CLINICAL CHEMISTRY LAB

LECTURE 1: GLASSWARES, PLASTICWARES

AND MEASURING VESSELS
Prof. Bea Angelli Laude, RMT
August 16, 2021
For updates and corrections → @mar4rii on Twitter

Clinical chemistry is the area wherein we are going to measure the GLASSWARE
different analytes that are present in the blood sample of the patient.
They are usually metabolites of the body which are quantified to Advantages
identify the disorders the patient is facing.
(Some) heating
LABORATORY GLASSWARE & PLASTICWARE Longer storage of some chemicals
● Equipments in the laboratory can either be made of glass
or plastic Regardless of the design, glasswares in the laboratory
○ If they are made up of glass, it is actually a must fall into 2 types:
specific type of glass that’s why they are ● Class A glasswares
correctly termed as glasswares ● Class B glasswares
○ Nowadays, the use of plastic equipments are These classes will be identified or labelled in glasswares
rampant, however plastics tend to have if they are able to satisfy certain tolerances of accuracy
limitations = they cant support heat
■ That's why we still continue to use Class A Glasswares
glasswares
Main Functions of Glasswares and Plasticwares
Why use glasswares/ plastic wares in the lab?
● Storage
○ In the laboratory, we tend to store both the
samples as well as the reagents
○ That’s why it's necessary that we make use of ● Found in professional laboratories
glasswares and plasticwares, that has a National Institute of Standards and Technology
specific quality such as: Class A Glasswares
■ Ability to withstand extreme ● Class B tends to have twice the tolerance limits of Class
temperature; A
■ And the corrosive effects of strong ● Class B are much more durable
acids and strong alkaline solutions ○ Often found in student lab
● Measurement
● Containment Cleaning of plastic/ glassware
● Those in direct contact with biohazard material is usually
PLASTICWARE disposable
- Since the early times, majority of the laboratory ○ Biohazard material such as samples, be in the
equipments are actually made up of glass form of stool, urine, or blood must be disposed
- Since the discovery of plastics, it slowly replaced the after use
glasswares that we often use in the lab ■ Regardless if it's made of plastic or
- However, nowadays we still tend to continue using glass
glasswares mainly because it is able to fill in the ○ Advantage of plastic = since it’s very cheap,
limitations of plastics they are often in disposable form
Advantages Disadvantages ○ Unlike glass, they tend to be much more
expensive = reusable form
Evaporation through breathing
● If not disposable, follow proper decontamination protocol
of plastic
○ Immediate rinsing + washing with powder/
Cheaper - Plasticwares are not often
liquid detergent
utilize as storage vessels
■ Wash intensely to remove the
- They tend to be porous
remaining detergent (which can be a
More Durable contaminant) if you are going to reuse
- Much efficient in resisting that particular lab equipment
the corrosive effects of ○ Pre-soaking in soapy water
strong acids and strong ■ May contain bleach (usually small
alkali solutions amount) mainly for contamination
Evaporation of dyes, stains, and
Preferred for some analyses purposes
proteins
- Example:Testing for heavy ○ (read furthermore Bishop Chapter 1)
- They will evaporate in
metals (NO glass) ○ For reusable lab equipments, it must also be
plasticwares
- Some heavy metals that we rinsed properly
- Essential to be stored in
need to analyze could also ■ Perform multiple rinsing to ensure that
glasswares
be present in glasswares, there are no remnants of detergent
and they could be considered left = Remnants may act as
as a contaminant contaminants
- These heavy metals are not ○ One way of determining proper rinsing of lab
present in your plasticwares equipment, you have to check the water used in
the last rinse
1
 ○ Check the pH twice before rinsing and after ● Characteristics:
rinsing ○ Flexible or rigid
○ If after rinsing and pH is higher, we can ○ Chemical-resistant
determine that the detergents were removed ○ Can be autoclaved
(Detergents make the solution more alkaline) ● Uses:
■ Higher pH of water after rinsing ○ For cryogenic procedures
compared to pH before it was rinsed, ○ Specially formulated to withstand temp down to
means that the detergents are -190°C
removed ● Several tube designs
○ specimen tubes and test tubes
PLASTICWARE ○ Microwavable containers and folders

Major types of resins used in Clin. Chem Lab: C. Polyethylene

(Details on Henry, chapter 4, p. 60) ● Disadvantage:
A. Polystyrene ○ NOT suitable when using picric acid, stains,
B. Polyethylene dyes and proteins
C. Polypropylene ● Uses:
D. Teflon ○ test tubes
E. Polycarbonate ○ Bottles
○ graduated tubes
The type of plastic you will use will depend on the procedure you ○ Stoppers
are going to perform ○ Plastic balls (toys)
○ Plastic wraps
● Indicated with arrows = types of resins that are incontact ○ Insulators
with our food:
○ PET or PETE (polyethylene terephthalate) D. Polycarbonate
○ HDPE (high density polyethylene) ● Characteristics:
○ LDPE (Low density polyethylene) ○ Very strong plastic but NOT chemically
○ PP (Polypropylene) resistant
● Others cannot be used for food because once ingested, ○ Autoclavable but with limitations
they may produce harm ● Usable temp. range: -100°C to +160°C
● Uses:
A. Polystyrene ○ tubes for centrifugation
● Characteristics: ○ graduated cylinders
○ Rigid ○ Flasks
○ Clear type ○ Roofs of waiting sheds
○ NOT to be autoclaved
E. Teflon
● Characteristics:
○ Almost chemically inert - unable to react to
chemicals
○ Chemical-resistant
● Suitable work temp. @: -270 ⁰C to 255 ⁰C
● Uses: stirring rods, tubing, cryogenic vials, bottle cap
liners
● Used for storage or mixing equipments
● Pans for cooking and Teflon tapes utilized in water lines
at home

GENERAL CATEGORIES OF GLASS

1. Borosilicate (Kimax/ Pyrex)
2. High silica Glass
3. Aluminosilicate (Corex)
4. Acid & Alkali resistant (Vycor)
5. Low actinic (amber-colored)
6. Flint glass (soda lime)
7. Disposable glassware

1. Borosilicate Glass (Kimax/ Pyrex)

■ Can be reused with other means of ● Most common type of glass encountered in volume
disinfection but NOT AUTOCLAVING measurements
● Disadvantage: ● Composition:
○ Not resistant to most hydrocarbons, ketones ○ 80% silica
and ROH ○ 13% boric oxide - contains BORON
■ Thus not utilized as storage ○ 4% sodium oxide
● Uses: capped graduated tubes and test tubes ○ 2–3% aluminium oxide
● Can be used as a container - short time ● Characteristics:
● Ex. Styrofoam ○ High degree of thermal resistance and low
coefficient of thermal expansion
B. Polypropylene ■ Object has the ability to change its
● Primary constituent of pipet tips shape at a given temperature
○ Pipets are utilized to transfer and measure ■ Change is minimal
samples as well as reagents ○ Low alkali content
○ Reagents may be strong acids because the ■ Resistant to alkali corrosion
pipet tips can withstand extreme pH ○ Free of heavy metals
2
 ■ Free from Mg-lime-zinc group of
elements, heavy metals, arsenic and
antimony
● Uses:
○ Heating
■ Open flame or electric heating
elements such as hotplates
● Precautions:
○ Storing concentrated alkali solutions will
etch/destroy the calibration
○ Heavy-walled type of glass should not be
heated with direct flame or hotplate
■ Only the thin-walled type of
borosilicate glassware will be heated
in direct flame or hotplate
○ Avoid heating beyond its strained point
■ Strain point- the temperature where
the glass softens or will transform
from solid to liquid
■ Heating beyond strained point could
lead to breakage
○ In the case of volumetric glassware, heating
can destroy the calibration
● Popular Brands
○ Pyrex®
■ Strain point: 515
°Celsius (Henry)
○ KIMAX®
■ Strain point: 513
°C
4. Acid- Resistant & Alkali-Resistant Glass
● Boron-free
● A.k.a “soft glass”
● Vycor® (No. 7900)
○ Made up of fused silica
○ 2 in 1 characteristic
■ Heat-resistant (not as efficient as
borosilicate glass)
■ Chemically inert (a substance not
chemically reactive)
○ Unique characteristics:
■ Stable to all acids except hydrofluoric
types
- Hydrofluoric solutions corrode
glasses with silica
■ Superior in resisting corrosion by
alkali than borosilicate glass
■ Unlikely contamination by contact
with solutions
■ Relatively inert to acids and neutral
salts
■ Chlorine and acid gases do not affect
it at any temperature
○ Withstand high temperature (1200°C)
■ Softening temp.: 1500 °C
○ Withstand downshocks from this temperature to
ice water
○ Use:
■ ashing and ignition techniques

2. High Silica Glass

● Silica fused to quartz
● More expensive than borosilicate glass
● Use: 5. Low Actinic Glass
○ Spectrophotometer cuvettes ● Contain materials that usually impart amber to red color
to the glass
○ Reduces the amount of light transmitted to the
substance in the glassware disallowing
3. Alumina-Silicate Glass oxidation
● A.k.a Aluminosilicate glass ● Common uses:
● With aluminum oxide ○ For light-sensitive substances
● Strengthened chemically rather than thermally ■ Bilirubin
○ Can withstand very strong acids, bases, ■ Vit. A
hydrocarbons, and concentrated alcohols ○ Store control materials and reagents
○ Greater chemical durability and can withstand
higher operating temperatures 6. Soda-Lime Glass
○ Compared to borosilicate, aluminosilicates are ● A.k.a flint glass
more difficult to fabricate ● Composition:
● Has a wide thermal range but not as wide ○ Silicon oxide
as borosilicate glass ■ Soda = sodiuum oxide
● When coated with an electrically ■ Lime = calcium oxide
conductive film, aluminosilicate glass is ● Most inexpensive glass
used as resistor for electronic ○ Most common type of glass found outside the
circuitry laboratory
● High impact, extremely strong glass ● Readily made into variety of types of glassware
○ Common use: manufacture of ● Has a high expansion coefficient and a high degree of
calibrated centrifuge tubes thermal resistance
● Corex® (Corning, N.Y.) ○ At given temperature, it will exhibit huge
○ Characteristics: structural changee
■ Radiation-resistant ● Mineral can be leached from the glass into the stored
■ 6X stronger than borosilicate (outlast solutions
conventional ● Common uses:
glassware by 10-fold) ○ Volumetric flasks
■ Resist clouding and ○ Stirring rods
scratching better ○ Single-use pipette or test tubes
○ Uses:
■ High-precision MEASURING VESSELS
analytical work a. Graduated cylinder
■ Optical reflectors and b. Burets
mirrors c. Volumetric flask
■ Not used as general d. Pipet
type of glassware in
the lab

3
 A. GRADUATED CYLINDER A. Design = TC vs. TD (to contain or to deliver)
● Long, cylindrical tubes usually held upright by B. Drainage characteristics = Blow-out vs.
an octagonal or circular base with gradations Self-draining
along its length C. Type = Measuring vs. transfer
● Semi-accurate
● Extremely convenient for rapid measurement of
TABLE 1-4 PIPET CLASSIFICATION
liquid
● Should NEVER be heated especially if plastic
since it will destroy the calibration marks along
its body I.Design
A. To contain (TC)
B. BURETS B. To deliver (TD)
● Long cylindrical graduated laboratory glassware with
stopcock II. Drainage characteristics
● Type of buret used depends on the chemical A. Blowout
used B. Self-draining
○ Glass >> ACID
○ Rubber for >> ALKALI III. Туре
● Extremely accurate in dispensing aliquots of a A. Measuring or graduated
solution 1. Serologic
○ Mainly used to dispense known 2. Mohr
amounts of liquid reagents in 3. Bacteriologic
experiments where such precisions 4. Ball, Kolmer, or Kahn
are necessary 5. Micropipet
● Generally used for titration purposes only B.Transfer
1. Volumetric
C. VOLUMETRIC FLASK 2. Ostwald-Folin
● Round lower portion and a long, thin neck 3. Pasteur pipets
with an etched neck or calibration line 4. Automatic macropipets or micropipette
which is going to be the mark in
measuring a specific volume
A. Design
● Generally used for:
○ Preparation of standard solution 1.) To Contain (TC
○ Measuring liquid colume ● A.k.a. “Rinsed-out pipets”
accurately ● Able to hold a particular volume but is unable to dispense
the volume indicated.
● For u to dispense the exact volume, Must be refilled and
rinsed-out with the appropriate solvent after the initial
D. PIPETS liquid has been drained from the pipet
● Glass or plastic material in the lab used to transfer liquids ● Examples:
● Can either be reusable or disposable ○ Sahli-hemoglobin pipets
● Majority of pipette can hold only up to 20 mL of a specific ○ Long-Levy pipets
solution 2) To Deliver (TD
● Used to transfer measured volumes of liquid between ● Able to transfer the exact volume indicated in the pipette
containers ● Designed to drain by gravity
● Must be held vertically with the tip placed against the
side of the container and must NOT TOUCH the liquid in
it
● Vessel ang nakatilt
● Examples:
○ Mohr pipet
○ Serologic pipet
○ volumetric transfer pipet
NOTE:
● A TC pipet holds or contains a particular volume but does
not dispense that exact volume, whereas a TD pipet will
dispense the volume indicated.
B. Drainage Characteristics
1) Self-draining pipet

○ Example (pic above): you have 5 in 1/10 which

means that the capacity of the pipet in
measuring a particular volume is only up to 5
mL with 10 increments to the next demarcation
line
● Numbers are indicated at the top or lower portion of the
pipet which indicates the volume or total capacity for
measuring and the major divisions
● In order to properly measure a specific volume of a
particular solution, you have to look for the meniscus of
the aspirated fluid where it should be aligned to the
graduation line of your pipet ● with a single painted at the top
● Pipets are classified based on: (Bishop 7th ● Allow to drain by gravity
● edition, table 1-4 pg.13) ● No frost/ etched / double lines
4
2) Blown-out pipet Volumetric or Transfer
● Used to measure and transfer a predetermined volume of
liquid
● Kaya niya ma dispense or ma measure na volume of a
particular solution is only one volume
● If it is indicated that it is able to measure 10 mL, therefore
10 mL lang jud ang pede niya i measure kay isa lang
iyang etched line
● Dispense one volume without further subdivisions
○ Ostwald-Folin Pipet
○ Pasteur Pipet

● With double rings/ frost/ etched

● designed to be "blown out" by pushing a small amount of
air out of the pipet, completely emptying it
● For u to deliver the exact amount of the volume specified
by the pipette, it has to be blown out with the use of a
bulb.

Left: volumetric (transfer)

Right: oswald-folin

TOP portions of self-draining & blown-out pipet

Single colored line = self-draining
● Allow it to drain by gravity
Double ring or frosted/ etched ring= blown out pipet
● To deliver the exact amount of solution u have measured,
you have to introduce air to empty the pipet.

Pic above: illustrates the proper positioning of pipettes in aspirating

TIP portions of self-draining & blown-out pipet and dispensing solution.
Left: self-draining ● Top pic: held vertically upright with the tip slightly
- Graduation is much farther from the tip. submerged in the surface of your solution without
Right: Blown out pipet touching the side of the vessel.
- Graduation is up to the tip of the pipet. ● Bottom pic: dispensing solution
○ Tip is not supposed to touch the surface of the
C. Type solution contained in your vessel but rather the
1. Transfer pipet (or volumetric) tip is going to touch the side of the vessel for a
a. Volumetric smooth flow of the draining of your solutions.
b. Ostwald-Folin ● In both aspirating and in draining, the pipette must be in
c. Pasteur a vertical position.
d. Automatic macro- & micropipet ● Draining
2. Measuring or Graduated pipet ○ In order to maintain the vertical position of your
a. Serologic pipet and to allow the tip to touch the sides of
b. Mohr the receiving vessel, so u have to tilt the vessel
c. Micropipet to maintain its vertical position.

5
 Positive Displacement Pipette

● Act or move like a syringe

● Each tip contains a disposable plunger like a syringe
○ Plunger is disposable but not the tip
● The plunger directly displaces the fluid with no error
● These are the most accurate type of pipette
● They are also the most expensive

https://www.youtube.com/watch?v=_YTwxakpY1U

The volumetric pipets are always SELF-DRAINING

Ostwald-Folin pipets are BLOWOUT PIPETS
● Volumetric – to transfer aquaeous solutions
● Ostwald-Folin – to transfer viscous fluid
DIFFERENCE VOLUMETRIC OSWALD-FOLIN

Drainage self-draining Blowout pipette

characteristics

Top portion Have single ring double/frosted/

etched ring

Measuring or Graduated
● Calibrated to distribute fractional quantity of liquid and
principally used for measurement of reagents
● The volume that it can measure is not fixed but rather 1. Air displacement method (left)
measure different volumes of a - It is going to rely on a piston for creating a
particular solution suction to draw the sample into a disposable tip
● Examples: that must be changed after each use
2.a. Mohr pipet - The piston does not come in contact with the
2.b. Serologic pipet liquid but there will be unprotected air space so
there could be a possible aerosol contamination
Mohr pipet of the sample or the reagents that is going to be
● No graduations to the tip aspirated
● Self-draining pipet - Can ensure that there will be no carry over
● Single ring since u are going to replace the tip
- However, it going to be less accurate compared
Serologic pipet to positive displacement pipet
● Has graduation marks to the tip 2. Positive displacement method (right)
● Generally a blowout pipet - Going to operate by moving a piston in the
● Designated by a frosted or double pipet tip or barrel
colored ring at the top - The piston is in the form of a plunger in which it
is going to be changed once u are going to
dispense one liquid after another
Micropipet - Advantage of piston:
● With a total holding volume of less than 1 mL - There will be protected air space
● It may be designed as either a Mohr or serologic pipet - No aerosol
● Comes in 2 forms: - Problem: Since you are not going to change the
○ Automatic and Semi-automatic pipet tip, there will be a carry over of samples or
■ Commonly used in the laboratory regents to another type of solution
2 Types of micropipet:
1. Air displacement
● Disposable, polypropylene tip
2. Positive displacement
● Use of capillary tip (siliconized, glass, plastic)

6
 ANATOMY OF AIR DISPLACEMENT PIPET POSITIVE DISPLACEMENT PIPET

● No first stop or second stop

● Pull the plunger in order to aspirate your fluids
● In draining, simply push down your plunger and drain all
the contents

https://www.youtube.com/watch?v=NgosWmRjjAo

Semi-automatic & Automatic pipettes and

dispensers (Tietz)

● Push button
○ pushed in order to aspirate or dispense a
particular volume of sample or reagent
○ Not going push it but turn it bcs it is going to
● Semi-automatic pipettes
serve as a large volume adjustment knob
○ Although it allows us to have an easier
■ In order to adjust the volume to be
measurement of volumes of samples and
aspirated in much more larger
reagents, but the aspiration and dispensing of
amounts
such liquids can still be done manually
● Tip ejector button
● Automatic pipettes
○ Simply push this down to remove the pipette
○ It is attached to a machine and a tube is going
tips that is found at the bottom portion
to be submerged to a particular solution
● Thumbwheel (Fine volume adjustment ring)
○ The machine will be regulating the aspiration as
○ Used to adjust the volume to be aspirated in
well as the dispensing of a particular volume of
smaller amounts
a reagent ot sample
● Volumeter display
● Dispensers (Dilutors)
○ Where u are able to determine how much of the
○ Automatic pipettes that obtain the liquid from a
solution or what is the exact volume to be
common reservoir and dispense it repeatedly
aspirated, transferred or measured
○ May be bottle top, motorized, handheld, or
● Tip ejector
attached to a dilutor
○ Connected to the shaft
○ Simply push to move and lead to the removal of
your disposable tip

AIR DISPLACEMENT PIPET

● If you are going to push the push button, it has 2 stops:

○ First stop
○ Second stop: mainly for the draining of the
entire solution that was aspirated in the pipet tip

Prior to the submersion of your tip to the solution, it must be

pushed to the first stop
↓
Once it is submerged in the solution, you are going to release it.
By then you are able to aspirate a specific amount of the solution
↓
In transferring, push down the push button from the first stop all
the way to the second stop (mainly for the draining)
↓
Release