injection of the sample is entirely automated, and you wouldn’t be expected to know how this is done at this introductory level. Because of the pressures involved, it is not the same as in gas chromatography (if you have already studied that).
The time taken for a particular compound to travel through the column to the detector is known as its retention time. This time is measured from the time at which the sample is injected to the point at which the display shows a maximum peak height for that compound.
Different compounds have different retention times. For a particular compound, the retention time will vary depending on:
- the pressure used (because that affects the flow rate of the solvent)
- the nature of the stationary phase (not only what material it is made of, but also particle size)
- the exact composition of the solvent
- the temperature of the column
That means that conditions have to be carefully controlled if you are using retention times as a way of identifying compounds.
There are several ways of detecting when a substance has passed through the column. A common method which is easy to explain uses ultra-violet absorption.
Many organic compounds absorb UV light of various wavelengths. If you have a beam of UV light shining through the stream of liquid coming out of the column, and a UV detector on the opposite side of the stream, you can get a direct reading of how much of the light is absorbed.
The amount of light absorbed will depend on the amount of a particular compound that is passing through the beam at the time.
You might wonder why the solvents used don’t absorb UV light. They do! But different compounds absorb most strongly in different parts of the UV spectrum.
Methanol, for example, absorbs at wavelengths below 205 nm, and water below 190 nm. If you were using a methanol-water mixture as the solvent, you would therefore have to use a wavelength greater than 205 nm to avoid false readings from the solvent.
Interpreting the output from the detector
The output will be recorded as a series of peaks – each one representing a compound in the mixture passing through the detector and absorbing UV light. As long as you were careful to control the conditions on the column, you could use the retention times to help to identify the compounds present – provided, of course, that you (or somebody else) had already measured them for pure samples of the various compounds under those identical conditions.
But you can also use the peaks as a way of measuring the quantities of the compounds present. Let’s suppose that you are interested in a particular compound, X.
If you injected a solution containing a known amount of pure X into the machine, not only could you record its retention time, but you could also relate the amount of X to the peak
The area under the peak is proportional to the amount of X which has passed the detector, and this area can be calculated automatically by the computer linked to the display.
TB 500 is not a drug nor is it intended to treat, diagnose, cure or prevent any disease, nor is it meant for ANY cosmetic purpose.
It is RRV Teams understanding that TB 500 is a generic peptide not controlled by any known drug conventions. Our Peptides are exclusively distributed legally within the laws of safe harbor for research peptides. If any country’s governing bodies feel we are in violation of their laws regarding TB 500 and/or other peptides, we ask you contact us immediately via firstname.lastname@example.org so we may expeditiously rectify the situation.
All customers represent and warrant that through their own review and study that they are fully aware and knowledgeable about the following:
- Their government regulations regarding the importation, purchase, possession and use of TB 500 and other peptides.
- The health and safety hazards associated with the handling of TB 500 and other peptides in a research setting.
- That TB 500 and other peptides are NOT intended to be used as a food additive, drug, vitamin, supplement, cosmetic or any other inappropriate application. Such a sale would be otherwise denied.
What Can Growth Hormone Therapy Do for Me?
The growth hormone (GH) is responsible for slowing the aging process, which can aid you exponentially in your bodybuilding regimen. As you get older, the growth hormone naturally starts to decline in your body. After 30, the hormone declines about 25 percent every ten years. This means that when you’re 60, you have 25 percent of your original levels.
The pituitary gland naturally produces the growth hormone, and it stimulates cell reproduction and growth in the body. It also helps to regulate your body fluids, body composition, sugar and fat metabolism and muscle and bone growth, and it may also help with heart function. Growth hormone therapy can slow down the aging process, and this synthetic hormone was first developed in 1985 and is approved by the FDA.
Benefits of Growth Hormone Therapy
Growth hormone has many benefits for bodybuilders and those desiring a more chiseled physique. The hormone diverts the calories in food away from fat synthesis and toward protein synthesis. Other benefits include:
- promoting and increasing the synthesis of protein muscles, including those in muscle repair and recovery
- improving the duration and quality of your kidneys and heart
- metabolizing body fat as it converts to energy
- improving your sleep pattern, which can lessen your chance of waking up unintentionally and improve your REM-stage sleep
- building stronger bones
- possibly improving your sexual performance
- producing more energy
Growth hormone therapy may even be the better alternative to testosterone, as there are fewer side effects and it isn’t androgenic.
How to Stimulate the Human Growth Hormone
In addition to growth hormone therapy, there are certain things you can incorporate into your regimen that will naturally stimulate the growth hormone.
Intense workouts can stimulate growth hormone, as well as events that consume energy and having long stretches of physical exhaustion from exercise. Don’t train longer than 45 minutes, or an hour if you slowly increase the length of time over a period of months. After 45 minutes, GH tapers off and your production of cortisol begins.
Promote more REM cycles by regulating your sleep pattern. If you stay out late on the weekend, still wake up at your normal time instead of trying to sleep in. If you do sleep in, it may disturb your sleep pattern. Instead, it’s better to take a one-hour nap in the afternoon for three days to make up for the sleep you missed. However, the best course of action is to maintain steady sleep hours.
Amino acids are crucial for bodybuilding. In order for the aminos to have the most positive effect possible, make sure clean fats make up 15 to 20 percent of your diet.
Vitamin C, Vitamin B3 and antioxidants also help promote GH.
Small doses of growth hormone have many benefits. It’s possible to acquire growth hormone supplements over the counter when they are below a certain a concentration. Currently, the growth hormone is sold as a supplement that is suspended in alcohol. You absorb the supplement either under the tongue or through your nose. This small supplement yields high rates, and it won’t find its way into the blood through your digestive tract. The supplement can allow your muscles to recover faster, can decrease a buildup up lactic acid and can help you sleep better.
When taken responsibly, growth hormone therapy can have positive results in aiding your training and recuperation.
Benefits of Peptides for Muscle Growth
The first thing you need to know when investigating the benefits of peptides for muscle growth is, what exactly are peptides? Peptides are interesting compounds of two or more amino acids in which a carboxyl group of one is joined to an amino group of another. Simply put, peptides are basically just small proteins. Technically, anything with fewer than fifty amino acids is considered a peptide, but that's not etched in stone. While a dipeptide is made up of two amino acids joined by one peptide bond, tripeptides are three amino acids bound by two peptide binds, and so on, a polypeptide is a long, un-branched chain of amino acids joined by peptide bonds that is not complex enough to be referred to as a protein, even though proteins are made up of polypeptides. If that sounds confusing, just keep in mind “protein.”
Since peptides are made from amino acids (protein), dietary intake of complete proteins make available all the amino acids necessary for production of all the peptides needed for the body to work at its best.
Various amino acids come together to create peptides that cause a vast stream of effects on the body. Peptides have been used by bodybuilders in recent years to achieve gains in muscle mass and ripped conditioning. While it's true that bodybuilders also use steroids, it's important therefore to understand the difference between them. Why do they use both steroids and peptides? Why do some use peptides and not steroids? While peptides cause fewer side effects than steroids – in some cases no side effects at all - the important distinction between peptides and steroids is the difference between muscle hyperplasia vs muscular hypertrophy respectively. Steroids increase protein synthesis in muscle to help grow and repair muscle fibers of the muscle you already have. The muscle fibers just get bigger - hypertrophy. Peptides, on the other hand, stimulate satellite cells on the muscle fibers to grow new muscle fibers – hyperplasia. So, bodybuilders use peptides to increase the number of muscle fibers, they use steroids to help increase the size of those fibers. Some just use peptides to grow lots of new muscle.
While the increase in muscle gain from peptides is in fact slower than the gains made with steroids, peptides don't cause you to retain fluids. All the gain in mass from peptides is muscle, not water. You only gain lean muscle mass. Also, since peptides enhance metabolic function, increasing your metabolism and consequently your energy level, you train harder in the gym and burn more fat, which makes that new muscle harder and more defined.
Additionally, all the new muscle you gain with peptides is permeant muscle. It doesn't go away when you stop using them. The increase in mass with steroids does diminish when you stop using them, both because the water weight dissipates and, since protein synthesis in muscle is attenuated when steroid use ceases, you lose size. The muscle gained with peptides isn't going anywhere.
The end result is that the benefits of peptides for muscle growth include better gains than with steroids, with relatively no side effects, such as water retention, gynecomastia, and no down regulation of natural testosterone production negating the need for post cycle therapy (PCT). And, since peptides cause few if any side effects, do not aromatize to estrogen or dihydrotestosterone (DHT), don't down regulate natural testosterone production or insult vital organs such as the liver and kidneys, you can take peptides virtually indefinitely.
As you can see, the benefits of peptides for muscle growth extend far beyond actually building muscle. Peptides increase metabolism which helps burn fat and help keep the muscle defined; peptides don't aromatize or convert to estrogen or DHT; don't effect natural testosterone production; and are not kidney or liver toxic. Peptides can be used year round to help keep your muscles growing.