SAIT Technical Project Week - Beatriz Pregillano, 2015

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Fluorometric Determination of Waterborne Bitumen using a Universal Bitumen Standard

(with closer degree of similarity in emission and fluorescence spectra) METHOD VALIDATION

By

Beatriz Pregillano

SAIT CHEMICAL TECHNOLOGY PROGRAM

Submitted to:

Trang Trinh, B.Sc (Hons) Operations Manager Petroleum Testing Services

AGAT Laboratories 3650 21st Street N.E.

Calgary, Alberta CANADA T2E 6V6

March 13, 2015

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Introduction

Fluorescence Spectroscopy is a type of electromagnetic spectroscopy that measures the

intensity of photons that are being emitted from a fluorescent analyte. All molecules and atoms

seek to exist in their lowest possible state and when a molecule absorbs light (usually ultraviolet

light), it moves into a more excited energy state. Molecules and atoms will seek to lose the

energy, return to its ground configuration and emitting light. [2] The analysis can be made by

measuring the intensity of the fluorescence light emitted and using a standard graph to

determine the concentration of samples.

0.8 to 2.4 barrels of fresh water are required for every barrel of bitumen produced. This

requires vigilant water management vital to operations. [1] To test purity, testing residual

bitumen is imperative after cleaning.

There are currently four analytical methods to test for oil content in water. They are

Chromatographic (Polyaromatic Hydrocarbons, Headspace Analysis, etc.), Infra-Red

Spectroscopy (Oil and Grease), Gravimetric and Flurorescence Spectroscopy. In this report, the

amount of bitumen in water will be determined using fluorescence spectroscopy using a

universal bitumen standard.

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Acknowledgement

I would like to express my sincere appreciation to those who guided me throughout this

successfully completed project. Your guidelines and suggestions throughout this project helped

me a lot and left me great knowledge about being a chemical laboratory technologist. I am very

grateful to AGAT Laboratories for having me for this technical project week. I am particularly

thankful to my supervisors (Trang, Joanna and Adil) for their kindness and teaching immense

information throughout the entire week. I also would like to thank Andrea Wiseman for

reviewing this report and ensuring everything is in good format.

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Table of Contents Introduction ................................................................................................................................... 2 Acknowledgement ........................................................................................................................ 3 Procedure ..................................................................................................................................... 5 Data/Observations ........................................................................................................................ 5 Calculations .................................................................................................................................. 9 Results ........................................................................................................................................ 11 Conclusion .................................................................................................................................. 12 Discussion .................................................................................................................................. 12 APPENDIX 1 ............................................................................................................................... 13 FLUOROMETRIC DETERMINATION OF WATERBORNE BITUMEN ...................................... 14

1. Sample Requirements ...................................................................................................... 14 2. Equipment, Materials and Supplies .................................................................................. 14

2.0 Equipment ..................................................................................................................... 14 3. Materials ........................................................................................................................... 14

3.1 Glassware ...................................................................................................................... 14 3.2 Solvents ......................................................................................................................... 14

4. Procedure ......................................................................................................................... 15 Bibliography ................................................................................................................................ 18

Table of Figures Table 1 Initial concentrations of known samples and observations ______________________________________________ 5 Table 3 Average intensity of calibration standards ____________________________________________________________________ 7 Table 4 Intensity results of TE samples obtained from fluorometer __________________________________________________ 7 Table 5 Intensity results of unknown samples obtained from fluorometer ______________________________________ 8 Table 6 Average intensity of calibration standards _______________________________________________________________ 11 Table 7 Final calculated concentration of bitumen sample "13RE3205 6" ____________________________________ 11 Table 8 Percent Recovery of QC ___________________________________________________________________________________ 11 Table 9 Concentration and percent error of known samples ____________________________________________________ 11 Table 10 Preparation of calibration standard of bitumen using matrix MM63 ____________________________________ 16 Table 11 Preparation of quality control standards of bitumen using matrix MM63 _______________________________ 16 Table 12 Preparation of unknown sample using matrix AT21 and MM63 __________________________________________ 17

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Procedure

The procedure was performed on February 18, 2015 performed in AGAT Laboratories, supervised by Joanna Gurnham. Method reference deviations from ASTM International

See Appendix 1-4.0 for the procedure

Data/Observations

DATE: February 17, 2015, DAY 1 COMPANY: AGAT Laboratories ANALYST: Beatriz Pregillano Arrived at the workplace and successfully completed a WHMIS test. The orientation included the presentation of the technical project week topic and tour of the laboratory. DATE: February 18, 2015, DAY 2 COMPANY: AGAT Laboratories ANALYST: Beatriz Pregillano The second day included preparing standards and samples for the analysis. Began with making matrix1 MM63 and AT21. Continued with preparing stock solution for calibration standards and prepared more samples. Bitumen ID: 13RE3205 (6) Sample Weight: 0.0791 g Samples with known concentration were given for method validation. The initial concentration of the samples before a series of dilutions and observations can be found in Table 1.

Table 1 Initial concentrations of known samples and observations

SAMPLE Initial Concentration (mg/L) Observation TE 86 32260 The TE Samples were faint

yellow. They contain only toluene. Dilution factor: 3.33

TE 59 31120 TE 53 25480 TE 25 15440

1 See Appendix 1-4.0 Procedure page 15

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Nos. BLK Intensity Av Int. STD 1 Intensity Av Int. STD 2 Intensity Av Int. STD 3 Intensity Av Int. STD 4 Intensity Av Int. STD 5 Intensity Av Int.mg/L mg/L mg/L mg/L mg/L mg/L

1 0 0.005 0.002667 0.5 59.6 59.676 2.0 175.299 175.246 5.0 426.638 426.5333 8.0 614.595 616.5805 10.0 663.442 662.4412 0 0.002 0.5 59.76 2.0 175.175 5.0 426.354 8.0 614.275 10.0 663.883 0 0.001 0.5 59.668 2.0 175.264 5.0 426.608 8.0 618.886 10.0 661.002

6295324 6.73

Grease sample obtained from 2910 with known oil content

but in different method of analysis. Needed to follow sample dilution protocol to

prepare with a matrix of MM63

1 part sample + 9 part AT21 6295495 3.08

Unknown Sample Information Sample ID: fwko a water dump Sample Date: 09/18/2013 Sample Time: 11:10 am DATE: February 19, 2015, DAY 3 COMPANY: AGAT Laboratories ANALYST: Beatriz Pregillano Analyzed all samples in a spectrofluorometer and obtained data for the report. Spectroflurometer: RF-1501, AGAT Laboratories Signal to Noise Ratio: 377 Wavelength used: 459 nm-496 nm Intensity was measured in the spectrofluorometer. The data below was obtained during the analysis.

Table 2 Data obtained from excel for the calibration of 5 standards

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Table 3 Average intensity of calibration standards

Concentration mg/L Avg. Intensity

0.00 0.00267 0.50 59.7 2.00 175 5.00 427 8.00 616 10.0 662

Table 4 Intensity results of TE samples obtained from fluorometer

gravimetric fluorometric

Sample ID int. Average

Known Conc. Corrected Av SD RSD

mg/L conc. mg/L

TE - 86 630.549

635.3673 9.68 7.97

8.04 0.071 0.89 Duplicate TE - 86 634.194 9.68 8.02 Triplicate TE - 86 641.359 9.68 8.11

Sample int. Average


mg/L conc. mg/L

TE - 59 608.014

636.4293 9.34 7.68


Sample int. Average


mg/L conc. mg/L

TE - 53 499.192

522.4247 7.65 6.27


Sample int. Average


mg/L conc. mg/L

TE - 25 290.006

298.0707 4.63 3.55


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Table 5 Intensity results of unknown samples obtained from fluorometer

Sample Intensity Average Intensity Actual Concentration mg/L

Bitumen Conoco Bitumen 3.26 mg/L in

MM63 234.079

216.533 2.60 Conoco Bitumen 3.26 mg/L in MM63 209.14 Conoco Bitumen 3.26 mg/L in

MM63 206.38 1Ax10 (10-fold diluted) in

MM63 650.225

697.431 8.84 1Ax10 (10-fold diluted) in MM63 721.742 1Ax10 (10-fold diluted) in

MM63 720.326 1A (undiluted sample) in AT21 522.781

501.902 6.30 1A (undiluted sample) in AT21 481.178 1A (undiluted sample) in AT21 501.747

6295324 (6.73 mg/L) 635.406 641.2006667 8.11 6295324 (6.73 mg/L) 638.385

6295324 (6.73 mg/L) 649.811 6295495 (3.08 mg/L) 211.400

233.741 2.82 6295495 (3.08 mg/L) 234.291 6295495 (3.08 mg/L) 255.532

QCBit-1LLQC (1.0 mg/L) 84.428 84.635 0.88 QCBit-1LLQC Dup 84.842

QCBit-4-ICV (4.0 mg/L) 304.413 301.8975 3.7 QCBit-4-ICV Dup 299.382

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Calculations

Figure 1 Calibration curve of standards in Magic Mix 63

*Note: To maintain a correlation coefficient of >0.995, Standard 5 had to be taken out from the curve.

From the calibration curve above, the equation of the line obtained is y = 77.011x + 16.759. Therefore: Slope = 77.011 Intercept = 16.579 = 16.57977.011 Sample Calculation for the concentration of samples: 1Ax10 10 fold diluted in MM63= 16.57977.011 1Ax10 10 fold diluted in MM63= 697.431 16.57977.011

y = 77.011x + 16.579 R = 0.99549

0 100 200 300 400 500 600 700

0.00 2.00 4.00 6.00 8.00 10.00 12.00

Intens

ity (fsu

)

Concentration (mg/L Bitumen)

Analysis of Calibration Standards in MM63 Matrix

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1Ax10 10 fold diluted in MM63= . Sample Calculation for QCRB %QCRB = Measured ValueTheoretical Value100% Limit: 100% +10 %QCBit 4 ICV = 3.7mg L bitumen4.0mg L bitumen 100% % 4 = 93% Acceptable

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Results Table 6 Average intensity of calibration standards

Concentration mg/L Avg. Intensity

0.00 0.00267 0.50 59.7 2.00 175 5.00 427 8.00 616 10.0 662

Table 7 Final calculated concentration of bitumen sample "13RE3205 6"

SAMPLE 13RE3205 6 Final Calculated Concentration (mg/L) Bitumen in Water

1A (AT21 Matrix) 8.84 1A (MM63 Matrix) 6.30

Table 8 Percent Recovery of QC

QC %QCRB Acceptability Lower Limit QCBit-1LLQC 88.37 Unacceptable

QCBit-4-ICV 92.62 Acceptable

Table 9 Concentration and percent error of known samples

Sample ID Approx. Bitumen Content

Concentration (mg/L)

Actual Concentration average (mg/L) Percent Error

Conoco Bitumen in MM63

3.26 2.60 25.4%

6295324 6.73 8.11 17.0% 6295495 3.08 2.82 9.21% TE 86 9.68 8.04 20.4% TE 59 9.34 8.05 16.0% TE 53 7.65 6.57 16.4% TE 25 4.63 3.66 26.5%

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Conclusion

The bitumen content of sample 1A using MM63 in water is 8.84 mg/L. The bitumen content of

sample 1A using AT21 in water is 6.30 mg/L. The QC percent read back of lower limit QC

(QCBit-1LLQC) is 88.37% which is falls outside the limit 100%+10 while the initial calibration

verification QC, QCBit-4-ICV, falls within the limit.

Discussion

In this report, fluorescence spectroscopy was chosen to analyze bitumen content in water.

Based on the results, the method is valid and acceptable. The TE samples that was analyze

initially via gravimetric analysis gave very close similar results to fluorescence spectroscopy

therefore the concentration of the unknown samples can be confidently reported using this

method. An average of 18.7% percent error for the samples that has known concentration prior

to this analysis was obtained. The percent error value can be improved if there were less

interferences during the experiment. A possible source of error would be contamination in the

lab. Long series of dilution might have affected the accuracy of volume for the samples affecting

its concentration. The usage of matrix is helpful for this analysis (saturating a glass membrane

media by solvent extracted bitumen (which was extracted using toluene and in toluene matrix)

and then allowed to evaporate the solvent).

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APPENDIX 1

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FLUOROMETRIC DETERMINATION OF WATERBORNE BITUMEN

1. Sample Requirements This analysis requires a parent source with a volume of 50 mL of hydrocarbon. This is to be utilized to obtain reliable calibration standards.

2. Equipment, Materials and Supplies

2.0 Equipment In this fluorometric analysis, a spectroflurometer RF-1501 acquired from Shimadzu is used. This instrument performs quantitative and qualitative analysis with an outstanding sensitivity 300 and higher signal-to-noise ratio for the Raman lines of distilled water. 2.0.1 Spefication: (4) The RF-1501 uses a light source 150 W xenon lamp with ozone resolving lamp housing. (3) Monochromators : Ion-blazed holographic concave grating monochromator F/2.4 Wavelength scan range : 220-900 nm and 0-order light Wavelength range : 220-750 nm and 0-order light Slewing Speed : Approx. 30000 nm/min Sensitivity Selection : HIGH and LQW

3. Materials

3.1 Glassware 1. 2.5 L bottle (2) 2. 250 mL Vol. Flask 3. 100 mL Vol. Flask 4. 250 mL Schott bottle 5. 25 mL Vol. Flask 6. 40 mL glass vial (7+ as per requirements for all samples) 7. Measuring cylinder, 500 mL 8. Measuring cylinder, 100 mL 9. Pipette 10 mL 10. Pipette 1 mL 11. Pipette bulb

3.2 Solvents 1. Toluene (Purity 99.9%) 2. Isopropanol 3. Distilled water DI water (resistivity: > 18 4. Acetone

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4. Procedure 4.1 Prepare Magic Mix, MM63 = Maintaining a ratio of 1:6:3 using Distilled water, isopropanol

and toluene: Pour 750 mL of toluene into a 2.5 L bottle. Add 1500 mL of isopropanol followed by 250 mL of distilled water.

4.2 Prepare Binary Solvent System, AT21 = Maintaining a ratio of 2:1 using isopropanol and

toluene: Pour 1200 mL of isopropanol in a 2.5 L bottle. Add 600 mL of toluene into the same bottle.

Matrix solution Steps 4.1 and 4.2 refer to the matrix to be used in the analysis. The matrix refers to the components of the sample other than analyte of interest. The matrix MM63 will be used to build the calibration standards.

4.3 Prepare High Concentrate 500 mg/L Standard Stock Solution (HiBiSS-250) using Matrix MM63:

4.3.1 Obtain a bitumen sample. Record the sample ID

4.3.2. Weigh 0.1250 g of bitumen sample (actual concentration of the solution will be calculated therefore weight does not have to be exactly 0.1250 g) in a 250 mL volumetric flask

4.3.3. Add 75 mL of toluene and shake vigorously and to obtain dissolution of the bitumen 4.3.4. Add 150 mL of isopropanol and dilute to mark with distilled water 4.3.5. Calculate the actual concentration of the stock solution Actual Conc. mg L = 500 mg Ltransferred weight0.1250 g bitumen

4.4 Prepare a working standard stock solution with concentration of 250 mg/L of bitumen (WoSS-250) using Matrix MM63: 4.4.1. Obtain 50 mL of HiBiss-500 solution and transfer into a 100 mL of volumetric flask.

4.4.2. Pour partly MM63 matrix into same volumetric flask and shake vigorously until the solution has become homogenous and finally, carefully dilute to the mark.

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4.5 Prepare calibration standards and quality control standards of bitumen. 4.5.1. Using tables 10 below, create a 5-point calibration curve.

Table 10 Preparation of calibration standard of bitumen using matrix MM63

Sample ID

Type of Standard to be used for

diluting the sample

Volume of Standard (mL)

Final Volume after dilution using MM63 (mL)

Concentration of Working

Standards (mg/L)

CSBit-1 CSBit-5 1.25 25.00 0.5

CSBit-2 CSBit-5 5.00 25.00 2.0

CSBit-3 WoSS-250 0.50 25.00 5.0

CSBit-4 WoSS-250 0.80 25.00 8.0

CSBit-5 WoSS-250 2.00 50.00 10.0

4.5.2. Using table 11 below, create 2 quality control standards for the analysis. A lower limit QC, QCBit-1-LLQC, and a calibration verification QC, QC-Bit-4-ICV, are needed to be prepared for the analysis.

Table 11 Preparation of quality control standards of bitumen using matrix MM63

QC ID

Type of Standard to be

used for diluting the

sample

Volume of Standard (mL)

Final Volume after dilution using MM63

(mL)

Concentration of QCs (mg/L)

QCBit-1-LLQC CSBit-5 2.5 25.00 1.0

QCBit-4-ICV WoSS-250 0.4 25.00 4.0

4.6 Prepare unknown sample. Analyze the sample as quickly as possible. If filtration is needed for the suspended solids, let it settle down and obtain the sample by decantation.

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4.7 Following table 12, prepare the unknown samples using a 10-fold dilution and 4-fold dilution

Table 12 Preparation of unknown sample using matrix AT21 and MM63

4.8 Select any of the prepared unknown sample to run as duplicate.

4.9 Analyze the method using a spectrofluorometer

Unknown Sample ID Type of Matrix Used Volume of sample (mL) Final Volume (mL)

1A (undiluted sample) AT21 2.00 20.00

1AX10 MM63 5.00 20.00

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Bibliography [1] "SAIT D2L," SAIT, [Online]. Available: https://learn.sait.ca/d2l/le/content/76447/viewContent/2028123/View?ou=76447. [Accessed 20 February 2015]. [2] "Bitumen In Water," AGAT Laboratories, Calgary, 2015. [3] SHIMADZU, "Shim-pol," [Online]. Available: http://shim-pol.pl/pliki/produkty/Shimadzu/C125E006E_RF_1501.pdf. [Accessed 05 03 2015]. [4] LabExchange, "Labexchange," 2012. [Online]. Available: http://www.labexchange.com/nc/en/buy-devices/d/serial/13930/. [Accessed 1 March 2015].

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SAIT Technical Project Week - Beatriz Pregillano, 2015