What tests could be run to test the purity and type of HCG?

We are looking to purchase HCG from China but the purity and quality varies between labs, we are able to receive samples of the raw peptide and can use a contracted lab to perform tests but due to my academic background I have no clue which tests to request to determine the purity and type of the raw powder. The following is what we should be receiving from the labs in China.

Product Description CAS Number: 9002-61-3 pI = 2.951 Extinction Coefficient: EmM = 3.88 (278nm)2 Synonym: Choriogonin, hCG The molecular weight is approximately 37.9 kDa (with approximately 31% carbohydrate by weight). The theoretical molecular weight is 37.9 kDa based on the native form, which contains 2 subunits. The α subunit has a molecular weight of 14.9 kDa of which approximately 10.2 kDa is for the polypeptide and approximately 4.7 kDa for the carbohydrate. The β subunit has a molecular weight of 23 kDa of which approximately 16.0 kDa is for the polypeptide and approximately 7.0 kDa is for the carbohydrate.3,

To answer Mad Scientists Question, the peptide is "supposed" to be CGA (1081) for use on humans. This is why it is so important for us to find a test that can define the qualities of the raw peptide.

  • 2
    $\begingroup$ What exactly do you want to do with it afterwards? The level of purity and the kind of contamination that would be acceptable is very different if you do some in vitro experiments or if you do in vivo experiments in animals, not to speak of giving it to humans. $\endgroup$ Aug 31, 2013 at 7:24
  • $\begingroup$ A quick way could be a size analysis via a gel permeation chromatography, if you have access to this equipment. $\endgroup$
    – tyBee916
    Sep 8, 2020 at 10:51

1 Answer 1


A monograph from a pharmacopeia gives a good starting point for active pharmaceutical ingredient characterization. AFAIK there is no monograph for hCG nor your actual peptide of interest, but there is one for FSH (here for the european pharmacopeia monograph), which shares a subunit with CGA, meaning that modifications of it could suffice (though validating these modification will be a good amount of work, and will depend on the ultimate use for the peptide).

First, it is necessary to stress that even if the compound passes all assays, it would still be unsuitable for human use. It may be acceptable for animal experimentation. This is because the pharmacopoeia assumes a context where you have a non-adversarial relationship with your supplier, such that statement like "up to XX% remaining impurities" is an acceptable standard as the synthesis route & typical impurities are known. However, buying peptides from a random Chinese seller with no transparency on process is likely an adversarial situation.

Some keys to read the monograph:

  • Compound followed by the letter R ("water R", "glycine R"): these compounds have their own monograph. You could assay them accordingly, but for this use case understanding the R as "analytical grade" is possibly good enough.
  • "Follitropin CRS": chemical reference substance, a reference standard powder of follitropin obtained through reliable means (typically through EDQM for pharmaceutical purpose). Here also, substituting for an analytical grade standard from a reputable chemical seller could work.
  • Method followed with three numbers ("Bacterial endotoxins (2.6.14)"): reference a method monograph found in the given chapter/section. Apply methodology as described in the referenced text.

To adapt the assays to your peptide of interest, you need to understand which parts are specific to follitropin (and should be modified), and which applied to gonadotrophin-like peptides in general. For instance, for isoelectric focusing, the running conditions (pH, catholyte/anolyte) will be adequate, but the follitropin reference will be replaced with a CGA reference. On the contrary, the selective cleavage of peptide bonds assays is specific to the peptide, and will need extensive rework.


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