sunlight exposure is enough to aid the neonate in

Direct Absorbance Bilirubin processing bilirubin. Phototherapy treatment is

successful in treating hyperbilirubinemia in most
Spectrometer: Analysis of extreme cases; therefore, the key issue is timely
diagnosis and treatment.
Bilirubin Content Using Bilirubin concentration levels usually rise to
Direct Absorbance dangerous levels 3-4 days after birth. Many infants
and mothers are discharged from hospitals 2 days after
Spectroscopy birth, so hyperbilirubinemia is often not detected until
the first pediatric checkup, often 5-7 days after birth.
Andrea Wetteland, Kevin Landau, Bernardin This means more serious cases of hyperbilirubinemia
Dezius, Juan Gonzalez, Kevin Kuzius can go undiagnosed for days. The DABS project is a
Dept. of Electrical Engineering and device capable of quickly measuring bilirubin levels
from a small sample of whole blood and determining
Computer Science, Dept. of Optics and
whether the bilirubin levels are “low”, “borderline”, or
Photonics, University of Central Florida, “dangerous”.
Orlando, Florida, 32816-2450
II. GOALS AND OBJECTIVES
Abstract — Neonatal hyperbilirubinemia, or
jaundice, is a common but potentially dangerous DABS is designed for clinical use, so that a pediatric
condition in newborns wherein elevated concentrations clinic can provide worried parents with a quick,
of bilirubin causes yellow discoloration of the skin and inexpensive determination of the severity of their
eye whites. The Direct Absorbance Bilirubin newborn’s jaundice. DABS will be easy to operate
Spectrometer (DABS) is designed to determine whether and take under 5 minutes to calculate an output, so that
the bilirubin levels are threatening or safe by measuring an appointment would not be necessary for parents to
light absorbance through a sample. The absorbance is make an appointment. Given a sample, DABS will
calculated with the Beer--Lambert Law using currents determine if it is “safe”, “elevated”, or “dangerous”.
generated from a photodiode by incident light from two DABS will use natural pigments in place of real
monochromatic laser diodes transmitted through a hemoglobin and bilirubin but can be converted to
sample. The DABS device is capable of providing clinical use with real human blood serum with small
approximate results that can indicate whether bilirubin modifications and recalibrations.
levels are safe, elevated, or whether immediate medical
attention is recommended.
Index Terms — Central processing unit, optical III. BILIRUBIN ANALYSIS METHOD
wavelength conversion, diode lasers, data analysis,
temperature control, amplifiers. Bilirubin is a yellowish-orange and absorbs light at
440 nm. Hemoglobin also absorbs light at 440 nm but
absorbs light at 528 nm in equal measure. In neonates,
bilirubin absorbs light at 460 nm. No other substances
I. INTRODUCTION present in blood at this age absorb light at this
wavelength, besides hemoglobin. Hemoglobin also
Neonatal hyperbilirubinemia, or jaundice, is a
absorbs light at 460 nm and has an absorbance peak at
common condition in newborns wherein elevated
540 nm. The ratio of hemoglobin at both wavelengths
concentrations of bilirubin causes yellow discoloration
in full-term infants is constant enough so that a
of the skin and eye whites. Bilirubin is the product of
hemoglobin absorbance at 460 nm can be calculated
hemoglobin being broken down. When the neonate
liver is incapable of processing the bilirubin because from the hemoglobin absorbance measurement at 540
of increased production, decreased excretion, or other nm. [1]
impaired mechanic, an excess concentration can
occur. Levels resulting in visible discoloration occur
in almost 60% of all neonates. At higher levels (308
μmol/L), the bilirubin is toxic to the neonate brain, and
encephalopathy or brain damage (kernicterus) can
occur without treatment. For less extreme cases,
 IV. SAMPLE STANDARDS

In full-term neonates, bilirubin levels below 5.2 mg/dL

are considered low, levels between 5.2 mg/dL and 15
mg/dL are considered elevated, and levels over 15
mg/dL are considered dangerous and in need of
phototherapy [2]. Based on the molecular weight of
bilirubin and a 1 cm optical pathlength, this translates
to respective absorbance values of .5 and 1.4.
Curcumin standards with equivalent absorbance
values were developed by adding turmeric to isopropyl
alcohol or IPA until the desired absorbance value at
Fig. 1. Both substances contribute to the absorption taken 405 nm was obtained with respect to the intensity
at blue. The absorbance due to bilirubin can be determined transmitted through a blank sample and the Beer’s
by subtracting a projected absorbance value of hemoglobin Law equation in (1). Betalain solution was added to
at blue, based on its absorbance value at green. each standard in a 1-2 ratio. The absorbance values
Because this team does not have the resources to obtained through the MCU calculations, as shown in
meet the conditions for procuring, storing, and Table 1 and the calculated curcumin values became
handling human blood samples as dictated by code our standard absorbance values to deem whether
CFR 21 by the Human Research Protection Office, the unknown samples were “safe”, “elevated”, or
DABS project will not be measuring real bilirubin “dangerous”.
from real neonatal human blood serum samples.
Curcumin, a yellowish-orange pigment found in Table 1. Low and High threshold absorbance values were
turmeric, absorbs light at 405 nm. Betalains, reddish- calculated from curcumin concentrations.
purple pigmentation found in beetroot, absorbs light at Low High
both 405 nm and at 532 nm. The DABS device will Threshold Threshold
demonstrate the proof of concept by detecting
Measured Absorbance at 1.45016 1.70543
curcumin content from betalain samples using the
405 nm
same principle proposed for human blood serum.
DABS will determine the concentrations of Measured Absorbance at 1.16975 1.30103
curcumin by illuminating a blank sample with two 532 nm
light sources, one at 532 nm and one at 405 nm, and Calculated Betalain A at 1.0856 1.20749
recording this incident intensity. A blood sample or 405 nm
proxy will then be illuminated and the transmitted
intensity at each wavelength will be recorded. The Calculated Curcumin A .36452 .49794
absorbance at each wavelength will be determined at 405 nm
using the Beer Lambert Law equation.
The two standard curcumin solutions are depicted in
A=log(I0/I) ( 1) Figure 2 and have respective absorbance values of .5
The absorbance at 405 nm due to curcumin will be and 1.4. 1 mL of each solution was added to 2 mL of
determined by subtracting the betalain absorbance betalain solution, and the resulting absorbance values,
(405 nm), which was determined by its ratio to the .365 and .498 were coded as the high and low
betalain reading at 532 nm. threshold barriers for future samples, with readings
The DABS device will require a user to insert a below and up to .301 corresponding to the “safe”
blank cuvette filled with 100% isopropyl alcohol and output, readings above .369 and up to .498
corresponding to an “elevated” output, and readings
then a cuvette filled with a beetroot solution
containing unknown small concentrations of above .498 corresponding as “dangerous”.
curcumin. It will calculate the absorbance due to
curcumin in the sample, and deliver an output
recommendation of “low”, “borderline”, or
“dangerous”.
 beams to pass through the solution. This is a
satisfactory volume for the curcumin and betalain
solutions prepared for this project but would not be a
suitable sample volume to require of a neonate. Sub-
millimeter pathlength cuvettes are available but
expensive. If DABS were to go into production for
use in a clinical setting with real blood, it would be a
necessary expense.
The photodiode utilized by DABS is a BPW21R
Photodiode from Vishay Semiconductors. This
semiconductor has a slower response time when
compared to other semiconductors on the market, but
a higher sensitivity, which more closely meets the
Fig.2. The two curcumin solutions with need of the project. The goal of the DABS device is
absorbance values of .5 and 1.4 were used to determine the
to accurately measure varying unknown
low and high threshold values, respectively.
concentrations of turmeric in a betalain and turmeric
solution that acts as the blood proxy. Since the sample
V. OPTICAL SYSTEM is not changing at a relatively fast rate the response
time was not as big of a factor as the sensitivity to
The optical subsystem resides in its own light-
determine the varying concentrations. The spectral
containment compartment and comprise of the two
range of the BPW21R is 400 nm – 675 nm, which
light sources, a cuvette, and a photodiode. The light-
includes both wavelengths of the laser sources being
containment compartment sits inside the outer DABS
read. It also has the largest sensitive area allowing it to
structure. Both laser diodes are stationary and are
collect more light with a linear response to directly
aligned so that both beams directly hit the surface of
correlate to the Beer Lambert law to measure the
the photodiode.
The two laser diodes, 405 nm and 532 nm, each have correct concentrations of the samples.
5 mW output and a working voltage of 3-5 Volts. Both
laser diodes were purchased as modules with built-in VI. ELECTRICAL COMPONENTS
current drivers. The emission peaks were measured
with a spectrometer, and the blue and green diodes A. Transimpedance Amplifier
have 2 nm and 2.5 nm full width half max (FWHM) Since DABS measures the curcumin concentration
values, respectively. The narrow emission spectra is using direct absorbance (i.e., measuring the
important to determine the absorption values at absorbance of light), we used a circuit that links light
specific wavelengths. to a meaningful voltage value. This is the purpose of
the photodiode and transimpedance amplifier that was
incorporated into DABS, Figure 4 gives a schematic
illustration of this concept. For this, we used the OPA
170 by Texas Instruments.

Fig .3. The layout of the optical subsystem within its light-
containment housing.
The cuvettes are glass, as it was inexpensive and
provided the needed transmission at our two
wavelengths on the visible spectrum. The dimensions
of the cuvettes are 12 mm x 12 mm x 45 mm and
require a minimum 2.5 ml sample volume for the laser
 the lasers and sends an output voltage to an input pin
of the MSP430 microcontroller.
A quick summary of the LM35 is given below:

• Linear +10-mV/°C Scale Factor

• Temperature range of -55 °C -to 150°C
• Operates from 4 V to 30 V
• Less than 60- uA current drain
• 0.5°C ensured accuracy

For supplying cooled air to the lasers, two 5 V fans

are utilized at 13200 rpm. When the temperature is
measured from the temperature sensor and sent to the
microcontroller, the MCU sends an output voltage to
two bipolar junction transistors. Once the threshold
Fig. 4. Transimpedance amplifier. voltage is surpassed, the BJT will act as a short circuit
In this figure, the photodetector output is modelled and allow the fans to turn on from 5 Volts.
using the current source, the feed into the amplifier.
The feedback resistor, R1, amplifies the current giving
the circuit gain modeled in Figure 5. VII. MICROCONTROLLER SYSTEM

The microcontroller used in this project is the

MSP430FR6989 produced by Texas Instruments. It
acts as the coordinator of the system by mediating
when subsystems are activated, consuming power.
Such as not constricting the polling of touch response
to only when a user has touched the screen. And
powering down the display after a duration of idleness.
The microcontroller also handles the computation and
conversion of the analog readings from subsystems in
the module. More specifically the readings from the
touch display and the readings from the optical
Fig. 5. OPA170 Simulation Output. subsystem.
The photodiode will read the incident light, as it passes
through the sample, and then output a current that is VIII. DISPLAY/TOUCHSCREEN INTERFACE
dependent upon the sample concentration. This current The primary interface for user interaction is the
will then serve as the input to the amplifier, similar to touch-LCD display. It is a resistive 4-wire touch
the figure shown above. In order to produce a voltage interface and it allows for the user to operate the
that can be read, and interpreted by the ADC of the DABS module. It comes pre-compatible with the
microcontroller, we chose a resistor that gave a stable prototype board, so this display was a satisfactory
output over a relatively large range of inputs. Based on choice and gives the functionality needed.
simulations, using the TI-TINA schematic editor and The structure of the resistive touchscreen also
breadboard testing, we were able determine that a assists with the objective of making the module as
resistor value 4.75kOhms. This value gave us a useful energy efficient as possible. Instead of needing to run
voltage output that varies enough, over a range of input a constant loop that checks for a press-down the
current. display allows for the system to simply detect a change
B. Cooling system once the it’s been touched or un-touched. Regarding
its use in operation it is illustrated in Figure 6. At start
To measure the temperature of the optical system, up the screen will display some information regarding
the Texas Instruments LM35 centigrade temperature is its boot up. A touch calibration screen is a part of this
used to accomplish this. The LM35 was chosen as it is
the most common temperature sensor for small to mid-
level projects. The sensor measures the temperature of
boot up process but is only show after a full reset of
the system.

Fig. 7. Welcome screen on LCD touchscreen.

IX. POWER INPUT

The main power source is a lithium polymer-based

battery capable of outputting a voltage of 4.2 Volts
when fully charged and a current compacity at 2500
mAh The battery system will be able to recharged
using a USB Type-A connection to an external power
source either from a wall outlet using a USB adapter
or any computer system that has a USB connection
port.
The configuration shown in the schematic for the
charging circuit allows the device to be power by an
external power source while charging the battery
(Figure 8). So, two power source options will be
available for the functionality of the device.

Fig .6. Flowchart of the operation of the DABS module

device

Following the startup is the main menu where the

user has the option to start the optical analysis, go into
settings, put the device into sleep mode. Once the
reference sample is inserted and the analysis starts the
screen updates accordingly and indicate the running of
this analysis until the it is complete.
Following its completion, the next screen gives the
user the option to restart the process or continue and Fig. 8. Power input and battery charging circuit.
run a similar process for the next sample after the
blood sample has been placed in the device. From the schematic, a USB port was soldered on the
The final screen of the process gives back a PCB for a USB connection for optional power input.
summary of the results of the analysis along with any The main chip that will charge the battery is the
recommendation accordingly. Such as whether MCP73821 from Texas Instruments.
treatment or medical attention is required. Figure This chip was chosen for the reliability and safety of
below displays the welcome screen that displays the charging the main battery system at 4.2 volts and a
start button and settings for users to choose. The start programmable charge current rate of 500 mA. The
button begins the testing analysis. range of supply voltage to this chip is from 3.75 to 6
volts max which is safe operation since most USB type
connections are rated at 5 volts.
 The design also has 2-pin JST-XH connector that
was soldered on the PCB. This will allow the battery
to simply connect to the board and be secured tightly
and not have a loose connection. The battery can also
be replaced if the battery can no longer be functional.
The schematic also includes a Schottky diode that
connects between USB output to the drain pin on the Fig. 10. 5 V DC-DC converter.
P-Ch MOSFET. The diode is there to prevent any
leakage current from the device.
A P-Ch MOSFET is used to open the circuit from
battery to a DC-DC converter or a buck booster
whenever there is power coming from the USB
connection. When there is no power from an external
power source, the P-Ch MOSFET will close the switch
between source and drain and allow the battery to
provide power to the system.
For the low-powered microcontroller to function
properly, a voltage input of 3.3 volts is required as
Fig. 11 3.7 DC-DC converter.
specified the MCU datasheet (Figure 9) . A buck
converter is used to accomplish this feat. The input
voltage of this circuit will either be 4.2 volts from the X. PROCEDURE
lithium-polymer battery when fully charged or 5 volts
A betalain solution was created by soaking 1 oz of
from an external power source using USB connection.
diced fresh red beets (beta vulgaris) in 2 oz 100%
The IC used to provide 3.3 V to the controller is the
isopropyl alcohol (IPA). Various amounts of IPA
TPS6300x. This chip is rated to take an input from 1.8
to 5.5V and output 1.2 to 5.5 Volts at 1200 mA or less. were added to cuvettes with betalain solution to create
In our project, the IC is set to provide 3.3 V at 600 mA. multiple concentrations. The absorbance of each
solutions was recorded at 532 nm and 405 nm. A line
was fitted to the results, passing through the origin, to
give the absorbance of betalain at 405 nm as a result
of the absorbance at 532 nm.
A line was fitted through the datapoints and the
origin to give the absorbance of blue as a function of
betalain absorbance at green (Shown in Figure 12).
The curcumin concentrations were prepared by
submerging peeled turmeric into 10 ml of 100% IPA
for 5 seconds.

Fig. 9. 3.3 V DC-DC converter.

Two other DC-DC converters were used for a 5 Volt and
a 3.7 Volt output. The two converters and can be seen in
Figure 10 and Figure 11. All the DC-DC converters were Fig. 12. The absorbances of multiple betalain
created using Texas Instrument WEBENCH software tool. concentrations at 532 nm (green) and 405 nm (blue).
Three samples of this this solution with descending
concentrations were prepared by diluting subsequent
mixtures with 10 ml IPA. The absorbance of each
sample was measured at 405 nm for validation. After
the target values were recorded, 2 ml of betalain
 Using this methodology, the resulting curcumin
solution was added to 1 ml of each curcumin absorbances were consistent and matched values
concentration to form the 3 samples. expected based on the yellow coloration of the
The microcontroller calculated the absorbance of curcumin solutions. The resulting values maintained
each sample at 532 nm and at 405 nm by recording 4 the same relative scalars as the expected values, and
input values: the voltage values from the the correct “dangerous”, “elevated”, and “safe” values
transimpedance op amp generated by currents from the were identified, shown in Table 3. When curcumin
photodiode at each wavelength for the incident light was added to samples reading “safe” or “elevated”, the
through a blank cuvette containing pure IPA (I405nm measured absorbance went up and the output indicator
and I532nm) and the transmitted light through the sample changed accordingly.
(T405nm and T532nm). The absorbances were calculated
with the Beer’s Law equation Table 3. The results from three unknown solutions, and
A405nm =log(I405nm/ T405nm) ( 2) the output values printed for each.

A532nm =log(I532nm/ T532nm) ( 3) Calculated Curcumin Output

Absorbance
The absorbance at 405 nm for the betalain alone was .217 Safe
calculated using the absorbance at 532 nm and the .385 Elevated
equation from the fitted line in Figure 12, .857 Dangerous

AB405nm= A532nm*.9281 ( 4) XII.CONCLUSION

AB405nm was subtracted from the measured absorbance DABS successfully determined whether the
of the whole solution, A405nm. The remainder was the curcumin absorbance of a given sample was safe,
absorbance value attributed to curcumin, AC405nm. elevated, or dangerous in relation to the threshold
values input. Consecutive runs of the same sample
AC405nm = A405nm - AB405nm ( 5) contained some variance, but the output category for
each value remained consistent. There would be
XI. RESULTS clinical benefit if the concentration of each sample
could accompany the output, but the small
Three curcumin solutions with unknown discrepancies between consecutive runs would
concentrations were prepared and 1 mL each was translate into larger variance in concentration values
added to 2 mL of betalain solution, maintaining the and the results would be less meaningful. However,
same ratio as the standards. Table 2 shows the the primary objective of DABS was to provide a quick,
general reading so that parents could be reassured in
Table 2. The incident and transmitted inputs at 405 nm and cases of mild jaundice and be prompted to send further
532 nm and the calculated absorbances for each of the three
sample concentrations. samples to a lab for more accurate testing in the case
of dangerous levels. Given the appropriate threshold
A532nm AB405nm A405nm A405nm-
values, DABS successfully achieves the objective.
Sample # AB405nm
For DABS to be used to calculate the bilirubin
absorbance from real human blood serum, a few
1 1.1915 1.1059 1.3231 .217 modifications would be necessary. The current
2 1.1697 1.086 1.4706 .385 cuvettes require a minimum of 2 mL to ensure the laser
3 1.1154 1.035 1.8925 .857 beam passes through a sample. This is too high a
A532nm AB405nm A405nm A405nm-
volume to be taken from a newborn to meet the
Sample # AB405nm
objectives of safety and ease of use. Custom .05 mm
1 1.2578 1.1674 1.4154 .248 pathlength cuvettes are available that would require
2 1.2607 1.1701 1.6011 .431 only 4 or 5 drops of blood for a sample. Secondly, the
3 1.0345 .9601 1.9201 .960 laser modules used were inexpensive, but for more
consistent results, DABS would use higher-quality
measured absorbance logged for each sample at green laser modules with electrical attenuation to reduce
and blue, and the calculated absorbance due to noise in the signal. Readily available wavelengths 405
curcumin. Each sample was run two times in order to nm and 532 nm were sufficient to capture curcumin
verify that the values were similar for samples tested and betalain on blue and isolate betalain on green, but
twice in a row.
a 460 nm wavelength would be needed to capture the
absorption of bilirubin. Finally, blood standards Andrea Wetteland is graduating
would be used to recalibrate new threshold values. with a Bachelor of Science degree
With these three modifications, the DABS prototype in Photonic Science and
could successfully meet its goals and objectives in a Engineering from CREOL, The
pediatric clinic setting. College of Optics and Photonics at
the University of Central Florida.
She has been working as a laser
ACKNOWLEDGEMENTS
engineering intern at L3Harris
Throughout the project, there were several faculty Advanced Laser Systems
members that provided invaluable insight and Technology and plans to continue
guidance to completing a successful project. The team designing and testing laser
greatly appreciates and would like to thank Dr. Kyu resonators and optical systems
Young Han and Dr. Patrick LiKamWa from CREOL, upon graduation.
The College of Optics and Photonics, for their advice
on the optical system, Mr. Zotti of the CREOL Kevin Landau is graduating with
machine shop for taking the time to teach and assist us a Bachelor of Science degree in
with machining for the project, and to Dr. Richie, Dr. Photonic Science and Engineering
Lei Wei, and Dr. Hagan for their guidance and from CREOL, The College of
assistance throughout the year in helping us to meet Optics and Photonics at the
our project goals and milestones. University of Central Florida. He
plans to go into industry after
graduating with ASML which
BIOGRAPHY
works with photolithography
Bernardin Dezius is a graduating machines for the semiconductor
Computer Engineer at the industry. He also hopes to
University of Central Florida. He eventually return to school to
plans to go into the industry right obtain a master's degree in a STEM
after graduating and hopes to related field.
become a Software/Hardware
Consultant with a software firm. REFERENCES
He also hopes to eventually return
to school for a degree in one of
[1] A. Westwood, "The Analysis of Bilirubin
multiple stem fields that interest
in Serum," Annals of Clinical
him.
Biochemistry, vol. 28, pp. 119-130, Mar
1991.
Juan Gonzalez is a graduating
[2] S. Meites and J. W. Traubert, "Use of
Electrical Engineer at the
Bilirubin Standards," Clinical Chemistry,
University of Central Florida. He
vol. 11, no. 7, pp. 691-699, 1965.
plans to make his internship a full-
time job as a Control Systems
Engineer. He also plans to continue
his education by obtaining his
masters in a STEM related field at
the University of Central Florida.

Kevin Kuzius is a graduating

Electrical Engineer at the
University of Central Florida.
Currently a part of the College
Work Experience Program
partnership with Lockheed Martin.
He plans to enter the industry upon
graduation.