Easy 2-step sample preparation for LC-MS analysis on the ASSIST PLUS pipetting robot

  • Easy 2-step sample preparation for LC-MS analysis on the ASSIST PLUS pipetting robot

    LC-MS sample preparation

    Liquid chromatography mass spectrometry (LC-MS) is commonly used in analytical chemistry workflows to separate, identify, and quantify complex samples. It plays a vital role in areas such as environmental monitoring, cosmetics industry, and pharma. Varying pipetting and dilution steps during sample preparation, however, are time consuming and prone to error when performed manually.

    The VOYAGER adjustable tip spacing pipette on the ASSIST PLUS pipetting robot can access hazardous compounds from any tube format without exposing the user to danger. In addition, the ASSIST PLUS can take advantage of the D-ONE single channel pipetting module’s broad volume range and in combination with VIALAB worklist step, individual high volume sample dilutions are easier than ever. Reward yourself with reproducible results, additional hands-free time and profit from the versatility of the ASSIST PLUS.

  • Table of contents

    Liquid chromatography mass spectrometry (LC-MS) is commonly used in analytical chemistry workflows to separate, identify, and quantify complex samples. It plays a vital role in areas such as environmental monitoring, cosmetics industry, and pharma. Varying pipetting and dilution steps during sample preparation, however, are time consuming and prone to error when performed manually.

    The VOYAGER adjustable tip spacing pipette on the ASSIST PLUS pipetting robot can access hazardous compounds from any tube format without exposing the user to danger. In addition, the ASSIST PLUS can take advantage of the D-ONE single channel pipetting module’s broad volume range and in combination with VIALAB worklist step, individual high volume sample dilutions are easier than ever. Reward yourself with reproducible results, additional hands-free time and profit from the versatility of the ASSIST PLUS.

Key benefits

  • More walk-away time and reproducible results with automated sample preparation for LC-MS analysis on the ASSIST PLUS pipetting robot.
  • Risk-free automated compound handling on the ASSIST PLUS with the VOYAGER adjustable tip spacing pipette, because samples are reached from any tube or plate.
  • Efficient VIALAB parameter settings for pipetting speeds guarantee high precision when working with non-aqueous solutions. Adjustable mixing cycles help with the varying solubility of compounds.
  • Unbeatable very high volume sample dilutions with the D-ONE single channel pipetting module for the ASSIST PLUS. Two GRIPTIP sizes in one module enable high diluent and small sample volume pipetting without changing tools.
  • Foolproof liquid handling with automatic GRIPTIP® selection and liquid level detection (LLD), ensuring precision and accuracy when using the D-ONE on the ASSIST PLUS.
  • Effortless sample dilutions with VIALAB's worklist step allows the direct creation and import (e.g. LIMS) of .csv files. Instant worklist exchange without the need to change the program helps when adapting to different sample volumes.

Overview: How to make sample preparation for LC-MS analysis easy

The ASSIST PLUS pipetting robot operating a D-ONE single channel pipetting module

Downloads: App note and protocols for sample preparation for LC-MS analysis with the ASSIST PLUS pipetting robot

Experimental set-up

Experience reproducible results while having your hands free by using our easy sample preparation protocols for peptides, including high volume dilutions for LC-MS analysis (Figure 1).

Graphical representation of the ASSIST PLUS pipetting robot deck set-up with INTEGRA reagent reservoir, INTEGRA rack for microcentrifuge tubes and VOYAGER adjustable tip spacing pipette.
Figure 1: Experimental set-up of sample preparation for LC-MS analysis.

Step-by-step procedure

1. Dissolve samples in DMSO 

Dissolve freeze-dried peptides in 100 % DMSO

Place a 25 ml multichannel reagent reservoir filled with 10 ml of 100 % DMSO on deck position A (Figure 2, blue), and an INTEGRA rack with 1.5 ml microcentrifuge tubes containing freeze-dried peptides on deck position B (Figure 2, pink).

Select and run the VIALAB program ‘Dissolve_samples_in_DMSO'. With 1250 µl low retention, sterile, filter GRIPTIPS, the 8 channel 1250 µl VOYAGER adjustable tip spacing pipette aspirates 1 ml of DMSO from the reservoir (Figure 3a) and dispenses it into the 1.5 ml centrifuge tubes on deck position B (Figure 3b). Mixing 10 times at a gentle speed (6) ensures a homogeneous solution of peptides.

Tips:

  • Easy adjustment of mixing parameters in VIALAB helps mix reagents of different solubilities.
  • Accurate pipetting of DMSO with low retention GRIPTIPS and a reduced pipetting speed.
The ASSIST PLUS pipetting robot together with the 8 channel 1250 µl VOYAGER adjustable tip spacing pipette used for dissolving samples with DMSO from reagent reservoir in microcentrifuge tubes.
Figure 2: Deck set-up for dissolving samples in DMSO. Position A: Source – 25 ml multichannel reagent reservoir containing 100 % DMSO (blue). Position B: Target – INTEGRA tube rack for 1.5 ml centrifuge tubes with freeze-dried peptides (pink). Position C: Empty.
The ASSIST PLUS pipetting robot together with the 8 channel 1250 µl VOYAGER adjustable tip spacing pipette used for dissolving samples with DMSO from reagent reservoir in microcentrifuge tubes.
Figure 3: The VOYAGER transferring DMSO from a) a reservoir to b) microcentrifuge tubes.

2. Dilute samples in methanol

Worklist set-up for individual peptide dilutions

.csv files are created using the VIALAB worklist template (Figure 4) or imported directly into the VIALAB worklist step. The first worklist step includes the worklist for diluent distribution to analyze eight peptides in triplicates, and the second worklist step, the worklist for the different peptides.

Graphical representation of the VIALAB worklist template to create individual .csv files.
Figure 4: VIALAB worklist template for creating individual .csv files.

Tips:

  • Worry-free exchange of worklists for different sample dilutions without changing the VIALAB program.
  • Quick adaption to varying sample counts when deselecting samples in the worklist step.

Sample dilution with methanol in triplicates

A 25 ml multichannel reagent reservoir filled with 25 ml of 100 % methanol is placed on deck position A (Figure 5, blue), and dissolved peptides in 1.5 ml tubes with an INTEGRA rack on deck position B (Figure 5, pink). An INTEGRA rack with 24 empty HPLC vials is placed on deck position C (Figure 5, green).

Select and run the VIALAB program ‘Dilute_samples_with_methanol’. The D-ONE single channel pipetting module on the ASSIST PLUS pipetting robot with 1250 µl sterile, filter GRIPTIPS distributes diluent from position A to position C (Figure 6a) as indicated in ‘worklist_diluent’. An initial tip pre-wetting prevents dripping when pipetting volatile liquids by saturating GRIPTIPS with vapor. Decreasing the speed to 5 with a one second delay after pipetting followed by a tip touch against the side of the HPLC vial, increases accuracy when dispensing methanol. The D-ONE then individually dilutes peptides from position B to position C (Figure 6b) in triplicates using 125 µl low retention, sterile, filter GRIPTIPS, as indicated in ‘worklist_samples’. Every peptide is mixed five times with the full 125 µl GRIPTIP volume.

Tips:

  • Worry-free use of different sample aliquots with the D-ONE's LLD function.
  • Avoid dripping caused by vibrations with reduced movement speeds during pipette travel.
  • Risk-free methanol pipetting because the ASSIST PLUS fits easily in a fume hood.
Graphical representation of the ASSIST PLUS pipetting robot deck set-up with INTEGRA reagent reservoir, INTEGRA rack for microcentrifuge tubes, INTEGRA rack for HPLC vials and D-ONE single channel pipetting module.
Figure 5: Deck set-up for diluting samples in methanol. Position A: Diluent – 25 ml multichannel reagent reservoir containing 100 % methanol (blue). Position B: Source – INTEGRA rack for 1.5 ml centrifuge tubes with dissolved peptides (pink). Position C: Target - INTEGRA rack for 2 ml HPLC vials (green).
The ASSIST PLUS pipetting robot, together with the D-ONE single channel pipetting module, dilutes samples with methanol in HPLC vials.
Figure 6: The D-ONE pipetting a) methanol and b) peptides into HPLC vials.

Remarks

  • Low volume HPLC vials: Special low volume HPLC vials with inlets are compatible with the INTEGRA rack for HPLC vials and accessible by the D-ONE (Figure 7).
  • VIALAB software: The VIALAB programs can be easily adapted to your specific labware and protocols, for instance when partial plates are needed.
  • Partial plates: Programs can be adapted at any time to a different number of samples, giving laboratories total flexibility to meet current and future demands.
The D-ONE single channel pipetting module pipettes into 0.2 ml into low volume HPLC vials.
Figure 7: HPLC vial with inlet for low volume LC-MS analysis accessed by the D-ONE.

Conclusion

  • Remarkable dilution factors achieved with the D-ONE single channel pipetting module and the ASSIST PLUS pipetting robot, thanks to automatic selection of two different GRIPTIP sizes.
  • Save precious samples by using special low volume HPLC vials that are compatible with INTEGRA's tube rack and accessible by the D-ONE.
  • Reliable results with triplicate sample preparation and simplified individual dilutions using VIALAB's worklist step. The creation, import or exchange of .csv files is easier than ever providing fast adaptation to changing sample count.
  • Experience more flexibility when transferring liquids between different types of tubes and plates with the VOYAGER adjustable tip spacing pipette on the
    ASSIST PLUS.

Ask our expert. Leave a comment!

Write us if you have any questions regarding the application note or one of our instruments.

Any questions? I'm happy to help!

  • Rebecca Bodmer

    Application Specialist

    Email

Instruments and accessories

ASSIST PLUS, Pipetting Robot VOYAGER, 8 Channel, 1250 µl D-ONE Single Channel Pipetting Module, 0.5-300 µl Tip deck for D-ONE on ASSIST PLUS Rack for 1.5/2 ml microcentrifuge tubes Rack for 2 ml HPLC vials Multichannel Reagent Reservoir, 25 ml GRIPTIPS®, 125 µl, Sterile, Filter, Low retention GRIPTIPS®, 1250 µl, Sterile, Filter (for automation systems) GRIPTIPS®, 1250 µl, Sterile, Filter, Low Retention (for automation systems) Greiner Bio-One: Reaction Tube, 1.5 ml, PP, Natural, Attached Cap CARL ROTH: Sample vials ROTILABO®

ASSIST PLUS, Pipetting Robot

INTEGRA has developed the ASSIST PLUS pipetting robot to streamline routine pipetting tasks at an affordable price. Using INTEGRA electronic multichannel pipettes, the system:

  • automates pipetting tasks,
  • eliminates physical strain and
  • ensures superior reproducibility and
  • error free pipetting.

Part No. 4505

Visit product page
ASSIST PLUS Pipetting Robot

VOYAGER, 8 Channel, 1250 µl

VOYAGER pipettes allow the tip spacing to expand anywhere between 4.5 mm and 33 mm at the push of a button.

  • Single handed operation leaves the other hand free to handle labware.
  • On the fly access to up to 3 user-defined tip spacings.
  • These user defined tip spacings are saved. No need to memorize spacings of different labware formats.

Part No. 4724

See it work
VOYAGER, 8 channel series

D-ONE Single Channel Pipetting Module, 0.5-300 µl

The D-ONE single channel pipetting module offers hands-free transfers from individual tubes or wells on the ASSIST PLUS pipetting robot. This makes it the perfect add-on for labs that want to go beyond the possibilities of multichannel pipetting to access unlimited applications. It is designed to offer easy automation of normalization, hit picking and master mix preparation, freeing up your time while eliminating transcription errors.

Learn more about the D-ONE Single Channel Pipetting Module
D-ONE pipetting module 1ch 0.5-300ul

Tip deck for D-ONE on ASSIST PLUS

  • Supports accommodation of 1 x 12.5 µl/ 125 µl tip box and 1 x 300 µl/ 1250 µl tip box
  • Compatible with D-ONE Pipetting Module 0.5 - 300 µl and 5 - 1250 µl
Learn more about the Tip Deck
D-ONE tip deck

Rack for 1.5/2 ml microcentrifuge tubes

Hold up to 48 tubes on removable carriers for easy loading and storage.

Part No. 4540

Show all labware accessories
Rack for 1.5/2 ml microcentrifuge tubes

Rack for 2 ml HPLC vials

Part No. 4545

Find out more about accessories for ASSIST PLUS
Rack for 2 ml HPLC glass vials

Multichannel Reagent Reservoir, 25 ml

  • SureFlo™ anti-sealing array prevents pipette tip seal off which can cause liquid to pop into the tip, filter or even the pipette itself
  • Unique surface treatment spreads liquid evenly offering even lower dead volumes
  • Reusable base with bold, crisp, clearly visible graduation markings
  • Nest inside each other, making it possible to get twice as many reservoirs in half the space of other products on the market
  • Reducing inventory space requirements and shipping costs
  • Available as crystal clear polystyrene reservoirs or polypropylene reservoirs for improved chemical compatibility
Find out more about Multichannel Reagent Reservoirs
SureFlo™ anti-sealing array

GRIPTIPS®, 125 µl, Sterile, Filter, Low retention

INTEGRA uses a unique polypropylene blend to mold Low Retention GRIPTIPS® with heightened hydrophobic properties. Using Low Retention GRIPTIPS® prevents low surface tension samples from spreading out and “wetting” the inner wall of the tips, allowing them to bead-up for a maximum liquid recovery.

Part No. 6565

Learn more about low retention pipette tips
 Low Retention GripTips are offered in 6000 series racks as non-sterile, sterile and filter options.

GRIPTIPS®, 1250 µl, Sterile, Filter (for automation systems)

GRIPTIPS® pipette tips perfectly match the multi-lobe tip fitting, snapping firmly on during loading. This guarantees a perfect seal on every tip, preventing them from loosening, leaking or completely falling off. All tips are precisely aligned horizontally, enabling accurate touch-offs, even when pipetting with 384 tips.

Part No. 6445

Learn more about GRIPTIPS® pipette tips for Benchtop Pipetting Systems
Integra-Relaunch_Website-Output-Produktebilder-Product_Overview-GripTips.png

GRIPTIPS®, 1250 µl, Sterile, Filter, Low Retention (for automation systems)

GRIPTIPS® pipette tips perfectly match the multi-lobe tip fitting, snapping firmly on during loading. This guarantees a perfect seal on every tip, preventing them from loosening, leaking or completely falling off. All tips are precisely aligned horizontally, enabling accurate touch-offs, even when pipetting with 384 tips.

Part No. 6545

Learn more about GRIPTIPS® pipette tips for Benchtop Pipetting Systems
Integra-Relaunch_Website-Output-Produktebilder-Product_Overview-GripTips.png

Greiner Bio-One: Reaction Tube, 1.5 ml, PP, Natural, Attached Cap

  • Free of detectable DNase, RNase, human DNA
  • Non-Pyrogenic
  • High chemical and temperature resistance
  • Volume: 1.5 ml
  • With graduation and attached cap
  • Suitable for Eppendorf

Part No. 616201

Source: Website Supplier

Visit supplier website
Reaction tube, 1,5 ml, PP, natural

CARL ROTH: Sample vials ROTILABO®

Sample vials ROTILABO® with snap-on ring ND11, Clear glass, with labelling space, 1.5 ml

Part No. TY80.1

Source: Website supplier

Visit supplier website

Customer’s voice

Implementing this liquid handler in our molecular workflow has cut production time dramatically with the ease and reproducibility this instrument offers. Prep can be done in half the time and supports new lab personnel carry out multiple steps in our workflow with accuracy and confidence. The flexibility of the unit is also incredible with the ability to edit programs in real time or on the pc, reassign pipettes to base units freely as workflows require. This liquid handler is a staple in our laboratory.

Caitlin Osborn, Gentech Diagnostics