SAMPLING PLANS, SAMPLE
COLLECTION, SHIPMENT, AND
PREPARATION FOR ANALYSIS
OBJECTIVES
Assist sample collectors to obtain
representative samples of a food based on:
Appropriate sampling plan design
Prepare samples for shipment to a
laboratory in a condition that is
microbiologically
unchanged from:
The time of sampling
Prepare samples for the analysis
The first priority in the microbiological
examination of any food products is a
representative sample that is appropriately
collected, transported in the laboratory
and prepared for examination
The results and interpretation of the
laboratory is valid only when appropriate
samples have been examined
APPROPRIATE SAMPLE
Samples must:
Representative of the entire lot
material under evaluation
The proper type of sample for the
analysis to be conducted.
Protected against extraneous
contamination or improper handling
Packaging
Storage temperature
Other modes of preservation
GENERAL CONSIDERATIONS IN FOOD
SAMPLING
Packaging
Clean and clear container.
Original packaging
Sterile container or sampling bags.
Temperature:
To prevent the destruction of the
growth or
organisms in a sample, REFRIGERATION must
be
provided for holding and storage.
Unfrozen samples with high water
activity (aw
>0.64) must be refrigerated preferably between 0
to 4°C from the time of collection until the point
of
analysis
Samples of frozen foods should be
collected and
shipped solidly frozen.
A sealed eutectic coolant in the
shipment
containment is recommended to avoid
contacting the product with melting ice or
coolant.
When dry ice is used as shipping
coolant, the
containers should have tight closure to prevent
pH change in the samples caused by the
absorption of carbon dioxide.
As a general rule, samples should be
examined as soon as possible or within 36
hours after sample collection
Perishable samples cannot be analyzed
within 36 hours should be frozen or retained
inrefrigerated temperature up to 18 hours.
Unfrozen samples of shellfish should be
examined within 6 hours after collection
 Each sample or shipment must be
clearly and
completely identified by the following
information:
Sample description
Collector’s name
Name and address of food
manufacturing plant
Production lot number
Dealer or distributor
Date, place and time of collection
Reason for testing
EQUIPMENT, MATERIALS,
AND REAGENTS
Instruments for opening containers
Sterile scissors, knives, scalpels, and
can openers
Sample transfer instruments
Sterile single use spatulas, scoops,
spoons, triers,
forceps, tongue depressors, drills, auger
bits, corers,
dippers, metal tubes, and swabs.
Sample containers
non-toxic, leak proof, and pre-
sterilized polyethylene bags
Sterile plastic vials and bottles
Sterile vacuum packaging equipment
Sterile glass bottles are not advisable
to use due to high risk of breakage.
Thermometers: thermometers should be
using can measure between -20 to 100°C.
Microbiocide: Medium strength
(100mg/L) hypochlorite solution, 70% ethyl
alcohol,
and 71% isopropyl alcohol.
Labelling supplies: pressure sensitive
tapes and labels, tags of adequate size to hold
essential sample information, and indelible
marking pens.
Sample shipping containers:
For frozen or refrigerated products:
use insulated rigid metal or plastic
containers that are equipped
with a tight-fitting cover. Each container
should have an ample space for
refrigerant so that samples will remain at
the desired temperature until their
arrival at the laboratory.
Non-perishable samples: do not need
refrigeration. Containers should be made
from sturdy corrugated cardboard.
Balance: a calibrated balance between
2000 g capacity and a sensitivity of 0.1
g with a 200 g load is acceptable.
BALANCE
▪Abide to Good Weighing Practices
when operating with balances.
▪Calibration: is the act of comparing a
device under test (DUT) of an unknown value
with a reference standard of a known value.
▪Importance of calibration:
▪ The goal of calibration is to
minimise any measurement
uncertainty by ensuring the
accuracy of test equipment.
▪ Calibration quantifies and
controls errors or uncertainties
within measurement processes
to an acceptable level.
▪ To determine the accuracy of
instrument.
▪Verification: process that verifies
whether an instrument performs according to its
intended purpose, meeting predetermined
specifications and requirements consistently
. ▪Use of calibrated test weights
EQUIPMENT, MATERIALS,
AND REAGENTS
Blenders and mixers: use of
mechanical
blenders with several operating speeds or
variable speed control with sterile glass or
metal blending jars, and covers, a
stomacher.
Diluents:
Butterfield's phosphate buffer
0.1% peptone water
PRECAUTIONS DURING SAMPLE
COLLECTION/SAMPLING
▪The sampling operations must be organized in
advance with all necessary equipment and sterile
containers available.
▪Sampling instruments should be protected from
any source of contamination before and during
use.
▪When using sampling equipment to collect
samples, the sampling instrument should not be
passed over pre-sterilized instruments.
▪The sterile sampling containers should be open
sufficiently to admit the sample and then closed
and sealed immediately.
▪Do not touch the inside of the sterile container’s
lid.
▪Do not allow the open lid to become
contaminated.
▪Do not hold or fill a sampling container over
the
top of a bulk food container when transferring a
sample.
▪To prevent overflow and proper mixing of
sample in the laboratory, the sample container
should not be filled more than three-fourths full.
▪Do not expel air when folding or whirling
plastic
sample bags.
▪An empty sampling bags similarly open and
closed should be submitted in the laboratory as
control.
▪Samples of pre-sterilized gloves that were used
can be also tested.
▪The sample collectors must keep their hands
away from their mouth, nose, eyes, face, and
other body parts and other contaminating factors
in the environment.
▪Hands must be washed immediately before
beginning the sampling and washed during
sampling if they become contaminated.
▪Using sterile plastic gloves may prevent
contamination during the execution of the
procedure.
▪Contaminated sampling equipment must be
placed into proper containers for later disposal
or decontamination.
▪Direct label the container
SAMPLING PLANS
▪In 1923, engineers of Western Electric
Company first developed sampling plans.
▪Typical sampling points along food processing
continuum:
▪ Raw materials
▪ Production line
▪ Producer’s warehouse
▪ Retail storage or outlet
▪ International port-export or import
▪Sampling plan is developed so that the selection
of samples taken from a lot is
performed in a manner that ensures that each
sample has the same probability of
being selected for collection
TYPES OF SAMPLING PLANS
▪3 types of sampling plan used in food
microbiology
▪Single sampling attribute plan – can be
evaluated using hyper-geometric, binomial
or poison distribution. The choice of
distributions used to compute the probability
relationships depend on the number of units in a
lot.
▪ the two-class scheme, samples are
classified as acceptable or defective depending
on the test result.
▪ A sample is described as defective if it
is shown to contain more than a specified
number of organisms
or in cases where a presence or absence test is
applied, the target organism is detected.
▪Three class attribute plan – This plan introduce
a further category and divide
samples into three classes: acceptable,
marginally acceptable, and unacceptable. Use
of this extra classification of marginally
acceptable means that they are not used
with presence or absence tests but only with
microbiological count data.
▪Variables sampling plan – this plan is used
when the probability density function of a
measurement is known.
SAMPLING PROCEDURES
▪Finished products:
▪Consumer packages of foods should be
sampled from original unopened
containers of the target
processing lot.
▪ Processing information and product
code should be submitted on forms
together with the sample.
▪Bulk Liquid Materials:
▪ Before drawing a sample from the
container, aseptically mix the food mass
to ensure that the sample is
as homogenous as possible.
▪ If adequate mixing or agitation of the
bulk product is impossible, multiple
samples should be drawn
from the bulk container.
▪Disinfect the sampling port or opening
by microbiocide.
▪Aseptically transfer into sterile, leak
proof container with appropriate sterile
implants.
▪Do not fill sample containers over bulk
containers of food.
▪Carefully select sample containers that
have sufficient capacity to accommodate
the needed sample
volume when the container is three
fourths full.
▪Avoid using glass sample containers.
▪ Thermometers used in bulk food
containers should be sanitized before
and dried after use.
▪Cool perishable sample to 0-4.4°C
quickly, if they are not already
refrigerated.
▪Metal thermometers are preferred since
the breakage of a mercury thermometer
would be
contaminated the product with
hazardous and toxic materials.
▪When appropriate, a temperature
control sample should be prepared and
submitted with the samples
to be tested.
▪ If a temperature control sample is
needed for frozen samples, use a
container of ethylene glycol or
other suitable freezing point material.
▪ The sample container should be sealed
properly to avoid breakage, leakage, or
introduction of
extraneous contamination.
▪ If the sample is to be examined for a
regulatory purposes, the sample
container must be sealed so
that it cannot be opened without
breaking the seal.
▪An empty sterile sample container
should also be submitted as a container
control.
▪Bulk Solids or Semisolids:
▪Dry or semisolid foods should be
sampled by using sterile triers, spoons,
are spatulas.
▪ Sterile tongue depressors may be
substituted for spatulas.
▪ Portions from several areas of the food
under examination should be obtained to
ensure a
representative sample.
▪Carefully protect samples from excess
humidity.
▪Frozen Bulk Materials:
▪ Frozen bulk foods may be sampled
with sterile corers, auger bits, and other
sampling instruments.
▪A pre-sterilized auger bits or hollow
tube may be used to obtain sufficient
material for analysis.
▪ Frozen samples should be kept frozen
until their arrival at the laboratory.
▪Avoid thawing and refreezing samples.
▪A suitable procedure for obtaining test
units of frozen foods is to use ab electric
drill combined with
funnel.
▪ For large solid frozen or unfrozen food
samples, test units should be obtained
aseptically from several
areas by using sterile knives and
forceps.
▪Line Samples (In-Process Samples) of Liquids
▪ Sterile metal tubes or dippers may be
suitable sampling instruments at certain
plant locations for liquid
food samples.
▪Disposable pre-sterilized plastic
transfer pipets can also be used.
▪A special line sampling technique
involves using a disposable sterile
hypodermic needle and syringe.
▪ Sampling cocks on holding tanks and
product pipelines may be used.
▪Disinfection and material flowthrough
of the sampling cock must be
consequently be assured before
collecting the sample.
▪Line Samples of Solids
▪ Sampling of solid line samples may be
accomplished by using the same
equipment and procedures that
would be used for bulk solid products or
semisolid products.
▪Automatic sampling devices are
available for powdered products and
other solid products that do not
require refrigeration.
▪Nondestructive Sampling
▪When food products are sampled for
indicator organisms and/or pathogen,
non-destructive samples
may be preferred since product
appearance or quality cannot be
affected.
▪Special Purpose Samples (Foodborne Disease
Outbreaks and Consumer Complaints)
▪ Sampling for disease outbreak
investigations should involve collecting
samples from all suspect foods.
▪ If there are no leftovers foods, efforts
should be made to obtain sample of
items prepared in a similar
manner as the suspected food.
▪ Ingredients or raw items used in the
ingredients or raw items should be held
under suitable conditions
until an analysis of the attack rate data
and other epidemiologically gathered
data can aid in
identifying the suspect food or foods.
▪ The original containers in which the
foods were found should be collected,
labeled, and submitted for
examination.
▪Samples for Water Activity and pH
Measurement
▪ Samples intended for water activity
determination should be collected in
sealed vapor tight containers.
▪ Small, unopened, hermetically sealed
retail-seized packages should be
collected, if possible.
▪ Sampling from bulk containers should
be performed quickly to minimize
changes in the water content
of the product.
▪ Samples to be tested for their pH and
or titratable acidity should be collected
in tightly sealed
containers.
▪ If the material to be tested is
undergoing fermentation or some other
gas producing reaction, a vented
container or flexible plastic bag with
ample space for expansion should be
used
STORAGE, SHIPMENT, AND RECEIPT
OF SAMPLE
When it is necessary to store samples
before shipment, a storage area should
be available for frozen samples (-20°C)
and for refrigerated samples (0 –
4.4°C).
PREPARATION OF SAMPLE
HOMOGENATES
▪Samples should be examined promptly.
Nonperishable canned or low moisture
food samples may be stored at room
temperature until ready for analysis.
When the initiation of analysis must be
postponed, frozen samples should be
stored at - 20°C until they are ready for
examination.
▪To destroy microorganisms that may
later contaminating the sample, before
opening the container, wipe its exterior
area with 70% ethyl alcohol or other
appropriate disinfectant.
▪Frozen samples should be thawed at
refrigerator temperature (0 – 4.4°C) for
no longer that 18 hours in the original
container in which it was received.
▪The sample must be removed
aseptically from its original containers.
▪In thawing using higher temperature,
expose the frozen sample at <40°C for
15 minutes. This method prevent the
destruction of microorganisms present in
the sample.
▪Frozen samples can also be thawed
rapidly in circulated waterbath.
▪Liquid or semi-solid samples in
containers that have an airspace can be
mixed by rapidly inverting the sample
container 25 times.
▪Sample containers that are two-thirds to
threefourths full should be shaken 25
times in 7 seconds in 30 cm arc. The
interval between mixing and
removing the test portion should not
exceed by 3 minutes.
▪When no airspace is present in the
sample, aseptically open the container
and pour the product from the filled
container back and forth into a sterile
container three times.
▪Dry samples should be aseptically
stirred with a sterile spoon, spatula, or
other utensils to ensure a homogenous
sample.
▪Test portions of non-viscous liquid
products may be measured
volumetrically for dilution by using a
sterile pipette.
▪When measuring products having a
viscosity like milk, the last drop should
be blown from the pipette.
▪For viscous liquids, the test portion for
the initial dilution should be aseptically
weighed (10 ± 0.1g to 90 mL diluent or
11 ± 0.1 g to 99 mL diluent). This
serve as your 1:10 dilution.
▪Test portions of solid or semi-solid
foods should be weighed 50 ± 0.1g for
450 mL diluent.
▪A variety of diluent may be used
depending on the nature of the product:
▪Normal: Butterfield’s
phosphate buffer & 0.1%
peptone water
▪V. parahaemolyticus: 3%
sodium chloride solution
▪When analyzing fatty foods or lump
forming powder, wetting
agents/emulsifiers such as tergitol
anionic - 7 1% solution may be added to
diluent to promote emulsification.
▪The blending time may vary, depending
on the type of food. No more than 15
minutes should be elapsed from the time
the sample is blended until all dilutions
are prepared.
▪If the sample is not homogenous, weigh
50 g from a representative portions of
the package into a sterile, tared blender
cup or analyze each portions of food
separately.
▪Stomaching is an acceptable alternative
to blending when preparing a food
sample homogenate
▪In procedure of stomaching, food
sample (50 g) and diluent (450 mL) are
placed in a sterile bag filter.
The filter bag will be positioned
appropriately inside the chamber of
stomacher and pummeled for 2 minutes.
▪ Food with bones and other
sharp protruding objects should
not be prepared by stomaching
PREPARATION OF SAMPLES
FOR WATER ACTIVITY TESTING
▪The ideal preparation of samples for
water activity measurement is to grind
the material to a fine consistency before
testing.
▪Certain emulsions such as oil/water
emulsions may be difficult to measure,
unless the water can be separated by low
temperature cycling or centrifugation.
▪The prepared sample should be quickly
added to the test chamber of the water
activity meter and avoid an exchange of
moisture in the air
 ▪When performing an analysis, transfer
sample portions to the test instrument
sample holder and
follow the manufacturer’s instructions.
▪A reliable reading may take from 5
minutes to several hours, depending on
the type of instrument used.

PREPARATION OF
SAMPLES FOR PH
DETERMINATIONS

▪First, verify the pH meter using 3-point

calibration buffer and determine the
slope (>97%)
▪Many types of liquid samples require
very little preparation for pH
determination.
▪Semi-solid, mixtures of solids and
liquids, emulsions, and various types of
marinated foods in oil require special
preparation steps.
▪Semi-solid samples can be blended to a
thick paste and a small amount of
distilled water (<20 mL/100 g of
sample) is added to provide more fluid
test portion.
▪Mixtures of solid and liquid samples
can be blended to a paste consistency
and tested as is or filtered in a sieve size
#8 mesh wire.
▪For marinated samples in oil, separate
the oil from the product, blend the solid
portion into past consistency and
test as is.
▪When attempting to determine the pH
of emulsion, centrifuged the sample to
separate the water phase and test.
▪Environmental temperature should be
at 25°C to prevent fluctuations in pH
readings.
▪Temperature effects on pH electrode
and the actual hydrogen ion activity will
modify the pH readings from electronic
pH meters.