PhCh 136 Lec 2nd Exam
1st Trans
April 3, 2019
(Sorry guys hindi narecord ang konteng first part
ng IR Spectro kaya I will try my best to fill in the
first part based on information from references
such as Harvey or Skoog :(
Infrared Spectroscopy
- IR Region is from around 1 nm to
around 1 mm
- IR Spectro is typically associated only
with vibrational portion of IR (2.5 nm to
25 nm)
- Important in the identification
(qualitative) of both organic and
inorganic compounds because all
molecular species absorb IR (Except for
homonuclear molecules such as O​2​, N​2​,
etc)
- Important in quali also because every
molecule has a unique IR spectrum
(except for chiral molecules in the
crystalline state) (Skoog, et. al., 2013)
- Wavelength is in terms of
wavenumbers, which is the reciprocal of
the wavelength in cm
- Wavenumber is more preferred than
wavelength because it is directly
proportional to energy (higher
wavenumber corresponds to higher
energy)
IR Spectro for quantitative analysis?
- It is less satisfactory than its UV/Vis
counterpart since it offers less
sensitivity, less precision and higher
deviation from Beer’s Law
- IR radiation is less intense than UV/Vis,
therefore more prone to stray radiations
- However, IR MAY BE USED
quantitatively if precision requirements
are not that strict since it gives the
advantage of being quite selective
(Skoog, et. al., 2013)
Process of IR absorption
- IR is lower energy than UV/Vis,
therefore it can only affect the stretching
and bending of bonds, as compared to
UV/Vis wherein jumping of electrons to
a different energy state is concerned
Requirements for IR absorption
- Only the frequencies of IR that match
the frequencies of the natural vibration
of the molecule is absorbed
- Only bonds that have dipole moment
are able to absorb IR (hence the reason
why homonuclear molecules lack IR
absorption)
Types of vibrations involved in IR
1. Stretching Vibrations
- Symmetric
- Asymmetric
2. Bending Vibrations
- Twisting
- Wagging
- Scissoring
- Rocking
*Each of these types of vibrations will have a
signature signal in the IR spectrum
Another thing to look for in IR spectrum:
Intensity of the band
Intensity
- Tells the type of functional group
present
- ↑ dipole moment, ↑ intensity
- ↑ electronegativity, ↑ intensity
- ↑conjugation to polar double bond, ↑
intensity
Energy Level (which is in terms of
wavenumbers)
- Molecules with similar functional groups
will absorb photons of similar energies
- The higher the atomic mass difference,
the higher the energy of absorption
needed
- Double bonds are stronger than single
bonds and therefore absorb at higher
energy than single bonds
Is IR important in analytical chemistry?
- In certain monographs, IR spectra are
needed in the identification of these
certain compounds
- Not all compounds need IR spectra
- There is no pattern that will tell you
whether a compound will need an IR
spectra or not
- Most of the time, monographs require
only physical characteristics or physical
constants for the identification of
compounds; sometimes, UV spectrum is
needed
- IR is usually used for identification of
structure of compounds of unsure
identity
IR in QC
- IR is being looked at nowadays as a
potential QC test
 - This is because the sample preparation
in IR is as easy and fast as that of in
UV/Vis
- QC has to be as fast as possible
because it is a requirement in
manufacturing (wherein time is of the
essence)
2 types of IR instruments
- One on the left side of the ppt is
continuous or dispersive type of
instrument
- One on the right is the FTIR or the
Fourier Transform IR machine
Continuous IR
- Earliest type
- Gives less information as compared to
the FTIR
- Not that common nowadays
- Sa FTIR kasi mas clean ang data and
mas complete
- Continuous IR gives very general
information
How Continuous IR works
- IR radiation is given off by a filament of
zinc oxide or yttrium or thorium oxides
All of the wavelengths associated with the IR
region will go to the sample. After that kapag
nagkaroon na ng absorption (rotational to
vibrational energy levels), it will give out again
that light na icacapture ng monochromator one
wavelength at a time.
FTIR
● Still, the source will give out all the
wavelengths of that energy source
(zirconium oxide, yttrium oxide, or thorium
oxide) then it will bring it to the sample
simultaneously. The sample will give the
data simultaneously.
● Instead of getting the absorption data one
wavelength at a time, the moving mirror will
capture all of the information simultaneously
also.
● Mathematical equation of Fourier Transform
-- inaayos yung data na simultaneously
captured.
● There is no data loss in the process
because everything is captured
simultaneously. There is no selection or
filtering unlike that of continuous or
dispersed type. Pero essentially there is
filtering done by the monochromator.
● If the make up of the monochromator is not
as good, if the gratings are not as high (less
gratings = less resolution), they actually lose
some of the data. Ang nangyayari yung read
out from a dispersing type is a little cleaner.
Nawawala kasi yung ibang data from the
monochromator.
Source of radiation for IR: oxides of zirconium,
yttrium, or thorium
● Gives a wavelength from 2-15 micrometers
● Non-conducting at room temperature
● Alternative: glow bar → gives a less intense
type of radiation
Monochromator
● Will only appear in the continuous or
dispersive type of an IR instrument (older
type of the instrument)
● Most commonly used: grating type
● If the grooves are not as fine (mas
malalawak yung distances), data loss will
come in because hindi talaga nahihiwa yung
wavelength coming from the sample.
(Remember: 2-15 micrometers lang yung
range ng IR)
● It should be able to chop 2-15 micrometers
into as many wavenumbers or wavelengths
as possible.
● Yung one micrometer, malaki na yun. It can
differentiate a C-O absorption and a C-Cl
absorption.
● Kapag mas marami [ang grooves/gratings],
mas okay.
● Mas maganda ang FTIR because it will not
have this disadvantage of using a
monochromator. All of the data will be
captured. There is no filtering involved.
Sample cell
● Where the sample is placed
● If you have liquid sample, it should be as
much as possible anhydrous in nature. It
can be tested as neat (undiluted form of the
sample; no added liquids to deliver the
sample into the IR)
● It can also be diluted using carbon
tetrachloride or carbon disulfide in a
short-pathed cell.
● It’s still possible to have problems with
saturation → dilute!
● Carbon tetrachloride and carbon disulfide
should not give any interfering response to
the rest of the spectra

Solids
● KBr disc
○ KBr will just be mixed thoroughly with
the sample in a mortar and pestle
○ Sample is mixed in a particular ration
depending on the sensitivity of the
sample
○ E.g. 10 mg of the sample + 100 mg KBr
→ 10% lang ng buong preparation
○ standard
● NaCl
○ Alternative when KBr cannot be used
○ Preferred when the solid is in chloride
form so that halogen exchange will not
occur → kailangan pareho yung salt
form
○ If hindi nakasalt form, there is no
problem. Iniiwasan lang magkahalogen
exchange in the process.
Paano ba nagkakaroon ng halogen exchange?
● These materials are just mixed in a mortar
and pestle. When these are made in a
pellet, pressure is actually placed on the
sample.
● So, para kang gagawa ng waffle. Kapag
gumagawa ka nun may presser ka. So
ganun na ganun siya. Papaumbukin mo
siya. And then, you really have to press on
that so that it becomes a hard pellet.
● Once you placed pressure, the tendency is
you place enough energy for halogen
exchange to occur in the process.
● Yung iba naman, there are some who would
prepare the solid materials, as a dispersion
in mineral oil. The process is the same. It is
simply mixing the two components: mineral
oil and the sample and then placed it on the
sample cell.
● The other one, are the gasses where the
cells consist of a glass or metal body with
two iron transparent ending.
● For pharmaceutical purposes, we don’t
really encounter gasses. But for industry
who would still assay gasses in IR, then eto
yung ginagawa, so possible din siya.
● From the sample cell, we go to the portion of
the detector. There are so many detectors
right now that are available.
● After IR absorption of the sample, the
sample will either throw it to the
monochromator for further dispersion of light
or throw it to the moving mirror for the FTIR
type.
● From those mechanism, it will throw those
signals into the detector.
● There are several detectors right now, each
with an advantage.
● The earliest ones are the transmission type
of the detector or the courier transform
which is still the most widely used. It uses a
signal from there which relays the radiation
source in which individual can be monitor in
within a one second of radiation can be
monitor in a one second pulse of radiation.
● The important aspect here with courier
transform is how fast it can generate the (?)
● So pag sinabing one second pulse(?), all the
data are captured in a matter of seconds.
That’s why for FTRI instruments, the data
capture or the data generation will only
takes seconds.
● Pagkaprepare ng sample, paglagay dun sa
instrument, in a matter of second you can
get the results. Again, no filtering involved.
● But when a monochromator is used, the
process can take from 15-30 minutes,
before a data is generated.
● ·Madalas naman hindi siya nagiging
problem, just because you are placing
samples which are relatively stable inside
the instrument.
● There is no heating involved in this. The light
will just go through the sample, a low energy
light will just go through a sample. At one
point, data capturing lang ang magtatagal.
● With the FTIR, you’ll just simply get all the
data coming from the sample.
● For the other one, yung isa pang type ng
detector, Attenuated Total Reflectance IR. In
here, it occurs when a beam of radiation
enters from a more dense (with a higher
refractive index) into a less dense medium
(with a lower refractive index)
● So what happens is that, the sample, or the
light coming from the radiation source, it will
just heat the sample on one portion.
(potassium bromide).
● And then what happens is, kung isang
portion lang, paano mo mapapadami yung
signal that can reach the detector?
● So what happens is, it will try to heat the
sample on cellular points by placing the
crystals with different refractive index, which
means, it will bend the light again in this
type, on several points.
● So that, instead of just heating the sample
on one end, kapag binalik niya, natamaan
na yung ibang fractions.
● So ang magandang gawin ay is to get
signals from as many areas of the sample
as possible by just one beam of light.
● So instead of irradiating the whole pellet,
kumbaga marerecycle lang muna yung
portion na yun na nilight. Paulit-ulit lang siya
by the use of high refractive index crystals.
● So instead of irradiating the whole pellet,
kumbaga marerecycle lang muna yung
portion nay un na nilight. Paulit-ulit lang siya
by the use of high refractive index crystals.
● So yung sinasabing low dense medium, it
will now become a sample, yung
pangalawang media para magkaroon ng
bending of light.
● And then, the other one, which is the Diffuse
Reflectance IR. It is when light is reflected
off a matt surface, the light observed is of
the same intensity no matter what the angle
of observation.
● So what happens here is when the sample
prepared, as a powder, (potassium bromide
pellet), you don’t expect the surface of the
sample to be actually smooth.
● Unlike kanina sa hitsura nung attenuated
reflectance, it appears that the sample is just
a thin film on the sample cell but on
actuality, hindi siya nangyayari na ganun.
● During the preparation, as much as
possible, you try to level it properly but the
reality is magkakaroon talaga siya ng
rooms(?). So yan yung advantage na
inooffer ng diffuse reflectance.
● So, what happens is, instead of capturing all
these areas, ang mangyayari sa kanya is
isang region lang ang icacapture niya. Kaya
yung sinasabi diyan na no matter what the
angle of observation is macacapture niya
yung data.
● So yun ung mga common na ginagamit:
Diffuse, Attenuated Total Reflectance or the
most common FTRI.
● In terms of application, compound
identification is still the number application of
IR. – is still part of ID tests of sampling
compound.
● It is use as characterization of sample in a
solid and semi-solid for creams and tablets.
● If it is in dosage form, how will you able to
distinguish which signal is found from the
active ingredient, which one is coming from
all the other excipients?
● The technique here is to always have a
“baseline” data. Ibig sabihin may data na
dapat kung ano yung mga ginagamit na
excipients. Sila mismo may IR spectra na.
● Pwedeng may library. Pwedeng may
characteristic siya na hindi macacapture lalo
pag nagiiba yung source nung excipient na
yun
● So, what can be done in a laboratory is you
actually build your own library. So ang
gagawin lang kahit literature kasi maraming
pwedeng madownload for free is that to
check your own raw material.
● You then check it on your access library.
Kapag maganda yung matching edi good.
But then again, you should still keep your
own library and then again, label from what
manufacturer it is. Because that might spell
out the presence or absence of impurities.
So yun ung isang titingnan.
● So from there, kapag meron ka nang library
nung mga excipients, once you get your
sample, you can now subtract the baseline
or the background coming from the
excipients of the creams and the tablets.
● It can also be used as a method for
fingerprint test, films, coatings, and
packaging plastics.
● Parts also of the quality control, training
kasi dito sa college, we only do quality
control of dosage form. Part pa rin ng
process is actually doing quality control of
the packaging materials. Yung primary niya
for example are the films and coatings in
capsule. Or even halimbawa yung mga
tablets na nakablister pack. Lahat yun dapat
dumadaan sa quality control.
● Imagine kapag gagawa ka ng plastic. You
can not prepare it in a solution. Di mo yun
magagawa using the usual techniques here
in our college. Remember that these can be
prepared in a pellet with potassium
bromide. There is no dissolution going on. It
is just mixing there. Magkakaroon ngayon
ng response yung films and the coatings,
and even packaging plastics.
Another use of IR is detection of polymorph
formation
- Polymorph formation causes loss of
pharmacologic activity
- This may be detected only by IR (i.e.
UV/Vis, etc cannot detect
polymorphism)
Near Infrared Spectroscopy
- Emerging technique in analysis
- Wavelength range of 700-2500 nm
- It borders the visible region (which
technically extends up to 900 nm)
- Concept is still the same as that of
regular IR (vibrational or rotational
energy levels etc etc)
Applications of NIR
- Multicomponent analysis of dosage
forms in a quantitative or
semiquantitative manner
- Fingerprint check for the identity of a
drug in QC of complex excipients (Just
like normal IR)
- Determination of physico-chemical
properties of drugs and excipients
- Others in the ppt
Multicomponent analysis of dosage forms using
NIR
- Quantitative analysis may be done
without taking the drug out of its
packaging
- New technologies are being developed
such as NIR gun (portable NIR device
for easier and faster QC)
- NIR can pass through the layers of
packaging of a drug
- This may also be used for the QC of raw
materials that are still in their primary
containers (in big barrels, etc)
- Main advantage lies in the ease and
quickness kasi kapag kukuha pa ng
sample sa barrel, may mga procedure
pa (kelangan pa magrandom sampling,
etc)
- Currently being validated; not yet official
in compendia
- Sometimes used in the preliminary
testing of raw materials. If the raw
material fails the NIR gun, definitely may
something wrong sa raw material. BUT if
the substance passes NIR, it must be
further supplemented with other tests in
order to conclude that the substance is
of acceptable quality.
Determination of physico-chemical properties
using NIR
- Water content: The presence of water in
the drug product causes noise or
distortion in the fingerprint region
Advantages for NIR
- Good penetration properties which gives
it its ability to analyze substances while
in their original packaging (due to it
having higher energy as compared to
other wavelengths of IR)
- More intense radiation sources ( does
not use the oxides of metals like
thorium, yttrium and zirconium)
- Rapid analysis as was mentioned
previously
- Still on the validation phase
Limitation of IR
- Expensive since demand is still not high
- Requires more method development
(One of the reasons why is because
analysts are still trying to figure out how
to take into account the baseline
(parang blank) measurements for the
packaging materials. Different
manufacturers of packaging will
definitely give off different signals, along
with different materials and different
layers of packaging.)
Mass Spectroscopy
- Another technique used for structural
elucidation
Principle of Mass Spectroscopy
- Charged fragments from the parent
molecule (whose formation was caused
by bombardment of electrons) will move
across the device through the use of
electric or magnetic fields
- The difference in movement of these
fragments is primarily (not always) due
to the mass to charge ratio
● On some types of mass spec
instruments, it’s not just the difference in
the mass to charge ratios but also on
the kinetic energy that the molecule or
the fragments will carry along the
process.
● But on most instruments, puro mass to
charge ratio lang ang tinitignan for the
spectrometer to capture.
Parts of mass spec
 ● Inlet source - where the sample is
introduced. The final introduction of the
sample can be done by attaching to a
chromatographic instrument.
Continuous Infusion (sample
introduction) - it introduces the solution
continuously over a time. It uses a direct
insertion probe
-Yung pag drop niya, sobrang konti lang.
Why that small?
-As mentioned in the video, the sensitivity of the
mass spec goes as little as 10^-12 moles.
Sobrang liit niya. The instrument is highly valued
for trace analysis. For pharmaceutical purposes,
trace analysis of impurities or degradation
products. It’s even better on biological systems.
Examples: Therapeutic Drug monitoring,
ADMETox studies (Absorption, Distribution,
Metabolism, Excretion and Toxicology)
For direct insertion probe- There is really no
requirement except that the sample should be
delivered as liquid.
-Choose a liquid where it is soluble and the
readily volatilizes
-Limonene- Dissolved in dichloromethane;
Limonene is soluble in DCM; DCM has the
advantage of being easily volatilized.
How hot is the area where all of this volatilization
occurs?
-The sample introduction point can have a
temperature as high as 250 degrees Celsius
Do we expect degradation at 250 degrees
Celsius?
-Probably, it can have but to prevent
degradation, the residence time of the sample
introduction system should be as short as
possible.
How do you keep that short?
- Everything should be in in vacuum. From here
pagka heat up, diretso sa mass spec.
Millisecond residence time.
-Kailan, on the process, maibato kaagad yung
solute papunta doon sa mass spec
-In cases where buffers are use, what is
recommended is that the component of the
buffer should be volatile also.
- Bawal yung galing sa solid. Kasi if galing sa
solid, hindi completely ma volatilized. May mga
maiiwan na solid.
- If it has to be introduced as an aqueous
solution, Formic acid or Ammonium Formate
yung ginagamit.
If the sample is introduced as liquid, the choice
of solvent is that:
1.) Solubility
2.) Volatility
3.) No solid should be deposited on the
sample introduction system
Some of the sample will not go through the
mass spec.
Pag may deposition of particles, cleaning is
performed by placing isopropyl alcohol and the
surface is scrubbed using a microparticle
sandpaper. Parang Velvet using feel niya. All
solid particles are scrubbed off so that in the
next introduction of the sample, hindi masama
yung solids na yun.
Another requirement should be the injection
should be in microliter quantities. For every
eluate drop, it should be in microliter qualities.
● Excess volume of liquid from the
chromatographic system will be
vacuumed out so that only a
representative amount will be put into
the mass spectrometer
● Atmospheric pressure chambers make it
so that some ion sources in some cases
do not need an initial vacuum
environment; however, for the most part
most ion sources are in a vacuum
● It is essential to force the sample to
enter the mass spec ASAP to prevent
degradation
● Mass specs use several vacuum pumps
to keep the environment, well, in
vacuum; takes time to establish a
vacuum eg 3 whole days
● In sample introduction system,
fragmentation of molecules does not
occur even with subjection to high heat;
fragmentation occurs in ionization
chamber—may be initial as some mass
specs are connected to one or even two
other mass specs in one system
 ○ Why should there be more
ionizations?
■ Higher resolution
■ To be able
differentiate APIs from
impurities and other
degradation products or
just compounds
general; one mass spec
will not be able to
differentiate well - low
resolution mass spec
Ionization methods
1. Gas phase
- sample is vaporized then
ionized
- Restricted to thermally stable
compounds unless you know
the amount of degradation
products present.
- Small molecules (MW < 10 kDa)
because gas phase is often
associated with hard ionization
sources
2. Desorption
- Sample (solid or liquid) is
converted directly to gaseous
ions
- simultaneous ionization and
vaporization
- sample is usually placed in a
matrix since the samples are
often hard to volatilize without
degrading them
- Large molecules because
desorption is associated with
soft ionization
Gas Phase Instruments or sources
Desorption phase sources
to
in
Different methods use different energy sources
to bombard the sample/ionizing agent for
desorption. The only way for the sample to be
fragmented is to place enough energy that the
electron or the functional group will be knocked
off in the process.
How will you know if the compound is
undergoing degradation?
It is indicated in references
Hard ionization
- Many molecules/fragments are created
- Impart sufficient energy on the analyte
to leave the molecule in an excited state
in which relaxation results in the rupture
of bonds
- Not good for large molecules (as high as
100 kDa) because the spectra will be
too complicated
- Molecular ion peak may be completely
gone or negligible
Soft ionization
- Cause very little fragmentation and the
spectra have few, if any, peaks besides
the molecular ion (M) peak
If you have a reference standard or if you know
the associated impurities will probably not have
the same molecular weight then field ionization
may be used. Otherwise, use harder ionization
sources.
Electron Impact - earliest ionization source for
mass spec
- Still very much in use
- Hardest of the hard ionization sources
- Bombardment with a beam of energetic
electrons
- 70 electron-volts (eV) produced by a
radium or tungsten filament which are
accelerated towards a positive target or
the anode
 - Sa video earlier, the electron passes
perpendicular to the movement of the
sample so that there is maximum impact
- Isipin niyo pag parallel lang sila
may impact naman pero mas
less likely kasi they are all
moving in a single direction
- Hindi siya yung magwawander
from one side to the other ‘tong
mga electrons na to
- The stream coming from the
chromatographic instrument (inlet) and
the stream from the ion source will
impact
- At 70 eV, you immediately get to knock
off one ​electron​ from the sample
- As long as it has not yet
reached the anode, it can be
reused (since it still has high
energy) to impact the fragments
resulting from the initial
fragmentation hanggang sa
magkaroon ng stable na
fragment
Chemical Ionization - hard ionization source
- Gaseous atoms of the sample are
ionized by collision with ions produced
by particle bombardment of an excess
of a reagent gas
- Yung chemical na gagamitin for
bombardment dapat mataas yung
energy kasi walang mangyayari pag
normal reagent lang
- Kailangan maglagay ng excess energy
sa reagent gas so there should be an
initial collision of high energy going
towards the reagent gas
- 2 types: Positive Ion Chemical Ionization
(PICI) and Negative Ion Chemical
Ionization (NICI)
- Nag-iisa lang ang true chemical
ionization (dun dun duuuuuun)
Positive Ion Chemical Ionization (PICI)
(Methane gas reacting with high-energy
electrons)
- It is hard to imagine how to destabilize a
methane molecule (looks very stable
kasi simple molecule lang siya) but if
you place enough energy (e.g. electron
impact ionization), you get to knock off
one electron from methane which
means nag-impart ka ng energy
papunta doon sa methane
- Methane’s natural progression is to
stabilize itself
- Magkakaroon ka ng charged
methane and a proton
- Yung charged methane can
impact or can bombard another
methane.
- The initial collision of the
electron with the methane gas
will give enough energy to
unionize methane
- May excess energy to form additional
ions
- If a sample hits a charged methane,
magkakaroon ng fragmentation
- Question: Bakit di na lang yung electron
impact since electron din naman yung
ginagamit pang bombard doon sa gas?
- Answer: If the analyst would
want lesser fragmentation,
pipiliin mo yung chemical
ionization. [T/N: pero pwede rin
naman ako ;) ] Ayaw mo lang
yung sobrang taas na energy
bombarding the sample.
- If the need is to have an energy
less than 70 eV, Chemical
Ionization na yung gagamitin mo
Negative Ion Chemical Ionization (NICI) - hindi
chemical yung nambobombard sa sample
- Fake chemical ionization :(
- You still have the collision of the
electron with the reagent gas but the
result is instead of the reagent gas
colliding with the sample, the electron
which collided with the reagent gas will
collide with the sample.
- When the electron collides with the
reagent gas, some of the energy carried
by the electron will be transferred to
this(?)
- Yung electron sa NICI, since it is
decreased in energy, it will be the one
that will bombard the sample
- These low-energy thermal
electrons are then captured by
molecules with high affinity for
electrons
- Yung difference niya sa electron impact
is the amount of energy carried by the
electron
- Naging chemical ionization ang NICI just
because you need a chemical to reduce
the energy carried by the electron.
 Otherwise, it is not actually the chemical
that bombards the sample
- If the ion source makes use of a gas
(e.g. methane), dadagdagan yung
diagram ng gas tank para masabi na
yun yung ion source
Comparing the Hard Ionization Sources
(Psoralen - furocoumarin for vitiligo mga chong
ko na nag-1-128)
Electron Impact - maraming fragments
PICI and NICI - less fragments
Lahat naman ay gumagamit ng electrons
Electron Impact - directly utilized by the sample
PICI - utilized by the reagent gas then reagent
gas bombards the sample
NICI - electron has reduced energy by collision
with reagent gas then lower energy electron will
bombard the sample
Field Ionization sources
- the answer for under the influence of a
large electric field
- have 10 to 20 kilovolts of energy applied
to 10 micrometer diameter wire where
little fragmentation occur
Thermospray
- the most commonly used ionization
source is the electrospray ionization
sources or ESI
- A charged aerosol is generated at
atmospheric pressure and then the
charged molecules are drawn into the
mass spec by electrostatically charged
weights.
- In the diagram, this ionization is usually
in tandem with liquid chromatography
technique. -
- So anong nangyayari?
● From the capillary outlet of liquid
chromatography, nakadirect siya
papunta sa mass spec. In itself, the
capillary is already subjected to a high
voltage of 3 kV (3000 volts).
● Charged up na yung sample palabas pa
lang siya. Since 3000 volts siya,
kailangan niya ng sariling electrical line
*And this is just the ionization source,
wala pa ng ibang energy from the other
parts of the mass spec*
● Th 3000 volts is just for the initial
ionization
● At this rate, the sample will already be
charged. Fragmentation has not yet
started. Bakit? Probably the sample is
already charged, but the real
fragmentation occurs as it passes
through the plates which are also
electrostatically charged.
● And then the charged molecules are
drawn into the mass spec by
electrostatically charged plates, where
further fragmentation occurs
● Nitrogen gas placed on this portion for
drying purposes. Kasi kung titingnan,
while there is heat involved, to further
remove the solvent, dry nitrogen gas is
also continuously infused to the
electrospray ionization chamber.
Continuous infusion means it will get the
nitrogen from the air, purify that, pass
the nitrogen going to the machine, and
then the rest is carbon dioxide and
oxygen.
● There is a hose at the back where
carbon dioxide and oxygen is released
➢ Why not just use a nitrogen
tank?
Because if you are doing a
continuous analysis,continuous
drying is also involved. Even if
there is no equipment plugged,
nitrogen flow is continuous. 12
liters per min ang rate ng
infusion of nitrogen gas, which
ensures removal of solvent. So
ayun, mabilis lang ang buhay ng
nitrogen gas; on a long term
basis, it is not economical.
● Same mechanism applies with your TLC
applicator. There is a nitrogen tank on
the side, but not running at 12 L/min; it’s
a lot less. Its purpose is to dry the
solvent off the sample.
● So there is nitrogen gas for the sample,
and then there is nitrogen gas again if
the solvent is not completely vaporized
in the process.
● “Electro” portion- the electric field given
by the 3000 volts
● “Spray” portion- as soon as the sample
gets out of the capillary, parang mist na
siyang lalabas papunta sa
electrostatically charged plates.
 * Nitrogen gas has 2 purposes: to
remove the solvent, and to create a fine
mist of the sample
- If you have instruments which involve
high heat and high volatilization, its is
really expected that some solids get
deposited on any of these parts.
- To clean this:
● If the nitrogen gas is not stable anymore
in terms of blowing the solvent off,
remove sonication device
● A needle at the tip will cause the spray
effect. It is easy to remove the needle
but difficult to put back because there is
the danger of bending the needle if it
hits a metal surface.
- If you have an instrument which involves
high heat and volatilization, it is
expected that some solids get deposited
in any of these parts.
Matrix -assisted Laser Desorption Ionization –
also placed in a matrix that should absorb UV
radiation
- the energy from the laser is transferred to the
matrix, the excess energy ibibigay niya sa
molecule which is mixed with the matrix and the
transfer of energy will create fragments
For more information, you can read Mass
Spec for Ionization Techniques for Pharmacy
Students (2​nd​edition) by David Watson. For
the IR and other parts of Mass Spectroscopy,
read Pharmaceutical Analysis: A Textbook
for Pharmacy Students and Pharmaceutical
Chemists also by David Watson (Same ang
nasa ppt ni ma’am sa mga nakalagay sa
book na to).
- end -

Atmospheric Pressure Ionization – similar to ES,

but run at usual LC flow rates and used for less
polar molecules
- meron ka pa ring nitrogen gas to help vaporize
the solvent and to create a fine mist of the
sample and the solvent but yung creation of the
fine mist sa second source nangyayari
- the added nitrogen gas helps in treating the
less polar molecules
- at high liquid chromatography flow rates
kailangang maraming nitrogen to remove the
solvents kasi mataas ang volume load and
kailangan minimal amount lang ang makarating
sa mass spec para hindi masaturate ang mass
spec
- this occurs at atmospheric pressure
- the 3000 kilovolts is still there but it’s not a part
of the LC capillary and is a separate
mechanism; hindi simultaneous because of the
volume load

Thermospray – the volatilization is done by

heating the capillary

Fast Atom Bombardment – the sample should

be first run with a matrix
- for samples who do not easily volatilize
- one of the classic example of desorption phase
- the sample is mixed with glycerol to hold it
- the source of energy is a fast atom