BLLE PROCEDURES
Apparatus & Reagents
BLLE Rotisserie
Inversion Chair
Funnelcock
Pesticide grade Methylene Chloride
HPLC grade Ethyl Ether (Ethanol preserved or unpreserved)
Pesticide grade Isooctane
Pesticide grade Methanol
Baked (400°C for 4 hours) Sodium Sulfate
Baked (400°C for 4 hours) Sodium Chloride
6N NaOH
12N H₂SO₄
METHOD 608/8081/610/8100/8310 (etc.) - BLLE Neutral Extraction
1) Remove bottle containing sample water to be tested from cold storage and allow to equilibrate to room temperature. (You may want to protect the bottle label with a strip of packaging tape to prevent it from rubbing off during the rotating process to follow).
2) The bottle will need approximately 100 mLs of free headspace. (Ideally a 'fill-mark' is provided the sampler), If the bottle is overfilled, shake briefly to homogenize then discard up to 100 mLs. (BLLE may accommodate 5% or more particulate matter). Sufficient volume must be available to enable the addition of 80 mLs of solvent. The discarded sample volume (if any) may be used to check initial sample pH and residual chlorine content). Mark the meniscus of the water level on the side of the bottle for a later accurate determination of the water volume.
3) Assure that the pH of the sample is neutral. Adjust if necessary.
4) Fortify the samples with surrogate and batch QC samples with appropriate solutions prepared in a water soluble solvent .
5) Add 60 mL of methylene chloride (bottom layer) to each bottle followed by 20 mL of ethyl ether (top layer). Do not shake. Tightly cap and rotate 12 hrs. (Layers will unite upon rotation).
6) Remove the bottle from the rotisserie, uncap and wipe clean the neck and lid of the bottle with methanol. Insert a funnelcock (be sure stopcock is closed) onto an inversion chair, and, while holding the funnelcock firmly in place, overturn the inversion chair onto the bottle. With a steady, sharp motion, invert the apparatus once again. (You may want to practice this function using a bottle of tap water to learn to totally avoid spillage) Discharge the solvent into a suitably sized, pre-cleaned container (250 mL amber, glass recommended). Add approx 10g of baked sodium sulfate to the discharged solvent, cap tightly and shake vigorously.
7) Allow the extract to remain in contact with the sodium sulfate for at least one hour. Shake periodically. If the sodium sulfate forms clumps, add additional sodium sulfate. Ultimately the sodium sulfate should be free flowing - an indication of complete water adsorption.
8) Decant the extract into a Kuderna-Danish apparatus, Turbovap tube or other solvent concentration device being careful not to dislodge the sodium sulfate from the bottle. Rinse the extract bottle two or three times with DCM to assure a quantitative transfer.
9) Proceed with normal concentration/solvent exchange (if necessary) procedure. (Yields generally exceed 80%).
METHOD 625/8270 - BLLE Base/Neutral (BN) Extraction
1) Remove bottle containing water to be analyzed from cold storage and allow to equilibrate to room temperature. Cover the bottle label with a strip of packaging tape.
2) If the bottle is too full, shake briefly to homogenize then discard up to 100 mLs. (Sufficient room must be available for 9 mLs of base and 80 mLs of solvent to be added). This discarded sample volume (if any) may be used to check initial sample pH and residual chlorine content). Mark the meniscus of the water level on the side of the bottle for a later determination of the exact water volume.
3) Fortify the samples with surrogate and batch QC samples with appropriate solutions.
4) Adjust the pH of the sample with 9mL of 6N NaOH. Check that pH is ≥ 12 with wide range pH paper. Add additional NaOH if necessary.
5) Add 60 mL of methylene chloride (bottom layer) to each bottle followed by 20 mL of ethyl ether (top layer). Do not agitate. Tightly cap and rotate 12 hrs.
6) Remove the bottle from the rotisserie, uncap, clean the neck and lip of bottle with a methanol wetted Khemwipe and overturn onto a funnelcock using the inversion chair. Discharge the solvent into a suitably sized, pre-cleaned container (250 mL amber, glass generally). Add approx 10g of baked sodium sulfate to the discharged solvent, cap tightly and shake vigorously.
7) Allow the extract to remain in contact with the sodium sulfate for at least one hour. Shake periodically. If the sodium sulfate forms clumps, add additional sodium sulfate. Ultimately the sodium sulfate should be free flowing.
8) Decant the extract into a Kuderna-Danish apparatus, Turbovap tube or other solvent concentration device being careful not to dislodge the sodium sulfate from the bottle. Rinse the extract bottle two or three times with DCM to assure a quantitative transfer.
9) Proceed with normal concentration procedure.
METHOD 625/8270 - BLLE Acid/Neutral (AN) Extraction
1) Remove bottle containing sample water to be analyzed from cold storage and allow to equilibrate to room temperature. (You may want to cover the bottle label with a strip of packaging tape at this stage to avert the possibility of it rubbing off).
2) Shake the bottle briefly to homogenize and discard up to 100 mLs. (Sufficient room must be provided for 3 mLs of acid and 80 mLs of solvent to be added. Ideally, a fill mark is provided for the sampler to eliminate this step. Discarded sample volume (if any) may be used to check initial sample pH and residual chlorine content). Mark the meniscus of the water level on the side of the bottle for a later determination of the exact water volume.
3) Fortify the samples with surrogate and batch QC samples with appropriate solutions.
4) Adjust the pH of the sample with 3 mL of 12N H2SO4. Check that pH is less or equal to 2 with wide range pH paper. Add additional H2SO4 if necessary.
5) Add 60 mL of methylene chloride (bottom layer) to each bottle followed by 20 mL of ethyl ether (top layer). Do not agitate. Tightly cap and rotate 12 hrs.
6) Remove the bottle from the rotisserie, uncap, clean the lip and neck of the bottle with methanol and overturn onto a funnelcock using an inversion chair. Discharge the solvent into a suitably sized, pre-cleaned container (250 mL amber, glass recommended). Add approx 10g of baked sodium sulfate to the discharged solvent, cap tightly and shake vigorously.
7) Allow the extract to remain in contact with the sodium sulfate for at least one hour. Shake periodically. If the sodium sulfate forms clumps, add additional sodium sulfate. Ultimately the sodium sulfate should be free flowing.
8) Decant the extract into a Kuderna-Danish apparatus, Turbovap tube or other solvent concentration device being careful not to dislodge the sodium sulfate from the bottle. Rinse the extract bottle two or three times with DCM to assure a quantitative transfer.
9) Proceed with normal concentration procedure.
METHOD 625/8270 - BLLE Base/Neutral/Acid (BNA) Extraction
1) Remove bottle containing water sample to be analyzed from cold storage and allow to equilibrate to room temperature. Cover the bottle label with a strip of packaging tape to prevent the sample ID from wearing away during the rotation process.
2) If the bottle is overfilled (ideally a fill mark is provided to the sampler to avoid this) shake briefly to homogenize and discard up to 100mLs. (Sufficient space in the bottle must be available to accommodate subsequent additions of solvent and salt. If some sample volume is discarded it may be used to check initial sample pH and residual chlorine content). Mark the meniscus of the water level on the side of the bottle for a later determination of the exact water volume.
3) Fortify the samples with surrogate and batch QC samples with appropriate solutions.
4) Adjust the pH of the sample with 3mL of 12N H2SO4. Check that pH is less or equal to 2 with wide range pH paper. Add additional H2SO4 if necessary.
5) Add 60 mL of methylene chloride (bottom layer) to each bottle followed by 20 mL of ethyl ether (top layer). Do not agitate. Tightly cap and rotate 12 hrs.
6) Remove the bottle from the rotisserie, un-cap and overturn onto a funnel cock using an inversion chair. Discharge the solvent into a suitably sized, pre-cleaned container (250 mL amber, glass generally). Add approx 10g of baked sodium sulfate to the discharged solvent, cap tightly and shake vigorously.
7) Rotate the inversion chair restoring the bottle to the upright position. Add 100 g of baked (400C) sodium chloride and agitate to dissolve.
8) Readjust the pH of the sample with 9mL of 12N NaOH. Check that pH is greater than or equal to 12 with wide range pH paper. Add additional NaOH if necessary.
9) Add 60 mL of methylene chloride to each bottle, tightly cap and rotate 12 hrs.
10) Remove the bottle from the rotisserie, un-cap and overturn onto a funnel cock using an inversion chair. Discharge the solvent adding it to the previous base/neutral extract in the 250 mL amber, glass container containing sodium sulfate.
11) Allow the combined extracts to remain in contact with the sodium sulfate for at least one hour. Shake periodically. If the sodium sulfate forms clumps, add additional sodium sulfate. Ultimately the sodium sulfate should be free flowing.
12) Decant the extract into a Kuderna-Danish apparatus, Turbovap tube or other solvent concentration device being careful not to dislodge the sodium sulfate from the bottle. Rinse the extract bottle two or three times with DCM to assure a quantitative transfer.
13) Proceed with normal concentration procedure.
METHOD 615/8151 Herbicides - BLLE Automated Hydrolysis, Preparation, and Extraction
1) Remove bottle from cold storage and allow to equilibrate to room temperature. Cover the bottle label with a strip of packaging tape.
2) Shake to bottle briefly to homogenize then discard approximately 250 mLs. Mark the meniscus of the water level on the side of the bottle for later accurate determination of the water volume.
3) Add 190g of baked (400C) sodium chloride and shake to dissolve.
4) Adjust the pH of the sample to with 13mL of 6N NaOH. Check that pH is ≥ 12 with wide range pH paper. Add additional NaOH if necessary.
5) Fortify the samples with surrogate and batch QC samples with appropriate solutions.
6) Cap bottle tightly, place on rotisserie and spin for 2 hr at 3 rpm.
7) Following this 2 hr period, add 100mL DCM to the bottle, re-cap and turn for an additional 4 hrs.
8) Remove the bottle from the rotisserie, uncap and overturn onto a funnelcock using an inversion chair. Discard the lower DCM layer and return the water to the original bottle.
9) Readjust the pH of the water to to ≤ 2 (check with pH paper) using ~9 mLs 12N sulfuric acid
10) Add 120 mL of unpreserved diethyl ether to the water returned to the original bottle, tightly cap and rotate 12 hrs.
11) Remove the bottle from the rotisserie once more, un-cap and overturn onto a funnel cock using an inversion chair. Discard the water layer and collect the ethyl ether layer into a polyethylene plastic or 10% sulfuric acid washed 250mL bottle containing approx 10g of baked, acidified sodium sulfate.
12) Allow the extract to remain in contact with the sodium sulfate for at least two hours. Shake periodically. If the sodium sulfate forms clumps, add additional sodium sulfate. Ultimately the sodium sulfate should be free flowing.
13) Decant the extract into an acid washed Turbovap tube being careful not to dislodge the sodium sulfate from the bottle. Rinse the extract bottle two or three times with diethyl ether to assure a quantitative transfer.
14) Concentrate approximately 2 mL DO NOT ALLOW TO GO BEYOND 2 mL and protect the turbovap tube from water vapor as much as possible
15) Dilute the extract with 1 mL of isooctane and 0.5 mL of methanol. Bring to a final volume of 4 mL with diethyl ether. Proceed with normal derivatization procedure.
PROCEDURE For Discharging Solvent from Bottles Using Funnelcock and Inversion Chair
1} Seat the funnelcock securely into the inversion chair cradle (stopcock end down) and close the stopcock.
2) Place the bottle containing the sample and solvent to be discharged on a flat surface next to the inversion chair and remove the cap.
3) Invert the inversion chair with the funnelcock in it and carefully seat on top of the bottle.
4) Tilt the apparatus slightly to insert your fingers of one hand under the bottle.
5) With the other hand, hold the funnelcock in place snuggly against the bottle and flip the entire apparatus with one rapid, deliberate and smooth motion.
6) Place an appropriately sized retaining bottle under the apparatus (typically 250 mL) to capture solvent to be discharged.
The funnelcock must be flush up against the bottle and the motion rapid to avoid spillage. Practice with water and a small amount of DCM prior to handling actual samples.