Attachment 09 - NIH Particle Distribution.1714658465646.docx

Attachment 09: NIH Particle Distribution

Objective of the Analysis: The testing protocol was developed based on ISO 21501 – 2: Determination of particle size distribution – single particle light interaction methods. The testing protocol describes the calibration and verification method for light scattering liquid borne particle counter (LSLPC), which is used to measure the size and particle number concentration of particles suspended in liquid. The typical size range of particles measured is between 0.1 micron to 10 micron in particle size.

Procedure of Sample Collection and Storage NIH shall collect samples, and the Contractor shall pick up the samples from NIH Bethesda, Building 11. The Contractor shall provide NIH with sampling kits for collecting each sample.

A. In the event that the Contractor collects the sample, they shall perform the following:Operational settings of side stream filtration systems during sample collection

a. Ensure that the side stream filter is in service mode with the system chiller running

b. Ensure sample valve is opened and left to run to avoid disturbing flow once it is established through the sample point and dislodging any material

c. Sample bottle shall be rinsed three times with system water and then filled to the top

d. Sample bottle top shall be rinsed, filled, and then screwed on to the bottle to ensure no air is present in sample B. Procedures for sample shipping and delivery to the analytical lab

a. Sample bottles shall be prepared for shipping the day of sampling

b. System labels shall be applied indicating which tests to be run

i. Basic Analysis plus bio count

ii. Bio identification

iii. Particles

c. Bottles shall be taped to ensure caps do not loosen in transit

d. Samples shall be placed in a waterproof clinical pack, then packaged in priority shipping box and shipped overnight for 10:00 AM deliver to the analytical laboratory.

C. Calibration processes of the instrument Comment by Hughes, Casey (NIH/OD/ORF) [E]: What do you mean by “Lab staff typing this up”? Comment by Gomes, Andrew (NIH/OD/ORF) [E]: These are elaborated in the text.

a. The lContractorab shall provide to NIH their calibration procedure upon request. will come out with their own calibration procedure and will share with NIH for their approval.

D. data analysis procedures for the particle size distribution including count and volume.

a. The Contractor shall provide to NIH their data analysis procedure upon request.The lab will come out with their own data analysis procedure and will share with NIH for their approval.

Sample Preservation and Storage Water samples shall be placed on ice and reach the Laboratory as soon as possible after collection as testing must be completed within 24 hours.

Method Of Shipping and Duration and Receiving The Contractor shall place the sample into a cooler with ice for immediate delivery or shipment to the laboratory. The sample shall be received with a chain of custody (COC). The COC shall have the name of the sample collector, date and time of sample collection, signature or initial of the sample collector, location of the sample and type of sample to be analyzed.

Analysis Method and Protocol:

Instrument: The Contractor shall perform the analysis with the HIAC 9703+ Pharmaceutical Particle Counter or equivalent instrument. The instrument shall be used for light obscuration in accordance with USP 787 or 788 standards.

Size setting error: The maximum permissible error (MPE) for size setting in the minimum detectable particle size and other sizes specified by the manufacturer of an LSLPC is 15% of the specified size. Pulse height analyzer (PHA) obtains a pulse height distribution for a sample liquid in which calibration particles are suspended. The lower voltage and upper voltage limits determine the median voltage of the pulse height distribution and is assigned to the certified size of the calibration particles.

Counting efficiency: Counting efficiency shall be defined as the ratio of the concentration measured by a light scattering liquid borne particle counter to that measured by a reference instrument for the same sample. The counting efficiency shall be within 20% to 80% for calibration particles with a size close to the minimum detectable size, and it shall be within 70% to 130% for calibration particles with the particle size 1.5 to 3 times larger than the minimum detectable particle size. When calibration particles with the same size as the minimum detectable particle size are not available, particles whose size are within ±5% of the minimum detectable particle size may be used, and the diameter of the calibration particles shall be reported.

Size resolution: The size resolution is the measure of the ability of an instrument to distinguish between particles of different sizes. It shall be less than or equal to 10% of the specified particle size when it is evaluated using calibration particles of a certified average size specified by the manufacturer.

False count: The false count refers to an apparent count per unit volume of sample liquid when a sample liquid contains no measurable particles. The false count per liter of volume and its 95% upper confidence limit shall be determined. The 95% upper confidence limit shall be less than or equal to the value specified and reported by the manufacturer of the LSLPC.

Maximum particle number concentration: The maximum measurable particle number concentration shall be specified by the manufacturer. The coincidence loss (reduction of particle count caused by multiple particles passing simultaneously through the sensing volume and by the finite processing time of the electronic system) at the maximum particle number concentration of an LSLPC shall be less than or equal to 10%.

Calibration interval: The calibration of the LSLPC shall be conducted at an interval equal to or shorter than one year. The instrument requirements shall be met during the calibration interval.

Calibration particles: The calibration particles shall be monodisperse spherical particles with a known mean particle size (polystyrene latex particles, for example), where the certified size is traceable to the SI units. Relative standard uncertainty needs to be equal to or less than 2.5%, and a refractive index is approximately 1.59 at the wavelength of 589 nm.

Procedure A light scattering liquid-borne particle counter (LSLPC) is used to measure the size and particle number concentration of particles suspended in liquid. The Contractor shall use the light scattering method described in ISO 21501-2:2007 which is based on single particle measurements. The typical size range of particles measured by this method is between 0.1 µm and 10 µm in particle size. Instruments that conform to ISO 21501-2:2007 shall be used for the evaluation of the cleanliness of pure water and chemicals, as well as the measurement of number and size distribution of particles in various liquids. The measured particle size using the LSLPC depends on the refractive index of particles and medium, and the measured particle size is equivalent to the calibration particles in pure water.

ISO/IEC 17025 specifies requirements for the competency of the entity to carry out tests and/or calibrations, including sampling. It covers testing and calibration performed using standard methods, non-standard methods, and laboratory-developed methods.

Maintenance Plan for the Instrument The Contractor shall maintain equipment according to ISO 21501 protocol.

Calibration Procedure

1. The Contractor shall perform calibration on associated equipment according to ISO 21501 protocol. Identification and statement of metrological traceability of all reference equipment and calibration particles used shall be noted.

2. Relevant environmental conditions (temperature, air pressure, and humidity) under which the calibration was performed shall be noted.

3. A stated uncertainty for each result with reference to the calculation method

4. The false count per volume in litre and its 95 % upper confidence limit (UCL) shall be determined according to ISO protocol. The evaluation of false particle count shall be obtained at the size channel corresponding to the minimum detectable particle size for a certain volume of particle free liquid to the LSLPC under test.

5. The counting efficiency of the LSLPC shall be determined by two populations of calibration particles as mentioned to the ISO protocol.

6. Sample volume error shall be evaluated by measuring the sampling volume by weighing the pure water with the balance and converting to volume, or measure the volume by means of a calibrated graduated cylinder.

Reporting of Test and Calibration Results The particle size distribution report shall contain the following information:

The calibration certificate shall include:

1. Identification and statement of metrological traceability of all reference equipment and calibration particles used.

2. Relevant environmental conditions, such as temperature, air pressure and humidity, under which the calibration was performed.

3. A stated uncertainty for each result for size setting error and counting efficiency with reference to the ISO calculation method

4. A stated false count at a 95% confidence limit.

Quality Assurance and Quality Control The Contractor shall follow quality control procedures according to ISO 21501 protocol.

The lab runs particles on a Hiac 9703+ counter that is paired with a MC05 laser that utilizes a light scattering laser diode. Samples are diluted and dispersed in ultra-filtered lab deionized water and are drawn through the laser using a glass syringe. All samples shall have a blank run prior to each analysis to verify the cleanliness of the dilution water, and the software shall subtract the background of the dilution water off the results. Each sample shall be run ten times and averaged to allow for data to be normalized.

Data Acquisition The data shall be collected according to the ISO 17025 Protocol.

Fate of Sample (Destruction or Return) After analysis, the sample shall be discarded by the lab unless otherwise instructed.

References ISO 21501-2, Determination of particle size distribution, https://www.iso.org/standard/75048.html ISO 17025, Testing and calibration laboratories, https://www.iso.org/ISO-IEC-17025-testing-and-calibration-laboratories.html