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UV-HPLC separation

Determination of liquid chromatographic pattern

Do you need to determine the liquid chromatographic pattern of the proteins in your drug sample?

We offer to separate your drug sample by a standard 30 min UV-HPLC gradient. This results in a chromatogram. The graph shows you the liquid chromatographic pattern of a protein mixture.

What does the ICH Q6B say? (Test Procedures and Acceptance Criteria for Biotechnological and Biological Products). ICH Q6B requires you to determine the liquid chromatographic pattern by UV-HPLC.

Our standard services include:

  • Standard or customized UV-HPLC separation of your sample.
  • Fractions can be collected for further analysis.
  • Can be combined with ESI-MS analysis for mass determination of each peak.

You can order our standard services online.

Need to set up a custom UV-HPLC method?

We can also develop a customized method meeting your specific needs. For instance an in-house method, which you want us to set up in our laboratory. We can combine it with high resolution Mw determination by mass spectrometry.

You can send us a request so we can meet your specific needs.

Services/Products
Protein analysis by Alphalyse

UV-HPLC analysis of proteins in drug sample

  • UV-HPLC separation of proteins on C8 reverse phase
  • Standard 30 min gradient
  • Quantification of all peaks >1%

 

$900.00

World Class Service
Quality analysis, presented in easy-to-understand reports
Info / Order
Request quote
Protein analysis by Alphalyse

Customized UV-HPLC separation of proteins in drug sample

  • UV-HPLC separation of proteins
  • Customized method meeting your needs
  • Agree on which fractions to collect
  • Price depends on customization

World Class Service
Quality analysis, presented in easy-to-understand reports
Info / Order
Request quote

Technical details

The analysis is conducted on one of our Agilent 1200 HPLC systems with UV detection and reverse phase column.

Standard analysis on 30 min gradient:
The  analysis is a 30 minutes gradient with a flow of 0.2 ml/min on a 5 cm Poroshell  300 SB-C8 column.

All the major peaks with an area above 1% will be identified by retention time and quantified by integrating the area below the peak.

Custom designed analysis:
The custom designed analysis is based on HPLC setting from clients including column, temperature and gradient settings. The analysis can be coupled to MS instrument to provide accurate mass information on all peaks.

Sample requirements

Sample Preparation Guidelines
Protein samples can be submitted in liquid or as lyophilized material.
Avoid detergents and keep buffer concentration at a minimum. HPLC  analysis can be done on samples containing small amounts of salts, urea or detergent, but the best result are obtained with low buffer strength in volatile solvents without detergents.
Minimum amount of material ~ 20-50 micrograms

Put the sample into microcentrifuge tube
Use Eppendorf Safe Lock tubes, or similar tubes from your lab
Lyophilize or speed vac the sample to a solid to ensure protein stability during shipment
Alternatively, freeze or refrigerate to +4°C for cold shipment of the liquid sample

 

Sample shipment

shipping option
Ship your protein samples by a courier company of your own choice

Place your samples into micro tubes and pack in a suitable transport box.

  • Contact a courier company of your choice (FedEx, UPS, DHL, TNT, World Courier etc.). Use the following sentence to describe the content: “Protein material – Non-toxic, non-hazardous, for research purpose only“.
  • Use cool packs or cooling units if your samples should be stored below 5°C/40°F
  • Ship the samples to the office closest to you (Canadian customers, please ship to Europe):
Customers in the USA
Alphalyse, Inc.
200 Page Mill Road, Suite 100
Palo Alto, CA, 94306
USA
Att: Jette Williams
Tel: (650) 543-3193
Customers outside of USA
Alphalyse A/S
Unsbjergvej 4 D
DK-5220, Odense
DENMARK
Att: Anette Rasmussen
Tel: +45 6310 6500
shipping option
Order a prepaid FedEx Air Waybill < 1 lb./ 500g

A prepaid FedEx Air Waybill can be used for fast and easy shipment of samples in packages up to 1 lb./ 500g.
We will generate a prepaid Air Waybill, prefilled with the correct information, and send it to you by e-mail (PDF-format) within 24 hours.
Then follow these simple instructions:

  • Place your samples into micro tubes. Pack in suitable transport box.
  • Print the PDF document and stick it on the transport box or just hand it to your FedEx officer upon pickup.
  • Call to request a pickup from your local FedEx office.

If you plan to send your samples stored at room temperature, this option is usually the right choice. If you fill your box with cooling packs, the weight will most often exceed 1 lb./500g.

shipping option
Order a prepaid FedEx Air Waybill < 10 lb./5kg (e.g. for packages with cooling packs)

This prepaid FedEx Air Waybill can be used for shipment of samples in packages up to 10 lb./ 5kg, with or without cooling packs.
We will generate a prepaid Air Waybill, prefilled with the correct information, and send it to you by e-mail (PDF-format) within 24 hours.
Then follow these simple instructions:

  • Place your samples into micro tubes. Pack them in a suitable transport box. Fill with cooling units/cold packs.
  • Print the PDF and stick it on the transport box or just hand it to your FedEx officer upon pickup.
  • Call to request a pickup from your local FedEx office.

If you plan to fill your transportation box with cooling packs, select this shipping option.

shipping option
Get a quote for shipment by World Courier

The World Courier service can be used for shipment of samples of all types. If you select this option, we will contact you to provide a quotation based on location and shipping conditions. You will receive a prepaid Air Waybill which will be invoiced together with your analysis results. This is usually a good option if FedEx does not service your location.
With the prepaid World Courier service you get:

  • GxP compliant processes to ensure maximal security and traceability
  • Door-to-door solution on a dedicated vehicle
  • Possibility of providing qualified packaging, temperature monitoring and dry ice
  • Shortest possible transit-time through “next flight out”-procedure
  • Customs clearance and help with paperwork

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