Volume 34 • Winter Edition, 2011 | Analytical Sciences

HAPPY NEW YEAR

2012
Fresh VOLUME 34 • Winter Edition, 2011

WHAT’S Dear customers and readers,

Fresh inside...
We have come to the end of one more year. The year
2011 was full of action on business, economic, climatic
and environmental fronts. For many industries it was a
Articles Pg. no. year of mergers and acquisitions.

• Solvent of recrystallization in pharmaceutical. . . . . 3 For PerkinElmer this year was a year of new products
introduction as well. We brought new FTIR platform
• Raman Spectroscopy – A powerful tool to with Spectrum Two and Frontier early this year. The
differentiate pharmaceutical polymorphs. . . . . . . . 5 introduction of application packs and Spectrum Touch
are for the markets who demand the ease of use and
• Analysis of impurities in Melatonin by knowledge.
LC-TOF MS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Informatics products like ELN (Electronic note book),
™ ™
ChemDraw and ChemBio office are for the
• The chromatography of sunshine . . . . . . . . . . . . . 9
customers from pharmaceutical, Bio pharmaceuticals
• Determination of oil content in membranes and various other regulated environments.
used in compressed air sampling by infrared In the middle of the year PerkinElmer introduced new
spectroscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 PinAAcle™ Atomic absorption spectrometer and Optima
8x00 ICP spectrometer to meet the demands and
• E-Notebook for formulation scientists . . . . . . . . . 14 applications need of various industries.

New Clarus SQ8 the most sensitive GCMS in the range

and AxION™ TOF mass spectrometer were introduced to
leap in to the research areas.

Apart from these various consumables and preventive

maintenance kits were brought in to the market by
PerkinElmer; but the most important was the solid
phase extraction-SPE cartridges for high throughput
labs and industries.

PerkinElmer also focused on corporate social

responsibilities in various parts of the world including
Fukushima nuclear disaster in Japan and Garib Nagar
fire in India. In this issue of ‘Fresh’ we are incorporating
the articles and applications for pharmaceuticals and
allied chemicals using various techniques from
PerkinElmer.

We wish you a Merry Christmas and a promising

New Year.

Merry Christmas Team Marketing

& Very Happy New Year PerkinElmer (India) Pvt. Ltd.
Fresh Analytical Sciences

Solvent of
recrystallization
in pharmaceutical

Evolved gas analysis: A high sensitivity study

of a solvent of recrystallization in a pharmaceutical
Typical applications of TG-MS pharmaceutical industry, materials Instrumental setup
include: often show weight losses associated All of the TGA systems supplied by
with the loss of solvent/water, PerkinElmer (Pyris™ 1 TGA, STA 6000
• Detection of moisture/solvent
desolvation or decomposition of the and TGA 4000) can be easily
loss from a sample (e.g. loss on
sample. This information is then interfaced to MS systems.
drying or dehydration of
used to assess the purity and PerkinElmer can supply systems with
a pharmaceutical)
stability of the material and its either the PerkinElmer Clarus® MS or
• Thermal stability (degradation) suitability for use. The TGA gives a with a Hiden Analytical MS. In this
processes quantitative measure of mass lost study, a Pyris 1 TGA interfaced to a
• Study reactions from the sample, but it does not Hiden Analytical HPR-20 MS was
(e.g. polymerizations) provide information on the nature of used.
the products that are lost from the
• Analysis of trace volatiles in sample, and this information is often
a sample (e.g. volatile organic required for complete
compound (VOC) testing) characterization
Introduction Coupling a mass spectrometer (MS)
Thermogravimetric analysis (TGA) of to a TGA allows evolved gases to be
materials is commonly used to analyzed and identified giving this
measure weight loss as a sample is additional valuable information. Figure 1. Pyris 1 TGA is shown interfaced
heated or held isothermally. In the to a HPR-20 MS

3
Fresh VOLUME 34 • Winter Edition, 2011

the capillary. This single feature

prevents hours of down time and The thermogram in Figure 2 shows
cleaning with no loss of vacuum in the TGA trace collected for the
the mass spectrometer. sample. An extremely small weight
• Soft ionization so the system can loss of just 1.2 µg (0.026%) was
be optimized to reduce the found, indicating the outstanding
splitting of ions leading to a performance of the Pyris 1 TGA.
Figure 1. Clarus 600 MS simpler spectrum for analysis. The gases evolved during this very
Experiment small amount of weight loss were
The HPR-20 MS is optimized for the In the following study, a Pyris 1 TGA analyzed. The background partial
analysis of the evolved gases from was interfaced to a HPR-20 MS. pressure of mass 49 was observed at
the TGA and offers the following The sample was heated to 200 °C at a 2 x 10-11 bar and a slight peak to
benefits: 10 °C/min with helium purge gas at 5 x 10-11 bar observed in the weight
30 mL/min. A sample weight of loss region as shown below
• The ability to use either Faraday 6.1374 mg was used. (Figure 3). Despite the very small
cup or secondary electron amount of volatile loss and the
multiplier (SEM) detectors, correspondingly small increase in
depending on the sensitivity levels partial pressure, the MS was clearly
required. The SEM offers the ability able to detect this small change.
to detect very low partial pressures This trace clearly indicates the loss of
of evolved gases below 10-8 bar, dichloromethane from the sample,
allowing identification of very low as suspected, and shows the power
levels of contaminants. The system of this technique to identify very
is able tooperate in air, posing no small amounts of residual
issues for sample changing. Figure 2. Weight loss due to the loss of
solvent from the sample. Data has been contaminants in a sample.
• Optional foreline and bypass expanded to show the weight loss clearly.
vacuum pumping permits the most
efficient pumping of helium which A sample of a pharmaceutical was
is normally used as a purge gas found to exhibit an unusual
since it produces no interfering recrystallization behavior over a
mass ions. period of time. This behavior is often
• A very flexible heated transfer line associated with the loss of a solvent
allows simple interfacing to the from the sample. A method was
TGA systems. The lightweight required to confirm the suspected
identity of the solvent which had Figure 3. Weight loss curve and mass 49
heated line can be moved around (dichloromethane) in the 10-11 bar region.
in use without damage, making been used in the production of the
the system very robust. Gas sample.
The application of TG-MS to low-
transfer uses a wide-bore fused- 6.1374 mg of the sample was run in level contaminants that would be
silica capillary which gives the TGA; the HPR-20 MS was used undetectable by TG-IR is an example
16 mL/min flow to the mass in multiple ion detection (MID) mode of the power of this technique. For
spectrometer for highest using the high-sensitivity SEM more information, please contact
sensitivity. detector. The mass ion chosen to your sales representative.
• An anti-blockage filter has been identify the presence of the solvent
added to catch high-molecular- was 49 amu (atomic mass units) for
weight material that might block dichloromethane.

4
Fresh Analytical Sciences

Raman Spectroscopy –
A powerful tool to differentiate
pharmaceutical polymorphs

™
There is a constant need in the (damage to the esophagus caused photo-stability, than Omeprazole
pharmaceutical industry to prepare by stomach acid). In addition, it may form B. Form C is highly crystalline
stable crystalline forms of the active also be given together with and may be synthesized by a simple
pharmaceutical ingredient (API) in a antibiotics to treat gastric ulcers cau high yield process compared to
simple and cost-effective way within sed by Helicobacter pylori infection. A and B. As for all pharmaceutical
™
strict quality and regulatory Omeprazole is a racemate. It formulations, it is necessary to know
specifications. In particular, contains a tricoordinated sulfur atom the form present in formulation to
processing so that the proportion of in a pyramidal structure and can ensure patentability issues are
residual solvents, degradation therefore exist in equal amounts of addressed. In addition to the study
products and other impurities is kept both the S and R enantiomers. and detection of polymorphs,
within low limits. Pharmaceutical production processes Raman spectroscopy can also be
manufacturing generic drugs used to study the presence of
The compound 5-methoxy-2-[[(4- normally produce compounds in enantiomers in racemic mixtures. In
methoxy-3,5-dimethyl-2- various racemic forms. this study, our aim is to identify the
pyridinyl)methyl] sulfinyl]-1H- form A present in form B using the
™
benzimidazole, has the generic It has been found that Omeprazole technique of Raman spectroscopy.
™
name Omeprazole . It is known can exist in more than one
as a proton pump inhibitor which polymorphic form (typically forms A,
inhibits gastric acid secretion. B and C) as well as an amorphous
™ ™
Omeprazole is used for the form. Omeprazole form A is a
treatment of gastric acid related crystalline form exhibiting
diseases in man, as well as in other advantageous properties, such as Omeprazole™
mammals. It is used to treat the being well-defined and less
symptoms of gastro-esophageal hygroscopic than Omeprazole form
™
X-ray powder diffraction as well as
reflux disease and other conditions B, especially at room temperature. FT-IR may be used to differentiate
™
caused by excess stomach acid. ™
Omeprazole form A also shows Omeprazole form A from other
It is furthermore used to promote a better chemical stability, in crystalline and non-crystalline forms
™ ™
healing of erosive esophagitis particular, thermostability and of Omeprazole . Omeprazole forms

5
Fresh VOLUME 34 • Winter Edition, 2011

A, B and C are characterized by the 95-3500 cm-1 at high resolution in pure polymorphic forms or
positions and intensities of the peaks a single acquisition, removing the mixtures can be identified
in the X-ray powder diffractogram, need for scanning and stitching, (not described in this report).
as well as by the unit cell typical in other raman instruments. Conclusion
parameters. Another technique, An industry standard NIR 785 nm Raman spectroscopy is a very quick
Raman spectroscopy, can help laser is used for fluorescence and accurate tool to identify
identify amorphous forms too if rejection. The RamanStation 400 is polymorphs in pharmaceutical
present along with crystalline forms. equipped with a motorized XYZ compounds. In this case even
stage, allowing for routine analysis though FT-IR and XRD are used to
Experimenal of single or multiple samples. detect such polymorphs, Raman is
™
In this study, Omeprazole form A is considered as confirmative tool to
obtained upon slow crystallization Using the Raman spectrometer, we detect form A in form B as
™
and Omeprazole form B is obtained can easily identify the polymorphic mentioned in a U.S. Patent. This
from fast crystallization. ™
forms of Omeprazole . Raman work is being carried out to quickly
™
Omeprazole form A may be spectroscopy presents the advantage identify the forms created during
prepared by reaction crystallization compared with other techniques on crystallization process.
™
or recrystallizing Omeprazole of any the speed of analysis (few seconds)
form, or mixtures of any forms, in an and it does not require sample
appropriate solvent, for instance preparation. Those are strong
methanol, at room temperature and arguments on fast and accurate
for a prolonged time period from identification in a pharmaceutical
2 hours, to several weeks. production batch or the
™
counterfeiting analysis on
Omeprazole form C is prepared by Commerical drugs. Figure 1. Pure form A
™
dissolving crude Omeprazole,
Results and discussion
obtained in the reaction between
Figures 1 and 2 show the Raman
5-methoxy- 2-[[(4-methoxy-3,
spectra of pure form A and form B.
5-dimethyl-2-pyridinyl)methyl]thio]-
They present a zoomed portion of
1Hbenzimidazole and 3-
the spectra.
chloroperoxybenzoic acid at room
™
temperature first in a solvent or a Omeprazole form A is easily charac-
mixture of solvents in which terized by the absence of Figure 2. Pure form B
™
Omeprazole is freely soluble. a band at 1364 cm-1, which is
™ ™
Omeprazole form C is precipitated observed for Omeprazole form B,
at room temperature with a solvent and by the ratio of the relative inten-
in which it is poorly soluble. By this sities of the 842 and 836 cm-1 bands.
process pure crystals of The ratio (intensity of 842 cm-1
™
Omeprazole are precipitated while band/intensity of 836 cm-1 band) is
impurities remain in solution. The <1 for Omeprazole form A, while Figure 3. Overlay form A and form B
precipitated crystals are filtered off the ratio is >1 for Omeprazole
™

and washed. form B.

Similarly, Figure 4 shows that the
In this study, a PerkinElmer
™ peak ratio of 842 cm-1 to 836 cm-1
RamanStation 400 model was
for Form A is less than one, whereas
used. The unique echelle
for Form B it is more than one
spectrograph and 2D CCD detector
indicating identification of individual
offers complete spectral coverage, Figure 4. Peak ratio of form A and form B
polymorphs. In the same way, other

6 *We recognize Omeprazole brand of Zydus Cadilla Health Care Ltd.

Fresh Analytical Sciences

Analysis of impurities in Melatonin

by LC/TOF-MS
What is melatonin? When considering the use of advantages of melatonin may not be
Melatonin has been used to ease herbal/health supplements, known. Herbal/health supplements
insomnia, combat jet lag, protect consultation with a primary health should be purchased from a reliable
cells from free-radical damage, care professional is advisable. source to minimize the risk of
boost the immune system, prevent Additionally, consultation with a contamination.
cancer, and extend life. practitioner trained in the uses of
Important information about
herbal/health supplements may be
melatonin
beneficial, and coordination of Melatonin is a hormone secreted by
treatment among all health care the brain to help regulate sleep.
providers involved may be Since melatonin is found in certain
advantageous. foods ,it can be sold as a dietary
“over-the-counter” (OTC)
Melatonin is naturally produced in supplement. Unlike pharmaceuticals,
the body in response to the OTC drugs are not regulated by
perception of light. agencies such as the Food and Drug
Administration (FDA). Hence,
The use of melatonin in cultural and
Melatonin has not been evaluated impurities present in OTCs are not
traditional settings may differ from required to be characterized,
by the FDA for safety, effectiveness,
concepts accepted by current quantified, or reported. Impurities
or purity. All potential risks and/or
Western medicine.

7
Fresh VOLUME 34 • Winter Edition, 2011

found in OTC drugs can be harmful What are the possible side C13H16N2O4 (m/z=265.1183). This
and even cause death. For instance, effects of melatonin? provided the confirmation that a
in1989 an epidemic referred to as Although uncommon, allergic previously published structure
eosinophilia-myalgiasyndrome (EMS) reactions to melatonin have been
proposed from data provided by a
resulted in the death of thirty people reported. Stop taking melatonin and
and affected as many as 1500 seek emergency medical attention if triple quadrupole MS was incorrect
people. The epidemic was attributed you experience symptoms of a for a specific melatonin impurity.
to consumption of the OTC serious allergic reaction including
2.The significant improvement in
supplement tryptophan which was difficulty breathing; closing of your
manufactured by a single chemical throat; swelling of your lips, tongue, sensitivity gained from operating the
company. or face; or hives. AxIONTM 2 TOF in trap mode along
with the mass measurement
The synthesized tryptophan had at Other less serious side effects have accuracy allowed the identification
least six impurities which could have also been reported. Talk to your of additional melatonin impurities.
caused the on set of EMS. Since doctor or pharmacist if you These includedN1-acetyl-N2-formyl-
melatonin has structural similarities experience headache; drowsiness; 5-methoxy kynurenin (AFMK) as well
to tryptophan we have used the or stomach upset. Side effects other as melatonin dimmers not observed
high resolution and accurate mass than those listed here may also in previously published work.
capability of time-of-flight mass occur.
Courtesy:
spectrometry to identify and
Analysis of OTC Melatonin can be Sharanya Reddy, Shida Shen, Eugene
characterize impurities in OTC Davidov, Adam Patkin
easily and accurately carried out
melatonin tablets. PerkinElmer, Inc., 940 Winter Street, Waltham,
using PerkinElmer AxION™ 2 TOF MA USA
The use of melatonin derived from with Flexar FX 10 pump. C18
animal pineal tissue may carry the Brownlee Supra column. A well
risk of contamination or the means dissolved and centrifuged OTC DIRECT SAMPLE ANALYSIS SYSTEM
of transmitting viral material. The Melatonin tablet in Mili Q water was
synthetic form of this medication subjected to analysis with
does not carry this risk. PerkinElmer Ultraspray™ 2 (Dual ESI
source). Various method parameters
Melatonin has not been evaluated
were optimized using internal
by the FDA for safety, effectiveness,
celebrants. (Please ask for detail
or purity. All potential risks and/or
analysis conditions)
advantages of melatonin may not be
known. Additionally, there are no Operating the TOF MS in trap
regulated manufacturing standards (vs.pulse) mode provide da 3 to 5
in place for these compounds. There fold improvement in sensitivity. The
have been instances where increased sensitivity allowed
herbal/health supplements have additional impurities to be detected
been sold which were contaminated
with toxic metals or other drugs.
and measured in an OTC melatonin
sample that were not observed in
AxIONTM 2 TOF MS
Herbal/health supplements should pulse mode. • Non destructive direct sampling of
solids, liquids and gases
be purchased from a reliable source
1. The accurate mass measurement • No sample preparation required
to minimize the risk of
contamination. capability of the AxIONTM 2 TOF • Enclosed source ensures reproducibility
could easily distinguished between • Automated for rapid screening of
samples
elemental compositions
C14H20N2O3 (m/z=265.1547) and
REVOLUTIONALY MS CHARACTERIZATION

8
Fresh Analytical Sciences

The chromatography of sunshine

The “sunshine vitamin”, vitamin D, valuable tool in identification and D3 (400, 800, 1000 and 2000
is the only vitamin to be management of treatment regimes. IU/capsule).
manufactured by the human body, Vitamin D, see Figure 1, refers to
being formed on exposure of the vitamin D3 (cholecalciferol), the form When patients are being treated
skin to sunlight. Even people living in specifically with vitamin D2 it is
that is synthesized in the skin by
the sunniest climates may not make exposure to sunlight and to vitamin highly desirable to be able to
enough vitamin D. This deficiency D2 (ergocalciferol), which is obtained differentiate between the two forms
can contribute to a whole host of of the vitamin to determine the
from the ultraviolet irradiation of
health problems including rickets in efficacy of the dosage. In the body
plant materials and is the most
children, osteoporosis, the vitamin is tightly bound to a
common form of vitamin D used in
cardiovascular disease, diabetes, transport protein so sample
prescription supplements. Natural
osteoarthritis and many forms of preparation must be able to separate
sources of vitamin D3 include oily fish
cancer. Vitamin D deficiency is 25(OH)D from its carrier proteins.
(such as salmon or mackerel), cod
increasingly common amongst the liver oil and fortified food (such as Fast column separation
general population, which is leading milk, orange juice, butter, cheese Advances in column technology
to a significant growth in the need and breakfast cereals). Prescription make UHPLC a very practical and
for determination of 25-hydroxy vitamin D preparations contain reliable technique for the specific
vitamin D, also known as 25(OH)D, ergocalciferol, vitamin D2 (50 000 analysis of vitamin D2 and D3. Recent
in blood serum. As such, the ability IU/capsule), while over-the-counter developments in LC column packing
to determine vitamin D levels in supplements contain cholecalciferol, make use of sub 2 µm particle size
patients rapidly and reliably is a

9
Fresh VOLUME 34 • Winter Edition, 2011

materials. This enables higher mobile samples as the sample volumes

phase velocities without required are small, typically 50 to
compromising the separation 150 µL.
efficiency that is achieved using
columns packed with larger Capillary Entrance
Vo ltage
Capillary Exit
Voltage Hexapole
Quadrupole

HO HO Skimmer

particles. [M+H] +
M 1+

Vitamin D2 (Ergocalciferol) Vitamin D3 (Cholecalciferol) M 2+

M 3+

These sub 2 µm column materials Figure 1: Structures of vitamin D, Fragment lons

are typically packed into 2.1 mm i.d. (ergocalciferol) and vitamin D, (Cholecalciferol).
columns. By using smaller particles
Figure 2: Fragmentation zone on the Flexar
however, elevated operating The acceptance of UHPLC–MS into SQ 300 MS where CID is performed to
pressures are required, which brings obtain detailed structural information.
the clinical testing laboratory is in
its own challenges in pump and part due to developments in Analysis of Vitamin D in human
injector designs, but with software and instrument control that serum
technological advances these micro allows the seamless integration of Patient samples and human serum
particle columns can be used at clear chromatographic separation control samples were prepared by
pressures as high as 18 000 psi using with the powerful detection and taking 150 µL serum, adding 50 µL
commercially available systems such measurement capabilities of mass internal standard 25-hydroxyvitamin
D3-d6, then vortexed. 150 µL of
as the PerkinElmer FX15 LC. The spectrometry. Equipped with an
0.2 M ZnSO4 was then added and
columns are packed at high pressure electrospray ionization ion source the sample again vortexed. Finally,
giving exceptionally tight particle (Ultraspray) charged sample 750 mL hexane was added to the
size distribution that ensures column molecules are directed towards the mixture and centrifuged at 8000
stability for UHPLC applications. The entrance of the mass spectrometer, RPM for 7 min. The top layer–the
small particle size and rugged where they are sampled from the organic phase–was removed and
construction combines to deliver edge of the sample plume. As the blown down under nitrogen. The
sample was reconstituted in 100 µL
both high column resolution and droplets are smaller in this region
of MeOH/H2O.
high sample throughput. they require less energy for
ionization, enabling weak molecular Following sample preparation the
Routine clinical analysis using
structures to be preserved for samples were analysed using a
UHPLC with MS detection
Whilst chromatographic separation analysis. PerkinElmer Brownlee HRes
of the samples is a key element of analytical DB C18 column (2.1 mm×
Established sample preparation
the analysis, unequivocal 50 mm, 1.9 µm). The mobile phase
identification of the sample Liquid-liquid extraction (LLE), solid-
consisted of A: water containing
components is essential. Single phase extraction (SPE) and protein
2 mM ammonium acetate and
quadrupole mass spectrometry is an precipitation (PPT) are commonly
0.1% formic acid and B: methanol
excellent detection method because used sample preparation methods to
of its speed, ease of use and containing 2 mM ammonium
be found in bioanalytical
specificity. The resulting UHPLC–MS acetate and 0.1% formic acid.
laboratories. The samples are
combined technique is ideal for The analysis was run under isocratic
vitamin D determination because of precipitated out to remove high
conditions with a flow-rate of 0.3
its ability to differentiate between molecular weight substances, the
mL/min.
25(OH)D2 and 25(OH) D3, which has solid phase is extracted, the eluate
been highlighted as particularly evaporated with nitrogen and The advantage of this UHPLC–MS
important when monitoring the suspended in a mobile phase ready technique for determining vitamin D
vitamin D levels of patients being for injection through the HPLC is the specificity of the analysis,
treated with vitamin D2.
column. UHPLC–MS lends itself to allowing differentiation between
the rapid analysis of biological vitamins D2 and D3. Thanks to an

10
Fresh Analytical Sciences

efficient chromatographic separation 250 quadrupole mass spectrometry

combined with the soft ESI 200
detectors (MS) and sophisticated
ionization technique, the system can 25-Hydroxyvitamin
R t=0.99 5
D3 data systems is now leading to

Counts Per Second

150 25-Hydroxyvitamin D2

distinguish between the various R t=1.02

UHPLC–MS systems becoming
25-Hydroxyvitamin D3-d 6

forms of vitamin D. Its quadrupole 100

Rt=0.99 5
commonplace in the bioanalytical
offers exceptionally fast scanning 50
laboratory.
acquisition (10 000 u/s) making it 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0
Sean daugherty is currently mass
ideal for UHPLC analysis in Time (min)

Figure 3: Patient sample with separation and spec detection technical sales
simultaneous full scan mode as well
detection of 25-hydroxyvitamin D2, (blue trace), specialist at PerkinElmer. He received
as in selected ion monitoring (SIM). 25-hydroxyvitamin D3, (Red trace) and
his degree in analytical chemistry at
Whereas liquid chromatography 25-hydroxyvitamin D3, -d6, (purple trace).
the renowned kemikum at Uppsala
with tandem mass spectrometers
University, Sweden while working
(LC–MS–MS) is frequently used, the monitoring of ions specific to
with mass spectrometry. He has
single quadrupole system with vitamins D2 and D3. The conditions
since continued his work in
collision induced dissociation (CID) used here for routine vitamin D chromatography and mass
often allows the required specificity testing by UHPLC/SQ MS resulted in spectrometry in a wide breadth of
to be achieved. Figure 2 shows highly symmetric peaks with a run countries and industries including
where the capillary exit voltage is time of 6 min, demonstrating the pharmaceutical, biotech and clinical
modulated to ensure effective CID speed of UHPLC–MS compared with where his work has kept him at the
and reproducible fragmentation of other techniques. Ease and speed of cutting edge of chromatography and
the sample. The single quadrupole analysis do not compromise the mass spectrometry. Sean brings a
analyser then mass filters these levels at which trace analytes can be unique and insightful perspective to
molecular fragments enabling users detected. Patient samples at below sample analysis with LC–MS
to define a particular species to be 4 ng/mL 25 hydroxyvitamin D were
detected. able to be detected and quantified VERIFY. QUANTIFY. SIMPLIFY.
Controlled flexibility with confidence. According to
The Chromera data system enables current industry definitions, severe
seamless control of both the UHPLC vitamin D deficiency is defined as
and MS components of the < 6 ng/mL, deficiency as
instrument and was specifically < 16 ng/mL, insufficiency as
designed for chromatography < 20 ng/mL and normal as
applications. Users can select exactly > 20 ng/mL. Therefore, UHPLC–MS
the type and style of information is a robust technique for detecting
they want displayed for simpler, and quantifying vitamin D levels
clearer, more relevant results. Tuning within the clinically significant range.
can be performed across 3000 mass Conclusion FlexarTM SQ 300 MS
units or limited to a user-specified The power of chromatography to
mass range for superior speed and separate very similar components in • Innovative ionization sources
results. complex mixtures has long been • A unique, angled, fully
proven and exploited for the adjustable probe
Figure 3 shows the clear peak
determination of vitamin D levels in • Most effective desolvation and
separation from samples of 25 ionization
hydroxyvitamin D3, 25 human serum. The combination of
improved column design, fast
hydroxyvitamin D3-d6 and 25
chromatographic separations For more information log on to
hydroxyvitamin D2 enabling SIM www.perkinelmer.com/flexarsq300ms
(UHPLC) with robust single

11
Fresh VOLUME 34 • Winter Edition, 2011

Determination of oil content in membranes used in

compressed air sampling by infrared spectroscopy
Compressed air sampling is very tool for the monitoring of oil
essential from an environmental content. The methodology followed
point of view and quality of such air for the analysis by FT-IR is reported
used in laboratories, production and in BIS (Bureau of Indian Standard)
instrumental purpose where it is and have also tested for its
required to know all the parameters ruggedness, spike recovery, linearity
such as SOx and NOx. and detection limits. Air filtration or
These parameters are very important Air filters are used and designed into
to determine the air quality. gas/air flow systems to remove
Membranes used in compressor Spectrum Two FT-IR instrument particulates from the air flow. Air
should be oil free. Hence filters and filtration techniques are
The test method for the quantitative used for wide variety of purposes,
quantitative estimation of oil
analysis of aerosol oil and liquid oil some of which include:
trapped in the membranes being
typically present in the air discharged
used in air compressor is very • Improvement in the breathable
from compressors and compressed
important. FT-IR studies can be very environment for occupant
air systems. The method is rapid,
effective in calculating the oil • Elimination or minimizing fire
sensitive and cost effective and
content in membranes to a very low hazards
shows the FT-IR can be an effective
level as well.

12
Fresh Analytical Sciences

• Removing airborne bacteria from membranes. It does not require

controlled environments complex sample preparation. Spiking
This method is applicable when an studies can be studied using the
additional sampling probe is inserted sample solutions and with three
to maintain isokinetic conditions levels of C1, C2 and C3.
from the main pipe flow. For The instrument, Spectrum Two gives
sampling from high flow systems, us the flexibility to run the samples
isokinetic sampling may be used Standards spectra with 1,1,2,2 tetrachloroethane in liquid phase with 50 mm path
when wall flow is not present. solvent length cells. This application is useful
Condensate may also be collected in environmental segment wherein
at the sampling filters. When high Set Resolution 4 cm-1 with 8 scans. we need to monitor the quality of air
levels of condensates are present, The critical part of the analysis is compressors in terms of oil being
collection rates increase and larger sample preparation where trapped. This technique is very useful
collecting containers are required. Membrane was first accurately for the air compressor manufactur-
weighed on a balance. Further it was ers and users of it in the critical envi-
The liquid collected consists of
kept for extraction of oil in 25 mL ronments like pharmaceutical pro-
water, oil/water emulsion and oil.
carbon tetrachloride/1,1,2,2 cesses and laboratories.
Depending upon the type of oil,
tetrachloromethane solvent for
separation of the oil/water emulsion Courtesy:
30 mins. The solution was decanted
may occur, allowing the water to be Avdhut L. Maldikar, (Ph.D)
into a 25 mL standard flask and the PerkinElmer Pvt. Ltd.
drained off and oil to be measured.
membrane was washed thoroughly. Kasarvadavali, Thane (West), India.
Making use of reagents like Carbon All the washings were collected in
tetrachloride (CCl2) and 1,1,2,2, the same 25 mL standard flask. APPLICATION & RESOURCE PACKS
tetrachloroethylene (C2Cl4) AR Finally volume was made up using
Grade, used as solvent. Preparation carbon tetra chloride/1,1,2,2
of stock and calibration solutions tetrachloroethane. FT-IR spectrum
should be done as per the procedure was recorded using the same FT-IR
provided in detail application note. parameters which were used for
PerkinElmer Spectrum Two™ FTIR standard calibration plot. The
spectrometer with slide holder and Absorbance (A) was calculated as FT-IR Pharmacopoeia
transmission cell with Spectrum 10 per the graph and reported as Compliance Resource Pack
software to be used for unknown sample concentration.
The FT-IR Pharmacopoeia
measurements and calibration The experiment was repeated and Compliance Resource Pack
between Scan Range 3400 cm-1 to checked for reproducibility of the assures absolute compliance to
IR testing. The Pack is compatible
2500 cm-1 method. The data demonstrates the
with seven major global
repeatability and reproducibility of pharmacopoeias. (USP, BP, EP, JP,
the method. The working range for IP, CP & IntPhr)
the method was found to be 1 ppm
• FT-IR Sampling Methods
to 70 ppm provided that a 50 mm
• FT-IR Pharmacopeia Identification
path length cell has been employed. Tests
Conclusions • 21CFR Part 11 Technical
The developed method is sensitive Compliance
and cost effective and shows that
FT-IR can be an effective tool for the For more details log on to
Calibration plot with 1,1,2,2 tetrachloromethane
monitoring of oil content in
www.perkinelmer.in
solvent

13
Fresh VOLUME 34 • Winter Edition, 2011

E-Notebook for formulation scientists

The E-Notebook formulation module • Multi-step formulations in a single view, expand and collapse
provides all the tools that • Screening experiments allowing steps and that supports out-of the
formulation scientists working in for quick set-up of multiple box specialized calculations such as
pharmaceutics, chemical, flavor and formulations in a single calculation of non-granular
fragrance, food and beverage and experiment excipients needed to complete
other industries need to effectively a formulation.
document their experiments and to Additionally, it is possible to include
make the most value out of other formulation as ingredients and
knowledge gained from previous still calculate the amounts of primary
experiments. Specialized tools for ingredients. Other essential features
formulation scientists include: include an auto text capability that
• Pre-defined and user-definable allows users to insert pre-defined
calculations supporting the specific Figure1. Detailed formulation setup
and user definable common phrases
calculations and units such as that include numbers linked to
%w/w and mg/ml needed by Key to the formulation set up form information in the data tables such
formulationists is a user-customizable spreadsheet that if a number is changed in data
• Quick access to commonly used with special capabilities for tables then that change is reflected
excipients and other ingredient formulationists including the immediately in the auto text. Spectra
databases capability to set up multiple steps and other analytical data may be

14
Fresh Analytical Sciences

easily linked with the formulation over a formulation series to information and quickly analyze
composition to make recording of determine its effect on a property of results. All of these capabilities are
the total experiment quick for the the formulation, or the length of provided on top of the E-Notebook
author and easy to follow for one operation such as heating time framework ensuring that all
colleagues. might be investigated. For these intellectual property and 21 CFR
purposes, a powerful summary Part 11 requirements are satisfied.
spreadsheet capability is provided The combination of detailed
that allows a formulation scientist to workflow support, including
set up multiple formulations in one specialized calculations and units,
experiment. Moreover, users may along with analysis, reporting and
drill-down to the underlying general ELN capabilities results in
formulation to record information, dramatic productivity and
including analytical data per compliance benefits. Users will
formulation, in as much detail as wonder how they ever worked in
Integral to formulation science is the
if the experiment were carried out paper or Excel before.
need to vary one or more
individually. This capability even
independent variables in a series of For more information about
extends to multistep formulations.
related formulations in order to best E-Notebook, please send email to
understand the impact of these The E-Notebook Formulation [email protected]
changes on relevant dependent Module provides formulation
Courtesy:
variables. For example, a scientist scientists with all the tools they need Michael W. Swartz
might vary one or more ingredients to record experiments, organize PerkinElmer Informatics, USA

The desktop productivity toolkit for scientists

CHEMBIO • ChemBio3D: Prediction of IR Raman and UV VIS data • Modeling within PowerPoint using ChemBio 3D

OFFICE • ChemBioFinder – Chemical Database Management System

• TLC Plate, Table Tools, Arrows
• Highest Presentation Quality Graphics
• ChemBioDraw Calculates Reaction Stoichiometry
• Expand Generic Structures

For more information log on www.cambridgesoft.com

*CambridgeSoft is now PerkinElmer Informatics

15
© 2009 PerkinElmer, Inc. 400161_09. All trademarks or registered trademarks are the property of PerkinElmer, Inc. and/or its subsidiaries.

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