Introduction - Resources

Technology Basics | Required Materials | Test Kit Selection | General Tips | Resources

Product & Technology Information

The Science of ATP Monitoring - this document provides more details on how the ATP test works.
Evolution of the ATP Test - View a presentation given at the 2nd Society of Industry Microbiology Recent Advances in Microbial Control conference.
The Myths and Truths about ATP Monitoring - this document provides answers to some of the most common perceptions about rapid ATP testing technology.
2^nd Generation vs. 1^st Generation ATP - this document discusses in great detail the fundamental differences between 2^nd and 1^st Generation ATP tests, complete with validation data for 2^nd Generation technology.
2^nd Generation ATP vs. Other Microbiological Tests - the definitive comparison of 2^nd Generation ATP testing versus other microbiological test methonds can be found in this document.
Surveying Microbiological Test Methods - View a presentation done at GlobalSpec's Water Quality & Water Reclamation online conference comparing various microbiological test methods.
Going Green with ATP - View our webinar on how 2^nd Generation ATP testing can assist in your efforts to make your process more environmentally-friendly!

Troubleshooting, Diagnostic & Other References

Establishing the Detection Limit of ATP Assays - This document explains how we have established the detection limits of our off-the-shelf test methods, and how you can modify the detection limit if you wish.
Frequently Asked Questions - Some of the most common questions and answers regarding our product line are covered here.
Frequently Asked Questions - Luminase - This document builds on our basic FAQ, with specific questions and answers regarding the Luminase family of reagents.
UltraCheck and the Benefit of ATP Standard Measurements - This explains why the UltraCheck family of ATP standards are paramount to quantitative ATP analyses.
Diagnostics and Troubleshooting - This document provides a matrix for troubleshooting common issues with 2^nd Generation ATP testing.
Troubleshooting Focus: dATP > tATP - This document deals specifically with what to do when results indicate a higher Dissolved ATP concentration than Total ATP.
Effects of High-Intensity Light on Cuvettes - Occasionally, high blank readings are obtained from cuvettes used to conduct ATP analyses. This document provides an overview of this phenomenon and provides guidance on how to overcome it.
ATP vs. CFU Part 1 - This powerpoint presentation complete with notes explains why results from 2nd Generation ATP test methods are different from those generated with classical culture methods.
ATP vs. CFU Part 2 - This powerpoint presentation complete with notes build on Part 1 (above) and explains why there can often be unexpected variation between 2nd Generation ATP and CFU/mL trends.

► 1 femtogram of ATP is equivalent to 1 E. Coli-sized bacteria

This is a frequently cited conversion and has been observed by many researchers, including LuminUltra. There are a variety of ATP-to-cell number correlations that have been done by various researchers using pure cultures.

The reference citing the above conversion can be found in an October 2001 Application Note by Lee and Deininger entitled “Rapid quantification of viable bacteria in water using an ATP assay”. The precise citation is as follows on the second page:

It is known that the average ATP content in one bacterial cell is about 1 fg.

The reference cited as the source of this conversion: Crombrugge J, Waes G. ATP method. In: Heeschen W, ed. Methods for assessing the bacteriological quality of raw milk from the farm. Brussels: International Dairy Federation, 1991: pp. 53–60.

► 1 part ATP is equivalent to 250 parts biomass carbon

The reference citing this conversion can be found in a July 1978 paper by Holm-Hansen entitled “Chemical and biological characteristics of a water column in Lake Tahoe”. This reference states:

ATP (in micrograms per liter) was converted to organic carbon in living cells by multiplying by 250 (Holm-Hansen 1970); this carbon value was then multiplied by 10 to give biomass wet weight ( microgram liter-l).

The reference (Holm-Hansen 1970) is listed as:

Holm-HANSEN, 0. 1970. ATP levels in algal cells as influenced by environmental conditions. Plant Cell Physiol. 11: 689-700.

► Dry biomass is approximately 50% carbon

This is a very common conversion found in many industry reference books. You can find it in the Metcalf & Eddy handbook of wastewater treatment. In the 4th Edition, it can be found on page 558 in table 7-4.

Glossary

Sample: The sample from the process to be tested for ATP concentration.

Diluted Sample: A portion of sample diluted in a stabilizing buffer for dATP measurement.

Extract/ATP Extract/Sample Extract: The mixture produced when a sample is treated with an ATP extraction agent (i.e. UltraLyse).

Diluted Extract: The UltraLyse-treated portion of sample that is diluted prior to the assay step.

Assay (or ATP Assay): Measurement of the light produced when an ATP-containing mixture is mixed with Luminase and placed in a luminometer.

Luminometer: Instrument responsible for measuring the RLU emitted during an assay.

RLU: Relative Light Unit. This is the instrument display (i.e. the amount of light output) on a luminometer after an assay has been completed.

ng/mL: Nanogram per milliliter. This is the standard unit for ATP concentrations in wastewater treatment applications, and is equivalent to "parts per million" (ppm).

pg/mL: Picogram per milliliter. This is the standard unit for ATP concentrations, and is equivalent to "parts per billion" (ppb).

fg/mL: Femtogram per milliliter. This is the standard unit for ATP concentrations in single cells, and is equivalent to "parts per trillion" (ppt).

Dilution Factor (DF): The factor by which the user must multiply the standardized assay RLU to compensate for the dilution of the original sample (to remove inhibition).

Total ATP (tATP): The total amount of ATP in the sample.· Intra-cellular + Extra-cellular.

Intra-cellular ATP: The portion of sample ATP associated with (i.e. contained within) microorganisms.

Cellular ATP (cATP): LuminUltra's terminology for Intra-cellular ATP measurement or quantification.

Extra-cellular ATP: The portion of sample ATP not associated with (i.e. contained outside) microorganisms.

Dissolved ATP (dATP): LuminUltra's terminology for Extra-cellular ATP measurement or quantification.

Biomass Stress Index (BSI): The ratio of Extra-cellular to Total (Intra-cellular + Extra-cellular) ATP.· Put simply, it is the proportion of ATP in the sample that is outside of living cells.· A higher BSI is indicative of greater stress.

Floc-Bulking ATP (fbATP): A measurement of ATP associated with bulking floc, achieved by pre-screening the sample and measuring the Total ATP retained on that screen.

Specific Floc-Bulking ATP (s-fbATP): The ratio of fbATP to tATP; in other words, the specific proportion of ATP associated with bulking floc relative to the total amount.

Total Suspended Solids (TSS): The total concentration of suspended matter in a given sample.· Includes all particles, both inorganic and organic, a subset of which is living biomass.

Volatile Suspended Solids (VSS): The concentration of organic suspended matter in the sample.· Includes only organic particles, a subset of which is living biomass.

Active Volatile Suspended Solids (AVSS): The·concentration of living, suspended matter in the sample.· Calculated by converting the cATP from ng ATP/mL to mg Biomass/L using third-party correlations.

Active Biomass Ratio (ABR): The ratio of AVSS to TSS; in other words, the proportion of total suspended matter that is living biomass for a given sample.

Colony Forming Units (CFU): A conventional unit of measure for culture-based microbiological test methods. Each colony that is counted after a specific time period is 1 CFU.

Microbial Equivalents (ME): A conversion of 1 fg ATP into 1 E. Coli sized cell. Used for putting ATP results on the same scale of measurement as CFU.

Special Protocols & Validation Resources

QGA Validation Package - this document outlines initial experiments you can run to validate the QGA test kit for your application.
QGO-M Validation Package - this document outlines initial experiments you can run to validate the QGO-M test kit for your application.
QG21W Validation Package - this document outlines initial experiments you can run to validate the QG21W test kit for your application
QG21I Validation Package - this document outlines initial experiments you can run to validate the QG21I test kit for your application
QG21S Validation Package -this document outlines initial experiments you can run to validate the QG21S test kit for your application
Biocide/Kill Study Fundamentals - this document provides fundamental guidance on how 2^nd Generation ATP testing can be used to augment/compliment existing methods for the purpose of evaluating microbiocidal process performance (e.g. chemical treatment, pasteurization, filter sterilization, etc). For a more descriptive methodology on setting up a comprehensive microbiocidal evaluation, consult ASTM Standard E2783 (Standard Test Method for Assessment of Antimicrobial Activity for Water Miscible Compounds Using a Time-Kill Procedure).

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