ISBT Bev Grade Nitrogen QualityGuidelines Analytical Methods Ref

Beverage Grade
Nitrogen
(Cryogenic Source)
Quality Guidelines
And
Analytical Methods
Reference
2015
Beverage Grade Nitrogen (Cryogenic Source)
Quality Guidelines
And
Analytical Methods Reference
Copyright © 2015 by the International Society of Beverage Technologists (ISBT).
All rights reserved.
Portions of this document may be used in other publications by including the following statement:
‡ Original Source: ISBT Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines and Analyt-
ical Methods Reference © 2015
This document may not be reproduced in its entirety, except for the purpose of translating into an-
other language. Non-English translations must include the following statement:
‡ Original Source: ISBT Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines and Analyt-
ical Methods Reference. The ISBT is not responsible for the accuracy of this translation from original
English source text. It is up to the user of this translated document to ensure that it corresponds with
the most current version of the ISBT source document.
Complete English-language documents may be obtained online from the ISBT at

www.bevtech.org.
Revision # Release Date Revision Notes

0 – Original March 2015 Original: Beverage Gas Committee
International Society of Beverage Technologists

14070 Proton Rd., Dallas, TX 75244-3601
P: 1-972-233-9107 x 208 F: 1-972-490-4219
Email: Office@Bevtech.org
Webpage: www.bevtech.org
Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines

Issue Date: March 2015 Page 2 of 72
Beverage-Grade Nitrogen (Cryogenic Source)
Quality Guidelines
And
Contents
Part 1 Beverage-Grade Nitrogen Quality Guidelines Page
1) Introduction ---------------------------------------------- 4
2) Quality Guidelines Table ---------------------------------------------- 5
3) Glossary and Definition of Terms ---------------------------------------------- 6-10
4) Beverage Plant LIN Operational Factors ----------------------------------- 11-12
5) References ---------------------------------------------- 12
6) Quality Control Sampling & Testing Frequencies -------------------------- 13
7) N2 Certificate of Analysis / Compliance -------------------------------------------- 13
8) Acknowledgements --------------------------------------------------------- 14
9) Appendix --------------------------------------------------------- 14
Part 2 Analytical Methods for N2 Analysis
N2 Analysis – Preamble -------------------------------------------------------- 15

Equivalent Analytical Methods -------------------------------------------------------- 15
Sampling and Testing Equipment -------------------------------------------------------- 16
Analytical Methods and Container Suitability for N2 Analysis (Table Summary) 17
LIN Sampling Safety Considerations ---------------------------------------------------- 18
Part 3 Method List Method # (Pages)
Sampling Using Cup-In-Metal Cylinders --------------------------------------------- 1.0N (19-23)

Sampling Using High Pressure Metal Cylinders ----------------------------------- 1.1N (24-27)
Sampling Using Small, Low Pressure Metal Cylinders -------------------------- 1.2N (28-31)
Sampling Using Polymeric Sampling Bags ----------------------------------------- 1.3N (32-35)
Identification ------------------------------------------------------------------ 2.0N (36-41)
Trace Oxygen ------------------------------------------------------------------ 3.0N (42-48)
Water Vapor ------------------------------------------------------------------ 4.0N (49-53)
Total Volatile Hydrocarbons ------------------------------------------------------- 5.0N (54-57)
Carbon Monoxide ------------------------------------------------------------------ 6.0N (58-63)
Odor ------------------------------------------------------------------ 7.0N (64-68)
% Purity Calculation ------------------------------------------------------------------ 8.0N (69-71)
Analytical Method Bibliography ---------------------------------------------------- 72

1) Introduction
This quality guideline focuses on defining the purity grade of nitrogen (N2) appropriate for use in
associated beverages along with providing recommended practices for its processing, safe handling,
transport, storage, sampling, sample shipping and analytical testing. All efforts were made to harmonize this
revision with Compressed Gas Association (CGA) and European Industrial Gases Association (EIGA)
standards. An expert international committee comprised of beverage manufacturers, N2 producers, supply
chain manufacturers, analytical service providers, and filter suppliers developed these guidelines according to
best available practices.
This document is intended to serve as an educational resource for N2 suppliers and beverage manufacturers.
CGA Publication G-10.1 “Commodity Specification for Nitrogen” and the EIGA Document IGC Doc 126/04/E
“Minimum Specification for Gases used in Food Applications” also provide data concerning N2 use in
beverages.
The nitrogen used in non-carbonated beverage production is predominantly produced through the liquefac-
tion of ambient air via cryogenic cooling, followed by fractional distillation and selective recovery. This docu-
ment does not address quality issues associated with the production of N2 from other, non-cryogenic pro-
cesses (ex. pressure-swing or membrane-enrichment). Trace impurities remaining in cryogenic-sourced N2
are largely dependent on the details of the manufacturing process.
Nitrogen is primarily used in the beverage industry for non-carbonated beverage production. It is also used
for bottle rigidification, purging, inerting, de-aerating, and undercover gassing as a substitute for carbon diox-
ide.
Once processed to beverage-grade level, the N2 final product must then be properly transported and stored
to prevent contamination. All applicable principles of safety and good manufacturing practices for food addi-
tives, ingredients, technical and processing aids should be considered throughout this supply chain from pro-
duction through final point-of-use. N2 quality management is essential for ensuring customer satisfaction.
It is the responsibility of the gas supplier to identify, measure, and control the levels of impurities listed in this
ISBT guideline to produce beverage-grade quality N2. Similarly, beverage manufacturers have an
independent responsibility to ensure that their suppliers consistently meet this desired purity level.
This guideline is not intended to be a comprehensive quality document. The impurity parameters listed are of
key concern to all non-carbonated beverage manufacturers as they are known to negatively impact the char-
acter of a non-carbonated beverage. All guideline limits are voluntary, achievable and measurable.
The International Society of Beverage Technologists (ISBT) does not warrant the efficacy, accuracy or com-
pleteness of these guidelines.

2) Quality Guidelines for Beverage Grade N2
Parameter Guideline Limit Rationale1
N2 Purity2 99.99 % v/v min Process
Oxygen (O2)3 10 ppm v/v max Sensory
Total Volatile Hydrocarbons4 (THC) as

Sensory /
Methane (CH4) 5 ppm v/v max
Process
Process /
Carbon monoxide4 (CO) 10 ppm v/v max
Regulatory
Odor No foreign odor Sensory

Note 1: Rationale definitions:
Sensory: Any attribute that negatively impacts the taste, appearance, or odor of a beverage.
Process: Any attribute that defines a key parameter in a controlled process and an important
consideration in the beverage industry.
Regulatory: Any attribute whose limit is set by a governing regulatory agency and related to food
safety.
Note 2: Definition of N2 Purity: Unless otherwise required by the customer, the % N2 value typically includes
minor quantities of inert argon (Ar) and neon (Ne) not removed during cryogenic nitrogen production.
Calculated N2 % Purity = 100.00% v/v minus the % v/v sum of all target list impurities (See Method 8.0N).
Note 3: O2 content is typically measured in real time using an on-line process analyzer during the N2 produc-
tion process.
Note 4: The likelihood of high THC and CO impurity levels from a cryogenic source is considered negligible;
however, any trace CO that may be present under normal ambient conditions will accumulate in the nitrogen
during production. The supplier’s risk assessment program must account for this impurity so periodic CO test-
ing, while considered optional in most applications, is advised.
Most beverage products will benefit from using ISBT beverage-grade N2 as described above. Bever-
age manufacturers should evaluate the sensitivity of their products to the trace levels of impurities listed
above. For some very sensitive beverage products, lower maximum allowable levels of one or more of these
parameters may be required. Such limit modifications shall be defined and agreed-to between a supplier and
end-user before cryogenic nitrogen is brought on-line.

3) Glossary and Definition of Terms
Acronyms
The following section of the Guideline explains the acronyms used and defines the technical terms refer-
enced herein. Wherever these acronyms and defined terms appear in this document they will be printed in
italicized type.
ANSI: American National Standards Institute (U.S.)
ASME: American Society of Mechanical Engineers (U.S.)
CFR: Code of Federal Regulations (U.S.)
CGA: Compressed Gas Association (U.S.)
DOT: Department of Transportation (U.S.)
EEC: European Economic Community
EIGA: European Industrial Gases Association
EU: European Union (includes 25 member states)
FAO: Food Additives Organization
FDA: Food and Drug Administration (U.S.)
GHS: Global Harmonization System (UN)
IATA: International Air Transport Association
ISBT: International Society of Beverage Technologists
ISO: International Standards Organization
JECFA: Joint FAO / WHO Expert Committee on Food Additives
NFPA: National Fire Protection Association (U.S.)
NIST: National Institute of Standards and Technology (U.S.)
OSHA: Occupational Safety and Health Administration (U.S.)
TC: Transport Canada
TPED: Transportable Pressure Equipment Directive (Europe)
UN: United Nations
WHO: World Health Organization

Definition of Terms
Asphyxiant: Any gas that can cause suffocation by displacing some or all of the oxygen normally present in
ambient air. Note that some gases (such as carbon dioxide) may be hazardous at concentrations lower than
that required to suffocate by simple displacement of oxygen.
Air Separation Unit (ASU): A process for liquid nitrogen production that employs ambient air as a feed gas,
then cryogenically liquefies the air, fractionally distills it into its component gases (ex. N2, O2, Ar and other
minor gases), collects and purifies each fraction and stores them as liquefied pure gases.
Batch: In a continuous process, a batch corresponds to a defined portion of the production based on time or
quantity.
Beverage Grade: A minimum N2 quality and purity designation for non-carbonated beverage applications
based on the ISBT publication guideline “Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines
and Analytical Methods Reference.”
Boiling Point: The temperature that a liquid’s vapor pressure equals the prevailing external pressure (ex.
boils). N2 will boil at well below room temperature; therefore it exists only in the gas phase after equilibration
under ambient conditions.
Certificate of Analysis (CoA): An official (signed) document typically verifying the actual analytical results
obtained on a specific container of N2 product at a specific point in the delivery process.
A CoA provides traceability of the quality for the parameters analyzed. Normally, a CoA is only provided by
the supplier if specifically required by contract, or if requested by the customer.
A CoA issued at a manufacturing plant may be used as a reference to generate a Certificate of Compliance
(CoC) for deliveries from that location.
Certificate of Compliance (CoC): (ex. also described as a Certificate of Conformance). An official document
that certifies a N2 product meets a previously agreed-upon quality specification based on quality procedures
and handling of the material; but does not include actual data from analysis of the shipping container.
Compressed Gas: Any material (or mixture) that exerts in the packaging an absolute pressure of 40.6 psia
(280 kPa) or greater at 68°F (20°C) or any flammable or toxic material that is a gas at 68°F(20°C) or less
and 14.7 psia (101.3 kPa) of pressure.
Confined Space: An area that is:
1.) large enough and so configured that an employee can bodily enter and perform assigned work.
2.) has limited or restricted means for entry or exit.
3.) is not designed for continuous employee occupancy.
Improper N2 storage and usage in confined spaces can result in potentially hazardous situations.
Contaminant / Contamination: An undesirable component found in N2 which is introduced after the N2 has
been tested and certified by the supplier. Contaminants may originate from some type of leak, mishandling of
the product, or mistakes occurring during transport, intermediate storage (cross-contamination), delivery or
bottling process. Contamination can be avoided by following proper procedures and quality guidelines
throughout the entire delivery and bottling process.

Cup-in-Cylinder Sampler (ex. Cosmodyne® type): A specialized cylinder (ex. 4 L) designed for safely and
directly sampling the liquid phase of various cryogenic liquids including LIN. A liquid phase sample is allowed
to flow through this cylinder, which involves the chilling and filling of a small internal “cup” with the liquid. The
cylinder body itself is isolated and does not fill with liquid. The final step in this process involves cutting off the
flow of liquid stream and opening an internal valve which allows the cup-trapped liquid to vaporize and fill the
sample cylinder body. In this manner a safe level of pressure is generated (typically 300 – 500 psig [2,070 –
3,450 kPag]).
Flame Ionization Detector (FID): An instrument component common to gas chromatographic and total hy-
drocarbon analyzers which consists of a mini-hydrogen (H2) / air burner and associated electronics. An FID
produces an electronic signal in the presence of many volatile organic hydrocarbon (VOC) class impurities.
This signal is used to quantify the amount of either individual VOC impurities, or as a “Total” amount versus a
methane (CH4) standard.
Flammability Rating: The NFPA, (U.S.) rating for N2 is zero (0), which indicates the material will not burn.
This rating is found in the top (red) box of the NFPA Diamond (U.S.).
Flash Vaporization: A thermally-assisted process designed to instantly convert an enclosed volume of liquid
N2 sample along with its impurities into a homogenous, representative gas phase sample for subsequent
analysis.
Gas Cylinder / Cylinder: An uninsulated, cylindrically shaped, pressure-containing device (see Figure 1)
with a water capacity not greater than 120 U.S. Gal (454 L) designed to withstand an internal pressure great-
er than 40 psia (175 kPa, abs) as defined by DOT, TC, UN or TPED (see References Section 5).
Figure 1 – Gas Cylinder
Hazard Analysis Critical Control Points (HACCP): A systematic approach to the identification, evaluation,
and control of food safety hazards.
Hazard Analysis and Risk-Based Preventative Control (HARPC): A systematic approach to establishing
standards for conducting hazard analysis, documenting hazards, implementing preventive controls, and doc-
umenting the implementation of the preventive controls.
Hazard Class (Division): A universal parameter (ex. see CFR - 49 (U.S.), IATA and GHS references) used
in classifying materials designated as dangerous goods for their safe transport and storage. Liquefied, refrig-
erated liquefied and compressed N2 gases are designated a Hazard Class of “2.2” which indicates that they
are considered “non-flammable, non-toxic gases” by definition. Rectangular hazard labels associated with
this class are colored green and “2.2” marked.
Hazardous Materials (Hazmats) / Dangerous Goods: Those substances that have been listed in, for
example, CFR - 49 (U.S.), IATA and GHS regulations (see Reference section) that because of their chemical
and / or physical properties must be handled to avoid health risks, injury or physical damage during their
transport and storage. Liquefied, refrigerated N2 and compressed N2 gas are listed as “hazardous materials.”
Note: Non-compressed (less than 29 psig [200 kPag] @ 20oC [68oF]), gaseous N2 is NOT classified as a
Hazmat.

Headspace Gas (Vapor): This term refers to the gas phase region above a contained charge of liquefied gas
(ex. LIN). Headspace gas is produced by the direct evaporation of the liquid phase below it.
Health Rating: The health rating for a material is found in the left (blue) box of the NFPA Diamond (U.S.).
The NFPA and GHS rating for liquefied cryogenic N2 is three (3), which indicates that the material can, upon
short-term exposure, cause serious temporary or residual injury. The NFPA and GHS rating for gaseous N2
are zero (0), which indicates that the material poses no health hazards upon exposure.
Impurity: An undesirable component that may be found in N2, coming from either the initial feed gas source
or from the manufacturing process. Processing and purification of the feed gas N2 by the supplier should re-
move all impurities of concern before it is designated as finished product for delivery to customers. Undesired
components that may be accidentally introduced to the product after this point are considered contaminants.
Liquefied Compressed Gas: A gaseous component or mixture converted to the liquid phase by cooling or
compression; also, any gas which remains partially liquid when placed in a container at 68ºF (20ºC).
Liquefied Nitrogen (LIN): A cryogenically cooled and refrigerated liquid form of nitrogen (N2). LIN is pro-
duced at ASU facilities and, for efficiency, bulk shipments of N2 are typically transported and stored as LIN.
Local Regulations: For the purposes of this document, local regulations include relevant local municipality,
city, state, federal, national, or international requirements that must be followed at the location of use.
Low Pressure Metal Cylinders (LPMC): A low pressure cylinder designed for obtaining N2 samples from a
flash vaporized and pressure controlled liquid N2 source or a compressed N2 cylinder.
Nitrogen (N2): Nitrogen (N2) is a colorless, odorless, non-flammable, relatively inert gas. One liter of N2 gas
(28.0134 g/mol) weighs approximately 0.8 grams at 0°C. When chilled to a very low (cryogenic) temperature
(-196 °C), N2 converts into a volatile, colorless liquid state (LIN). Bulk N2 is transported and stored in its lique-
fied state within specially insulated containers. Only small orders of N2 are typically shipped as compressed
gases in high pressure cylinders. The primary function of N2 use in a packaged non-carbonated beverage is
to act as an inert gas blanket which protects the beverage from any contact with O2. This protection helps to
preserve the sensory character of the beverage and also increases its shelf life. The absence of O2 in N2
blanketed beverages also helps to protect them from various forms of microbial contamination. Compressed
N2 gas is also used in line purging and liquid pushing operations.
Non-Hazmat Shipment: Shipments of non-compressed N2 samples to an analytical testing lab. A variety of

sample containers including polymeric sampling bags and low pressure metal cylinders typically comprise a
non-hazmat sampling / shipping kit. Non-compressed samples of gaseous N2 do not meet the definition of a
“hazardous material” and therefore can be properly and legally shipped as “non-hazardous goods of no
commercial value”.
Polymeric Sampling Bag (PSB): A low pressure, balloon-like sample storage container of typically 1 - 5 L
capacity comprised of cleaned, inert gas-impermeable, multilayer polymeric film and 1 or two valves. These
containers are primarily used for non-hazardous shipping of N2 samples to an analytical testing lab. PSB’s
used for sampling beverage-grade N2 must not outgas any target impurities into the sample, exhibit oxygen
and moisture impermeability and be capable of maintaining the integrity of all impurities and contaminants to
be tested to within approximately 90% of their original concentration for at least 15 days (see Sampling Meth-
od 1.3N for more details). Single valve equipped PSB’s are not recommended for performing trace oxygen,
moisture or % N2 purity tests. Dual-valved PSB’s designed for thorough and efficient sample flushing and
comprised of gas impermeable films may be used for all guideline tests.
ppb: Trace concentration level expressed as parts per billion (parts per 109) by volume of impurity per vol-
ume gas (v/v) unless otherwise stated.
ppm: Trace concentration level expressed as parts per million (parts per 106) by volume of impurity per vol-
ume gas (v/v) unless otherwise stated.

Proper Shipping Name: A universal parameter (ex. see CFR – 49, IATA and GHS references) used in clas-
sifying materials designated as dangerous goods for safe transport and storage. LIN is properly identified on
all shipping containers as “Nitrogen, refrigerated liquid” and compressed nitrogen gas cylinders must be des-
ignated as “Nitrogen, Compressed”. Associated with the “proper shipping name” is a unique “UN” number
which also linked to the proper shipping name. For example, UN1977 is linked to “Nitrogen, refrigerated liq-
uid”, and UN1066 is linked to “Nitrogen, compressed” for hazardous shipping activities.
Regulator (Pressure): A mechanical device used in a gas delivery system to reduce the internal pressure of
the system from a higher (source) pressure to a lower (delivery) pressure via an internal mechanism such as
a diaphragm. Regulators may be made of many types of materials (brass, stainless steel, etc.), and have
varying designs (ex. single-stage, two-stage) based upon their intended usage. For N2 sampling and testing
applications the regulator should be equipped with a metal diaphragm.
Relief Valve (also Pressure Relief Valve, Safety Relief Valve): A safety device actuated by inlet static
pressure and designed to open during an emergency or any abnormal conditions to prevent a rise of internal
fluid pressure in excess of a specified value.
The valve may be either a single-use type such as a rupture disc, or a re-usable (spring-loaded) type. If a
single-use type relief valve is over-pressurized, it will vent the entire contents of the source container, and will
require replacement before the system can be put back into service.
A re-usable relief valve such as a spring-loaded device will vent excess pressure until an acceptable level is
reached, at which point the valve will re-seat, allowing the system to function normally until another over-
pressure condition occurs.
All relief valves should be properly sized and piped to vent outside the facility away from egress, structures,
and personnel.
Residual Pressure Valve: A gas cylinder safety device that prevents a cylinder from being completely emp-
tied, ensuring there is always a small amount of positive pressure kept in the cylinder. This means that when
an empty cylinder is stored outside awaiting collection, moisture cannot ingress from the atmosphere and
contaminate the cylinder. Without a residual pressure valve, empty cylinders could be left with the valve fully
open and atmospheric contaminants could enter the cylinder, contaminating the contents when the cylinder is
refilled.
Safety Data Sheet (SDS): A summary of relevant technical data related to the physical and chemical proper-
ties, toxicity, safe handling, routine use practices and appropriate emergency response actions associated
with a specific chemical, commercial product or agent. AnSDS is typically issued by the manufacturer of the
material.
Specific Gravity: The ratio of the density of a substance to the density of water or air (unit less). The specific
gravity of N2 =1.0 which indicates that it is roughly equivalent to normal room air (which is approximately 78%
N2).
Vapor Pressure: The pressure that a liquid and its vapor are in an equilibrium state at a given temperature.
Liquefied gases (ex. LIN) with high vapor pressures evaporate very rapidly.
Vaporized Liquid Phase Sample: This refers to a gas sample generated by direct vaporization of a “metal-
cup-in-cylinder” contained cryogenic liquid sample which is in turn taken from a “below surface” point in a
Bulk LIN tank. This type of gaseous sample should be considered uniquely different from a “headspace gas”
sample taken directly from the gas phase within a LIN storage tank.

Classification: LIQUID N2 OPERATIONAL PRACTICES Revision Date: 03/2015
BULK RECEIVERS AND PRESSURE BUILDING VAPORIZERS Page 1 of 2
4) Beverage Plant LIN Operational Factors
All LIN storage vessels and associated hardware at a beverage plant should comply with common
industry standards for construction and safety.
On a Regular Basis
 Inspect valves and pressure relief devices for leaks. Service as needed.
 Check pressure gauges and liquid level.
 Check for unusual build-up of ice on the storage vessel.
 Inspect tank and distribution lines for leaks and any signs of corrosion. Repair as needed.
Annually
 Schedule external preventive maintenance and safety inspection with manufacturer.
Typical Maintenance Procedures for LIN Bulk Receiver Operations
Introduction
The following procedures are provided as a guide for the routine maintenance of bulk tanks. A supplier can
provide the specific training required for maintaining your system.
An annual LIN system inspection should be scheduled with your supplier.
Purpose
LIN in bulk quantity is typically stored in vacuum insulated storage tanks or receivers at tank pressures typi-
cally in the range of 20-150 psig (130-1,000 kPag), (higher storage pressure is also possible).
Important: Beverage plant processes may be configured to draw N2 from the liquid phase, gas phase, or
both.
Internal Cleaning Procedures for Bulk LIN Storage Vessels
Important Note:
LIN vacuum-insulated storage tanks are manufactured to ASME, TPED or other standards. The internal sur-
face is made from materials suitable for contact with food and beverage applications.

Classification: LIQUID N2 OPERATIONAL PRACTICES Revision Date: 03/2015
BULK RECEIVERS AND PRESSURE BUILDING VAPORIZERS Page 2 of 2
No internal cleaning procedures are required under normal operating conditions. If there is concern
for tank contamination, the customer should work with the supplier to return the tank to beverage
standard.
Caution: Attempting to conduct internal cleaning is not recommended.
Companies producing LIN for beverage applications should have a risk based, food safety program* de-
signed to address product quality in a manner that ensures that the beverage gases it manufactures and
supplies as a food or beverage ingredient, additive, or processing aid can be safely consumed in the final
food or beverage product.
*Note: Examples of risk-based food safety programs include HACCP and HARPC.
5) References
Compressed Gas Association (CGA): P-1 ---- 2008 “Safe Handling of Compressed Gases in Containers”
Eleventh Edition
Compressed Gas Association (CGA): G-10.1 ---- 2008 “Commodity Specification for Nitrogen” Seventh Edi-
tion
Compressed Gas Association (CGA): P-9 ---- 2008 “The Inert Gases Argon, Nitrogen and Helium” Fourth
Edition
Compressed Gas Association (CGA): P-12 ---- 2009 “Safe Handling of Cryogenic Liquids” Fifth Edition
European Industrial Gases Association (EIGA): IGC 115/12/E “Storage of Cryogenic Air Gases at Users
Premises”, 2012
European Industrial Gases Association (EIGA): IGC 44/09/E “Hazards of Inert Gases and Oxygen Depletion”,
2009
European Industrial Gases Association (EIGA): IGC 114/09/E "Operation of Static Cryogenic Vessels", 2009
European Industrial Gases Association (EIGA): IGC 126/011/E “Minimum Specifications for Food Gas Appli-
cations”, 2011
European Union: Regulation (EC) No. 1333/2008 of the European Parliament and of the Council of 16 De-
cember 2008. “Regulations for the Specification of Food Additives”, Official Journal of the European Union
31.12.2008, L 354.COMMISSION REGULATION (EU) No 231/2012 of 9 March 2012 laying down specifica-
tions for food additives listed in Annexes II and III to Regulation (EC) No 1333/2008 of the European Parlia-
ment and of the Council
Joint FAO/WHO Expert Committee on Food Additives (JECFA): 53rd JECFA Meeting (1999), “Compendium
of Food Additive Specifications”: Addendum 7, Issue 52.
US Department of Transportation. Hazardous Materials Regulations (HMR), Code for Federal Regulations
(CFR), Title 49 Transportation, Parts 100-185, Oct. 2013.
International Air Transport Association (IATA), “Dangerous Goods Regulations”, 54th Ed. effective January
2013.
United Nations Economic Commission of Europe (UNECE) “Globally Harmonized System of Classification
and Labeling of Dangerous Goods”, (GHS 5th Rev.) Ed. 2013.
United Nations Economic Commission of Europe (UNECE) “Recommendations on the Transport of Danger-
ous Goods”, 17th Rev. Ed. 2011.
Classification: N2 SAMPLING & TESTING FREQUENCIES Revision Date: 03/2015
Page 1 of 1
6) Quality Control Sampling & Testing Frequencies
The nitrogen producer should conduct an initial qualification and periodic quality verifications by sending
samples to a properly accredited laboratory annually.
7) Example - Certificate of Analysis (CoA) and Compliance (CoC)1
Nitrogen - Certificate of Analysis & Compliance Date of Issue

mm/dd/yyyy
Distributor:
Distributing Equipment (tank truck, container, rail car):
Delivery Date (yy/mm/dd):
Lot or Delivery Number:

Truck Number:
Manufacturing Plant:
ISBT
Parameter Guideline Limit Actual Load Result
N2 Purity 99.99 % v/v min _____________________
Oxygen (O2) 10 ppm v/v max _____________________
Date and Time of Analysis:
Report Summary: The delivered product meets the purity grade specification as agreed to between suppli-
er & customer.
Authorized Signature: _______________________ Date: ________________
Printed Name: ______________________________ Title: ________________
Note 1: Considerable flexibility concerning the contents of a CoA /CoC document exists in practice. All beverage manu-
facturers should evaluate these CoA / CoC recommendations and adjust them as needed to match their own quality pro-
gram objectives and circumstances of use. In all cases, a N2 producer and bottler must clearly define and agree to an
acceptable “CoA / CoC” program as well as the details of document issuance and application.

8) Acknowledgements
The ISBT would like to thank the members from the following companies and associations who participated in
and contributed to the publication of this ISBT Beverage Grade Nitrogen (Cryogenic Source) Quality Guide-
line:
- Airborne Labs International, Inc.

- Airgas
- Air Liquide
- Air Products and Chemicals, Inc.
- Analytical Science & Technology
- Atlantic Analytical Laboratory, LLC
- Dr Pepper Snapple Group
- Linde
- MicroPure Filtration
- NuCO2
- Parker Hannifin
- PepsiCo
- Praxair
- Reliant Holdings
- The Coca-Cola Company
- Unisensor Sensorsysteme GmbH
9) APPENDIX - Hazards Associated with N2

The section that follows was transcribed by permission from the EIGA IGC Doc 66: “Refrigerated N2 Storage
at Users Premises”.
HAZARDS OF N2
LIN / Gaseous N2 Handling Safety
Refer to the current edition of the N2 Safety Data Sheet as provided by the gas supplier for information re-
garding the hazards associated with LIN and gaseous N2.

ANALYSIS - PREAMBLE
The procedures and recommended “Best Practice / State-of-the-Art” analytical test methods that fol-
low are the collaborative works of ISBT members, which include recognized international analytical experts
from N2 producers, commercial laboratories and beverage manufacturers. Every method was written with the
intent of recommending several appropriately verified analytical measurement systems that have been suc-
cessfully employed by many beverage industry groups for the required application. In most cases, acceptable
equivalent methods are also noted in the method’s “Discussion and Notes” section. Appropriately verified,
equivalent alternate methods may be substituted upon agreement between the N2 supplier and end-user.
Phase sampling recommendations are included for each impurity parameter which must be carefully
followed. The vast majority of test methods require that the liquid N2 phase be flash-vaporized and sampled.
A representative sample of N2 is absolutely essential to obtain for either on-line analyzers or grab samples
taken in containers for laboratory testing. Recommendations concerning the location and frequency of sam-
pling are offered in various sections of this document.
Any laboratory or on-site instrumentation group intending to apply these methods should be experi-
enced in N2 testing and adhere to basic “good laboratory practices (GLP’s)” which include recommended
safety and result confirmation practices. Key safety and test result diagnostic / confirmation issues are thor-
oughly discussed in all ISBT methods.
The information contained in the “Performance Criteria” section of all methods defines the key per-
formance features needed for each impurity measurement. This includes recommendations for measurement
range, accuracy, precision, detection levels, interferences that may be encountered and other related figures-
of-merit. When appropriate, it is suggested that the method user obtain assurances from the analyzer manu-
facturer that their systems can achieve these ISBT performance guidelines. All methods also discuss a rec-
ommended calibration and verification program involving certified gas standards obtained from industry-
recognized, reputable manufacturers (ex. NIST traceable if appropriate).
Equivalent Analytical Methods
It is recognized that many types of analytical test methods can be used to properly measure a given
impurity in beverage grade, liquid N2 (LIN). The intent of the ISBT N2 Quality Committee, through publication
of "Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines and Analytical Methods Reference" is
not to restrict or limit the methods or technologies that can be used for N2 quality assurance. Our primary ob-
jective is to offer the worldwide beverage industry some examples of reliable, practical, appropriately verified,
state-of-the-art test methods and procedures that can be successfully used to ensure the quality of their N2.
The objective of any equivalent method (EM) is to correctly identify and measure a given target impu-
rity or class of impurities in the concentration range of interest, with sufficient accuracy and precision under all
circumstances. "Equivalency" indicates that a method has been demonstrated to be statistically similar (or
superior) in reliability and performance to a known, accepted ISBT method.
The general criteria for defining an EM would include, for example:
-The method's measurement range (working range) relative to the typical concentration range found for that
impurity in beverage-grade N2 as well as the guideline limit for that parameter.
-The ability of test data obtained from the method to reliably correlate with an existing, accepted method.
-The level of method freedom from interference that could be expected from other impurities typically found in
LIN.
- The method’s limit of detection relative to the maximum allowable limit.
- The complexity of use and safety issues associated with the EM relative to existing, accepted methods.

Sampling & Testing Equipment
Because of the stringent impurity guidelines shown in the “Guideline Impurity Limit Table”, sample in-
tegrity issues such as impurity losses and exclusion of artifact air during sampling operations and by sample
container wall permeation are primary concerns. Therefore, to obtain accurate N2 impurity data, users should
perform a review of all hardware materials used throughout the entire N2 measurement system (ex. transfer
lines to in-line analyzers including any sample containers and analyzer internal hardware). This review should
be designed to ensure that only acceptable, gas impermeable materials are employed and that good flow
design elements are practiced.
The table “Analytical Methods and Container Suitability for N2 Analysis” related to N2 sampling and
testing materials is offered for user review (page 17).

ANALYTICAL METHODS AND CONTAINER SUITABILITY FOR N2 ANALYSIS
Table Summary
Phase Method
Parameter & Suitable Container Guideline Limit Sampled or Tested Number
Liquid phase N2 sampling with
Direct liquid phase sampling
special cup-in-cylinder containers na 1.0N
ONLY
(ex. Cosmodyne®-type)
Pressurized headspace gas

Gas phase N2 sampling with high above bulk liquid or cylinder gas
na 1.1N
pressure metal cylinders sampling ONLY - (NOT for liq-
uid phase sampling)

Gas phase N2 sampling with low
above bulk liquid or cylinder gas 1.2N
pressure metal cylinders na
sampling ONLY - (NOT for liq-
uid phase sampling)

Gas phase N2 sampling with poly- above bulk liquid or cylinder gas
na 1.3N
meric bags sampling ONLY - (NOT for liq-
uid phase sampling)
N2 Identification + N2 Purity: Positive ID + 99.99 %

Bulk liquid phase, headspace or
Metal cylinder or dual-valved, gas- v/v min. Purity (sub- 2.0N + 8.0N
cylinder gas sample
impermeable sampling bag tractive method)
Oxygen (O2):
Metal cylinder or dual-valved, gas- 10 ppm v/v max 3.0N
cylinder gas sample
impermeable sampling bag
Water Vapor (H2O):

Metal cylinder or dual-valved, gas- na 4.0N
cylinder gas sample
impermeable sampling bag
Total Volatile Hydrocarbons

(THC, as methane [CH4]): 5 ppm v/v max 5.0N
cylinder gas sample
Metal cylinder / polymeric sampling
bag
Carbon monoxide (CO):
Metal cylinder / polymeric sampling 10 ppm v/v max 6.0N
cylinder gas sample
bag
Odor of vicinal headspace:

Metal cylinder / polymeric sampling No foreign odor 7.0N
cylinder gas sample
bag

LIN Sampling Safety Considerations
1.) Review all SDS information relating to N2 and all sampling instructions before attempting to perform any
sampling activity. Carefully inspect all containers and hardware for cleanliness and integrity.
2.) Personnel trained in handling cryogenic liquids and compressed gases must perform sampling in a well-
ventilated area. Appropriate safety equipment including face shields, safety glasses, cryogenic safety gloves,
ear protection, steel-toed shoes and other equipment should be used as needed.
3.) High pressure or low pressure metal cylinders must not be used to directly sample cryogenic liquid phase
N2. Use of such cylinders may lead to containment of a dangerously high amount of liquid load which
can result in excessive cylinder over-pressurization upon liquid vaporization which can result in
burst disc failures and/or catastrophic cylinder failure.
ONLY SPECIALIZED CUP-IN-METAL CYLINDER TYPE CONTAINERS (ex. COSMODYNE® TYPE 4 L

CYLINDERS) AND TRANSFER TUBING DESIGNED FOR DIRECT SAMPLING OF CRYOGENIC LIQUID
PHASE N2 SHOULD BE EMPLOYED WHENEVER LIQUID N2 PHASE SAMPLING IS REQUIRED.
4.) High pressure metal cylinders containing a N2 sample charge above 29 psig (200 kPag) must be properly
shipped as dangerous goods by a hazmat certified staff member.
5.) Low pressure metal cylinders and polymeric sample bags that contain an N2 sample charge below 29 psig
(200 kPag) can be properly shipped as non-dangerous goods (non-hazmat).

INTERNATIONAL SOCIETY OF BEVERAGE TECHNOLOGISTS
Classification: Cryogenic Liquid N2 Sampling Method 1.0N

Revision Date: 03/2015 Page 1 of 5
LIQUID NITROGEN (N2) SAMPLING

USING
CUP-IN-METAL CYLINDERS
ABSTRACT
This method describes how to directly sample the liquid phase of a cryogenic liquid nitrogen (LIN)
vessel using cup-in-metal (CIM) sample cylinders (ex. Cosmodyne® type). CIM cylinders are not intended for
either headspace gas or compressed N2 cylinder sampling. Using a CIM sample cylinder, a small volume of
liquid phase sample is collected and allowed to vaporize to generate a safe level of N2 pressure (typically 300
- 500 psig [2,070 - 3,450 kPag]) for subsequent sample transfill operations into non-hazmat kits or for direct
analytical testing. High pressure N2 samples taken in a CIM cylinder must be shipped as a hazardous materi-
al (hazmat).
SAMPLING AND PHASE
As many impurities in beverage-grade LIN accumulate in the liquid phase charge, this procedure is
designed to obtain a representative sample of this cryogenic liquid phase. Only cryogenic-liquid rated cylin-
ders, valves, transfer lines and other hardware should be used for obtaining liquid phase samples using this
procedure.
SAMPLING EQUIPMENT
1.) A 4 - 5 L stainless steel (ss) or alloyed cup-in-metal (CIM) type sampling cylinder that is approved by the
user's local regulatory agency* (ex. 4B500 DOT-rated [USA] or European Council TPED 1999/36/EC) cup-
containing and equipped with a minimum of one (1) 600 psig (4,140 kPag) burst disc. A 4 L cylinder size
is adequate for containment of a sufficient amount of vaporized, LIN sample (ex. between 300 - 500 psig
[2,070 - 3,450 kPag]) for performance of all ISBT beverage-grade N2 purity tests. CIM cylinders should
display a decal to easily indicate their Cup-filling "Inlet" and "Outlet" valves and also the vaporized liquid
sample "Outlet" valve.
*Note: All users should review and comply with any local regulations concerning approved cylinder construction materials
and design specifications needed for sample transport within their geographic region. The cylinder employed must be
designed with appropriate materials to minimize wall adsorption effects involving trace O2 and H2O.
2.) A short (ex. 3- 6 ft [1 - 2 m]), flexible length of foam-jacket insulated, high pressure, 1/4" (6 mm) ID ss
transfer tubing with appropriate end port fittings should be employed between the N2 source valve and CIM
cup-assembly inlet valve. This tubing should be rated for operation at cryogenic LIN temperatures and a
minimum 3,000 psig (20,700 kPag) pressure. Flexible, insulated transfer tubing allows for the convenient
orientation of a CIM sample cylinder to a LIN source and facilitates rapid, efficient CIM cup chilling and fill-
ing steps.
3.) "Sacrificial" ss adapter fittings should be connected to transfer line ports for connections to a LIN source
valve and for routine mating onto the CIM Cup-assembly inlet port (typically a 1/2" [12.7 mm] flare [male]
37o port). Use of sacrificial port adapters protects expensive, insulated transfer lines from fitting wear is-
sues that can be encountered in rough field sampling use.
4.) All valves and fittings should be rated for cryogenic liquid N2 service to minimize chances of leakage or
improper valve on / off performance.



USING
WARNINGS AND SAFETY CONSIDERATIONS
Before sampling, review all Safety Data Sheet (SDS) information on LIN handling. Sampling should
be performed in a well-ventilated area by personnel trained in handling cryogenic liquids and pressurized,
compressed gases. Face shields; insulated, long sleeve cryogenic-safety gloves; steel-toed shoes and other
necessary safety equipment must be worn. Ensure that the CIM cylinder's maximum allowable hydrostatic
test period has not expired*. DO NOT use the cylinder if its last hydro-test date is exceeded. The LIN source
sampling valve must be installed at a location that allows for safe, convenient LIN sampling. It should be fitted
with a capped, compression-type fitting (ex. 1/4" size [6 mm]) and be capable of a controlled LIN flow into a
CIM sample cylinder.
*Note: This hydrostatic re-test period typically varies from 5 years (US) to 10 years (some European regulations). Users
should review and comply with any local regulations concerning cylinder hydrostatic re-testing.
WARNING: DO NOT ATTEMPT TO OBTAIN A CRYOGENIC, LIQUID PHASE N2 (LIN) SAMPLE USING A
NON-CUP CONTAINING CYLINDER AS THEY ARE NOT DESIGNED FOR THIS PURPOSE AND COULD
RESULT IN VALVE FAILURE OR A DANGEROUS OVERCHARGE OF LIQUID N2.
A LIQUID N2 OVERCHARGE WILL RESULT IN EITHER A RUPTURED BURST DISC AND/OR A

CATASTROPHIC CYLINDER FAILURE THAT MAY RESULT IN INJURY OR DEATH.
AFTER LIQUID EVAPORATION, THE COMPRESSED N2 GASEOUS SAMPLES CONTAINED IN THESE

CIM CYLINDERS MUST BE PROPERLY SHIPPED AS DANGEROUS GOODS (HAZARDOUS
MATERIALS [HAZMATS]) BY A HAZMAT-QUALIFIED STAFF MEMBER.
PROCEDURE
1.) Check the last Hydrostatic test date stamped on the cylinder body to ensure that the maximum allowable
period has not elapsed. Record all sample ID data on an ID Tag and remove both cylinder end caps (lan-
yard attachment is recommended).
2.) Connect a short (ex. 3 - 6 ft [1 – 2 m]), flexible, foam-insulated, gas-impermeable transfer line to the LIN
source sampling valve. To facilitate rapid CIM filling, activate an appropriate "pressure building" valve on
the N2 source vessel (optional).
Note: The sample cylinder should not be connected to the transfer line during this step.
3.) Remove the top case section of the CIM cylinder's storage / shipping case and uncap the cup-assembly
inlet and outlet flare caps on the sides of the cylinder body.



USING
4.) Attach a section of insulated 1/4" (6 mm) ID ss tubing (or equivalent) to the CIM cup “outlet" exhaust port
(typically a 3/8" [9.5 mm]) flare fitting [male]). This tube should be bent at an approximate 90o angle at its
outlet to direct LIN flow to a safe area away from all personnel or equipment in the near vicinity (typically
this bent outlet exhaust tubing is pointed toward the ground at a sufficient distance away from sampling
personnel). The purpose of this exhaust port tubing is to prevent LIN from spraying out great distances
during its exit from the CIM cylinder and also to prevent LIN from dripping into the CIM's lower container
section*.
Note: This lower case unit is often used with commercially available (ex. Cosmodyne® type) samplers as a CIM
cylinder support base.
5) Securely connect the insulated sample transfer line to the CIM cup-assembly "Inlet" port (typically a 1/2"
[12.7 mm] flare [male] 37o fitting).
6.) Ensure that the "cup-filling" valve handle (located at the top of the CIM cylinder) is in its fully closed posi-
tion and also the "Sample OUT" needle valve (located at the bottom of the CIM assembly) is fully closed at
this point*.
Note: This ensures that the internal sample cup will not be filled during the initial cup chilling step.
7.) Slowly, carefully open the LIN source sampling valve to start a steady LIN flow through the transfer line
and through the CIM - internal cup assembly. This step is designed to chill the outer walls of the internal
cup, so that no distillation / impurity accumulation effects will occur during the subsequent "cup-filling"
step*. Typically this initial cup-chilling process may take between 5 - 15 min to accomplish as the initial
phase of this step results in only liquid N2 vaporization and gas flow through the CIM body until the trans-
fer lines and CIM cup-chilling areas have cooled sufficiently to maintain N2 in its liquid state.
*Note: LIN Sample does not enter the cup interior or cylinder body during this cup-chilling step.
8.) When liquid phase N2 begins to continuously "stream" from the CIM outlet tube, the cup-chilling phase has
been successfully completed and the next step involving "cup-filling" can proceed.
9) Upon observing a steady stream of liquefied N2 streaming from the CIM outage tube, turn the "cup-filling"
control valve (located on top of a typical CIM unit) to a totally open position. This will allow LIN to enter and
begin filling the cryo-chilled internal sample cup.
10.) Allow the cup-filling valve to remain open for about 20 seconds. This is typically enough time for the cup
to appropriately fill without overfilling and spilling liquid into the internal cylinder-body areas. Totally close
the cup-filling valve after this 20 sec cup-filling period.
11.) Wearing appropriate cryogenic safety gloves and equipped with proper tools, shut off the LIN source
valve. In some cases, this valve may be difficult to close and appropriate channel-wrench type tools may
be needed to fully close this extremely chilled valve. Full valve closure will be signaled by a stoppage of
liquid N2 flow from the CIM outlet tube.



USING
12.) Disconnect the inlet transfer line and outlet tube from the CIM assembly, then quickly re-cap the CIM
inlet / outlet ports to prevent water vapor incursion and ice formation from occurring within the cylinder's
chilled interior regions.
13.) Slowly, carefully disconnect the inlet transfer line from the LIN source sampling valve. Ensure that no
liquid N2 remains within the tubing, then cap both end ports to minimize the intake of water vapor and in-
ternal ice formation. This tubing assembly should be stored within a clean container during standby condi-
tions.
14.) Attach the upper casing body to the CIM assembly. Carefully transport the liquid- charged sample cylin-
der to the testing area and securely store it in a "flipped" position with bottom casing pointing upward with-
in a properly vented gas cabinet. Storage with the lower casing section pointing upward is recommended
as this allows the evaporating liquid charge to be quickly forced out of the cup area and into the 4L internal
body area, which facilitates more rapid evaporation and quicker sample readiness for testing. As the liquid
charge evaporates, the CIM pressure gauge will rise.
15.) A properly charged CIM cylinder typically results in cylinder pressures between 300 - 500 psig ( 2,070 -
3,450 kPag), which is more than adequate for performing all needed ISBT N2 quality tests. Total liquid
evaporation typically takes between 30 min and 1 hour to achieve.
16.) If the sample must be shipped, follow all carrier regulations associated with hazardous materials
transport. For proper Hazmat (Dangerous Goods) shipments, the staff member responsible for shipping
must be “Hazmat Certified” which involves successful attendance in one or more recognized “Hazmat
Training” courses for desired ground or air shipments.
17.) When sample shipment is required, securely package the CIM cylinder within its sturdy, rugged metal or
plastic case that is designed for CIM cylinder shipments. Follow all appropriate HAZMAT shipping regula-
tions including, for example*: affixing the UN code (UN1066), shipping name "Nitrogen, compressed",
green "NON-FLAMMABLE GAS" diamond label + “OVERPACK” label to the outer shipping box. Complete
all courier paperwork including: Hazmat Class or Division = “2.2”, Subsid. Risk “--”, Packing Group “--”,
Packing Instr. “200”. Note: the net wt. (kg) of N2 shipped is both cylinder size and internal pressure de-
pendent*.
*Note: Always refer to the latest hazardous shipping regulations (ex. CFR - 49 (US), IATA). The information supplied
above is to be used only as an example of hazmat regulations applicable at the time of document production.



USING
CALCULATIONS
Not applicable.
DISCUSSION, NOTES AND EQUIVALENT METHODS
1.) After the completion of all testing activities, CIM sample cylinders should be blown-down to just above at-
mospheric pressure and stored under a slightly positive N2 pressure (ex. 20 psig [138 kPag]) to prevent in-
ternal contamination by air and moisture. Alternatively, after a sampling session, it is often desired to fill
the cylinder with helium (He) and perform a leak test before continued use. CIM evacuation and refilling to
a slight positive pressure with beverage-grade gaseous N2 is recommended before continued LIN service.
2.) All transfer line equipment and supporting hardware should be capped when stored to prevent contamina-
tion.
3.) If Hazmat shipments of N2 samples are not feasible or desired, consideration should be given to N2 transfill
sampling procedures using low pressure metal cylinders (LPMC) and polymeric bags (PSB) for proper
Non-Hazmat shipment.
4.) Alternate CIM-type cylinders may be employed if appropriately verified by the manufacturer for this appli-
cation and approved by the appropriate regulatory authorities for hazmat shipping (if necessary).
REFERENCES
CGA G-6.3-2002, IATA Hazardous Shipping Regulations 2014, 49-CFR Hazardous Materials Shipping Regu-
lations 2014.

Classification: Cryogenic N2 Quality Sampling Method 1.1N

GASEOUS NITROGEN (N2) SAMPLING

USING
HIGH PRESSURE METAL CYLINDERS
ABSTRACT
This procedure describes how to use a high pressure rated, metal cylinder (HPMC) to obtain a sam-
ple from pressurized headspace gas above a liquid nitrogen (LIN) vessel, a compressed gaseous nitrogen
(N2) cylinder, or transfill from a CIM sample cylinder (see Method 1.0N). A HPMC is not intended for direct
liquid phase N2 sampling. Compressed N2 samples taken in a HPMC must be shipped as a hazardous mate-
rial (hazmat) when applicable.
SAMPLING AND PHASE
This method is designed to obtain a representative sample of vaporized, liquid phase N2. Brass or
stainless steel hardware and gas-impermeable transfer tubing should be used for sampling operations. All
materials used must be gas impermeable to prevent errors when performing trace O2 and H2O analysis.
SAMPLING EQUIPMENT
1.) A 0.5 -1.0 L, stainless steel (ss) sampling cylinder that is approved by the user's local regulatory agency*
(ex. DOT3A1800 rated [USA] or TPED "π" mark rated [Europe]), dual-ended, dual-valved and equipped
with one 1800 - 1900 psig (12,400 – 13,110 kPag) burst disc is recommended.
*Note: All users should review and comply with any local regulations concerning approved HPMC construction materials
and design specifications needed for sample transport within their geographic region. The cylinder materials em-
ployed must meet ‘materials suitability’ requirements, to minimize trace O2 and H2O adsorption effects.
2.) A short (ex. 1-3 ft [0.3 – 1 m]), flexible length of high pressure transfer tubing should be used between the
N2 source valve and HPMC inlet valve. This tubing should be rated for a minimum 3,000 psig (20,700
kPag) pressure, low temperature service and have a narrow internal diameter no larger than about 1/4" (6
mm). Flexible transfer tubing allows for the convenient orientation of a sample cylinder, which facilitates
rapid, efficient sample flushing and filling.
3.) When sampling N2 at pressures greater than 29 psig (200 kPag), a high pressure gas regulator must be
used to safely reduce the sample pressure. When sampling from compressed gas cylinders, the pressure
regulator shall be equipped with the appropriate CGA inlet fitting. When sampling from high pressure
sources other than high pressure cylinders, appropriate matting fittings shall be used. The outlet fitting of
the pressure regulator shall adapt safely to the high pressure transfer tube.



USING
Before sampling, review all Safety Data Sheet (SDS) information on liquid and gaseous N2. Sampling
should be performed in a well-ventilated area by personnel trained in handling pressurized, compressed, cry-
ogenic gases. Safety glasses; insulated, long sleeve cryogenic safety gloves; steel-toed shoes and other
necessary safety equipment must be worn. Ensure that the cylinder's maximum allowable hydrostatic test
period has not expired*. DO NOT use the cylinder if its last hydro-test date is exceeded. The N2 source valve
must be installed at a location that allows for safe, convenient sampling of headspace N2 gas. It should be
fitted with a capped, compression-type fitting (ex. 1/4" [6 mm]) and be capable of a controlled flow of gaseous
N2 into a sample cylinder.
*Note: This hydro-retest period typically varies from 5 years (U.S.) to 10 years (some European regulations). ISBT users
should review and comply with any local regulations concerning cylinder hydrostatic re-testing.
HPMC AS THEY ARE NOT DESIGNED FOR THIS PURPOSE AND COULD RESULT IN VALVE FAILURE,
AND/OR A DANGEROUS OVERCHARGE OF LIQUID N2.
A LIQUID N2 OVERCHARGE WILL RESULT IN EITHER A RUPTURED BURST DISC OR A

THE COMPRESSED N2 GAS SAMPLES CONTAINED IN A HPMC MUST BE PROPERLY SHIPPED AS
DANGEROUS GOODS (HAZARDOUS MATERIALS [HAZMATS]) BY A HAZMAT-QUALIFIED STAFF
MEMBER.
PROCEDURE
1.) Check the last stamped Hydrostatic test date to ensure that the maximum allowable period has not
elapsed. Hydrostatic test dates are typically stamped on the cylinder body near one of the valve ends.
Record all sample ID data on an ID Tag and remove both cylinder end caps (lanyard attachment is rec-
ommended).
2.) Sample Source Connections
a. Prior to attaching the pressure regulator, carefully open the source valve briefly in order to remove any
debris, particles or foreign matter from the fitting.
b. Attach the pressure regulator to the N2 Source. Cycle purge the regulator assembly by:
(1) Open the source valve and pressurize the regulator. Close the source valve.
(2) Vent the regulator by increasing the delivery pressure. Allow the regulator to drain completely. Di-
al back the regulator to close the outlet.
(3) Repeat steps 1 and 2 at least five times to remove trace contamination. When complete, proceed
to Step #3.
3.) Connect a short (ex. 1 ft [0.3 m]), flexible, pressure rated, gas-impermeable transfer line to the pressure
regulator outlet. Hold the tubing near its free end, and pointing this end away from you, open the N2
source valve and slowly increase the outlet pressure of the regulator. This will allow a controlled flow of ni-
trogen to purge the transfer line. Allow the system to purge for 1 minute.



USING
4.) Immediately after hardware purging, connect the HPMC's inlet fitting to the transfer line tubing’s outlet
port. Tighten then slightly loosen the cylinder connection to this transfer line, so that N2 will leak from the
1/4” (6 mm) compression fitting to help remove entrapped air.
5.) Slowly increase the outlet pressure of the regulator, and allow N2 gas to purge from the regulator, through
the transfer line and escape at the HPMC inlet fitting. This will displace any oxygen and other contami-
nants trapped in the sample line. After purging, tighten the HPMC inlet fitting. Set the required sample
pressure on the pressure regulator.
6.) Slowly, carefully open the HPMC's inlet valve fully, then slowly crack open the HPMC’s outlet valve par-
tially to start gentle, steady sample cylinder body purging. When a proper purge rate is set, the exiting N2
stream should not make an annoyingly loud sound, nor should it be a quiet whisper. Systematically cycle
the HPMC’s outlet needle valve to a wider open setting – then back to ideal purge flow position several
times to help remove any entrapped air from the outlet valve stem areas. Continue this cylinder body
purge for at least 3 – 5 minutes*.
*Note: Failure to properly flush an HPMC may lead to an erroneously high trace O2 or H2O result. If transfilling from a 4 –
5 L CIM sample cylinder, a slightly shorter purge time may be needed to prevent running out of sample yet still con-
taining an adequate HPMC internal pressure and sample volume needed for subsequent analytical tests.
7.) Securely close the HPMC's outlet valve*. Wait about 10 sec to ensure full cylinder pressurization, then (in
this sequence) securely close the HPMC's inlet valve then close the N2 source valve. When performed
properly a cylinder flush / fill step takes about 5 - 10 minutes.
*Note: DO NOT OVERTIGHTEN valves as this can cause internal damage and leakage.
8.) Dial back the sample regulator, disconnect the transfer line and remove the pressure regulator from the N2
source. Store all sampling components in a clean dry location.
9.) Check for any audible gas leaks from the HPMC. DO NOT TEST OR SHIP a HPMC if any leaks are de-
tected.
10.) Re-cap the HPMC ports. Apply PTFE tape to any NPT threads before re-capping.
11.) Carefully transport the sample-filled HPMC to the testing area and securely store it in a proper, vented
cabinet until testing is scheduled. If the sample must be shipped, follow all carrier regulations associated
with hazardous materials transport. For proper Hazmat (Dangerous Goods) shipments, the staff member
responsible for shipping must be “Hazmat Certified” which involves successful attendance in one or more
recognized “Hazmat Training” courses for desired ground or air shipments.



USING
12.) When sample shipment is required, securely package the HPMC (using bubble-wrap only – do not use
shipping peanuts) in a sturdy, weight-stamped fiberboard box or an ATA certified shipping case. Follow all
appropriate HAZMAT shipping regulations including, for example*: affixing the UN code (UN1066), proper
shipping name "Nitrogen, compressed", green "NON-FLAMMABLE GAS" rectangular label +
“OVERPACK” label to the outer shipping box. Complete all courier paperwork including: Hazmat Class or
Division = “2.2”, Subsid. Risk “--”, Packing Group “--”, Packing Instr. “200”. Note: the net wt. (kg) of N2
shipped is cylinder size and internal pressure dependent*.
*Note: Always refer to the latest hazardous shipping regulations (ex. CFR - 49 [US], IATA). The information supplied
above is to be used only as an example of hazmat regulations applicable at the time of document production.
13.) Ship the sample properly using any hazmat-qualified courier.
CALCULATIONS
Not applicable.
1.) After the completion of all testing activities, HPMC should be blown-down and stored under a slight posi-
tive N2 pressure (ex. 20 psig [138 kPag]) to prevent internal contamination by air and moisture. Alterna-
tively, after a sampling session, it is often desired to fill the cylinder with helium (He) and perform a leak-
test before continued use. A slight positive He pressure should be maintained to prevent external contam-
ination.
2.) Proper choice and minimal use of sample cylinder valve stem lubricant should be practiced to prevent lub-
ricant leaching and sample contamination. Use of inert / soft seat valve stem tips with a wide surface area
is recommended for long / reliable, leak-free needle valve service life.
3.) All transfer line equipment and supportive hardware should be capped when stored to prevent contamina-
tion.
4.) If hazmat shipments of N2 samples are not feasible or desired, consideration should be given to N2
sampling using low pressure metal cylinders (LPMC) and polymeric bags (PSB) for proper Non-Hazmat
shipment. Refer to Sampling Methods 1.2N and 1.3N for details concerning non-hazmat N2 sampling and
shipping.
REFERENCES
CGA G-6.3-2002 IATA Hazardous Shipping Regulations 2014, 49-CFR Hazardous Materials Shipping Regu-
lations 2014.



USING
SMALL, LOW PRESSURE METAL CYLINDERS
ABSTRACT
This procedure describes how to obtain samples from either pressurized headspace gas above a liq-
uid nitrogen (LIN) vessel, compressed gaseous N2 cylinders or transfill operations involving cup-in-metal
(CIM) sample cylinders using low pressure (29 psig [200 kPag]) metal cylinders (LPMC). A LPMC is specifi-
cally designed to allow a non-cryogenic N2 sample to be properly sampled and shipped as non-hazardous
material. This cylinder design is not intended for direct liquid phase N2 (LIN) sampling. All ISBT N2 Guideline
tests can be performed on LPMC samples (see ANALYTICAL METHODS AND CONTAINER SUITABILITY
FOR N2 ANALYSIS table on page 17). When employing an LPMC for Non-Hazmat shipping programs it is
recommended that all analytical tests should be performed as soon as possible.
SAMPLING AND PHASE
This method is designed to obtain a representative, low pressure sample of vaporized, liquid phase
N2. Brass or stainless steel hardware and gas-impermeable transfer tubing should be used for sampling op-
erations. All materials used must be gas impermeable to prevent errors when performing trace O2 and H2O
analysis.
SAMPLING EQUIPMENT
1.) A sample vessel constructed of suitable and compatible materials must be used. The LPMC and associ-
ated sampling equipment must be designed to safely capture a sample at 29 psig (200 kPa) or less with-
out introducing atmospheric contamination.
Note: All users should review and comply with any local regulations concerning appropriate LPMC construction materials
and design specifications needed for sample transport within their geographic region. The cylinder materials em-
ployed must meet ‘materials suitability’ requirements to minimize trace O2 and H2O adsorption effects.
2.) When sampling N2 at pressures greater than 29 psig (200 kPag), a high pressure gas regulator must be
used to safely reduce the sample pressure. When sampling from compressed gas cylinders, the pressure
regulator shall be equipped with the appropriate CGA inlet fitting. When sampling from high pressure
sources other than high pressure cylinders, appropriate mating fittings shall be used. The outlet fitting of
the pressure regulator shall adapt safely to the high pressure source.



USING
Before sampling, review all Safety Data Sheet (SDS) information on liquid and gaseous N2. Sampling
should be performed in a well-ventilated area by personnel trained in handling pressurized, compressed, cry-
ogenic gases. Safety glasses; insulated, long sleeve cryogenic-safety gloves; steel toed shoes and other
necessary safety equipment must be worn. Ensure that the LPMC’s maximum allowable hydrostatic test peri-
od has not expired*. The N2 source valve must be installed at a location that allows for safe, convenient sam-
pling of pressurized headspace N2 gas. It should be fitted with a capped, compression-type fitting (ex. 1/4" [6
mm]) and be capable of a controlled flow of gaseous N2 into the LPMC.
*Note: This hydrostatic re-test period typically varies from 5 years (U.S.) to 10 years (some European regulations). ISBT
users should review and comply with any local regulations concerning cylinder hydrostatic re-testing.
LPMC AS THEY ARE NOT DESIGNED FOR THIS PURPOSE AND COULD RESULT IN VALVE FAILURE,
AND/OR A DANGEROUS OVERCHARGE OF LIQUID N2.
A LIQUID N2 OVERCHARGE WILL RESULT IN EITHER A RUPTURED BURST DISC OR A

THE NON-COMPRESSED, N2 SAMPLE CONTAINED IN THESE LPM CYLINDERS SHOULD BE

PROPERLY SHIPPED AS NON-DANGEROUS GOODS (NON-HAZARDOUS MATERIALS [NON-
HAZMATS]). STAFF ASSOCIATED WITH SAMPLE SHIPPING DO NOT NEED TO BE HAZMAT
CERTIFIED.
PROCEDURE
1.) Check the last stamped Hydrostatic test date to ensure that the maximum allowable period has not
elapsed. Hydrostatic test dates are typically stamped on the cylinder body near one of the valve ends.
Record all sample ID data on an ID Tag and remove both cylinder end caps (lanyard attachment is rec-
ommended).
2.) Sample Source Connections
a. Prior to attaching the pressure regulator, carefully open the source valve briefly in order to remove any
debris, particles or foreign matter from the fitting.
*Note: Any N2 vessel with a gas pressure above about 500 psig [3,450 kPag] requires a high pressure regulator.
b. Attach the pressure regulator to the N2 Source. Cycle purge the regulator assembly by:
(1) Open the source valve and pressurize the regulator. Close the source valve.
(2) Vent the regulator by increasing the delivery pressure. Allow the regulator to drain completely. Di-
al back the regulator to close the outlet.
(3) Repeat steps 1 and 2 at least five times to remove trace contamination.



USING
3.) Connect a short (ex. 1 ft [0.3m]), flexible, gas-impermeable transfer line to the pressure regulator outlet.
Hold the tubing near its free end, then while pointing this end away from you, open the N2 source valve
and slowly increase the outlet pressure of the regulator. This will allow a controlled flow of nitrogen to
purge the transfer line. Allow the system to purge for 1 minute
4.) Immediately after hardware purging, connect the LPMC's inlet fitting to the transfer line tubing’s outlet port.
Tighten then slightly loosen the cylinder connection to this transfer line, so that N2 will leak from the 1/4” (6
mm) compression fitting to help remove entrapped air.
5.) Slowly and carefully open the N2 source, adjust the pressure regulator and allow gas to enter the LPMC.
Consideration of the specific design of the LPMC should be made at this point, following the manufactur-
er’s specific instructions for filling the vessel.
6.) For LPMC’s equipped with an inlet and outlet valve, fully open the LPMC’s inlet valve*, then fully open the
LPMC’s outlet valve to start a steady sample purge through the cylinder body. Cycle purge the cylinder by
repeatedly opening and closing the outlet valve. Cycle purge the cylinder 10 times over 5 minutes.
*Note: Failure to properly flush an LPMC may lead to an erroneously high trace O2 or H2O result. If transfilling from a 4 –
5 L CIM sample cylinder, a slightly shorter purge time may be needed to prevent running out of sample yet still con-
taining an adequate LPMC internal pressure and sample volume needed for subsequent analytical tests.
7.) While still purging, securely close the LPMC’s outlet valve. Wait about 10 sec to ensure full LPMC pres-
surization (at or below 29 psig (200kpa)), securely close the LPMC’s inlet valve* then close the N2 source
valve.
*Note: DO NOT OVERTIGHTEN a needle valve as this can cause valve damage and leakage.
*Note: For containers with a single inlet valve prepared for direct filling or a non-traditional sample
valve, follow the manufacturer’s recommendations for sample purging.
8.) Dial back the sample regulator, disconnect the transfer line and remove the pressure regulator from the N2
source. Store all sampling components in a clean dry location.
9.) Check for any audible gas leaks from the sample cylinder. DO NOT TEST OR SHIP the LPMC if any
leaks are detected.
10.) Re-cap the LPMC’s end ports. Apply PTFE tape to any NPT threads before re-capping.
11.) When sample shipment is required, securely package the LPMC (using bubble-wrap only – do not use
shipping peanuts) in a sturdy fiberboard box or preferably an IATA-compliant sample shipping container.
Non-compressed N2 samples obtained with this procedure can be properly shipped as non-
dangerous goods and should be described in shipping papers as a “non-hazardous industrial
sample of no commercial value”*.
*Note: Always refer to the latest hazardous shipping regulations (ex. CFR - 49 [U.S.], IATA). The information
supplied above is to be used only as an example of hazmat regulations applicable at the time of doc-
ument production. Non-compressed N2 samples are exempt from IATA hazmat shipping regula-
tions.



USING
12.) Ship the sample as a non-hazmat using any qualified courier.
CALCULATIONS
Not applicable.
1.) After the completion of all testing activities, an LPMC should be stored under a slightly positive N2 pres-
sure (ex. 20 psig [138 kPag]) to prevent internal contamination by air and moisture. Alternately, after a
sampling session it is often desired to flush and fill the LPMC with helium (He) and perform a leak-test be-
fore continued use. A slightly positive He pressure should be maintained to prevent external contamina-
tion.
2.) Proper choice and minimal use of sample cylinder valve stem lubricant should be practiced to prevent lub-
ricant leaching and sample contamination. Use of inert / soft seat valve stem tips with a wide surface area
is recommended for long / reliable, leak-free needle valve service life.
3.) All sampling equipment should be capped when stored to prevent contamination.
4.) When using an LPMC, specific analytical test equipment and methods must be employed due to the low
volumes and pressures of the sample gas. Ensure the samples taken and equipment used are compatible
with the laboratory procedures employed.



USING
POLYMERIC SAMPLING BAGS
ABSTRACT
This method describes the use of inert polymeric sampling bags (PSB) and associated equipment for
obtaining low pressure gaseous samples from pressurized headspace gas above liquefied N2 (LIN) storage
vessels, compressed gas cylinders or transfilled from cup-in-metal (CIM) sample cylinders. This container
type allows N2 samples to be properly shipped as non-hazardous goods. Some tests such as trace water
(H2O) and oxygen (O2) cannot be performed using a standard 1-valved PSB but can be if 2-valved, gas im-
permeable sampling bags are employed (see ANALYTICAL METHODS AND CONTAINER SUITABILITY
FOR N2 ANALYSIS table on page 17). When employing PSBs for non-hazmat shipping programs it is rec-
ommended that all analytical tests be performed as soon as possible and within the timeframe that the lab
recommends for maintaining the sample’s integrity. Test results obtained beyond this storage interval should
be duly-noted for re-sampling / re-testing considerations.
SAMPLING AND PHASE
This sampling method can be employed on either pressurized headspace gas above LIN, high pres-
sure, compressed gas N2 cylinders or CIM cylinders. A 1 to 5 L size polymeric film bag equipped with an inert
plastic valve port is recommended. For beverage-grade N2 sampling programs, a minimum of three one-liter
bags or two two-liter bags of gas sample is suggested. Brass or stainless steel hardware and gas-
impermeable transfer tubing should be used for sampling operations to prevent contamination which will ad-
versely affect trace O2 and H2O analysis.
SAMPLING EQUIPMENT
1.) Bag Requirements and Quality Testing: Chemically inert, multi-layer barrier (MLB) films are highly
recommended as a bag material. Other types of film materials are also acceptable if they have been
appropriately verified for beverage-grade N2 use. While many single film materials have been shown to be
adequate for performing CO, THC and odor tests on N2 samples, most are not a suitable material for tests
associated with H2O or permanent gases such as O2 (although some H2O and O2 impervious MLB film
bags of dual valve-based design can be used if appropriately verified for trace H2O and O2 testing). Care
must be exercised in choice of bag grade, as many standard commercially available PSB’s are of
insufficient purity which can result in foreign odors and / or excessive THC artifacts from bag
contaminants.
2.) Valve / Septum / O-Ring Materials / Assembly Aid / Sample Label Design: The recommended inlet
valve material and valve port is polypropylene which is vacuum-baked and packaged with a polytetrafluo-
roethylene (PTFE) or other inert plastic port cap. The port cap prevents contaminants from entering the in-
terior valve during shipment / storage. Commercial valves designed from polypropylene have been found
to be physically rugged, reliable, versatile, and chemically inert when conditioned as described above.
Other valve port materials can be used if appropriately verified for this application. Recommended valve
designs include both a septum sampling port and side-port stem (ex. for attachment of 3/16” [4.7 mm ID]
flexible tubing). The septa employed in the valve assembly should be vacuum-baked and septa material
selected for minimal organic impurity / sulfur impurity outgassing (ex. COS / H2S / CS2). O-ring materials
used for obtaining a leak-tight mating of a valve-port to bag film should be made of a synthetic elastomer
(ex. Viton® or equivalent) to minimize or prevent sulfur and aromatic impurity outgassing which will nega-
tively affect N2 test result accuracy. O-ring assembly aids should be high molecular weight synthetic



USING
agents (ex. poly-alpha-olefin) to ensure leak-tight sealing and not outgas any odiferous, volatile impurities.
Paper-based sample ID labels used must not be directly affixed to the film surface as impurities from the
label’s adhesive can slowly leach through the film and cause false positives on the impurity test results.
Likewise, no pen labeling should ever be made on the bag film body.
NOTE: Other types of plastic or metal connective materials are acceptable if they have been appropriately
verified for beverage-grade N2 use. Care must be exercised in alternate tubing selection as plasticizers
and residual oxygenated solvents such as methanol are common contaminants in certain types of plastic
tubing. This may impart a foreign odor to a bag-contained N2 sample, or introduce volatile, THC-
generating impurities. Transfer lines must always be kept as short as practical.
3.) T-Assembly for Bag Filling: To assist in the safe / controlled filling of a PSB, a hose adapter / T-fitting
assembly consisting of a brass or ss hose-barb coupled to the center port of a 3/16” (4.7 mm) polypropyl-
ene T-fitting using a short (ex. 5” [13 cm]) section of inert, clean 3/16” (4.7 mm) ID PTFE-FEP tubing. The
other 2 ends of the T-fitting are connected respectively with one 2” (5 cm) length of PTFE-FEP tubing (for
connection to a PSB valve port) and one 2” (5 cm) length of flexible Tygon® tubing (ex. beverage-grade
Formula R-3603) for finger-pinch control of the bag-filling process. These tubing materials have been ap-
propriately verified for use in beverage-grade N2 applications.
4.) 2-Valve Bag Filling Assembly: If employing 2-valved, MLB (H2O / O2 impermeable) film bags, employ
appropriate swaged hardware connectors equipped with a 0.33 - 5 psig check valve between the N2
source valve and dual-valved bag inlet. Dual-valved bags mimic low pressure metal cylinders (LPMC- see
method 1.2N) in principle and allow for thorough bag purging prior to outlet valve closing and bag filling to
the inlet check valve cracking pressure.
Before sampling, review Safety Data Sheet (SDS) information on liquid and compressed gaseous N2. Sam-
pling should be performed in a well-ventilated area by personnel trained in handling compressed gases and
cryogenic liquids. Safety glasses; insulated, long sleeve cryogenic-safety gloves; steel toed shoes and other
necessary safety equipment must be worn.
DO NOT ATTEMPT TO OBTAIN A LIQUID PHASE N2 SAMPLE USING A PSB.

DO NOT OVERFILL A PSB AS THIS CAN CAUSE A VIOLENT BAG RUPTURE. DO NOT FILL BAGS
MORE THAN TWO-THIRDS (2/3) FULL TO PREVENT OVEREXPANSION AND SEAM-STRESS
RUPTURE DURING AIR TRANSPORT.



USING
PROCEDURE
1.) Equipment Pre-check: Before sampling, ensure that the shelf-life of the PSB has not been exceeded, all
sample bags are clean, fully evacuated, port capped, and all valves securely closed. All transfer tubing,
pressure regulators, metering valves, and connective fittings should be inspected for cleanliness and kept
stored in a sealed plastic bag until needed. Check that the proper size and number of bags are filled.
*Note: If employing dual-valved PSBs refer to the sampling and shipping procedures offered by the man-
ufacturer.
Single Valve PSB Applications (ex. for THC, CO and Odor tests ONLY)
2.) Sample Source Connections: When sampling N2 at pressures greater than 29 psig (200 kPag), a high
pressure gas regulator must be used to safely reduce the sample pressure. When sampling from com-
pressed gas cylinders, the pressure regulator shall be equipped with the appropriate CGA inlet fitting.
When sampling from high pressure sources other than high pressure cylinders, appropriate mating fit-
tings shall be used. The outlet fitting of the pressure regulator shall adapt safely to a PSB “T” assembly.
3.) Transfer Hardware Purging: Allow the entire system to purge for 1 minute, then loosely connect the
PSB side port to the PTFE - FEP end of the T-Assembly. Allow 30 seconds for valve stem purging, then
securely attach the bag’s valve port and open the PSB’s valve.
4.) Bag Purging / Final Sample Filling: Using finger pressure, pinch-close the flexible tube on the T-
assemblies free-end to direct and precisely control the gas stream into the sample bag. When the bag
reaches the desired 2/3’s full, securely close the PSB's valve and remove it from the T-assembly. Next,
open the bag valve and gently press all sample gas from the bag. Repeat this fill / purge process, then fill
the bag 2/3’s full with sample and securely close the PSB’s valve. After final sample filling and disconnect-
ing the bag from the T-assembly, close the N2 source valve and allow all line pressure to slowly escape
before removing the transfer hardware.
5.) Leak Check / Sample ID: Carefully observe the sample-filled PSB for several minutes to ensure that no
leaks are evident. Label the PSB with appropriate sample identification information.
6.) All sampling hardware should be stored in a clean, sealed container to prevent contamination.
Sample Bag Storage and Shipment
A sample-containing PSB should be stored in a cool, dark, secure cabinet until testing is performed. All
testing should be completed within the timeframe that the lab recommends for maintaining the sample’s in-
tegrity. If sample shipment is required, follow manufacturer recommended packaging instructions. As the
pressure inside a bag sample is essentially atmospheric (0 psig, [0 kPag]) and therefore non-compressed,
PSB-contained N2 samples are IATA exempt and should NOT be defined as hazardous / dangerous goods
and can be properly shipped by ground or air as non-hazardous goods (ex. "non-hazardous industrial sam-
ple of no commercial value").



USING
CALCULATIONS
The approximate total volume of PSB contained N2 gas sample required for a specific analytical test
program = Sum of all test gas volume requirements (cc) x 1.5.
PSB re-use is not recommended.
REFERENCES
IATA Guidelines 2014
CFR - 49 Code of Federal Regulations Section 49 Guidelines 2014
Viton® is a registered elastomer trade name of DuPont de Nemours
Tygon® is a registered elastomer trade name of Saint-Gobain Co

Classification: Cryogenic N2 Quality Analysis Method 2.0N

NITROGEN (N2) IDENTIFICATION

BY GAS CHROMATOGRAPHY
ABSTRACT
This method is designed to establish the positive identification (ID) of N2 as the primary gas in bever-
age-grade nitrogen (N2) samples. A gas chromatograph (GC) equipped with a thermal conductivity detector
(GC/TCD) is employed. Positive N2 ID is based upon both a retention time and peak area count match
against a 100% v/v pure N2 gas standard. This method is capable of positively distinguishing pure (99+ %) N2
from other potential commodity gases including air, argon (Ar), oxygen (O2), nitrous oxide (N2O), carbon diox-
ide (CO2), helium (He), hydrogen (H2) and many common gas mixtures containing N2. This identification re-
sult is reported as either “Pass” or “Fail”.
SAMPLING AND PHASE
The required impurity data for this test should be obtained from a sample collected using Sampling
Methods 1.0N, 1.1N, 1.2N or 1.3N (only if employing dual-valved MLB film bags). A single-valve polymeric
sampling bag (PSB) cannot be used for this method. Refer to each individual ISBT analytical sampling meth-
od for associated details.
EQUIPMENT, STANDARDS AND REAGENTS
1.) GC System with Electronic Data Handling: The recommended instrumentation system includes a GC
equipped with helium (He) carrier gas, gas loop injector, column backflush and 6-port column-to-detector
selection valves, high resolution capillary or passivated packed column bank, a TCD (or equivalent) detec-
tor and an electronic integrator or computer-based GC data-handling system for peak retention time (tr)
and peak area count data acquisition. A GC loop control manifold is recommended for handling low pres-
sure / low volume samples, but not required. The GC column bank must adequately time resolve the major
N2 peak from all other potential gas impurities. The most critical separations are N2 from Ar / O2 but evi-
dence must be shown that the GC can resolve N2 from other gases. A Haysep type "pre-column" (or
equivalent) 60/80 mesh packed 1/8" x 7' Ni or Sulfinert® passivated ss column can be used to separate out
molecular sieve-deactivating impurities such as CO2, N2O and H2O that may be in a N2 sample. One or
more high resolution molecular sieve type columns (ex. a Molsieve 13X, 45 / 60 mesh packed Ni or Sulfin-
ert® passivated ss 1/8" ID column x 7 ft length connected beyond the pre-column) are recommended for N2
peak separation from Ar / O2. Other column types can be used if appropriately verified for this ID applica-
tion. With the proper column-to-TCD switching valve sequencing, all potential impurity gases such as CO2,
N2O and other non-condensable gas (NCG) analytes can be identified based upon a peak retention time
match vs. external pure gas standards or high percentage mixtures in He.
2.) Equipment for High Pressure Cylinder Samples: A 2-stage pressure regulator (PR) equipped with a
metal diaphragm and an outlet ball valve is recommended when sample pressures above 500 psig (3,450
kPag) are involved. A fine metering valve attached downstream of the ball valve is also recommended for
controlled injector loop filling whenever an inlet manifold system is not employed.
3.) Inlet Manifold System for GC Loop Filling: A GC inlet manifold system designed to provide the GC in-
jector loop with an air-free, representative sample of either calibration gas or sample at a precisely con-
trolled inlet pressure (ex. 760 ±0.5 torr) is optional, but highly recommended. It should consist of VCR ss
hardware, on-off VCR diaphragm valves, fine metering valves, He-purged sample / calibration gas selec-
tion valve, pressure sensor system (ex. 0.0 – 1000.0 torr range), vacuum pump system (for manifold air



removal), 1/16” (1.6 mm) sample transfer line, inlet on-off / fine metering valve assembly plus appropriate
mating fittings for accommodating cup-in-metal cylinders (CIM), high pressure metal cylinders (HPMC) or
low pressure metal cylinders (LPMC). This inlet manifold is ideally suited for laboratory testing applications
when sample volumes and pressures are limited, such as when LPMC samples are collected for "non-
hazmat" sample shipping.
4.) Helium (He) Carrier Gas: O2-scrubbed high purity He of approximately 99.99+ % purity is recommended
as a carrier and TCD reference cell gas.
5.) Pure Gas Standards or High % Mixture Standards: It is recommended that pure gases or certified cali-
bration gas standards including N2, H2, Ar, O2, N2O, CO2, and air be used for routine retention time calibra-
tion and for demonstrating the critical resolution of N2 from Ar / O2 GC peaks.
6.) Zero (Blank) Gas Standard: A certified, 99+% He standard is required for system blank runs.
ANALYTES
Nitrogen (N2, MW = 28)
Potential Target Non-Condensable Gas (NCG) Analytes
N2 – major sample matrix gas - requiring positive identification
Synthetic or Compressed Air - potential interferent to a positive, pure N2 Identification
H2 - potential interferent to a positive, pure N2 Identification
Ar - potential interferent to a positive, pure N2 Identification
O2 –potential interferent to a positive, pure N2 identification
N2O - potential interferent to a positive, pure N2 Identification
CO2 - potential interferent to a positive, pure N2 Identification

Note: Molecular sieve deactivating impurities such as CO2, N2O and H2O vapor are prevented from contacting this col-
umn by use of a pre-column (ex. Haysep) which strongly retains CO2 , N2O, H2O and any C2+ organics. Upon molsieve
column elution / TCD detection of all target analytes including Ar, O2, N2, the GC back-flush and column switching valve
systems are activated which reverses carrier flow through the pre-column which then sends CO2, N2O, C2+, etc directly to
the TCD for peak retention time and peak area evaluation.



MEASUREMENT RANGE
Not applicable – this is an ID test only, not a quantification method.
The retention time (tr) and GC peak area counts of pure N2 and each target analyte are required to be meas-
urable by a graphical means - for positive ID of pure N2 only. Other ISBT methods (ex. 8.0N) are employed
to precisely determine the % v/v N2 present in the sample by subtractive methods.
PRECISION
Typically 1 - 2 % relative range for retention times and major peak area count repeatability is achiev-
able for N2 and other targeted non-condensable gases (NCG) in the high % v/v range of interest.
LIMIT OF DETECTION
A minimum limit of detection (LOD) of approximately 0.1 % v/v or lower is typically achievable for de-
tecting the target impurities of interest*.
*Note: Ar can be present up to a few thousand ppm v/v (typically less than 1% v/v) in cryogenic N2 sources.
INTERFERENCES
For pure, beverage grade N2 ID, no interferences from other potential major gases should be encoun-
tered based upon the GC method employed. This method depends on the GC system adequately resolving
and identifying pure N2, from other common commodity gases such as air (which is approx. 77% N2, 21% O2).
Review all Safety Data Sheet (SDS) information on liquid N2 and the target air gas impurities deter-
mined in this method. Wear safety glasses and other necessary protective equipment as chemical, pressur-
ized gas and thermal hazards are present. Perform all tests in a well-ventilated area. Carefully follow all op-
erational instructions provided by the equipment manufacturers.
PROCEDURE
A.) GC CALIBRATION
1.) Calibrate the GC using a He blank, 100% N2 (and other target gas analyte) standards according to GC
manufacturer instructions using the same GC loop purging, filling procedure and pressure as used with N2
samples (ex. 760.0 ±1 torr recommended - see "Sample Analysis” section B, below).
2.) Determine the retention times (tr), resolution of critical peaks and system "Response Factor" values of N2
(Rf = peak area counts / 100% v/v pure standard concentration) and all other desired target analytes (ex. Ar,
O2, CO2, etc.).



B.) SAMPLE ANALYSIS (High Pressure & Low Pressure Sample Source Options)
Follow GC manufacturer’s instructions regarding optimal He carrier / TCD reference cell gas flow, column
temperature program, instrument settings, pre-column backflush / column-switching times, and overall oper-
ating conditions.
1.) High Pressure N2 Analysis (CIM or HPMC Samples)
- If a CIM cylinder is involved, attach a metering valve assembly to the CIM’s sample outlet valve to control
delivery flow.
- If an HPMC containing 500+ psig (3,450 kPag) of sample is involved, orient the sample cylinder for conven-
ient attachment of a 2-stage PR* with an outlet ball valve assembly to the HPMC’s outlet port. Carefully
open the cylinder valve and set the PR to the required pressure (ex. 2 - 5 psig [13.8 – 34.5 kPag]) with the
outlet ball valve closed. If an inlet manifold is not employed, also install a metering valve for injector loop
purge / fill control.
*Note: Small, low dead-volume PR’s are easier to purge of any entrapped internal O2 or H2O vapor.
Inlet Manifold System Purge / Fill Procedure
1.1) Slowly, carefully open the outlet ball valve and allow the PR to purge for 10 - 20 sec, then close the
valve. Next, connect the PR-ball valve to the GC inlet manifold valve / transfer line assembly. This mani-
fold inlet system should consist of inert, small diameter (ex. 1/16” [1.6 mm]) ss tubing and associated on
/ off and fine metering valves for precise flow control and line evacuation.
1.2) Evacuate the manifold line up to the previously purged and closed PR-ball valve, isolate the vacuum
source and check for any leaks by monitoring any pressure increase in the line.
1.3) Close the manifold inlet line valves, carefully re-open the PR-ball valve and then slowly, carefully open
the manifold inlet line valves to fill the inlet manifold and GC loop with sample gas to approximately 1000
torr. Next, shut off the sample outlet valve and evacuate the manifold inlet / GC loop again. Repeat this
evacuation / purge step several times to ensure that the manifold and GC loop are filled with a repre-
sentative sample.
1.4) Using the appropriate manifold valves, fill the inlet manifold GC injector loop with the sample to the de-
sired system pressure for analysis (ex. approx. 760 torr).
1.5) Immediately inject the sample and establish the tr and peak area signal for N2 and any other target impu-
rities present. If necessary, an additional repeat of this analysis can be performed to verify results.



Non-Manifold System Purge / Fill Procedure
1.1) Slowly, carefully open the outlet ball valve and metering valve (slightly) and allow the PR assembly to
purge for 10 - 20 sec, then close the outlet ball valve. Next, connect the PR-metering valve to the GC
transfer line / loop assembly – this transfer line should consist of inert, small diameter (ex. 1/16” [1.6
mm]) ss tubing and the loop outlet port should be attached to a small flowmeter to monitor purge flow
through the injector loop.
1.2) After line/loop purging, close the ball valve and when the outlet flowmeter ball reaches zero, immediately
start the GC run. This process ensures that the loop GC injector loop will be filled with a representative
N2 sample to the desired system pressure (ex. approx. 760 torr).
2.) Low Pressure N2 Analysis (LPMC Samples* - For GC Manifold Systems Only)
2.1) Securely orient the LPMC so that a sample can be obtained from its outlet fitting.
2.2) Connect sample inlet manifold valve / transfer line assembly to the outlet port of the LPMC.
2.3) Evacuate this line assembly up to the LPMC’s outlet port, isolate the vacuum source and check for any
leaks by monitoring any pressure increase in the line.
2.4) Close the manifold inlet line valves, carefully open the LPMC’s outlet valve and then slowly, carefully re-
open the manifold inlet line valves to fill the inlet manifold and GC loop with sample to approximately
1,000 torr. Next, shut off the LPMC’s outlet valve and evacuate the manifold inlet / GC loop. Repeat this
evacuation / purge step several times to ensure that the manifold and GC loop are being filled with a
representative N2 sample.
2.5) Fill the manifold / GC loop to the same sample pressure used for GC system calibration (ex. approx. 760
torr).
2.6) Immediately inject the sample and establish the tr and peak area signal for N2 and any other target impu-
rities present. If necessary, an additional repeat of this analysis can be performed to verify results.
*Note: An equivalent line manifold connection / line evacuation – purge and precise pressure GC loop fill pro-
cedure is employed if dual-valved MLB film bags are employed for this test.
CALCULATIONS
GC Calibration: The average GC/TCD response factor (Rf (i)) for N2 and other non-condensable gas impuri-
ties in N2 is calculated by the following general formula:
Rf (I) = Peak area counts / 100% = (area counts per % units)
* Where i = pure N2, Ar, O2 etc.
For example, if a pure N2 standard produced 900,000 area counts:
Rf(N2) = 900,000 / 100% = 9,000 counts / %



Sample Analysis: The amount of N2 or other target air gas impurities detected in a N2 sample is calculated
from its average peak area count and Rf value using the following formula:
Analyte Concentration (% v/v) = (Peak Area Counts ÷ Rf(i))
Example: If a N2 sample produced a 890,000 area counts, and if the Rf(N2) = 9,000 counts / %, then the ap-
proximate % N2 = 890,000 counts ÷ 9,000 counts / % = 98.9%
Retention Time (tr) Match Analysis: An acceptable tr match for a positive N2 identification is reached if the
tr of the sample's major peak is within ± 2% relative to the tr of the pure N2 standard.
Example: If the tr of a pure N2 standard = 3.60 min, the tr of the sample's major peak should be within ± 0.07
min (2% relative) of this 3.60 min reference peak to be positively identified as N2.
1.) If the N2 GC peak area counts from a vaporized liquid phase N2 sample agree within 2% relative to the GC
peak area counts of a 100% N2 standard and the retention time (tr) of the sample’s major GC peak agrees
with a 100% N2 standard within 2% relative to each other, then no other ID testing is required. The sample
is positively identified as pure N2 (ID test "Pass").
2.) If either the peak area counts or peak retention time of the sample’s major GC peak do not agree within
2% relative to the standard then source re-sampling and repeat testing on a fresh, independent sample are
recommended to ensure the consistency of test results and to check for any sampling errors. Analyzing a
He blank standard through the same PR / flow path as the sample can establish the overall "air-free status"
of connecting PR hardware.
3.) Proper bulk N2 source sampling techniques and adequate cylinder / transfer line purging is essential to
prevent entrained artifact air from influencing the positive identification of pure N2 in the sample.
4.) Fresh, independent, representative samples of pure N2 gas should result in repeatable retention time and
peak area count test matches against a pure N2 standard. If adequate tr and peak area count agreement
cannot be established against a pure N2 standard, alternate equivalent methods should be used to investi-
gate the cause.
5.) Alternate N2 identification methods should be aligned with recognized standards such as USP and CGA.


TRACE OXYGEN (O2) IN NITROGEN (N2)

AND OTHER SELECTIVE ANALYZERS
ABSTRACT
This method determines the trace oxygen (O2) content in beverage grade nitrogen (N2). A gas chro-
matograph (GC) equipped with a pulsed discharge ionization detector (PDID), mass spectrometer or fuel cell-
/ zirconia cell-based voltametric analyzer can be employed. For GC based methods, other non-condensable
gas (NCG) trace impurities such as argon (Ar), hydrogen (H2), krypton (Kr), xenon (Xe), methane (CH4) and
carbon monoxide (CO) can also be measured. An NCG testing capability is useful for performing required %
N2 purity calculations (see Method 8.0N). A guideline limit of 10 ppm v/v O2 is recommended.
SAMPLING AND PHASE
1.) GC System with Electronic Data Handling: The recommended instrumentation system includes a GC
equipped with Helium (He) carrier gas, pressure controllable (ex. 0 – 1,000 torr) GC loop-filling VCR man-
ifold, gas loop injector (He purge-chamber housed), high resolution capillary or passivated packed col-
umn bank, GC oven with cryogenic-cooling (by liquid CO2) / temperature programming capability, auto-
mated column back-flushing / peak heart-cutting switching valves, a PDID (or equivalent) detector, and a
chromatographic control / data-handling system. For some acceptable column systems, cryogenic oven-
cooling may be needed to achieve the desired peak resolution. The column bank selected must resolve
oxygen (O2) from all other target air gas impurities. The most critical separation is O2 from argon (Ar). In
this method, a Haysep or Porapak® type "pre-column" (or equivalent) can be used to separate molecu-
lar sieve deactivating impurities such as CO2, N2O and H2O that may be in the N2 sample. A high resolu-
tion molecular sieve-type column connected beyond the pre-column is recommended for Ar / O2 separa-
tion. Other column types can be used if appropriately verified for this application.
The GC detector employed for this application must have sufficient sensitivity to measure O2 and other
air gases at very low or sub ppm levels. O2 and other NCG analytes are identified based upon a peak re-
tention time (tr) match vs. an external low ppm v/v gas standard in He. An electronic integrator or PC-
based chromatographic data handling system is recommended for GC peak data quantitation.
2.) Soft Ionization Mass Spectrometric (SIS) System with Electronic Data Handling: An SIS system em-
ploying a xenon (Xe) soft ionization beam and set to monitor the O2 molecular ion peak (m/e = 32) has
been appropriately verified for this application. This method is highly selective for O2. CH4 can also be
easily measured with this analyzer, but other NCG components such as H2, Ar, N2, and CO cannot be
measured due to their high ionization potentials.



3.) Fuel Cell Voltametric / Zirconia Cell Analyzer: This type of analyzer employs an O2-selective fuel cell
(galvanic cell) or zirconium (Zr) - catalyzed galvanic cell that produces a voltage signal related to the
sample’s O2 content. These types of analyzers are limited to the measurement of O2 only.
4.) Equipment for Cylinder Samples: A 2-stage metal diaphragm pressure regulator (PR) with an outlet ball
valve and also a fine metering valve (if not using an inlet manifold system for GC sample loop filling).
5.) Inlet Manifold System for GC Loop Filling: An optional manifold system designed to provide the GC
injector loop with an air-free representative sample of either calibration gas or sample at a precisely con-
trolled inlet pressure is highly recommended. It should consist of appropriate mating fittings for accom-
modating cup-in-metal, high pressure metal or low pressure metal sampling cylinders; diaphragm and fi-
ne metering valves; 1/16” (1.6 mm) sample transfer lines; sample / calibration gas switching valve (He
purged); loop pressure sensor system (ex. 0.0 – 1000.0 torr range); ss hardware plus a vacuum pump /
manifold isolation system for manifold air removal and loop system sample purging. This inlet loop mani-
fold is ideally suited for use when sample volumes and pressures are highly limited such as when
LPMC’s are employed.
6.) Helium Carrier Gas: Helium (He) carrier gas of 99.9999% purity and further purified with a heated O2
“getter” filter is recommended for most GC-based O2 analyzer applications. A slightly lower purity grade
of He can be used for GC valve blanketing.
7.) Carbon Dioxide GC Coolant: Cylinder of liquefied carbon dioxide gas of commercial-grade purity
(99.9%) equipped with an internal dip-tube for direct cryogenic liquid delivery to the GC oven-cooling in-
let.
8.) Span Gas Standard: A certified, compressed gas standard of approximately 2 - 40 ppm v/v O2 (and other
NCG gas analytes if desired) in N2 or He balance gas is typically recommended for calibration of analyz-
ers used for this application. Carefully follow manufacturer’s guidelines for selection of span gas stand-
ards and ensure that all standards are properly stored (ex. avoid storage in extreme temperature envi-
ronments) and within their marked expiration date.
9.) Zero (Blank) Gas Standard: A certified, O2 filtered N2 or He gas standard which is certified to contain
less than approximately 0.25 ppm of O2 and other target NCG impurities is recommended. Follow all
manufacturer instructions for selection of an appropriate zero gas.



ANALYTES
Primary Analyte
Oxygen (O2, MW = 32)
Optional NCG Analytes* (measurable by GC systems and useful for N2 test programs)
Hydrogen (H2)
Nitrogen (N2) – sample matrix gas
Methane (CH4) – recommended
Carbon Monoxide (CO) - recommended
Neon (Ne)
Argon (Ar) - key potential interferent to accurate O2 measurement by GC
Krypton (Kr)
*Note: O2 and other trace air gas impurities are separated from molecular sieve deactivating impurities such as CO2, N2O
and H2O vapor by use of a pre-column (ex. Haysep® or Porapak). The pre-column used should strongly retain
CO2, N2O and H2O. Upon elution of all target analytes including Ar, O2, N2, Xe, CH4 and CO from this pre-column,
the GC back-flush valve system is activated which reverses carrier flow through the pre-column to vent, thereby
removing all CO2, N2O and H2O impurities from the pre-column. This procedure protects the analytical molecular
sieve column(s) from CO2, N2O and H2O poisoning. In addition, a “heart-cut” valving system is recommended,
which diverts the major N2 peak from entering the final mole-sieve column and sensitive PDID detector. This keeps
the GC system in a more stable, robust state and allows easier measurement of late eluting CH4 and CO peaks.
MEASUREMENT RANGE
An overall linear working range of 0 - 50 ppm v/v for O2 and other NCG analytes is desired. This con-
centration range brackets the acceptable O2 limit for ISBT beverage grade N2 (10 ppm v/v). The manufactur-
er should provide sufficient validation data to ensure that their analyzer can routinely achieve this recom-
mended measurement range.
PRECISION
Typically a 5 - 10% relative range for O2 replicate measurements around the acceptable 10 ppm v/v
level should be attainable. This precision typically decreases to approximately 10 - 15+% for O2 and other
NCG analyte levels below 2 ppm v/v. The manufacturer should provide sufficient validation data to ensure
that their analyzer can routinely achieve this recommended precision.
LIMIT OF DETECTION
A minimum limit of detection (LOD) of 1 ppm v/v for O2 and other potential NCG impurities is desired
for beverage grade N2 applications. The manufacturer should provide sufficient validation data to ensure that
their analyzer can routinely achieve this recommended LOD.



INTERFERENCES
For beverage grade N2 applications, no interferences from impurities other than Argon (Ar) should be
encountered.
Argon can be present at levels up to a few thousand ppm v/v in typical cryogenic N2 sources. This method
requires that the GC system be capable of adequately resolving Ar from O2 peaks. Most GC/PDID in-
strument systems are very sensitive to contamination from external air due to fitting leakage, trace impurities
in carrier gas, and build-up / bleed off of highly retained impurities such as trace CO2 and H2O. Therefore,
leak-tight instrument systems typically involving VCR fittings (or equivalent), careful control of valve switching
parameters, use of high quality carrier gas/ scrubber (“getter”) systems and periodic column re-conditioning
are required. It is highly recommended that a PDID detector bypass step be performed after the elution of any
trace O2 peak and before elution of the major N2 peak. In this manner the N2 is kept off of the final column
bank and is not sent to the PDID detector – but is instead vented so as not to overwhelm the detector. After
the N2 peak has been vented (heart-cut), the PDID bypass valve can be returned to the PDID position for elu-
tion / detection of the CH4 and CO peaks, if desired.
Review all Safety Data Sheet (SDS) information on all pure gases and gas mixtures used, including
liquid and gaseous N2. Wear safety glasses and other necessary protective equipment, as chemical, pressur-
ized gas and thermal hazards are present. Perform all tests in a well-ventilated area. Carefully follow all op-
erational instructions provided by the equipment manufacturer.
PROCEDURE
A.) CALIBRATION
Calibrate the O2 analyzer on-use using an appropriate, O2-scrubbed blank and a certified O2 span gas stand-
ard according to manufacturer instructions. Any additional O2 and/or NCG working standards employed for
calibration or verification purposes should be carefully and freshly prepared and bracket the critical ISBT
limit for O2 (10 ppm v/v). Verify all certified cylinder standards are within their marked expiration date.
For GC-PDID systems, the following calibration method is typically employed:
1) Run a certified He blank and O2 standard (and, if desired, other target gas analyte standards) according to
GC manufacturer’s instructions using the same GC loop fill pressure and method conditions as the sample
(ex. Approx. 760.0 torr).
2) Determine the retention time (tr) and response factor (Rf)* of O2 (and any other desired NCG impurities)
based upon the net signal area measurement obtained from the span gas and zero standards.
*Note: R(f) (net area counts per unit concentration) = (span area counts – zero area counts) / concentration of
span gas.



B.) SAMPLE ANALYSIS - GC Systems (High Pressure & Low Pressure Sample Source Options)
Follow GC manufacturer instructions regarding sample loop pressure, carrier gas flow, column temperature
program, instrument settings, pre-column back-flush time, N2 peak heart-cut time and overall signal detection
and display conditions.
If a CIM cylinder is involved, attach a metering valve assembly to the CIM’s sample outlet valve to control
delivery pressure. If an HPMC containing over 500 psig (3,450 kPag) is involved, orient the sample cylinder
for convenient attachment of a 2-stage PR* with an outlet ball valve assembly to the HPMC’s outlet port.
Carefully open the cylinder valve and set the PR to the required pressure (ex. 2 - 5 psig [13.8 – 34.5 kPag])
with the ball valve closed. If an inlet manifold is not employed, also install a metering valve for injector loop
purge / fill control.
*Note: Small, low dead-volume PR’s are easier to purge of any entrapped internal O2 or H2O vapor.
1.1) Carefully open the sample cylinder valve and set the PR to the required pressure (ex. 2 - 5 psig [18.8 –
34.5 kPag]) with the outlet ball valve closed.
1.2) Slowly, carefully open the ball valve and allow the PR assembly to purge for about 10 - 20 sec, then
close the valve. Next, connect the PR-ball valve to the GC inlet manifold valve / transfer line assembly.
This manifold inlet system should consist of inert, (ex. 1/16” [1.4 mm]) ss tubing and associated VCR on
/ off and fine metering valves for flow control and line evacuation functions.
1.3) Thoroughly evacuate this line up to the purged and closed PR-ball valve, isolate the vacuum source, and
check for any leaks by monitoring any pressure increase in this line.
1.4) Close the manifold inlet line valves, carefully open the PR-ball valve and then slowly, carefully open the
manifold inlet line valves to slowly fill the evacuated inlet manifold and GC loop with sample gas to ap-
proximately 1,000 torr. Next, shut off the sample outlet valve and re-evacuate the manifold inlet / GC
loop. Repeat this purge / evacuation step several times to ensure that the cylinder outlet / inlet manifold /
GC loop system has been adequately purged of any entrained air.
1.5) Next, using the appropriate manifold inlet valves, slowly fill the inlet manifold / GC injector loop with the
sample to the desired system injection pressure (ex. approx. 760.0 torr).
1.6) Immediately inject the sample and establish the tr and peak area signal for O2 and any other target impu-
rities. If necessary, an additional repeat of this analysis can be performed to verify results.
2.) Low Pressure N2 Analysis (LPMC Samples)*
*Note: This low pressure analysis option is limited to GC systems due to the limited amount of sample gas available.
2.1) Securely orient an LPMC so that a sample from its outlet fitting can be obtained.
2.2) Connect sample inlet manifold valve / transfer line assembly to the outlet port of the LPMC.



2.3) Evacuate this line assembly up to the LPMC’s outlet port, isolate the vacuum source, and check for any
leaks by monitoring for any pressure increase in the manifold.
2.4) Close the manifold inlet line valves, carefully open the LPMC’s outlet valve and then slowly, carefully
open the manifold inlet line valves to fill the inlet manifold and GC loop with sample to approximately
1,000 torr. Next, shut off the sample outlet valve and re-evacuate the manifold inlet / GC loop. Repeat
this evacuation / purge step several times to ensure that the manifold and GC loop are air-free and pre-
treated with the N2 sample.
2.5) Upon completion of several inlet manifold / GC loop purge / fill cycles, slowly fill the manifold / GC loop to
the same sample pressure used for GC system calibration (ex. approx. 760.0 torr).
2.6) Immediately inject the sample and establish the tr and peak area count signal for O2 and any other target
impurities present. If necessary, an additional repeat of this analysis can be performed to verify results.
Verification Runs
It is recommended that periodically, the certified O2 span gas should be analyzed as a “sample” to
check that the analyzer is functioning under good calibration control. Alternatively, a verification check should
be made with a second, independent, certified O2 span gas standard. This “2nd confirmation” check helps to
ensure that the primary O2 “span” gas standard has not undergone any degradation or been contaminated
with external air. When applicable, system response “control chart” monitoring is also recommended. Verifi-
cation data should fall within ± 2s (s = experimental standard deviation) of the expected ppm v/v result, oth-
erwise analyzer re-calibration and data review since the last known good calibration should be performed.
CALCULATIONS
Most voltametric and mass spectrometric systems provide direct read values once properly calibrated
and require no further calculations. For GC analyzers, the following calculations may be required:
GC Calibration Calculations: An average GC / PDID response factor (Rf (I)) for O2 and other non-
condensable gas impurities in N2 is calculated using the following general formula:
Rf (i) = (Cal. Std. Conc. [ppm v/v] x 10,000*) / Peak Area Counts
Where i = O2, N2, H2, Ar, Kr, CH4 or CO

*10,000 factor is simply used to minimize / prevent calculator truncation errors
GC Sample Calculations: The amount of O2 or other target air gas impurities present in an N2 sample is cal-
culated from its average peak area count and its Rf value using the following formula:
Analyte Concentration (ppm v/v) = (Peak Area Counts x Rf) / 10,000



1.) If the O2 value obtained for a vaporized liquid N2 sample is below the maximum limit for beverage-grade
N2 (10 ppm v/v), no further testing is required.
2.) If the O2 value obtained for a vaporized liquid N2 sample is above the maximum limit for beverage-grade
N2 (10 ppm v/v), then source re-sampling and testing on a fresh, independent sample are recommended to
ensure the accuracy of the test result and to check for any sampling artifact errors. Connection of a certified
He blank standard through the same regulator / flow path as the sample can establish the overall "O2-free
status" of connecting hardware.
3.) Proper bulk N2 source sampling technique and adequate cylinder / transfer line purging is essential to
prevent an erroneously high O2 measurement. Experience indicates that for beverage grade N2 applica-
tions, in the majority of cases, erroneously high O2 measurements are related to air contamination intro-
duced by improper sampling and / or analytical techniques.
4.) Confirmation of an O2 result should be performed by equivalent methods.
5.) A GC-based analyzer is typically required to determine trace O2 and % v/v N2 Purity (by subtraction meth-
od) when a low volume, “non-hazmat” type LPMC and associated analytical testing program is employed.
Under these conditions, the GC system must be capable of measuring all of the listed air gas impurities
down to an LOD of 1 ppm v/v or lower. Use of duplicate LPMC cylinders is recommended to help confirm
O2 and % v/v N2 Purity results.


WATER VAPOR (H2O) CONTENT IN NITROGEN (N2)

BY ELECTROMETRIC AND
OTHER SELECTIVE ANALYZERS
ABSTRACT
This method determines the water vapor (H2O) content in beverage grade N2. Appropriate electro-
metric analyzers, infra-red (IR) spectrometers, soft ionization spectrometers (SIS) or dew cup systems can be
employed. Results are reported in ppm v/v H2O. There is no maximum guideline limit for H2O content.
SAMPLING AND PHASE
1.) Electrometric Moisture Analyzer: Analyzer systems utilizing an aluminum oxide (Al2O3) dielectric-
constant (capacitive) cell detector (CD)1 or an amperometric compensation cell detector (AD)2 comprised
of phosphorus pentoxide (P2O5) are commonly employed. Other types of electrometric analyzers can be
selected if appropriately verified for N2 use.
Note 1: This type of H2O analyzer employs a thin film sensor of Al2O3 in a dielectric cell configuration. Water vapor from
the sample gas stream passing through this cell is adsorbed by the Al2O3 which causes the dielectric constant of
this sensor cell to change. The amount of cell capacitive-change produced is electro-physically related to the
amount of H2O vapor present in the N2 sample. The changing electrical signal produced is processed and dis-
played as either ppm v/v water vapor or the corresponding dew point temperature (oF or oC). Charts are available
for ppm v/v to dew point unit conversion.
Note 2: This type of H2O analyzer employs a thin coating of P2O5. A fraction of the water vapor from the gas stream pass-
ing over this coating is chemi-sorbed to form phosphoric acid (H3PO4). This acid is then electrolyzed by the cell to
generate oxygen and hydrogen, which also regenerates the P2O5 coating. The electrolytic cell current needed to
regenerate this coating is fundamentally proportional to the amount of water vapor contained by the sample gas.
The electrical signal produced is processed and displayed as either a ppm v/v water vapor value or the corre-
sponding dew point temperature (oF or oC).
2) Infra-Red (IR) Spectrometer Systems with Electronic Data Handling: A 10M Gas Cell equipped, Fou-
rier Transform IR (FTIR) spectrometer equipped with cell pressure / temperature control, inlet manifold
system and quantitative data handling package has been appropriately verified for this application. The
peak absorbance signal of a selected H2O IR band at fixed temperature and pressure is measured and
converted to ppm v/v H2O using the Beer-Lambert Law.
3.) Soft Ionization Mass Spectrometric (SIS) System with Electronic Data Handling: An SIS system em-
ploying a xenon (Xe) soft ionization beam and set to monitor the H2O molecular ion peak (m/e = 18) has
been appropriately verified for this application. This method is highly selective for H2O.



4.) Sampling Equipment and Connective Hardware: 1/16 to 1/4” (1.6 – 6 mm) inner diameter gas imper-
meable transfer tubing; pressure regulator (PR) with an appropriate delivery pressure range; CIM, HPMC,
or LPMC containers and non-permeable connective transfer hardware.
5.) Sample Line “Bypass” System: It is highly recommended that some form of sample line bypass flow be
employed between the sample source and the analyzer inlet to ensure that a fresh, representative N2
sample is always presented to the analyzer.
6.) "Blank" Gas Standard: System blanks of H2O adsorbent cartridge-scrubbed N2 are recommended for
H2O analyzer zero-point calibration. Follow all manufacturer instructions for selection of a valid zero-gas
standard. This zero gas can also be used as a standby "dry purge" gas. Whenever the analyzer is not in
use, a dry N2 purge flow is required to maintain instrument sensor cell dryness and allow for immediate
start-up of sample testing when needed.
7.) Calibrated Electrometric Cells (Manufacturer-based): Because both CD and AD types of electrometric
detectors are based upon sound, well-defined electrochemical principles, they can be physically calibrated
for their specific electronic cell behavior against known H2O vapor levels, and therefore do not require rou-
tine external H2O span gas standard calibration checks. Follow manufacturer instructions for recommend-
ed sensor cell re-calibration schedules (typically every 12 months), along with their proper storage and
use.
8.) Span Gas Calibration Gas Standards / Working Standards*: Desired span and working H2O standards
in N2 can be prepared to nominal concentrations using volume-calibrated, 1 – 4 L compressed gas prepa-
ration equipment and precise pressurization to a maximum of 500 psig (3,450 kPag). The exact ppm v/v
H2O levels prepared by these units are then precisely measured by cell-calibrated electrometric-type ana-
lyzers. These freshly analyzed H2O standards can then be used to calibrate IR, SIS and other alternate
types of analyzers that require periodic calibration / verification with an external span gas standard. H2O
calibration standards prepared in this manner must be used within 8 hours of preparation. Follow analyzer
manufacturer instructions for recommended frequency of calibration and sensitivity validation checks. Al-
ternatively, H2O permeation tube-based standard generators can be employed for preparation of fresh
span and working H2O standards if desired. Careful attention to manufacturer instructions for proper use of
such permeation-tube standard generators is essential.
*Note: Currently, no certified, stable, commercially available gas standards of H2O in N2 or other matrix gases have been
appropriately verified for use in this application.
ANALYTE
Water (H2O, MW = 18)
MEASUREMENT RANGE
An overall linear working range of 0 - 10 ppm v/v for H2O is desired. The manufacturer should pro-
vide sufficient validation data to ensure that their analyzer can routinely achieve this recommended range.



PRECISION
Typically a 5 - 10% relative range (wr) for H2O replicate measurements at the 5 ppm v/v level should
be attainable. This precision typically decreases to approximately 10 - 15+% for H2O levels below 2 ppm v/v.
The manufacturer should provide sufficient validation data to ensure that their analyzer can routinely achieve
this recommended precision.
LIMIT OF DETECTION
A minimum limit of detection (LOD) of 1 ppm v/v for H2O is desired for beverage grade N2 applica-
tions. The analyzer manufacturer should provide sufficient validation data to ensure that their analyzer can
routinely achieve this recommended LOD.
INTERFERENCES
For beverage grade N2 applications, the analyzer manufacturer should provide appropriate interfer-
ence study data to indicate that their analyzer is free from potential measurement errors due to other N2 im-
purities or other influences. For all H2O analyzer systems, transfer line and hardware leaks are the primary
cause of erroneously high H2O data. Likewise, great care needs to be taken during N2 sampling to ensure
that all environmental air has been thoroughly flushed from the sampling train and that all sample containers
used for sample storage are leak-tight. Proper storage of sampling cylinders with a slightly positive residual
pressure is also important to ensure no external moisture enters during extended storage periods. For many
types of analyzers cell temperature, pressure and sample flow rate will influence H2O response; therefore,
careful control of these parameters is essential.
Review all Safety Data Sheet (SDS) information on liquid and gaseous N2. Wear safety glasses and
other necessary protective equipment, as chemical, pressurized gas and thermal hazards are present. Per-
form all tests in a well-ventilated area. Carefully follow all operational instructions provided by the equipment
manufacturer.
PROCEDURE
A.) CALIBRATION
Calibrate the H2O analyzer using appropriate blank and H2O span gas standards (when applicable)
according to manufacturer instructions. Any additional H2O working standards employed for calibration
purposes should be freshly prepared. When employing an electrometric-based analyzer ensure the detec-
tor cell is within its calibration period. If a dew point cup system is employed, ensure the temperature / dew
point sensing device is calibrated and within its calibration period. For SIS-type analyzers, an appropriate
indirect calibration “span” standard should be used along with an H2O-cartridge-scrubbed zero-grade N2
blank.



B.) SAMPLE ANALYSIS (High Pressure & Low Pressure Sample Source Options)
If a CIM type sampling cylinder is involved, attach a metering valve assembly to the CIM’s sample outlet
valve to control delivery pressure.
If an HPMC containing 500+ psig (3,450 kPag) of sample is involved, orient the sample cylinder for conven-
ient attachment of a ss, 2-stage PR* and outlet ball valve assembly to the HPMC’s outlet port. Carefully
open the cylinder valve and set the PR to the required pressure (ex. typically 2 - 5 psig [13.8 – 34.5 kPag])
with the ball valve closed.
*Note: Small, low dead-volume PR’s are easier to purge of any entrapped internal O2 or water vapor.
1.1) Follow all manufacturer instructions for analyzer flushing, sample delivery and measurement conditions.
1.2) Allow the analyzer display reading to stabilize before recording the H2O ppm v/v value. This may take
several minutes with some systems. Use of a sample inlet bypass system is recommended to quickly
flush the sample transfer line of internal moisture and expedite a stable, accurate H2O ppm v/v meas-
urement.
2.) Low Pressure N2 Analysis (LPMC Samples)
2.1) Low pressure, gaseous N2 contained in a LPMC can be employed. Careful attention to the manufactur-
er’s sampling instructions should be followed.
2.2) For FTIR systems equipped with a cell evacuation / pressure control manifold or other appropriate ana-
lyzer systems, connect the LPMC to the analyzer’s inlet system using a port connector and flow control
hardware.
2.3) Briefly flush the transfer line / analyzer inlet hardware according to the manufacturer’s instructions and
initiate the analysis. If necessary, an additional repeat of this analysis can be performed to verify results.
Verification Runs
It is recommended that a H2O span gas should be periodically analyzed as a “sample” to check that the ana-
lyzer is functioning under good calibration control. Alternatively, a rapid verification check can be made with a
second, independent H2O span gas standard. This 2nd confirmation check helps to ensure that the primary
H2O span gas standard employed has not undergone any degradation or been contaminated with external
air. When applicable, system response control chart monitoring is also recommended. Verification data
should fall within ±2s (s = experimental standard deviation) of the expected ppm v/v result, otherwise analyz-
er re-calibration and data review since the last calibration period should be performed.



CALCULATIONS
The identification of H2O in a sample is based upon the presence of a positive electrometric cell read-
ing, a characteristic IR absorption band or analytical mass peak of 18 amu. In all cases, the amount of H2O
present is determined from the sample’s signal area ratioed against a direct or indirect calibration span
standard or internally stored electrical property conversion table. Most H2O analyzers are direct read with all
sample calculations performed by the instrument. Conversion charts are available for ppm v/v to dew point
units if needed. For beverage-grade N2 applications, ppm v/v units are preferred for describing water vapor
impurity levels.
1.) If the H2O content of the vaporized liquid phase is below a desired limit for beverage grade N2, no other
testing is required.
2.) If the H2O content of the vaporized liquid phase is at or above a desired limit, source re-sampling and re-
peat testing are recommended. Connection of an H2O cartridge-scrubbed blank standard through the same
regulator / flow path as the sample can establish the overall integrity of the pressure regulator and connect-
ing hardware from external air leakage or wall diffusion. True H2O contamination in N2 will result in repeata-
ble test values when taken from fresh, independent samples.
3.) Proper bulk N2 source sampling techniques and adequate cylinder / transfer line purging are essential to
prevent erroneously high water vapor measurements. Experience indicates that for beverage-grade N2 ap-
plications erroneously high water vapor measurements are most often related to (environmental) H2O con-
tamination introduced by improper sampling and / or testing technique.
4.) Confirmation of high water vapor content should be performed using alternate, equivalent test methods.
5.) A traditional dew point cup apparatus can also be employed as a back-up analyzer when appropriate.
6.) A standard PSB sample should not be used for water vapor testing. Dual-valved multilayer sampling bags
may be used if appropriately verified for this application by the manufacturer.


TOTAL VOLATILE HYDROCARBONS IN NITROGEN (N2)

BY THC ANALYZER
ABSTRACT
This method determines the total volatile hydrocarbon (THC) content in beverage grade N2, which is
typically reported as "ppm v/v methane" equivalent. A Total Hydrocarbon Analyzer utilizing a flame ionization
detector (FID) and requiring a constant sample gas flow is employed. This THC test provides quantitative in-
formation about the overall amount of carbon and hydrogen containing impurities in a vaporized, liquid phase
N2 sample. A THC guideline limit of 5 ppm v/v is recommended.
SAMPLING AND PHASE
Methods 1.0N, 1.1N, 1.2N or 1.3N. Refer to each individual ISBT analytical sampling method for associated
details.
1.) THC Analyzer: Any THC analyzer employing a FID can be employed if it has been appropriately verified
for N2 use.
2.) THC Analyzer Support Gases: As applicable to specific THC analyzers: UHP Hydrogen (H2) and zero
grade air with THC less than 0.1 ppm v/v should be used. Alternately a 40% H2 in balance helium (He) or
nitrogen (N2) can be employed as a fuel gas for an extended measurement range if desired.
3.) Sampling Equipment and Transfer Line Hardware: Pressure regulators (PR) of appropriate delivery
capability, 1/16 to 1/4” (1.6 – 6 mm) ID transfer tubing and connective transfer hardware are recommend-
ed throughout the entire analyzer system from the sample source to the THC detector.
4.) Polymeric Sample Bags (PSB): 1 - 5 L PSB samples require either short transfer tubing or a gas tight
syringe interface to a pressurization chamber or inert pump to supply a constant, controlled sample flow to
a THC analyzer.
5.) Polymeric Bag Pressure Chamber Assembly (PCA): A pressure chamber of appropriate size, equipped
with a chamber PR (0 – 25 psig [0 – 173 kPag) / pressure relief valve, sample outlet ball / metering valves
and capable of meeting THC analyzer sample delivery requirements (for use with 1 – 5 L PSB samples).
6.) Inert, Pressure Controlled Pump (PCP): An electrically powered pump comprised of all inert, passivat-
ed, gas-wetted parts, pressure regulator (0 – 25 psig [0 – 173 kPag] and flow control hardware capable of
meeting THC analyzer sample delivery requirements (for use with 1 – 5L PSB samples).
7.) Span Gas Standard: A certified compressed gas standard of approximately 10 - 20 ppm v/v Methane
(CH4) in N2* and appropriate PR is required for THC analyzer calibration. Carefully follow all manufactur-
er guidelines for selection of this span gas standard and ensure that it is within its marked calibration date
and properly stored (ex. avoid storage in extreme temperature environments or use of long transfer
lines).
*Note: Do not use THC span gases with any other balance gas except nitrogen as other gases will produce a different
matrix response than N2 samples in a THC analyzer.
8.) Blank Sample Gas Standard: Zero-grade N2 with a THC content less than 0.1 ppm v/v and appropriate
PR with a 0 - 25 psig (0 - 173 kPag) delivery range is required for THC zero point calibration.



BY THC ANALYZER
ANALYTE
THC analyzers employing a FID will primarily detect signals from organic impurities containing car-
bon and hydrogen. This includes measurement of: C1-C6+ volatile aliphatic, olefinic, and aromatic hydrocar-
bons, volatile oxygenates, and volatile organo-sulfur impurities. The response of a THC analyzer is different
for individual organic impurities, but tends to be correlated with the number of carbons and hydrogen atoms
per molecule. Oxygenated impurities however, tend to give lower response*. For descriptive, comparison
purposes, THC test results are reported as "ppm v/v CH4 equivalence."
*Example: A 1 ppm v/v methanol (CH3OH) impurity generates a much smaller THC signal than a 1 ppm v/v
methane impurity. A 1 ppm v/v ethane (C2H6) impurity generates approximately double the THC
signal of a 1 ppm v/v methane impurity.
MEASUREMENT RANGE
An overall linear working range of 0 - 50 ppm v/v THC as CH4 is recommended. This concentration
range brackets the acceptable THC limit for ISBT beverage grade N2 (5 ppm v/v). A display range of 0.1 – 50
ppm v/v (as CH4) is ideal for this application. Lower ranges can be used if desired. The manufacturer should
provide sufficient validation data to ensure that their analyzer can routinely achieve this range.
PRECISION
Typically a 5% relative range for replicate measurements at the critical 5 ppm v/v THC limit should be
attainable. This precision typically decreases to 5 - 10% for analyte levels below 1 ppm v/v. The manufacturer
should provide sufficient validation data to ensure that their analyzer can routinely achieve this recommended
precision.
LIMIT OF DETECTION
A minimum limit of detection (LOD) of 0.5 ppm v/v or lower for all potential THC impurities is desired
for beverage grade N2 applications. The manufacturer should provide sufficient validation data to ensure that
their analyzer can routinely achieve this recommended LOD.
INTERFERENCES
All THC analyzers are flow-rate and matrix gas sensitive so careful control of these parameters is
required. All calibration and sample gases should be delivered to the THC analyzer at the same pressure.
Follow the manufacturer’s guidelines for proper calibration, including the required use of N2 as a balance
gas in all calibration standards. No chemical interferences are typical for beverage grade N2 applications.
Review all Safety Data Sheet (SDS) information on liquid and gaseous N2. Wear safety glasses and
appropriate hand protection when performing this test, as potential chemical, pressurized gas and thermal
hazards are present. Perform all tests in a well-ventilated area. Carefully follow all instructions provided by
the equipment manufacturer.



BY THC ANALYZER
PROCEDURE
A.) CALIBRATION
1.) Calibrate the THC Analyzer using the zero and span gas standards according to the equipment manufac-
turer’s instructions. The span gas standard used must match or exceed the maximum allowable THC
ppm v/v limit for beverage grade N2 (5 ppm v/v).
B.) SAMPLE ANALYSIS
1.) High Pressure N2 Analysis (CIM, HPMC or LPMC Samples)
- If a CIM type sampling cylinder is involved, attach a metering valve assembly to the CIM’s sample outlet
valve.
- If an HPMC containing more than 500 psig (3,450 kPag) is involved, orient the HPMC for convenient at-
tachment of a 2-stage PR* and outlet ball valve assembly to the HPMC’s outlet port. Carefully open the
cylinder valve and set the PR to the required pressure (ex. 2 - 5 psig [13.8 – 34.5 kPag]) with the ball
valve closed.
*Note: Small, low dead-volume PR’s are easier / quicker to purge of any entrapped internal O2 or H2O vapor.
1.1) Follow all manufacturer instructions for analyzer flushing, sample delivery requirements and final meas-
urement conditions.
1.2) Allow the analyzer display reading to stabilize before recording the THC ppm v/v measurement value.
This may take several minutes with some systems. Use of a controllable sample inlet bypass system is
recommended to flush the sample transfer line and expedite the achievement of a stable, accurate THC
ppm v/v result.
2.) Low and Atmospheric Pressure N2 Analysis (PSB Samples)*
2.1) Vaporized liquid nitrogen samples contained in a 1 – 5 L PSB need to be placed into a PCA or attached
to an inert pump with pressure / flow control hardware to establish a constant, controlled sample inlet flow
for a stable THC measurement.
2.2) Allow the analyzer’s display reading to stabilize before recording a THC measurement value. If neces-
sary, an additional repeat of this analysis can be performed to verify results.
Verification Runs
It is recommended that the certified THC span gas should be periodically analyzed as a sample to
verify that the analyzer is functioning under good calibration control. Alternatively, a rapid verification check
should be made with a second, independent, certified CH4 span gas standard (ex. 10 ppm v/v CH4 in N2 is
recommended). This confirmation check helps to ensure that the primary CH4 span gas standard has not un-
dergone any degradation. When applicable, system response control chart monitoring is also recommended.
Verification data should fall within ± 2s (s = experimental standard deviation) of the expected ppm v/v result,
otherwise analyzer re-calibration and data review since the last known good calibration should be performed.



BY THC ANALYZER
CALCULATIONS
Most THC analyzers are direct read; therefore, after calibration with zero and span gas standards the
analyzer will display THC content directly in “ppm v/v as CH4” units with no additional calculations required.
1.) If the THC content of the sample is below the guideline limit for beverage grade N2 (5 ppm v/v), no other
2.) If the THC content of the sample is at or above the guideline limit, source re-sampling and repeat testing
are recommended. Confirmation of high THC content should be performed using alternate, equivalent
test methods.
3.) If a non-FID type THC analyzer is employed the THC results obtained will NOT directly correlate with data
obtained from a FID-type analyzer unless:
3.1) The only organic impurity present is CH4.
3.2) Accurate relative “FID-response” factors are applied to all organic impurities identified and meas-
ured in the sample. Manufacturers of non-FID based analyzers should provide a sufficient list of
relative “FID-response” factors for accurately calculating the equivalent THC value that would be
obtained by an FID-equipped THC analyzer. Appropriate manufacturer-sourced validation data for
non-FID-based analyzers used for this application should be made available. ISBT THC limits are
based on FID-based analyzer data as a reference.
4.) Gas chromatographs (GC) employing an FID detector are useful for obtaining THC measurements when
LPMCs or non-hazmat sampling programs are involved or PCA equipment or THC analyzers are not
available. Like all non-FID methods, every GC speciated C1 - C6+ organic impurity observed must be
identified, quantified against appropriate gas standards and each ppm v/v value multiplied by its THC-
equivalent "conversion factor" and summed to equal a THC analyzer result in "ppm v/v as CH4" units.
5.) PSB samples should be analyzed as soon as possible and within the timeframe that the lab recommends
for maintaining the sample’s integrity. Avoid bag exposure to light during storage.


CARBON MONOXIDE (CO) IN NITROGEN (N2)

BY COLORIMETRIC DETECTION AND
ABSTRACT
This method determines the CO content in beverage grade N2. Colorimetric detection (detector tube
[DT]) systems, gas chromatography (GC) with selective detectors or Infra-Red (IR) based analyzers can be
employed. CO results are reported in ppm v/v units. A guideline limit of 10 ppm v/v CO is recommended.
SAMPLING AND PHASE
details.
1.) Colorimetric Detector Tube (DT) Based Systems: Several CO detector tubes employing either a hand
pump or gas manifold system are commonly used. If gas a manifold is employed, it must be capable of
precise sample flow and time control in order to deliver the required volume of gas through the DT for an
accurate CO measurement. The DT models (with appropriate color-producing chemistry in a N2 matrix)
and test conditions listed in this method have been appropriately verified for this application; other tube
models can be used if appropriately verified for N2 use.
2.) GC System with Electronic Data Handling: GC systems equipped with a sample inlet manifold, helium-
blanketed gas loop injector, column backflush system, cryogenic column cooling and ionization discharge
type detector (ex. PDID, DID, HID) are commonly employed. The column system selected must be capa-
ble of backflushing any trace CO2 (to protect the main analytical column) and baseline-resolving O2 from
other air gases such as Ar and N2 in an N2 matrix. A multi-column bank consisting of: Haysep T (pre-
column for CO2 adsorption), plus Molsieve 13X analytical columns (all in Sulfinert-passivated 1/8” tubing)
run under either isothermal or temperature program conditions has been appropriately verified for this
application. Alternate columns and temperature programs can also be used if properly validated. Tem-
perature programmed elution conditions and high load / efficient columns that produce a sharper CO
peak are recommended for maximum sensitivity. CO is identified by a peak retention time (tr) match
against a certified gas standard. Alternately, a GC system equipped with a gas loop injector, column
backflush / CO2 peak bypass capabilities, a ruthenium methanizer (for converting CO into CH4) and flame
ionization detector has also been validated for this application. An electronic integrator or GC-software
package is required for signal processing and final result calculation.
3.) GC Carrier Gas: Helium (He), Nitrogen (N2) or Hydrogen (H2) carrier gas of 99.999% purity is recom-
mended for most GC-based CO applications.
4.) IR Based Absorbance Spectrometers: A high resolution IR spectrometer in combination with a tempera-
ture and pressure controlled long path cell with an optical path length greater than 5 meters is recom-
mended. The spectral range for the detection of the IR absorbance for CO is 2000 – 2300 cm-1. The ab-
sorbance to concentration algorithm should be based on the Beer-Lambert law. To prevent ambient CO
from interfering with the measurement an inert gas system purge, vacuum or adequate mechanical purg-
ing arrangement is required.



5.) Sampling Equipment and Connective Hardware: 1/16 to 1/4” (1.6 – 6 mm) ID gas-impermeable trans-
fer tubing, small pressure regulators (PR), CIM, HPMC, LPMC or 1 – 5 L PSB sample containers and ap-
propriate connective transfer hardware are recommended throughout the entire analyzer system from the
sample source to the detector.
6.) Blank Gas Standard: Zero-grade N2 with a CO level less than 0.1 ppm v/v is recommended for instru-
ment zero-point calibration. Follow all manufacturer instructions for selection of a valid blank gas stand-
ard.
7.) Span Calibration Gas: A certified compressed gas standard of approximately 20 - 50 ppm v/v CO in N2
balance gas is recommended for analyzer calibration. The gas delivery system should include a pressure
regulator and short transfer lines. Carefully follow all manufacturer guidelines for selection of this span
gas standard (including matrix gas selection) and ensure that it is within its marked calibration date and
properly stored (ex. avoid storage in extreme temperature environments or use of long transfer lines).
ANALYTE
Carbon Monoxide (CO, MW = 28)
MEASUREMENT RANGE
An overall linear working range of 0 - 50 ppm v/v for CO is desired. This concentration range brack-
ets the acceptable CO limit for ISBT beverage grade N2 (10 ppm v/v). A display range of 1 – 50 ppm v/v is
ideal for this application. The manufacturer should provide sufficient validation data to ensure that their ana-
lyzer can routinely achieve this recommended range.
PRECISION
Typically a 5 - 10% relative range for replicate CO measurements around the critical 10 ppm v/v level
should be attainable. This precision typically decreases to 10 - 20% for CO levels below 1 ppm v/v. The
manufacturer should provide sufficient validation data to ensure that their analyzer can routinely achieve this
recommended precision.
LIMIT OF DETECTION
A minimum limit of detection (LOD) of 1 ppm v/v is desired for beverage-grade N2 applications. The
manufacturer should provide sufficient validation data to ensure that their analyzer can routinely achieve this
recommended LOD.
INTERFERENCES
For beverage grade N2 applications, the manufacturer should provide appropriate interference study
data to indicate that their analyzer can specifically identify and accurately measure CO in the presence of all
other potential N2 impurities. For DT-based analysis, consult the manufacturer’s information for potential in-
terferences associated with the DT’s colorimetric reaction chemistry.



Review all Safety Data Sheet (SDS) information related to all reagent chemicals and gases used in
this method. Wear safety glasses and appropriate hand protection when performing this test, as potential
glass, chemical, pressurized gas and thermal hazards are present. Perform all tests in a well-ventilated area.
Carefully follow all instructions provided by the equipment manufacturer. If employing detector tubes, follow
all manufacturer’s instructions for proper tube handling and ensure that all broken glass ends and spent DT
bodies are safely collected and disposed.
PROCEDURE
A.) CALIBRATION
1.) For GC-based analyzers, calibrate the system using appropriate blank and CO span gas standards ac-
cording to the manufacturer’s instructions. Any additional working standards employed for calibration pur-
poses should be freshly prepared and bracket the critical beverage-grade N2 limit (10 ppm v/v).
2.) When applicable, determine the retention time (tr) and response factor (Rf) values* of CO based upon the
net signal area measurements obtained from the span and zero gas standards.
*Note: R(f) (net area counts per unit concentration) = (span area counts – zero area counts) / concentration of span gas.
3.) For Colorimetric DT-based analysis, ensure that the detector tube model employed is calibrated by the man-
ufacturer using certified CO gas standards and manifold test conditions (when applicable). When following
this analytical protocol, no span or zero gas standards are required for routine use. Carefully follow all manu-
facturer instructions for either hand pump or manifold analyzer testing regarding:
3.1) Total number (n) of pump strokes required (for hand pump operations).
3.2) Controlled gas flow (cc/min) and test time (min) requirements (for gas manifold operations).
3.3) Ensure all hardware associated with the measurement has been properly leak-checked, sample
flushed and that all manifold flow / time measurement hardware has been properly calibrated
according to manufacturer instructions.
3.4) The DT model employed is appropriate for this application, has been properly stored and is not
past its marked expiration date.
4.) For Infrared Absorbance Spectrometers: For basic calibration of Infrared Absorbance Systems, a dataset of
different concentrations of CO have to be generated, calibration with quantitative Algorithms (in the relevant
spectral absorbance range), has to then be performed. Frequent baseline measurement has to be completed
after purging with Nitrogen or evacuating the measurement cell.
B.) SAMPLE ANALYSIS – GC Systems (High Pressure & Low Pressure Sample Source Options)
Follow GC manufacturer instructions regarding sample loop pressure, carrier gas flow, column temperature
program, instrument settings, pre-column back-flush time, N2 peak heart-cut time and overall signal detec-
tion / display conditions.



- If a CIM sampling cylinder is involved, attach a metering valve assembly to the CIM’s sample outlet valve to
control delivery flow.
- If an HPMC containing more than 500 psig (3,450 kPag) is involved, orient the HPMC for convenient at-
tachment of a 2-stage PR* and outlet ball valve assembly to the HPMC’s outlet port. Carefully open the cyl-
inder valve and set the PR to the required pressure (ex. 2-5 psig [13.8 – 34.5 kPag]) with the ball valve
closed.
*Note: Small, low dead-volume PR’s are easier to purge of any entrapped air and impurities.
1.1) Carefully open the cylinder valve and set the regulator to the required pressure (ex. 2 - 5 psig [13.8 –
34.5 kPag]) with the ball valve closed.
1.2) Slowly, carefully open the ball valve and allow the PR assembly to purge for about 10 - 20 sec, then
close the ball valve. Next, connect the PR-ball valve to the GC inlet manifold valve/transfer line assem-
bly. This manifold inlet system should consist of inert, small diameter (ex. 1/16” [1.4 mm]) ss tubing and
associated on/off and fine metering valves for flow control and line evacuation.
1.3) Thoroughly evacuate the manifold system up to the purged and closed PR-ball valve, isolate the vacuum
source, and check for any leaks by monitoring any pressure increase in this line.
1.4) Close the manifold inlet line valves, carefully open the PR-ball valve and then slowly, carefully open the
manifold inlet line valves to slowly fill the evacuated inlet manifold and GC loop with sample gas to ap-
proximately 1,000 torr. Next, shut off the sample outlet valve and re-evacuate the manifold inlet / GC
loop. Repeat this purge / evacuation step several times to ensure that the cylinder outlet / inlet manifold
/ GC loop system has been adequately purged of any entrained air.
1.5) Next, using the appropriate manifold inlet valves, slowly fill the inlet manifold / GC injector loop with the
sample to the desired system injection pressure (ex. approx. 760.0 torr).
1.6) Immediately inject the sample and establish the tr and peak area signal for CO and any other target im-
purities. . If necessary, an additional repeat of this analysis can be performed to verify results.
2.) Low Pressure N2 Analysis (LPMC Samples)
2.1) Securely orient an LPMC so that a sample from its outlet fitting can be obtained.
2.2) Connect GC sample inlet manifold valve / transfer line assembly to the outlet port of the LPMC.
2.3) Evacuate this line assembly up to the LPMC’s outlet port, isolate the vacuum source, and check for any
leaks by monitoring for any pressure increase in the manifold.
2.4) Close the manifold inlet line valves, carefully open the low pressure cylinder valve and then slowly, care-
fully open the manifold inlet line valves to fill the inlet manifold and GC loop with sample to approximate-
ly 1,000 torr. Next, shut off the sample outlet valve and re-evacuate the manifold inlet / GC loop. Repeat
this evacuation / purge step several times to ensure that the manifold and GC loop are air-free and con-
ditioned with a representative N2 gas sample.



2.5) Upon completion of several inlet manifold / GC loop purge / fill cycles, slowly fill the manifold / GC loop to
the exact sample pressure as used for GC system calibration (ex. 760.0 ± 1 torr).
2.6) Immediately inject the pressure-controlled sample and establish the tr and peak area count signal for CO
and any other target impurities present. . If necessary, an additional repeat of this analysis can be per-
formed to verify results.
3.) Low Pressure N2 Analysis (PSB Samples)
3.1) Vaporized liquid N2 contained in a 1 – 5 L PSB can be syringe transferred to a GC injector system (when
applicable) or directly sampled using appropriate transfer hardware and DT hand pump methods.
3.2) If necessary, an additional repeat of this analysis can be performed to verify results.
Verification Runs
It is recommended that periodically, the CO span gas should be analyzed as a “sample” to check that
the analyzer is functioning under good calibration control. Alternatively, a rapid verification check should be
made with a second, independent CO span gas standard. This “2nd confirmation” check helps to ensure that
the primary CO “span” gas standard has not undergone any degradation. When applicable, GC system re-
sponse “control chart” monitoring is also recommended. Verification data should fall within ± 2s (s = experi-
mental standard deviation) of the expected ppm v/v result, otherwise analyzer re-calibration and data review
since the last known good calibration should be performed. For DT-based analysis, whenever a measurable
CO value is obtained or if using a span gas standard, it is useful to employ an alternate DT model with a dif-
ferent reaction chemistry. This helps to ensure that no chemical interferences are present and that a tube’s
ppm scale accuracy is within acceptable tolerance.
C.) SAMPLE ANALYSIS – Colorimetric DT Systems
Manifold Systems
Required
Analyte & Color N2 Sample DT Scale Reading
Meas. Range Change Mfg./Part No. Volume (cc) Correction Factor
Carbon Monoxide White - Pale Gastec 1CO 500 none

2 – 50 ppm v/v Brown/Green
Carbon Monoxide White – Pale Gastec 1LK 500 x 0.5
Carbon Monoxide White - Drager 6733051 1,000 none
Carbon Monoxide Orange – Kitagawa 106SC 100 none
1 – 50 ppm v/v Red/Purple



Hand pump Systems
Analyte & Color Required DT Scale Reading

Meas. Range Change Mfg./Part No. Strokes (n) Correction Factor
Carbon Monoxide White - Pale Gastec 1LK 3 none

Carbon Monoxide White - Drager 6733051 10 none
Carbon Monoxide Orange – Kitagawa 106SC 1 none
1 – 50 ppm v/v Red/Purple
Consult manufacturer instructions for scale conversion factors if alternate sample volumes are used.
CALCULATIONS
The identification of CO in a sample is based upon either the presence of a characteristic color
change (ex. colorimetric DT), a retention time (tr) match vs. a CO standard (ex. GC) or by an absorption band
match (IR). Many GC and IR analyzers are direct reading with all sample CO calculations performed by the
data handling system. For DT-based analysis, follow all manufacturer instructions for any DT scale conver-
sions that may be needed to obtain the correct CO ppm v/v result.
1.) If the CO content of the sample is below the guideline limit for beverage grade N2 (10 ppm v/v), no other
2.) If the CO content of the sample is at or above the guideline limit, source re-sampling and repeat testing
are recommended. Confirmation of high CO content should be performed using alternate, equivalent
test methods.


ODOR IN NITROGEN (N2)

BY
VICINAL HEADSPACE SENSORY TESTING
ABSTRACT
This sensory method evaluates the presence of any foreign odor that may be detected in beverage
grade N2 obtained from a cryogenic liquefaction. A simple, gaseous “above” headspace odor evaluation using
a glassware-entrapped N2 sample is employed. This headspace method evaluates any sensory active
organic or inorganic impurities that can potentially be present either as an N2 production impurity or as a
contaminant that has been introduced during N2 load delivery or by an N2 distribution system. A pure, odor-
free N2 standard is used as a “control” in this test. It is suggested that this sensory test only be performed
after all other impurity tests have been completed and only if these tests have not detected any significant
levels of target impurities. This test is designed to allow the operator to safely access room air and normal O2
content throughout the entire test. Sensory test results are reported as a “Pass” = “no foreign odor” or a “Fail”
= “foreign odor detected”.
SAMPLING AND PHASE
details.
1.) Conical Glass Flasks: 2, glass 250 cc conical flasks calibrated at the 200 cc containment point and hav-
ing a 24/40 standard taper neck. One flask is employed for N2 sample gas collection and one used as a
"control" flask that is purges with pure, odor-free, pure N2 standard as an odor reference point.
2.) Impinger Heads: 2 glass impinger heads 24/40 taper with a delivery tube that extends to 5 mm above the
flask bottom and having a tapered tip with a 1 mm opening. The impinger head inlet and outlet ports are
recommended to be 1/4" (6 mm) for secure connections with standard inlet transfer tubing and plastic
end caps. One impinger head is inserted into to the Sample flask and one into the Control flask.
3.) Odor-Free Transfer Tubing and Sleeve Adapter: Chemically inert, sensory-neutral plastic tubing should
be employed between the pressure-reduced gas source and impinger inlets. A non-stick Norprene® (or
equivalent) rubber sleeve should be used for easy on / off transfer tubing connections to the glass im-
pinger heads.
4.) Manifold / Gas Delivery System: A pressure-regulated, flow-controlled and purged gas manifold system
comprised of suitable materials can be used if appropriately verified for sensory odor evaluation. Alter-
nately, a hardware system capable of reducing and controlling the flow of pure, odor-free N2 gas standard
or N2 sample gas into a glassware system at a maximum of 5 psig (34.5 kPag), and capable of metering
the gas flow supplied to test glassware (approximately 50 - 500 cc/min) is acceptable.
5.) Odor-free Gas Permeable Headspace Filter & Securing Band: A chemically inert, sensory-neutral
filter paper or polymeric filter membrane of wide porosity (ex. Whatman Grade xx or standard commercial
coffee filter material) comprised of a sufficient diameter that can be securely positioned over the wide-
mouth 24/40 taper port of a 250 mL conical flask and held in place with a rubber band or equivalent thin
tie strap.
6.) Odor-Free Testing Area: An indoor, temperature controlled area free of background environmental odors
is desired.



BY
7.) Sensory Trained and Prepared Personnel: As this test is purely sensory in nature, the personnel per-
forming this test must undergo a sensory training program in order to properly prepare for this test and to
objectively interpret test results.
8.) Zero-grade Sensory-Neutral N2 Standard: A certified, zero-grade N2 cylinder is recommended for control
testing and establishing a sensory-neutral reference point.
ANALYTES
This method evaluates the composite effect of all N2 impurities and contaminants that either individually or in
a synergistic manner impart an undesirable, detectable sensory effect with respect to odor in the N2 sample.
A positive sensory response is typically associated with the presence of volatile sulfur compounds, oxygen-
ates, unsaturated hydrocarbons, aromatics, acid gases and/or nitrogen-containing impurities. The measure-
ment range of this method is highly subjective. In some cases, however, the level of sensory response will be
clearly evident to trained sensory panel personnel.
MEASUREMENT RANGE
Not a quantifiable.
PRECISION
Not applicable.
LIMIT OF DETECTION
Not applicable.
INTERFERENCES
Room odors, improper cleaning of glassware (ex. residual detergent or use of scented detergent),
use of tobacco products, perfumes, ingestion of food / drinks, medications or exposure to any strong odors
less than 15 - 30 min before conducting this test can negatively influence a test result.
Review all Safety Data Sheet (SDS) information related to all reagent chemicals and gases used in
this method. Wear safety glasses and appropriate hand protection when performing this test, as potential
glass and pressurized gas hazards are present. Perform all tests in a well-ventilated area. Carefully follow all
instructions listed by the equipment manufacturers. N2 is classified by the United States Occupational Safety
and Health Administration as an asphyxiant. Odor determinations should never be made by direct, high vol-
ume inhalation of the N2 gas.
CAUTION: ISBT Method 7.0N glassware should NEVER be exposed to gas pressures exceeding 10
psig (69 kPag), as violent glassware rupture can occur resulting in serious injury or death. A pressure
regulator and/or 5 psig (35 kPag) relief valve system designed to allow only a very low, safe amount
(ex. 5 psig [35 kPag]) of delivery pressure into all glassware is required.



BY
PROCEDURE
Label one clean, dry 250-cc conical flask as "Sample" and one "Control". Install a clean impinger head
onto each flask. Impinger heads should also be labeled "Sample" and "Control".
1.1) Before N2 source attachment to the conical flask unit, purge approximately 1L of pure, odor-free N2
standard gas through the transfer line using a passivated, pressure-regulated, flow-controlled and 5 psig
[35 kPag] check-valve limited gas delivery system. This 5 psig [35 kPag] limited purge rate should be
about 250 cc/min for 4 min. After this purge period has elapsed, Shut off the N2 source flow, then quickly
remove the impinger head and install a clean, porous filter membrane over the conical flask’s tapered
glass port. Secure the position if this membrane “cap” with either a rubber band or plastic tie, so that any
odiferous vapors from the entrapped N2 sample must totally permeate through this porous filter “cap”.
This entrapped N2 standard gas sample is used as a sensory headspace “reference”.
1.2) Immediately sniff the vapors emanating just above (ex. 1” [2.5 cm]) the porous filter cap and use this ex-
perience as a reference point for subsequent “sample” testing. Use this sensory perception for N2 sample
odor comparisons.
1.3) Immediately attach the sample’s N2 delivery line to the sample impinger assembly inlet and maintain a
purge rate of about 250 cc/min for 4 min. After this purge period has elapsed, shut off the N2 source flow,
then quickly remove the impinger head and install a clean, porous filter membrane over the conical flask’s
tapered glass port. Secure the position if this membrane “cap” with either a rubber band or plastic tie, so
that any odiferous vapors from the entrapped N2 sample must totally permeate through this porous filter
“cap”. This entrapped N2 sample gas sample is now evaluated for any foreign odor against the sensory
headspace “reference” flask.
1.4) Immediately sniff the vapors just above (ex. 1” [2.5 cm]) the porous filter cap of this sample flask for for-
eign odors relative to the N2 reference standard flask. Use this sensory perception for N2 sample odor
comparisons. Record if any foreign sensory notes are detected.
1.5) Go to Step 3.
2.) Low Pressure N2 Analysis (PSB Samples)
2.1) Before performing this sensory test using 1 – 5 L PSBs contained N2 samples, initially fill a clean, virgin
polymeric bag about 80% full with the pure N2 standard gas.
2.2) Attach a short section of inert plastic tubing between the “Control” bag's outlet port and “Control” imping-
er inlet. Open the bag’s valve and slowly squeeze the bag to transfer its entire gas contents through the
“Control” sample flask over an approximate 2 min. period. After this purge period has elapsed, disconnect
the PSB, then quickly remove the impinger head and install a clean, porous filter membrane over the con-
ical flask’s tapered glass port. Secure the position if this membrane “cap” with either a rubber band or
plastic tie, so that any odiferous vapors from the entrapped N2 sample must totally permeate through this
porous filter “cap”. This entrapped N2 sample gas sample is now evaluated for any foreign odor against
the sensory headspace “reference” flask.



BY
2.3) Immediately sniff the vapors emanating just above (ex. 1” [2.5 cm]) the porous filter cap and use this as a
reference point for subsequent “sample” sensory testing. Use this sensory perception for N2 sample odor
comparisons.
2.4) Attach a short section of inert plastic tubing between the “Sample” bag's outlet port and “Sample” im-
pinger inlet. Open the bag’s valve and slowly squeeze the bag to transfer its entire gas contents through the
“Sample” flask over an approximate 2 min. period. After this purge period has elapsed, disconnect the PSB,
then quickly remove the impinger head and install a clean, porous filter membrane over the conical flask’s
tapered glass port. Secure the position if this membrane “cap” with either a rubber band or plastic tie, so that
any odiferous vapors from the entrapped N2 sample must totally permeate through this porous filter “cap”.
This entrapped N2 sample gas sample is now evaluated for any foreign odor against the sensory headspace
“reference” flask.
2.5) Immediately sniff the vapors just above (ex. 1” [2.5 cm]) the porous filter cap of this sample flask for for-
eign odors relative to the N2 reference standard flask. Use this sensory perception for N2 sample odor com-
parisons. Record if any foreign sensory notes are detected.
*Note: Do not place an impinger head on any contaminating surface and avoid handling its interior parts.
3.) Repeat the entire sensory test process on a fresh, independent sample, if the initial sensory comparisons
are inconclusive.
Verification Runs
Periodic blind checks of sensory personnel response should be made using a zero-grade sensory
blank to check for overall test equipment cleanliness and test consistency.
CALCULATIONS
No calculations are relevant. Pure, beverage-grade N2 is odorless. If a foreign odor is detected by

this sensory test it should be described as accurately as possible so that a follow-up investigation of the
chemical impurity / contaminant can focus on the most likely cause.
Foreign sidestream or flask headspace odors can be described as*:
- Oily - Solventy - Fruity - Rotten eggs - Sewer-like - Sulfurous - Cabbage-Like – Musty / Stale - Skunky -
Phenolic - Sour - Rubbery - Fishy
*Note: These sensory responses are usually linked to sulfur (ex. hydrogen sulfide, carbonyl sulfide), nitrogen
(ex. oxides of nitrogen), oxygenated organics (ex. acetaldehyde) or possibly unsaturated / aromatic im-
purities. Follow-up chemical analysis (ex. IR spectral profile) can be used to potentially identify and
measure the impurity / contaminant(s) causing the negative sensory response.
1.) Whenever a positive foreign odor test result is experienced, a replicate test using re-cleaned glassware,
alternate panel member(s) and fresh, independent N2 sample are required for result confirmation.



BY
2.) If a positive (failing) foreign odor test result is repeatable, other ISBT approved methods for identifying and
quantifying common odorous agents employing detector tubes, gas cell infra-red spectrometry, gas chro-
matography, or gas chromatography / mass spectrometry are recommended to investigate a failing senso-
ry test result.
3.) It is important to thoroughly clean all glassware soon after use, and to store all glassware in a hygienically
clean, secure area to prevent contamination from bacteria or chemicals. Overnight storage in a warming
oven just prior to use is recommended.
4.) All odor tests are highly subjective. It is strongly recommended that they only be performed after all other
required analytical tests have been completed and should not be performed at all, if any other N2 purity
test has resulted in a failing result.


NITROGEN (N2) % PURITY CALCULATION

BY ISBT TARGET LIST SUBTRACTION
ABSTRACT
This method determines the % purity (assay) of beverage grade nitrogen (N2). A subtraction calcula-
tion from 100% is employed for % purity reporting after all other ISBT target list impurities including trace ox-
ygen (O2), total volatile hydrocarbons (THC as methane), and carbon monoxide (CO) values have been
measured using appropriate ISBT analytical test methods. The impurities included are initially measured and
reported in ppm v/v units. For this % purity calculation, these ppm v/v values must all be converted into their
equivalent % v/v values for proper subtraction. % v/v N2 purity results are then reported to 6 significant fig-
ures (four decimal places). Calculation of % N2 purity is required, as no analytical test method is capable of
determining N2 purity at the high levels of precision required for this application. A N2 purity guideline limit of
99.99% v/v is recommended (maximum of 100 ppm v/v total of all impurities).
SAMPLING AND PHASE
The appropriate instrumentation systems and reagents called for in the following ISBT analytical test
methods should be employed:
ISBT 3.0N “Trace Oxygen (O2) in Nitrogen (N2) by Gas Chromatography and Other Selective Analyzers”
ISBT 4.0N “Water Vapor (H2O) Content in Nitrogen (N2) by Electrometric and Other Selective Analyzers”
ISBT 5.0N “Total Volatile Hydrocarbons in Nitrogen (N2) by THC Analyzer”
ISBT 6.0N “Carbon monoxide (CO) in Nitrogen (N2) by Colorimetric Detection and Other Selective Analyzers”
ANALYTES
Target Analytes (for % Purity Calculation)
H2 – (optional)
Ne – (optional)
Ar – (optional)
Kr - (optional)
O2 – (required analyte)
THC – (required analyte reported as CH4 equivalent value)
CO – (required analyte)



MEASUREMENT RANGE
Not applicable – this method is a mathematical calculation. Refer to individual ISBT test methods for
measurement ranges for each method.
PRECISION
precision for each method.
LIMIT OF DETECTION
limit of detection for each method.
INTERFERENCES
interferences for each method.
Not applicable – this method is a mathematical calculation.
PROCEDURE
Not applicable – this method is a mathematical calculation.
CALCULATIONS
1.) Obtain all sample ppm v/v data (reported to 2 significant figures) for all listed target analytes using the ap-
propriate ISBT methods.
2.) Convert all ppm v/v analyte results into their equivalent % v/v values to 2 significant figures.*
*Note: Significant figure conventions do not apply to number of decimal places. Significant figure includes all of those
digits known with measurement certainty plus the first uncertain digit.
3.) Add together all of the resulting target analyte results as % v/v (O2 + THC + CO) and report this sum to 2
significant figures (round off if applicable to 5 decimal places).
4.) Subtract this sum of all target impurities (less Ar and Ne) from 100.00000% v/v.
5.) Round-off and report (to four decimal places) the net % v/v N2 Purity value after this subtractive difference
calculation.



Example Calculation:
After all ISBT methods are completed, a beverage-grade N2 sample has the following impurity profile:
4.5 ppm v/v O2 2.0 ppm v/v THC 0.90 ppm v/v CO
A.) ppm v/v conversion to % v/v = ppm v/v X 0.00010
4.5 ppm v/v O2 impurity content = 4.5 ppm v/v X 0.00010 = 0.00045% v/v O2
2.0 ppm v/v THC impurity content = 2.0 ppm v/v X 0.00010 = 0.00020% v/v THC
0.90 ppm v/v CO impurity content = 0.90 ppm v/v x 0.00010 = 0.00009% v/v CO
B.) The sum of all target impurities = 0.00045 + 0.00020 + 0.00009% = 0.00074% (rounded off to 2 significant
figures)
C.) Calculation of % N2 Purity = 100.00000% v/v – 0.00074% v/v = 99.99926% v/v N2
D.) Final reporting value = 99.99% v/v N2 (rounded to four decimal places)
Note: Unless otherwise required this % N2 purity value includes trace, inert Ar and Ne impurities
1.) If the % N2 v/v value of a vaporized liquid phase sample is at or above the minimum % purity limit for
beverage-grade N2 (99.99% v/v) and a passing odor test is reported, then no other testing is required.
Currently, the recommended maximum ISBT allowable sum of all target list impurities is 100 ppm v/v.
2.) Proper bulk N2 source sampling technique and adequate cylinder / transfer line purging is essential to
prevent an erroneously high O2 and H2O measurements. Experience indicates that for beverage-grade,
cryogenic N2 applications, erroneously high O2 or H2O measurements are often related to environmental
air contamination introduced by improper sampling and/or testing techniques.

ANALYTICAL METHOD BIBLIOGRAPHY
CRYOGENIC N2 QUALITY SAMPLING METHODS
1.0N LIQUID NITROGEN (N2) SAMPLING USING CUP-IN-METAL CYLINDERS: CGA G-6.3-2002, IATA
Hazardous Shipping Regulations 2011, 49-CFR Hazardous Materials Shipping Regulations 2011.
1.1N GASEOUS NITROGEN (N2) SAMPLING USING HIGH PRESSURE METAL CYLINDERS: CGA G-6.3-
2002, IATA Hazardous Shipping Regulations 2011, 49-CFR Hazardous Materials Shipping Regulations 2011.
1.2N GASEOUS NITROGEN (N2) SAMPLING USING SMALL, LOW PRESSURE METAL CYLINDERS
1.3N GASEOUS NITROGEN (N2) SAMPLING USING POLYMERIC SAMPLING BAGS

2.0N NITROGEN (N2) IDENTIFICATION BY GAS CHROMATOGRAPHY
3.0N TRACE OXYGEN (O2) IN NITROGEN (N2) BY GAS CHROMATOGRAPHY AND OTHER SELECTIVE
ANALYZERS
4.0N WATER VAPOR (H2O) CONTENT IN NITROGEN (N2) BY ELECTROMETRIC AND OTHER
SELECTIVE ANALYZERS
5.0N TOTAL VOLATILE HYDROCARBONS IN NITROGEN (N2) BY THC ANALYZER
6.0N CARBON MONOXIDE (CO) IN NITROGEN (N2) BY COLORIMETRIC DETECTION AND OTHER
SELECTIVE ANALYZERS
7.0N ODOR IN NITROGEN (N2) BY VICINAL HEADSPACE SENSORY TESTING: CGA G-6-2-1994 Section
5.15.
8.0N NITROGEN (N2) % PURITY CALCULATION BY TARGET LIST SUBTRACTION


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Complete English-language documents may be obtained online from the ISBT at

Revision # Release Date Revision Notes

International Society of Beverage Technologists

Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines

Part 1 Beverage-Grade Nitrogen Quality Guidelines Page

Part 2 Analytical Methods for N2 Analysis

N2 Analysis – Preamble -------------------------------------------------------- 15

Part 3 Method List Method # (Pages)

Sampling Using Cup-In-Metal Cylinders --------------------------------------------- 1.0N (19-23)

Analytical Method Bibliography ---------------------------------------------------- 72

Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines

Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines

Parameter Guideline Limit Rationale1

N2 Purity2 99.99 % v/v min Process

Oxygen (O2)3 10 ppm v/v max Sensory

Total Volatile Hydrocarbons4 (THC) as

Odor No foreign odor Sensory

Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines

ANSI: American National Standards Institute (U.S.)

ASME: American Society of Mechanical Engineers (U.S.)

CFR: Code of Federal Regulations (U.S.)

CGA: Compressed Gas Association (U.S.)

DOT: Department of Transportation (U.S.)

EEC: European Economic Community

EIGA: European Industrial Gases Association

EU: European Union (includes 25 member states)

FAO: Food Additives Organization

FDA: Food and Drug Administration (U.S.)

GHS: Global Harmonization System (UN)

IATA: International Air Transport Association

ISBT: International Society of Beverage Technologists

ISO: International Standards Organization

JECFA: Joint FAO / WHO Expert Committee on Food Additives

NFPA: National Fire Protection Association (U.S.)

NIST: National Institute of Standards and Technology (U.S.)

OSHA: Occupational Safety and Health Administration (U.S.)

TC: Transport Canada

TPED: Transportable Pressure Equipment Directive (Europe)

UN: United Nations

WHO: World Health Organization

Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines

2.) has limited or restricted means for entry or exit.

3.) is not designed for continuous employee occupancy.

Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines

Figure 1 – Gas Cylinder

Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines

Non-Hazmat Shipment: Shipments of non-compressed N2 samples to an analytical testing lab. A variety of

Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines

Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines

4) Beverage Plant LIN Operational Factors

 Check pressure gauges and liquid level.

 Check for unusual build-up of ice on the storage vessel.

 Schedule external preventive maintenance and safety inspection with manufacturer.

Typical Maintenance Procedures for LIN Bulk Receiver Operations

Internal Cleaning Procedures for Bulk LIN Storage Vessels

Beverage Grade Nitrogen (Cryogenic Source) Quality Guidelines

Caution: Attempting to conduct internal cleaning is not recommended.

6) Quality Control Sampling & Testing Frequencies

7) Example - Certificate of Analysis (CoA) and Compliance (CoC)1

Nitrogen - Certificate of Analysis & Compliance Date of Issue

Distributing Equipment (tank truck, container, rail car):

Authorized Signature: _______ Date:

Printed Name: ______________ Title: