Protein Hydrolysis Kit

An innovative system to eliminate the bottleneck in amino acid analysis

The accuracy and precision of amino acid analysis is often determined by the quality of protein hydrolysates.
While state of-the-art HPLC instrumentation allows accurate analysis in less than 60 minutes, the widely used standard protocol (Stein and Moore, 1950) requires heating samples in 6N HCl at 110°C for 24 hours or more. Disadvantages associated with conventional methods of protein hydrolysis include:

- Time consuming: up to 96 hours
- Labor intensive
- Potential contamination from acids in liquid phase methods and during sample transfer steps
- Inaccurate control of hydrolysis conditions

Advantages

• Time Savings: hydrolysis time is reduced from 24-96 hours to 5-30 minutes, more closely matching sample preparation time to analysis time requirements.
• Uniformity of Temperature Conditions: all samples are processed under equivalent temperature conditions avoiding possible variations between samples and assuring precision of results.
• Inert, Anaerobic Processing: hydrolysis is performed under inert, anaerobic conditions to prevent oxidative degradation of amino acids.
• No Contamination: hydrolysis is performed in standard HPLC autosampler vials, eliminating sample transfer steps and reducing the possibility of contamination or losses.
• Easy Operation: all sample vials are loaded in a single operation with minimal handling required.
• Complete Documentation: using Milestone’s software, every microwave hydrolysis parameter can be stored and recalled for complete data validation to support chromatographic results.

The state-of-the-art microwave accelerated acid hydrolysis system

A vapor phase microwave hydrolysis system consists of a special 500 ml fluoropolymer vessel with connections for vacuum/ inert gas purging, temperature measurement and control.
Samples are introduced into autosampler vials and loaded in a 8 position rack. Hydrochloric acid (6N) is added to the vessel, where the vial tray is then placed.
The vessel is assembled and, by means of a 3 way valve, air is removed by vacuum evacuation and nitrogen gas is purged into the vessel.
This cycle is repeated and 1 atmosphere of Nitrogen gas is sealed in the vessel. Microwave heating raises acid temperature and acid vapor contacts and hydrolyzes samples inside the vials. Hydrolysis time varies from 5 minutes to 30 minutes for 8 samples, depending upon sample type.

The vapor phase microwave hydrolysis sequence

1 - System set up		2 - Vacuum/evacuate
3 - Purging with N2		4 - Hydrolysis