Comparative analysis method of C-organic in fertilizers by gravimetry and

spectrophotometry
Tri Esti Purbaningtias, Nursi Biwi Qayyumah, Puji Kurniawati, Bayu Wiyantoko, and Alfa Akustia Widati

Citation: AIP Conference Proceedings 2026, 020055 (2018); doi: 10.1063/1.5065015

View online: https://doi.org/10.1063/1.5065015
View Table of Contents: http://aip.scitation.org/toc/apc/2026/1
Published by the American Institute of Physics
 Comparative Analysis Method of C-Organic In Fertilizers
by Gravimetry and Spectrophotometry

Tri Esti Purbaningtias1a, Nursi Biwi Qayyumah2, Puji Kurniawati1, Bayu

Wiyantoko1, Alfa Akustia Widati3

1
Chemical Analysis Program, Faculty of Mathematics and Natural Sciences
2
Student of Chemical Analysis Program, Faculty of Mathematics and Natural Sciences,
Islamic University of Indonesia Jl. Kaliurang Km. 14.5, Sleman Yogyakarta, Indonesia 55584
3
Chemistry Department, Faculty of Science and Technology,
Airlangga University Kampus C Mulyorejo, Surabaya Indonesia 60115
a)
Corresponding author: tri.esti.p@uii.ac.id

Abstract. Comparison methods of C-organic determining in organic fertilizers by gravimetry (ash content) and UV-Vis
spectrophotometry (Walkley & Black) had been performed. The difference between the two methods was seen from the
validation parameters of each method. This research used precision, homogeneity, stability test, control chart and
measurement uncertainty as parameter of validation method. The result of method validation indicates that both methods
are not precision, but have homogeneous results. The stability test showed that C-organic analysis by gravimetry had
stable results while in the UV-Vis spectrophotometry method, the results were unstable. Control chart analysis showed
the results of C-organic analysis in a organic fertilizer with gravimetry method statistically controlled, while the C-
organic analysis of glucose solution as standard by UV-Vis spectrophotometry method was uncontrolled. The measured
uncertainty value of the gravimetry method of 0.0041% is much smaller than the UV-Vis spectrophotometry method of
2.1529%. The average percentage of C-organic content obtained from the gravimetry method was 18.654%, while the
average percentage of C-organic content produced using UV-Vis spectrophotometry method was 8.674%. These results
indicated that both methods are significantly different.

INTRODUCTION
One type of fertilizer commonly used in agriculture is organic fertilizer. Organic fertilizers play an important
role in improving the chemical, physical, and biological soil as well as a source of plant nutrients. The chemical
fertilizers except water is normally classified into three nutrient classes. The three groups are primary or major
nutrients consisting of nitrogen (N), phosphate (P2O5) and carbon (C); Secondary nutrients are sulfur (S),
magnesium (Mg) and calcium (Ca); and micro nutrients which have the smallest amount in fertilizer i.e. iron (Fe),
copper (Cu), zinc (Zn), manganese (Mn), boron (B) and molybdenum (Mo) [1]. The main specification in organic
fertilizer is the C-organic content when compared with other nutrients because of naturally-occurring organic carbon
forms are derived from the decomposition of plants and animals [2].The minimum C-organic content in organic
fertilizer is 15% [3].
The determination of C-organic in soil and sediment can be classified into two methods specifically semi-
quantitative and quantitative analysis. Semi-quantitative methods are based upon the indiscriminant removal of all
organic matter followed by gravimetry determination of sample weight loss via chemically or heat at elevated
temperatures. Organic matter content is calculated as the difference between the initial and final sample weights

2nd International Conference on Chemistry, Chemical Process and Engineering (IC3PE)

AIP Conf. Proc. 2026, 020055-1–020055-11; https://doi.org/10.1063/1.5065015
Published by AIP Publishing. 978-0-7354-1746-5/$30.00

020055-1
divided by the initial sample weight times 100%. All weights should be corrected for moisture/water content prior to
organic matter content calculation based on the assumption that organic matter contains 58% C-organic [2]. This
method could be used for analysis of total organic carbon (TOC) in surface sediments. Sediment sample was heated
at 60 °C for 24 hours and 450 °C for 3 hours. In this method, the TOC percentage were recorded at 0.03 – 4.02%
[4]. The scale of operation for precipitation gravimetry is limited by the sensitivity of the balance and the availability
of sample. As a consequence, gravimetry is usually limited to major or minor analytes, in macro or meso samples.
For a macro sample containing a major analyte, a relative error of 0.1–0.2% is routinely achieved [5].
The quantitative analysis of C-organic is a Walkley & Black method that combined with spectrophotometry
techniques [2]. The Walkley-Black procedure is widely used because it is simple, rapid, and has minimal equipment
needs [6]. Walkley & Black method is a process of wet oxidation for sample extraction. This method not only could
be combined with UV-Vis spectrophotometer but could also be combined with the titration method [7]. The
principle of Walkley & Black method using UV-Vis spectrophotometry i.e. organic carbon in the sample is oxidized
by dichromate in acidic conditions and produces chromium (III). The chromium (III) formed is measured by
spectrophotometry and is equivalent to the oxidized C-organic.
Typically gravimetry techniques are relatively easy to perform implementing good laboratory practices.
Spectrophotometry techniques require more skill in operating the instrument, interpreting the results, trouble
shooting problems of system should arise and not suitable for samples with high levels. This study aimed to compare
methods of C-organic analyzed in organic fertilizers by gravimetry and UV-Vis spectrophotometry through method
validation. The comparison results could be used as a reference for routine analysis in the laboratory. Validation
parameter of this research is data selection test using Dixon test, precision test with repitability, homogeneity test,
stability test, control chart and measurement uncertainty.

MATERIALS AND METHODS

Materials
The materials used in this research was organic fertilizers, H2SO4 98%, K2Cr2O7 1 N, standart glucose solution
5000 ppm, and aquadest. The main equipment was laboratory glassware, analytical balance (Ohaus, PAJ 2003),
oven (Memmert), furnace (Compact Benchtop Muffle), and UV-Vis spectrophotometer Single Beam (Genesys 20).

Methods
Organic Fertilizer Sampling Technique

Organic fertilizers collecting from Yogyakarta district in a polyethylene bottle container were mixed with stirring
with a spoon, then weighed as much as 10 grams in a porcelain dish. The same treatment was repeated 10 times.

Gravimetry (Ash Content)

Organic fertilizer samples were weighed as much as 10 grams in closed porcelain. Samples were put into oven
for 8 hours with temperature 105°C. The sample was cooled in a desiccator and weighed using an analytical balance
to determine the water content in the sample. The sample was ashing at 300°C for 1.5 hours and continued with a
temperature of 550-660°C for 2.5 hours. The resulting ash was cooled in the desiccator and weighed as mass ash. C-
organic content calculated with following formula:

C-organic (%) = x x 100 x 0.58
whereas, w : mass sample (g)
w2 : mass ash (g)
cf : correction factor from water content = 100/(100 - water content%)
0.58 : correction factor of organic matter into organic carbon

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 UV-Vis Spectrophotometry (Walkley & Black)
A total of 0.05 grams of solid organic fertilizer samples were carefully weighed and put into a glass beaker.
Reagents K2Cr2O7 1 N of 5 mL and 7 mL of H2SO4 p.a. were added to the sample. The mixture was boiled and
allowed to stand for 30 minutes. Thereafter, the mixture was diluted with distilled water up to 100 mL. The sample
solution was allowed to stay overnight. The absorbance of standart glucose solution and sample that has been
prepared and allowed to stand overnight was measured using a UV-Visible spectrophotometer single beam with a
wavelength of 561 nm. C-organic content calculated with following formula:

C-organic (%) = ppm calculated x x
x cf

RESULT AND DISCUSSION

Determination of C-organic in organic fertilizer

The result of C-organic analysis on organic fertilizer by gravimetry and spectrophotometry method could be seen
in Table 1. In Table 1 showed that the results of C-organic analysis with both methods had different values. The
average levels of C-organic fertilizer with gravimetry method at 18.654% while the spectrophotometry method had
an average C-organic by 8.674%. The average C-organic content difference between the two methods was 9.979%.
The t-test was used to prove that gravimetry and spectrophotometry methods gave significantly different levels of C-
organic in fertilizers. The independent sample t-test is a parametric statistical test method used to analyze the
comparison of two unpaired samples [8]. The C-organic results of the gravimetry and spectrophotometry methods
are the same as the null hypothesis of this study. The t test results showed that C-organic analysis with two methods
had significant differences, this is indicated by the value of t-calculated greater than t-table.
TABLE 1. Comparison of C-organic analysis results in organic fertilizer by gravimetry and spectrophotometry methods
C-Organic (%) The Difference
Repetition Spectrophotometry Between The Two
Gravimetry Method
Method Methods
1 21.664 9.125 12.539
2 16.337 8.122 8.215
3 22.636 10.378 12.258
4 13.718 11.757 1.961
5 16.337 8.247 8.09
6 20.352 8.247 12.105
7 15.720 7.245 8.475
8 16.812 6.242 10.57
9 24.135 8.749 15.386
10 18.824 8.624 10.2
Mean 18.654 8.674 9.9799
Standart Deviation 3.415 1.536 3.638
t-calculated* 8.427
t-table 3.250
( )
*t-calculated =

Validation Methods
The method validation in this study was based on the C-organic measurements on fertilizer samples by
gravimetry and spectrophotometry and the C-organic measurement of glucose standard 250 ppm by UV-Vis
spectrophotometry. The C-organic measurements data of glucose standard solution 250 ppm was shown in Table 2.
Validation parameters are precision, homogeneity, stability, control chart, and measurement uncertainty. Before
performing validation calculations, it is necessary to first test data selection. The data selection test used in this study
was Dixon test.

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 TABLE 2. The results of C-organic analysis in a standard solution of 250 ppm glucose by UV-Vis spectrophotometry
Repetition Concentration (ppm)
1 282.235
2 279.882
3 254.588
4 270.471
5 282.235
6 259.294
7 248.706
8 241.059
9 255.765
10 228.706
Mean 260.294
Standart Deviation 18.276

Dixon Test
The Dixon test was performed using 10 repeated C-organic measurements in the same day. The first step in the
dixon test is to sort the data from the smallest to the largest data to make the selection. Preparation of data from the
smallest to the largest aims to facilitate the identification of extreme data. Extreme data is data that does not exceed
the value in the Dixon test crisis table of each selection test. Extreme data can be from the largest or smallest data.
Data were tested by using Dixon test formula for 10 data as follows:

Smallest Data: < Largest Data:
<

Table 3 showed the results of the Dixon test analysis for gravimetry (fertilizer) and spectrophotometry (fertilizer
and glucose standard solution) testing.
TABLE 3. Data selection by Dixon test in gravimetry and spectrophotometry method
Dtable of 95%
Methods Data (%) Category Dstatistic Results
with n=10 (Dn)
13.719 Smallest 0.224 Inlayer
Gravimetry
24.135 Largest 0.178 Inlayer
6.242 Smallest 0.242 0.530 Inlayer
Spectrophotometry (Sample)
11.757 Largest 0.305 Inlayer
Spectrophotometry (Standart 228.706 Smallest 0.231 Inlayer
Glucose 250 ppm) 282.235 Largest 0.000 Inlayer

Based on Table 3, it can be concluded that all data analysis was acceptable or not including extreme data because
it had Dstatistic smaller than Dtable. Therefore all such data could be used in other statistical tests.

Precision Test
The precision test is performed to determine the proximity or compatibility between test results with each other
in a series of tests. The result of the C-organic content with 10 repetitions on the same day was used to determine the
precision of the measurement (precision) by calculating the relative standard deviation (% RSD) compared to the
variation coefficient (% CV Horwitz). If the value of % RSD < 2/3 % CV Horwitz then the data has a good
repitability precision. The calculation result of % RSD and %CV Horwitz could be seen in Table 4.

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 TABLE 4.The result of precision analysis of C-organic content by gravimetry and spectrophotometry method
Standard Relative Standard 2/3 x % CV
Methods Conclusion
Deviation (SD) Deviation (RSD)a) Horwitzb)
Gravimetry 3.415 18.308% 1.717% Not Precision
Spectrophotometry (Sample) 1.537 17,714% 1.926% Not Precision
Spectrophotometry (Standart G
18.276 7.0214% 4.618% Not Precision
lucose 250 ppm)
a)
%RSD = x 100%
b) ,
% CV Horwitz = 2

In Table 4 it could be seen that all analytical methods had an inaccurate repetition, indeed for C-organic analysis
for standard glucose solutions. The %RSD in samples in both gravimetry and spectrophotometry had almost the
same value that was in the 17-18% but the standard solution had the smallest %RSD. This suggested that the
precision of the standard solution was better than the sample, since the factors that affected only from the method
but on the sample, the factors that affected not only the method but also were influenced by the sample conditions.
Factors affecting the analysis by spectrophotometry method is wavelength accuracy and reproducibility, stray light,
resolution, photometric accuracy and reproducibility, noise, baseline flatness, stability, and linearity while in the
gravimetry method, the not gradual ignition caused burning of the sample or loss by swelling outside the crucible
[9].

Homogeneity Test
Homogeneity test conducted to determine the degree of uniformity on data obtained so that knowable the
homogeneity of sample. Homogeneity test can be obtained by calculating MSB (mean square between) and MSW
(mean square within) [10]. The homogeneity test results using data from five duplicate measurements were shown in
Table 5.
TABLE 5. The homegeneity test result of C-organic content on gravimetry and spectrophotometry method
2
No C1 (ai) C2 (bi) (ai+bi) [(ai+bi)- ( )] [(ai+bi)- ( )] (ai-bi) [(ai-bi)- ( )] [(ai+bi)- ( )]

Gravimetry
1 21.664 20.352 42.015 4.708 22.167 1.312 2.342 5.486
2 16.337 15.720 32.058 -5.250 27.558 0.617 1.647 2.712
3 22.636 16.812 39.449 2.142 4.586 5.824 6.854 46.977
4 13.719 24.135 37.854 0.547 0.299 -10.416 -9.386 88.098
5 16.337 18.824 35.160 -2.147 -2.147 -2.487 -1.457 2.123
X 37.307 -1.030
Σ 59.220 145.397
MSB 7.402
MSW 14.540
F 0.509
Spectrophotometry (Standart Glucose 250 ppm)
1 282.235 259.294 541.529 20.941 438.533 22.941 -4.235 17.938
2 279.882 248.706 528.588 8 64 31.176 4.000 16.000
3 254.588 241.059 495.647 -24.941 622.062 13.529 -13.647 186.242
4 270.271 255.765 526.235 5.647 31.889 14.706 -12.471 155.516
5 282.235 228.706 510.941 -9.647 93.066 53.529 26.353 694.478
Xhm 520.588 27.176
Σ 1249.55 1070.173
MSB 156.19
MSW 107.02
F 1.460

020055-5
 The results of calculated F on the C-organic analysis by gravimetry and spectrophotometry methods respectively
at 0.509 and 1.460, while the F table at 5.19. Based on this observations, it can be concluded that calculated F is
smaller than F table. These results indicated that the sample of organic fertilizer used homogeneously so that it could
be concluded that the sampling technique has been done accordingly.

Stability Test
The stability test conducted on samples to see the stability of the samples tested by long usage. A sample can be
said to be stable if the first data and second or second and third the data did not show a significant difference.
Stability test is done by comparing the average value of homogeneity test minus the mean value of the daily analysis
result with the standard deviation of the homogeneity test results. The |Xhm| can be obtained from the average C-
organic content at a time (repetition) , and |Xi| obtained from the average value of daily C-organic analysis and nIQR
is the standard deviation of homogeneity test. The calculation of the stability test was presented in Table 6.
TABLE 6. Calculation of the stability test from C-organic analysis by gravimetry and spectrophotometry method
Day C1 (%) C2 (%) Mean
Gravimetry

1 21.664 24.135 22.899

2 16.189 13.626 14.908
3 20.217 23.810 22.013
4 15.604 13.583 14.593
5 20.168 23.755 21.961
6 16.176 13.558 14.867
7 20.143 23.724 21.934
Xi 19.025
0.3 x nIQR 0.7921
|Xi-Xhm| 0.3716
Spectrophotometry (Standart Glucose 250 ppm)

1 229.294 231.059 230.176

2 206.941 204.588 205.765
3 228.118 244 226.059
4 226.353 244 225.176
5 232.824 229.882 231.353
6 230.471 227.529 229
7 225.765 219.882 222.824
Xi 224.336
0.3 x nIQR 3.532
|Xi-Xhm| 35.958

Based on Table 6 it could be seen that the C-organic analysis by gravimetry method showed stable results
because the |Xi-Xhm| value is lower than 0.3 x nIQR. This result contrasts with the analysis of C-organic in a
standard glucose solution by spectrophotometry. The value of Xi-Xhm on spectrophotometry method is greater than
0.3 x nIQR so it can be said C-organic analysis of standard glucose solution by spectrophotometry unstable.

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 Control Chart
Control chart has three main lines, ie the center line, the upper control limit (UCL) and the lower control limit
(LCL). The center line is the mean value ( ̅ ) of the measurement data, the upper control limit is the value of the sum
of the average value with a standard deviation multiplied by three ( ̅ + 3SD), and the lower control limit is the value
of the reduction in the average value with a standard deviation multiplied by three ( ̅ - 3SD) [10]. Control chart of
the C-organic analysis with gravimetry and spectrophotometry methods could be seen in Figures 1 and 2

UCL

CL

LCL

FIGURE 1. Control Chart of C-Organic Analysis in Fertilizer by Gravimetry for a week

UC

LC

FIGURE 2. Control Chart of C-Organic Analysis in Standart Glucose 250 ppm by Spectrophotometry for a week

The control chart of C-organic analysis by gravimetry method indicated that the process of determining the C-
organic content was controlled or the analysis process was in statistical control. The controlled analysis process was
characterized by all test results of C-organic content distributed within the control limits (UCL and LCL) showing
no particular tendency. The other hand, the standard glucose solution control chart using the spectrophotometry
method showed that the C-organic analysis process in standard glucose solution was out of control. This was

020055-7
indicated by the presence of the data that was outside the lower control limits (LCL) and all the data there was
nothing to cut the center line.

Measurement Uncertainty
Uncertainty measurement was performed to estimate the value of which may be lost or gained from the actual
values. There are four steps to estimation of measurement uncertainty i.e. specify measurands, identify uncertainty
sources, quantify uncertainty components, and calculate combined uncertainty [11]. The specify measurements in
this method is shown in the C-organic equation in the section methods. Sources contributor to measurement
uncertainty can be identified by making a cause and effect diagram (Ishikawa diagram). Through this diagram, it can
be seen the relationship of the contributors to the uncertainty and to prevent double counting [11]. The cause and
effect diagram of C-organic analysis by gravimetry and spectrophotometry method shown in Figure 3 and 4. Based
on the cause and effect diagram, it can be concluded that the contributor of measurement uncertainty on the
spectrophotometry method is more complex than the gravimetry method.

Wsample cf
calibration of calibration of
analytical analytical
balance balance

% C-organic
calibration of
analytical
balance
Wash
FIGURE 3. Diagram of Cause and Effect of C-Organic in fertilizer analysis by gravimetry method

ppm calculated Volume

dilution
calibration
calibration of calibration of curve calibration of
measuring pipette
expansion volume flask
volume flask factor
standart
concentration sample concentration

Calibration of Calibration of Spectrophotometry

% C-
calibration of calibration of organic
analytical analytical
balance
balance
Wsample cf
FIGURE 4. Diagram of Cause and Effect of C-Organic in fertilizer analysis by spectrophotometry method

The main factor contributing to the measurement uncertainty of C-organic by gravimetry is the mass. While in
the C-organic analysis by spectrophotometry, the cause of uncertainty of measurement is mass, volume, and ppm
calculated. The formula for determining the uncertainty is as follows:
 mass
The uncertainty from analytical balance derived from the mass weighing tolerance on the certificate
(independent parameter calibration, s) divided by the coverage factor (K = 2 in 95%).
m=

 volume

020055-8
 Volume uncertainty (µV) is composed of the independent parameters calibration (cal.) and uncertainty of
temperature (T). The calibration uncertainty (µcal) is derived from independent parameters calibration (s) devided by
coverage factor (K) while the temperature uncertainty (µT) is derived from the influence of the temperature on the
thermal expansions of liquid and volumetric device.
cal =
, /
T =
√
( V) = ( ) + ( )

 ppm calculated
The uncertaintyof ppm calculated (µppm) is affected by the calibration curve of standard series solutions.
Uncertainty of regression line is obtained by the formula:
∑( )
Sy/x=
where: Sy/x = deviation standard
Y = absorbance of standard glucose solution by measurement
Yi = calculate absorbance of standard glucose solution using linear regression equation
n = number of standard glucose series solutions

The result of measurement uncertainty calculation could be seen in Table 7. In order to determine the combined
uncertainty (µC), the relative standard uncertainty must be squared with the previously divided by its value. The
expanded uncertainty (µE) is obtained by multiplying the combined standard uncertainty with a coverage factor of 2
corresponding to level of trust of 95%. Based on Table 7 it shown that the expanded uncertainty of the gravimetry
method was much smaller than the spectrophotometry method. This indicated that the range of error values in
spectrophotometry was very large.
TABLE 7. Uncertainty measurement of C-organic by gravimetry and spectrophotometry method
Relative
Standard Percentage Combined Expanded
Value Standard
Parameters Unit Uncertain Uncertainty Uncertainty Uncertainty
(x) Uncertainty
ty (µx) [(µx/x)/ ]x100] (µC) (µE)
(µx/x)
Gravimetry
Initial sample
10 g 0.000388 3.88 x 10-5 20.779
mass
-5
Ash mass 6.97 g 0.000388 5.57 x 10 29.813
Sample mass
0.00206% 0.0041%
(correction 10 g 0.000388 3.88 x 10-5 20.779
factor)
Final sample
7.259 g 0.000388 5.35 x 10-5 28.627
mass
-5
∑ 18.69 x 10
Spectrophotometry (Standart Glucose 250 ppm)
Concentration 40.706 ppm 3.6117 0.0887 32.1173
Volume flask 100 mL 0.0471 0.0005 0.1707
Volume
measuring 5 mL 0.0303 0.0061 2.1967
pipette
Absorbance of
0.077 abs 0.004 0.0519 18.8043
50 ppm
Absorbance of 1.0765%. 2.1529%
0.177 abs 0.004 0.0226 8.1804
100 ppm
Absorbance of
0.245 abs 0.004 0.0163 5.9099
150 ppm
Absorbance of
0.354 abs 0.004 0.0113 4.0902
200 ppm
Absorbance of
0.429 abs 0.004 0.0093 3.3751
250 ppm

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 TABLE 7. Uncertainty measurement of C-organic by gravimetry and spectrophotometry method (Continued)
Relative
Standard Percentage Combined Expanded
Value Standard
Parameters Unit Uncertain Uncertainty Uncertainty Uncertainty
(x) Uncertainty
ty (µx) [(µx/x)/ ]x100] (µC) (µE)
(µx/x)
Spectrophotometry (Standart Glucose 250 ppm)
Absorbance of
0.0654 abs 0.004 0.0612 22.1397
sample
Sample mass 0.05 g 0.0004 0.0078 2.8115
Volume flask 100 mL 0.0471 0.00047 0.1707
Sample mass
(before heated, 10 g 0.0004 0.00004 0.0141
cf)
Sample mass
(after heated, 7.259 g 0.0004 0.00005 0.0194
cf)
∑ 0.27626

CONCLUSION
The determination of C-organic in fertilizers by gravimetry and spectrophotometry showed different results.
Stability testing and control charts on the gravimetry method gave stable results, whereas the spectrophotometry
method was unstable. The range of error values that occur during the test expressed by the uncertainty extends to the
gravimetry method is smaller than that of spectrophotometry. To obtain better yield validity, it is better to analyze
the level of C-organic in organic fertilizer using gravimetry method, because the C-organic content in organic
fertilizer is possible high level.

ACKNOWLEDGEMENT
The authors acknowledge the research grant provided by Chemical Analysis Program, Islamic University of
Indonesia, under research grant no 199a/KaPro/DIII-AK/20/VIII/2016 and Assessment Institutes for Agricultural
Technology of Yogyakarta.

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