0
0
Your basket is empty...
Chromatography
MetAmino FAQ
What is the type of membrane inside the spin filter?
- The membrane is made of NYLON material and its porosity is 0.22 µm. Its diameter is optimized for use with a given spin filter.
It is possible to expand the MetAmino® kit with other analytes.
- Yes, the MetAmino® kit can be further expanded. Contact us with detailed information.
May I use the kit for urine as well?
- Yes, the MetAmino® kit can be used with urine. The matrix should be without proteins. So, the sample has to be prepared accordingly (centrifugation, filtration).
Can we order the reagents separately?
- Yes, the entire set of reagents can be ordered under catalog number MAK-5857-L002.
What is the back pressure of the LC/MS column?
- The pressure is 380 bars during the beginning of the analysis, at the end it is 200 bars.
Are MRMs for amino acids given in the MetAmino® Kit were made after derivatization of the standards or before?
- The MRM transitions for AAs given in the kit manual are given as transitions of the AA derivatives and not of the native AA
Should the sample of feed be hydrolyzed before working with the MetAmino® Kit?
- Precipitation with Precipitation medium (PM) is not an essential step for successful derivatization of the sample. The kit was tested mainly for the analysis of biofluids, which often contain peptides. To avoid their precipitation during derivatization, the precipitation step was included in the sample preparation protocol.
- A total analysis of the free and peptide-bound amino acids is required. In this case, we would recommend hydrolyzing the sample and then drying the aliquot of hydrolyzed sample under a nitrogen stream or in a speedvac. We would then dissolve the dried residue in 25 µL of deionized water or 0.1M aqueous HCl (better solubility) and follow the procedure in the manual by adding 10 µL of the IS solution.
- As for the peptide hydrolysis, the additives (phenol, thiodiglycol) in the hydrolysis medium could also be derivatized (although they are probably not visible in the full scan). Therefore, we would simply recommend 6M HCl as the hydrolysis medium.
Can we order a MetAmino® GC/MS sample preparation kit for 400 samples?
- Not yet, we currently only have a MetAmino® GC/MS sample preparation kit for 100 samples. The kit can be ordered under catalog number MAK-5857-BA01.
There are 78 amino acids in table 1, some of which are not in the available mixes of standards - should we therefore rely on MRM transitions and look at the retention time or are these the standards that we should buy if we want to determine them?
- For quantification purposes the MetAmino kit contains - one bottle of SD1 standard solution included in the kit contains 33 amino acids: AAA, ABA, ALA, APA, ARG, ASP, BAIBA, CC, CIT, CTH, GABA, GLU, GLY, GPR, HIS, HLY, HYP, ILE, LEU, LYS, MET, 1MHIS, 3MHIS, ORN, PHE, PHP, PRO, SAR, SER, THR, TPR, TYR, VAL and bottle of SD2 with lyophilized mixture of 3 amino acids: ASN, GLN, TRP. However, the amino acid set can further be expanded to other compounds containing primary or secondary amino functional groups.
- The masses (m/z) reported in the manual are correct and represent M+H+ ions. The masses (m/z) and the retention times from the manual were obtained from the linear ion trap LTQ instrument however, the data from the Certificate of analysis were obtained from Q Exactive plus HRMS instrument. In both cases, the same column, flow rate, and mobile phase composition were used. The retention times (RT) are experimental values and the discrepancy especially for late eluting compounds would be caused by using different LC/MS instruments.
How can I predict how much to dilute a sample?
- It depends on the concentration of analyzed amino acids in the sample. The total amount of amino acids in the sample taken to the reaction should not exceed 1.2 μmoles. The amount of sample also depends on the LCMS instrument.
How much sample we need to take to hydrolisys with 6M HCl to use it with MetAmino® kit?
- You can take a higher amount of sample for AA hydrolysis (e.g. 50 micrograms in 300 microliters of 6M HCl) and finally take a few microliters (remove the acid in the speedvac or under the nitrogen stream) for the Metamino workflow.
In which point should the sample be diluted? After hydrolysis of the sample and before applying the kit? Or is it possible after the complete application of the kit (finished sample)?
- The reaction is designed to analyze aqueous samples, so if 25 μL of the sample gives an overloaded signal, I would recommend diluting the sample with deionized water. It is also possible to dilute the finished sample.
How much derivatizing reagent is used during derivatization process of the sample? How can I be sure that the reaction is 100%?
- The derivatization agent is sufficient to derivatize 1.2 μmol total amount of amino acids. The derivatization and the purification don´t always proceed at 100%. If you perform the derivatization of samples and calibration points in the same way, you should obtain the correct values. (Even if the effectivity of the derivatization and the micro spin purification is lower than 100%).
MetAmino Sample Preparation Kit Description
MetAmino® kits offer an easy sample preparation method for your LC-MS or GC-MS analysis. MetAmino® kits incude derivatization reagences and all clean-up accessories to prepare your sample for injection. They elimilate the time consuming sample preparation procedures.
New clean-up procedure uses a special material as the end-step. The other odvantage is that the derivatization procedure enables to extenbd the analyte list.
- 75 amino acids, polyamines, biogen amines and coensymes
- 25 minutes (sample preparation and analysis time)
- Easy sample preparation
- SPE pipetting step replaced by MSPE
- No heating needed in the procedure
- There is no need to use a freezer
- NIST library for GC/MS available
- MetAmino_Brochure_300dpi_EN
4015 kB
Overview of MSPE sorbents
Sorbents for the MSPE technique are chosen to cover the widest possible field of applications. MSPE SpeExtra C18 is a hydrophobic type of octadecyl silica gel with a special endcapping. It is suitable for a wide range of analytes, showing lower retention for polar compounds. MSPE SpeExtra C18-P is a polar modified monomeric octadecyl silica gel. It offers different types of interactions: dipole-dipole, π-π and hydrophobic. It is therefore suitable for aromatic and polar compounds. MSPE SpeExtra HLB polymer sorbent with high specific surface area and special endcapping. It has a hydrophilic and lipophilic modification ensuring universal use and a higher capacity than C18 silica gel.
| MSPE sorbent | Particle size [µm] | Specific surface area [m 2 /g] | ||
| C18 | 60 | 310 | ||
| C18-P | 60 | 310 | ||
| HLB | 30 | 850 |
Purification & testing for cannabis and hemp
Hemp contains hundreds of cannabinoids with Cannabidiol (CBD) being the most prevalent in the plant and Δ9-Tetrahydrocannabinol (THC) being the active ingredient causing psychotropic effects. However, many more compounds are formed by the hemp plant and have been investigated for their medical effects. This limit often requires THC remediation of the distilled hemp extract (starting material) and can be achieved using preparative scale chromatography such as the puriFlashR XL-Cannabis system. HPLC analysis of the starting material (third pass distillate), fractions collected during the remediation process, and the finished product can be performed using the Advion AVANT HPLC-UV analytical system. Both the purification and analytical processes are shown in this application note to form a complete solution for THC remediation in the hemp industry.
- puriFlah® L-Canabis instrument with the output up to 4.3 kg/day (columns up to 15 cm ID)
- puriFlah® XL-canabis instrument with the output up to 12.2 kg/day (columns up to 20 cm ID)
- Plate Express TLC Plate Reader
SPE Phases
This site encloses SPE phases overview includint their technical parameters. Further information about the products are available in the product catalogue.
Clean up SPE columns
Reverse phase hydrophobic
| Phase | Pore volume (cm3/g) | Pore Size (A) | Surface Area (m2/g) | Carbon Load (%) | End Capping | Feature |
|---|---|---|---|---|---|---|
| C2 Ethyl | 0.77 | 60 | 500 | 6.6 | YES/NO | |
| C4 n-Butyl | 0.77 | 60 | 500 | 8.5 | YES | |
| C8 Octyl | 0.77 | 60 | 500 | 11.1 | YES/NO | |
| C18 Octadecyl | 0.77 | 60 | 500 | 21.7 | YES/NO | |
| C30 Tricontyl | 0.77 | 60 | 500 | 20.0 | YES | |
| Cyclohexyl | 0.77 | 60 | 500 | 11.6 | YES/NO | |
| Phenyl | 0.77 | 60 | 500 | 11.0 | YES/NO |
Normal phase hydrophilic
| Phase | Pore volume (cm3/g) | Pore Size (A) | Surface Area (m2/g) | Carbon Load (%) | Feature | |
|---|---|---|---|---|---|---|
| Silica | 0.77 | 60 | 500 | N/A | ||
| Diol | 0.99 | 60 | 500 | 8.0 | ||
| Cyanopropyl | 0.77 | 60 | 500 | 9.0 | ||
| Florisil | 0.82 | 60 | 500 | N/A | ||
| Alumina, Acidic | 60 | 500 | N/A | |||
| Alumina, Basic | 60 | 500 | N/A | |||
| Alumina, Neutral | 60 | 500 | N/A | |||
| Carbon | N/A | 120/140 mesh |
Ion Exchange - Anion Exchange
| Phase | Pore volume (cm3/g) | pKa | Pore Size (A) | Surface Area (m2/g) | Carbon Load (%) | Exchange (meq/g) |
|---|---|---|---|---|---|---|
| Aminopropyl (1 amine) | 0.77 | 9.8 | 60 | 500 | 6.65 | 0.31 |
| N-2 Aminoethyl (1/2 amine) | 0.77 | 10.1; 10.9 | 60 | 500 | 11.1 | 0.32 |
| Diethylamino (3 amine) | 0.77 | 10.6 | 60 | 500 | 10.6 | 0.28 |
| Quarternary Amine Chloride | 0.77 | Always charged | 60 | 500 | 8.4 | 0.25 |
| Quarternary Amine Hydroxide | 0.77 | Always charged | 60 | 500 | 8.4 | 0.25 |
| Quarternary Amine Acetate | 0.77 | Always charged | 60 | 500 | 8.4 | 0.25 |
| Quarternary Amine Formate | 0.77 | Always charged | 60 | 500 | 8.4 | 0.25 |
| Polyimine | 0.77 | Always charged | 13.5 | 0.25 |
Ion Exchange - Cation Exchange
| Phase | Pore volume (cm3/g) | pKa | Pore Size (A) | Surface Area (m2/g) | Carbon Load (%) | Exchange (meq/g) |
|---|---|---|---|---|---|---|
| Carboxylic Acid | 0.77 | 4.8 | 60 | 500 | 9.2 | 0.17 |
| Propylsulfonic Acid | 0.77 | 1 | 60 | 500 | 7.1 | 0.18 |
| Benzenesulfonic Acid | 0.77 | Always charged | 60 | 500 | 11.0 | 0.32 |
| Benzenesulfonic Acid, High Load | 0.77 | Always charged | 60 | 500 | 15.0 | 0.65 |
| Triacetic Acid | 0.77 | 60 | 500 | 7.61 | Anion 0.17/Cation 0.06 |
Copolymeric phases
| Phase | Pore volume (cm3/g) | pKa | Pore Size (A) | Surface Area (m2/g) | Carbon Load (%) | Exchange (meq/g) |
|---|---|---|---|---|---|---|
| Aminopropyl + C8 | 0.77 | 9,8 | 60 | 500 | 12,3 | 0,163 |
| Quarternary Amine + C8 | 0.77 | Always charged | 60 | 500 | 13,6 | 0,160 |
| Carboxylic Acid + C8 | 0.77 | 4,8 | 60 | 500 | 2,5 | 0,105 |
| Propylsulfonic Acid + C8 | 0.77 | 1 | 60 | 500 | 14,62 | 0,114 |
| Benzenesulfonic Acid + C8 | 0,77 | Always charged | 60 | 500 | 12.3 | 0,072 |
| Cyanopropyl + C8 | 0,77 | N/A | 60 | 500 | 14,6 | 0,163 |
| Cyclohexyl + C8 | 0.77 | N/A | 60 | 500 | N/A | N/A |
GC Phases
On this page we provide an overview of the supplied stationary phases for gas chromatography (GC). Each is given details of its properties and the applications that are suitable for them. In the product catalog you can then choose a suitable quartz or metal capillary column for GC.
Stationary phases LION
Fused silica capillary columns
| Stationary phase | Temperature range | Composition | USP Phase |
|---|---|---|---|
| LN-1 | -60 to 370°C | 100% dimethyl polysiloxane | G2 |
| LN-1 MS | -60 to 370°C | 100% dimethyl polysiloxane | G2 |
| LN-1 HT | -60 to 430°C | 100% dimethyl polysiloxane | - |
| LN-5 | -60 to 370°C | 5% diphenyl/95% dimethyl polysiloxane | G27 |
| LN-5 Sil MS | -60 to 370°C | 5% diphenyl/95% dimethyl polysiloxane | G27 |
| LN-5 MS | -60 to 350°C | 5% phenyl - arylene - 95% dimethyl polysiloxane | G27 |
| LN-5 HT | -60 to 430°C | 5% diphenyl/95% dimethyl polysiloxane | - |
| LN-35 | 50 to 360°C | 35% diphenyl/65% dimethyl polysiloxane | G42 |
| LN-35 HT | -60 to 400°C | 35% diphenyl/65% dimethyl polysiloxane | G42 |
| LN-17 | 40 to 340°C | 50% diphenyl/50% dimethyl polysiloxane | G3 |
| LN-624 | -20 to 260°C | 6% cyanopropylphenyl/94% dimethyl polysiloxane | G43 |
| LN-FFAP | 40 to 260°C | Nitroterephthalic Acid Modified Polyethylene Glycol | G35 |
| LN-1701 | -20 to 300°C | 14% cyanopropylphenyl/86% dimethyl polysiloxane | G46 |
| LN-XLB | 30 to 360°C | Low polarity phases | - |
| LN-XLB-HT | 30 až 400°C | Low polarity phases | |
| LN-WAX | 40 to 260°C | Polyethylene Glycol | G16 |
| LN-WAX Plus | 20 to 260°C | Polyethylene Glycol | G16 |
UHPLC
Ultra-High Performance Liquid Chromatography a milestone in the evolution of LC in that columns packed with <2µm particles, used with instrumentation capable of handling the resulting high back pressures, make possible extremely fast and efficient separations. UHPLC is a very powerful tool for today’s practicing chromatographer, as it can significantly increase the efficiency of a chromatographic separation. In addition, the wider range of usable flow rates makes high speed separations possible.
- Instruments
- UHPLC Columns
- UHPLC Performance on any LC instrument
- Applications and support
- Reagents
Effect on efficience
The lower the particle size, the higher column efficience is (see graph below). With particle size decrease the column back pressure increases significantly. This leads to ultra high pressure of longer LC columns. It means that we cannot use 1,9µm UHPLC column the same lenth like 5µm column (eg. 250 mm long column). In the end we have lower or similar column efficieny as we have with standard HPLC columns. The main benefit of the UHPLC columns is the analysis run time, not the efficiency.
If you need to increase efficiency, look at phase chemistry first. In this case look at stationary phase list.
UHPLC Phases
Raptor - RESTEK
| Packing Material | Particle Size (µm) | Pore Size (Å) | Effective Surface Area (m2/g) | Carbon Load (%) | pH Range |
|---|---|---|---|---|---|
| Raptor ARC-C18 | 1.8 | 90 | 125 | proprietary | 1.0-8.0 |
| Raptor ARC-C18 | 2.7 | 90 | 130 | proprietary | 1.0-8.0 |
| Raptor ARC-C18 | 5.0 | 90 | 100 | proprietary | 1.0-8.0 |
| Raptor C18 | 1.8 | 90 | 125 | proprietary | 2.0-8.0 |
| Raptor C18 | 2.7 | 90 | 130 | proprietary | 2.0-8.0 |
| Raptor C18 | 5.0 | 90 | 100 | proprietary | 2.0-8.0 |
| Raptor Biphenyl | 1.8 | 90 | 125 | proprietary | 1.5-8.0 |
| Raptor Biphenyl | 2.7 | 90 | 130 | proprietary | 1.5-8.0 |
| Raptor Biphenyl | 5.0 | 90 | 100 | proprietary | 1.5-8.0 |
| Raptor Fluorophenyl | 1.8 | 90 | 125 | proprietary | 2.0-8.0 |
| Raptor Fluorophenyl | 2.7 | 90 | 130 | proprietary | 2.0-8.0 |
| Raptor Fluorophenyl | 5.0 | 90 | 100 | proprietary | 2.0-8.0 |
| Raptor HILIC-Si | 2.7 | 90 | 150 | n/a | 2.0-8.0 |
| Raptor EtG/EtS | 2.7 | 90 | 130 | proprietary | 2.0-8.0 |
Raptor maximum pressure: 1,034 bar (1.8 μm), 600 bar (2.7 μm); 400 bar (5 μm). For maximum lifetime recommended maximum pressure for 1.8 µm particles is 830 bar.
PINNACLE DB - RESTEK
| Packing Material | Particle Size (µm) | Pore Size (Å) | Surface Area (m2/g) | Carbon Load (%) | pH Range |
|---|---|---|---|---|---|
| Pinnacle DB C18 | 1.9 | 140 | 150 | 11 | 2.5-8.0 |
| Pinnacle DB Aqueous C18 | 1.9 | 140 | 150 | 6 | 2.5-8.0 |
| Pinnacle DB C8 | 1.9 | 140 | 150 | 6 | 2.5-8.0 |
| Pinnacle DB CN | 1.9 | 140 | 150 | 4 | 2.5-8.0 |
| Pinnacle DB PFP | 1.9 | 140 | 150 | 6 | 2.5-8.0 |
| Pinnacle DB Biphenyl | 1.9 | 140 | 150 | 8 | 2.5-8.0 |
| Pinnacle DB IBD | 1.9 | 140 | 150 | proprietary | 2.5-8.0 |
| Pinnacle DB Silica | 1.9 | 140 | 150 | n/a | 2.5-8.0 |
Troubleshooting
Many GC and LC problems can be avoided with routine preventive maintenance. If you are seeking the cause of the chromatography problem, go step by step. Never make more changes in your instrument at the same time, otherwise you will never find, what caused the problem.
Select your category of chromatography to read more about the troubleshooting: (GC troubleshooting is not available yet)
GC troubleshooting
LC troubleshooting
puriFlash
puriFlash® Flash Cartridges
Interchim developed new technique for flash chromatography - Ultra Performance Flash Purification (UPFP) using special flash cartridges. The flash cartridges are in the form of regular or irregular silica. UPFP enables to run purifications with high purity of the yield and less solvent use.
Flash column selection
Flash column celection is available at this page.
