Journal of Analytical Toxicology, Vol.
21, October 1997
A Simple SpectrophotometricAssayfor the
Measurement of Soluble Silica in Water
Barbara R. Manno 1, Imad K. Abukhalaf2,, and Joseph E. Manno 1,3
Departments of 1Psychiatry,2Medicine, and 3EmergencyMedicine, Louisiana State UniversityMedical Center,
School of Medicine, 1501 Kings Highway, P.O. Box 33932, Shreveport, Louisiana 71130-3932
Introduction
The National Committee for Clinical Laboratory Standards
(NCCLS, 1) recommends that Type I water used in clinical
laboratories must be virtually silica (SiO2) free (< 0.05 rag/L).
The recommendation has been adopted by the Collegeof Amer-
ican Pathologists (CAP) with the inclusion of a question
regarding soluble silicate in water in CAP Laboratory Accred-
itation Program checklists (2). These requirements are based
upon silica's direct interference with spectrophotometric
measurements at specific wavelengths of the light spectrum.
Therefore, its presence in water not only interferes with trace
metal and electrolyte analyses but also could adversely affect
many enzymatic determinations such as those procedures
using alcohol dehydrogenase and various phosphatases (1).
A commercial silica testing kit (catalog # 22550-00, Hach
Silica Test Kit, Hach, Loveland, CO) was evaluated. Kit specifi-
cations indicated that it measured silica concentrations in the
range of 0-1 mg/L. Measurement of the color intensity of the
resulting silicate-molybdate complex required visual compar-
ison of the developed color to a color disc directed toward an
artificial or natural light source and depended on color com-
parison by the naked eye. Hach Silica Testing Kit reagents were
used in the development of a modified procedure for the de-
tection and more accurate quantitation of low concentrations
(< 0.05 rag/L) of soluble silica in water.
Materials and Methods
Reagents were purchased individually from the Hach Com-
pany. Sodium metasilicate nonahydrate (Na203Sio9H20, min-
imum 98%, FW 284.2) used for the preparation of the stock
standard silicate solution was purchased from Sigma Chemical
(catalog # S-4392, St. Louis, MO).
It is important that use of borosilicate glassware is avoided
* PresentAddress:MorehouseSchoolof Medicine, Departmentof Pharmacologyand
Toxicology,720 WestviewDrive, Atlanta, GA 30310.
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TechnicalNote [
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with this procedure to prevent silicate contamination from
the glassware. Therefore, all solutions were prepared in plastic
containers (polystyrene or polypropylene). Polystyrene test
tubes were used in the preparation of standard curves and
testing of unknowns.
Sodium metasilicate was dissolved in water to give a stock
silicate concentration of 1.0 mg/mL from which a seven-point
standard curve covering a concentration range of 0-0.1 mg/L
was constructed (TableI). Standard solutions and water samples
were used at room temperature (approximately20-25~ To 5.0
rnL of each working silicate standard solution and unknown(s),
125 pL of Molybdate 3 Reagent (catalog # 1995-37, Hach Silica
Testing Kit) was added. After vortex mixing, the tubes were al-
lowed to stand for 4 rnin. Then 125 pL of Hach Citric Acid
Reagent (catalog# 22542-37) was added to each tube, and the re-
action was allowedto proceed for 1 rain. TWo-hundredand fifty
microliters of the Hach Amino Acid F Suspension (catalog #
22538-69, AAFS)was then added to each tube. The AAFSmust
be prepared immediately before use by suspending the contents
of one AAFSreagent pillow in 1.0 mL of distilled water. Prepa-
ration of one pillowaccording to these instructions providedsuf-
ficient reagent for four tubes for testing. Tubeswere immediately
vortex mixed and absorbance measured at 803 nm using a
Hitachi U-2000 spectrophotometer (San Jos~, CA) using a water
blank.
Final quantitative results are calculated using the following
formula (3):
Cu = (AuC,) + (As-Au)
Cu = Concentration of the unknown (mg/L)
Au = Absorbance of sample without added silicate
Cs = Concentration of the standard (rag/L)
As = Absorbance of sample with added silicate
Results and Discussion
Silica occurs in soluble and colloidal forms. Colloidal silica
removal from water may be achieved by either ion exchange
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 Journal of Analytical Toxicology, Vol. 21, October 1997

chromatography or more effectively by dis-

Table I. Details of the Standard Curve Construction for the Quantitation of tillation or the combination of both sys-
Soluble Silica in Water
tems. Soluble silica may be eliminated from
Concentrationof 0.1 mg/t Molybdate 3 Citric acid F Amino acid F
water by the use ion exchange and micro-
silica Water Sodium reagent reagent suspension* filtration or their combination.
(rag/L) (mL) metasilicate(mL) (pL) (pl) (pL) Both soluble silica and phosphates co-
exist in most primary water supplies. Both
Blank 5.0 0 0 0 0 react with ammonium molybdate at acidic
pH to form heteropolyacids. Colloidal silica
0.02 4.0 1.0 125 125 250
is extremely difficult to measure because it
0.04 3.0 2.0 125 125 250
is unreactive to mdybdate. Molybdophos-
0.05 2.5 2.5 125 125 250 phoric acid, the product of the phosphate-
0.06 2.0 3.0 125 125 250 molybdate reaction, is labile to citric acid
hydrolysis, but its silica counterpart, molyb-

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0.08 1.0 4.0 125 125 250
0.10 0.0 5.0 125 125 250 dosilicic acid, is resistant to this hydrolysis.
An obvious concern with the method would
Unknown 5.0 0.0 125 125 250
be the probability of phosphate interference
* Vortex mix the Amino Acid F suspension before adding it to the test tube. Vortex mix the test tube before in a spectrophotometric method. Phosphate
moving to the next one, and so on.
interference was negligible in this proce-
dure for two reasons. First, excess amounts
of citric acid were added to destroy the
0.04 molybdophosphoric acid. Second, because
this complex absorbs light in the yellow
X range of the spectrum and molybdosilicic
acid absorbs light in the blue range, reading
0.035 the absorbance of the reaction mixture at
803 nm would negate potential phosphate
interference.
A method referenced by NCCLS (1) for
0.03 X
the measurement of soluble silica in water
IlL is similar to the water silicate method de-
scribed in Standard Method for the Exami-
0.025
nation of Water and Waste Water (4). The
methods are tedious and time consuming
X
for a variety of reasons. Some of these
1
X 0 reagents have relatively short shelf lives, as
0.02 short as two weeks in some instances. The
use of at least seven chemicals and acids,
.<
X some of which have very specific storage
conditions, is required. Large amounts of
0.015 g
stock material are needed, and a series
of standards is specified, each in a volume of
100 mL of water. Because water testing is
conducted on a periodic basis, the com-
0.01
0 plexity of the method and the lability of the
g reagents translate to increased expenditure,
analyst time, specific chemical storage re-
0.005 quirements, and hazardous waste disposal.
Although the Hach Silica Test Kit with
visual color comparison may be useful for
field work for the estimation of high con-
centrations of silicates in water, its use at
0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09 0.1 0.11 low silicate concentrations required for
Soluble silicate concentration (rag/L) clinical laboratory applications was limited.
It was virtually impossible to reliably com-
Figure 1. Scatter plot of replicate absorbance readings (n = 5) at 803 nm wavelength at six con- pare the intensities of two blue colors with
centrations with the spectrophotometer set using a water blank. the naked eye at low silicate concentrations
(< 0.07 mg/L).

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Journal of Analytical Toxicology,Vol. 21, October 1997
Table !!. Exampleof ResultsObtained usingTwo Spectrophotometric
Silicate Methods
Before system cartridge
replacement
Sample source Silicate method I' Silicate method II r Silicate method I' Silicate method II t
(mg/mL) (mg/mL)
Tap water 295 300
PostMilli-Q system - 0.069
* Reference 3.
t Method described herein.
The modification resulted in a simple spectrophotometric
procedure for the determination of soluble silica concentra-
tions in water. Because soluble silica in the water supply of
certain geographical locations is high, CAP requires that labo-
ratories test their water soluble silica content to determine if it
exceeds the recommended limit of___0.05 mg/L.
The spectrophotometric method described was found to be
linear over the concentration range of 0-0.5 mg/L. Data, how-
ever, are only included in the range of 0-0.1 mg/mL because we
focused our work around the maximum allowable silicate con-
centrations of 0.05 mg/mL (Figure 1). Linear regression ana-
lysis of data (5) from replicate standard curves (n = 5) de-
scribed the curve ~ = m x + b) as absorbance = 0.3466X +
1.3286E-03 with a correlation coefficient of 0.996.
Derivation of the final quantitative results is accomplished by
calculations based upon measurement of small concentrations
of ions in solutions (standard addition method) as is done in
atomic absorption spectroscopy (3). This approach is necessary
because the water used in all solutions used in the procedure
as well as the testing of the source water is essentially from the
same source. It also accommodates the measurement of very
low concentrations of target analytes in solution.
Our laboratory waterpurification system consists of a Culligan
mixed bed ion exchange and carbon purification system (Cul-
ligan Technologies, Northbmok, IL) in tandem with a MilliQ
purification system consisting of one Super C and two ion
exchange cartridges and a 0.22-t~mfilter (Milli-Q,Bedford,MA).
Table II shows examples of results using both methods before
and after system maintenance. It was found that water produced
before replacement of the single Super C and two ion exchange
cartridges in the MilliQ System but after replacement of the
Culligan cartridges still passed resistivity and bacterial culture
testing to meet Type I specifications.
Replacement of all cartridges in the system
resulted in no measurable soluble silicate in
After system cartridge water from the purification system.
replacement Analytical issues associated with practical
quantitative analysis of water for soluble
silica at very low concentrations have been
(mg/L) (mg/L)
addressed. It was found that the Hach Silica
- _ Test Kit with visual color comparison was
not sufficiently sensitive at soluble silica
0.005 0.00o concentrations below 0.07 mg/mL. The
reagents supplied with the kit when used
with a spectrophotometer at a wavelength
of 803 nm rather than the visual filter
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reading device overcomes the limitations of the method. There
are several other issues, however, surrounding the NCCLS
guidelines themselves which are ambiguous and that must be
addressed. The NCCLS definition for "source water" was un-
clear, that is, water supplied to or water exiting the purification
system.
It is recommended that water that exits the final water
purification system be selected for soluble-silica testing be-
cause this will be the water that is used in the laboratory, and
laboratories that outsource their soluble-silica water testing
ascertain the analytical methodology used by the reference
laboratories, including the limit of detection and limit of quan-
titation of the method.
References
1. National Committee for Clinical Laboratory Standards. Prepara-
tion and Testingof Reagent Water in the Clinical Laboratory. 2nd
ed. Approved Guideline. NCCLS document C3-A2. NCCLS,
Villanova, PA, 1991.
2. College of American Pathologists (CAP), Laboratory Accreditation
Program, Northfield, IL, 1996.
3. I. Rubeska and B. Moldan. Atomic Absorption Spectrometry.CRC
Press, Cleveland, OH, 1967.
4. A.E. Greenberg, R.R. Trussell, and L.S. Clesceri, Eds. Standard
Methods for the Examination of Water and Waste Water, 16th ed.
American Public Health Association, Washington, D.C., 1985.
5. R.T.Tallarida and R.B. Murray. Manual of Pharmacologic Calcu-
lations with Computer Programs. Springer-Verlag, New York,
1981.
Manuscript received March 25, 1997;
revision accepted June 13, 1997.
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