Policies and Guidelines

Policies and Guidelines

The BAF Policies and Guidelines section provides important information relative to sample and data handling, good laboratory practices, payment and other topics for internal and external users. BAF strongly encourages users to read the guidelines and policies. Should you have any questions regarding these policies and guidelines, please contact Jean Christophe Cocuron (Jeanchristophe.Cocuron@unt.edu).

Policies

The BioAnalytical Facility is open to any researchers at the University of North Texas and external users. Open access to this pay per use service laboratory will be granted when the user will have been trained by a qualified mass spectrometry personnel. BAF policies are listed below, and we encourage users to read them. Please, contact Jean Christophe Cocuron (Jeanchristophe.Cocuron@unt.edu) for any questions related to these policies.

Equipment Access
Users are required to be trained (contact Jean Christophe Cocuron) by one BAF mass spectrometry personnel prior to accessing the mass spectrometer instrument(s). In order to be added to the Outlook calendar corresponding to the mass spectrometer instrument(s) of their choice, the users will receive an invitation by email from the facility's lab manager. At this point, users will be able to login and schedule a time slot on a specific device. Key card access to the facility will be given once the user's training is completed. Please, contact Brier Lee-Renken (Brier.Lee@unt.edu) for key card access.
Scheduling and Cancellation
Users need to make sure that they schedule time on the right Outlook calendar associated with a specific mass spectrometer at least 24-48 hours prior to the analysis of their biological samples. Instrument time scheduled more than 2 weeks prior to the analysis is not permitted. Moreover, reserved time slots have to be accurate in order to avoid any overlap and, consequently delay other users. In the event that you have to cancel a reservation, you have to give a 24 hour notice by sending an email to the BAF staff. In the case that you need to extend your run time, you will have to notify the BAF staff ASAP. Then, they will decide to grant or deny your request. Billing will reflect the time slot selected through the Outlook calendar, meaning users will be fully charged even if they do not use the total allocated time slot. Users who fail to show up for the utilization of instrumentation, will be charged automatically for the allocated time slot. If scheduling issues occur more than three times, the BAF staff has the right to temporarily or permanently deny any access to the facility.
Instrumentation and Core Usage
Users are required to bring their own column, guard column, solvents, and supplies. Users have to prepare samples and solvents in accordance with the General GC/LC-MS Preparation Guideline. The BAF personnel reserves the right to deny the use of an instrument if a facility user is not following guidelines. Careless users will be financially responsible for any mass spectrometer repairs arising from usage of unconventional solvents to "dirty" samples. Every time users want to run samples through a mass spectrometer, they will have to sign in on the logbook. Users are responsible for installing, equilibrating, and conditioning their own columns. The calibration of the mass spectrometer will only be performed by the staff facility. Users are encouraged to run a known test sample in order to ensure that the system is working properly. Users are not allowed to modify or overwrite methods that are not part of their own folder. On the other hand, users have to make sure that the instrument is working properly before they leave. In the case of a malfunction of an instrument, users are required to send an email immediately to Jean Christophe Cocuron (Jeanchristophe.Cocuron@unt.edu). Users have to leave a clean laboratory working area without any used or dirty accessories as well as vials in the autosampler. Users are not allowed to share key cards with any of their co-workers. The door of the facility has to be closed after 5:00 p.m.

Failure to comply to these policies can lead to a loss of access to a specific instrument, and even into the BAF.
Computer
Each computer is configured to pilot a specific mass spectrometer instrument, and has internet access. No one outside of the BAF staff is allowed to install new programs or alter/uninstall existing software. No web-based searching or processing of acquired data is allowed while samples are running: this can result in computer hang-ups, and consequently stop the acquisition. Internet connectivity should only be used to facilitate the transfer of data, and not for any other purposes. Any personal web searching or chatting is strictly prohibited.
Data
The data acquired using a specific mass spectrometer are stored for a period of six months in the host computer. Then, they are transferred to an external hard drive. Note that users are responsible for handling and storing their own data.
Confidentiality
Data generated from samples analyzed through the BAF are considered confidential and will not be discussed nor shared with a third party without a written consent from the project leader(s).
Sample Retention
Direct users of the facility are responsible for the storage and disposal of their own samples. Do not leave vials in the autosampler. For external users, samples can be returned upon request at their expense. Otherwise, once data has been analyzed and sent to external users, samples will be stored for a month before disposal
Facility Acknowledgement and Publications
If you include data generated through the BAF in your presentations and/or manuscript, the facility must be acknowledged using the following wording: "The authors acknowledge the BioAnalytical Facility at the University of North Texas for the support with mass spectrometry analyses during this work".

Co-authorship in publications should be considered when one BAF personnel is providing a substantial intellectual contribution or customized analyses (for instance: method development, statistical analysis of metabolomics data, labeling analyses and interpretation, etc.). Please, contact the BAF Director Ana Paula Alonso (Anapaula.Alonso@unt.edu) prior to your manuscript submission for any questions regarding our contribution, and whether it justifies a co-authorship.
Payment
For internal users, payment has to be made using an InterDepartmental Transfer (IDT) form upon completion of the requested work.

For external users, a PO number has to be provided to initiate the requested work. Payment by check must be received prior to the release of the data/results.
Grant Support
Do not hesitate to contact the BAF Director Ana Paula Alonso (Anapaula.Alonso@unt.edu) if you need a letter of support for your grant application.
Conflicts
Any issues arising from access to the facility, scheduling calendar, instrumentation and core usage, computers, data confidentiality, authorship, and service quality to payment has to be reported to the BAF Director and/or BAF staff. They will meet with the facility user to discuss and solve the problem in accordance with BAF policies. If both parties do not find an agreement, the BAF Users Committee will get involved and will be asked to resolve the issue(s).

BAF Policy Agreement form to be completed and signed

BAF Policy Agreement Form

Guidelines

Use the following guidelines as you prepare samples and prep for the use of BAF.

General GC/LC-MS Preparation Guideline

The following guidelines should be followed when you prepare your samples for mass spectrometry.

Should you have any questions on what may or may not be used, please contact the BAF manager (Jeanchristophe.Cocuron@unt.edu).

LC-MS/MS Sample Preparation Guideline

Rules for LC-MS sample preparation

  • Samples MUST BE filtered to a 0.22 micron filter or 3 kDa Amicon filtering device prior to the submission to BAF.
  • Samples MUST BE completely dissolved prior to the injection through the mass spectrometer or LC-MS system.
  • For ESI and APCI analysis, please submit at least 50 µL or up to 100 µmol/L of biological sample solutions, and at least 5 mg of material for dried samples (standard or biological sample).
  • The liquid samples should be submitted in polypropylene vials or deactivated glass vials with PTFE or polypropylene-lined screw caps. Use a small volume insert (300 or 500 µL) if a limited amount of sample is available. For dried material, submit samples in 2 mL screw cap tubes.
  • All sample tubes/vials MUST BE labeled with a sample ID, requester name, and must be accompanied by the sample submission form and a form (Word or Excel document) listing the samples that will be analyzed.

Rules for LC/MS Solvents and Modifiers

Safe solvents (volatile, low molecular weight protic):

  • Acetonitrile
  • Methanol
  • Ethanol
  • Isopropanol
  • Water

Safe mobile phase modifiers:

  • Acetic acid (up to 1.0 %)
  • Formic acid (up to 0.1 %)
  • Ammonium hydroxide (up to 0.1 %)
  • Ammonium formate (salt concentration = 10 mM or less)
  • Ammonium acetate (salt concentration = 10 mM or less)

DO NOT USE

  • Nonvolatile salts (Tris, phosphates, citrates, and HEPES) can deposit in the source and plug capillaries.
  • Surfactants/Detergents such as SDS, CHAPS, PEG, Tween, and Triton that suppress electrospray ionization.
  • Inorganic acids (Sulfuric, nitric, and hydrochloric acids), which are corrosive and destructive for the LC/MS system.
  • DMSO, DMF, THF, acelate, and glycerol.
  • Trifluoroacetic acid (TFA) suppresses positive-ion electrospray at levels exceeding 0.01 %. Commonly used alternatives to high concentrations of TFA are mixtures of either 1 % acetic or 0.1 % formic acid with 0.025% TFA.
  • Hydrocarbon solvents (hexane and benzene) cannot be used for ESI. Triethylamine (TEA) yields an intense [M+H]+ ion at m/z 102 and suppresses positive-ion electrospray of less basic ions.
GC-MS Sample Preparation Guideline

Rules for GC/MS Solvents

Safe solvents (high volatility) for sample preparation:

  • Acetone
  • Acetonitrile
  • Benzene
  • Ethers
  • Hexanes
  • Methanol
  • Methylene
  • chloride

DO NOT USE/INJECT

  • Water, DMSO, DMF because they will damage the inlet of the GC system as well as the column. Therefore, DO NOT RESUSPEND your samples in water, DMSO or DMF.
  • Metals, strong acids or bases, salts, oligomers, and polymers should not be injected through the GC-MS system. These compounds will damage the inlet of the GC, the column, and the EI source used for the ionization of your metabolites.

Rules for GC-MS sample preparation

  • Try to minimize the use of "plasticizers" (tips, tubes…) when you perform your sample extraction and derivatization. This will help in minimizing contaminant peaks on your GC-MS chromatogram. Pipette tips can be replaced by Hamilton syringes.
  • Samples MUST BE filtered to a 0.22 micron filter or a 3 kDa Amicon filtering device prior to the submission to BAF.
  • Samples MUST BE volatile and completely dissolved in a non-polar solvent prior liquid injection through the GC-MS system.
  • The molecular weight from your metabolite(s) of interest has to be under 1,100 amu.
  • For liquid injection analysis, please submit at least 50 µL or up to 1000 µmol/L of derivatized biological sample solution, and at least 5 mg of material for dried samples (standard or biological sample).
  • For headspace or solid phase micro-extraction (SPME) injection analysis, liquid, wet/dried samples can be prepared directly in 20 mL screw cap vials and run through the GC-MS system as they are.
  • The liquid samples should be submitted in polypropylene vials or deactivated glass vials with PTFE or polypropylene-lined screw caps. Use a small volume insert (300 or 500 µL) if a limited amount of sample is available. For dried material submit samples in 2 mL screw cap tubes.
  • All sample tubes/vials MUST BE labeled with a sample ID, requester name, and must be accompanied by the sample submission form and a form (Word or Excel document) listing the samples that will be analyzed.