vinmetrica SC-50 MLF User manual

1
SC-50 Manual v3.1
Vinmetrica SC-50 MLF Analyzer™
User Manual
The Vinmetrica SC-50 MLF (malolactic fermentation) Analyzer*is a simple and robust device that
provides high accuracy in determination of malic acid concentration levels in wine, an essential
parameter to control in the effort to make high quality wines.
Note: Check the Vinmetrica FAQ page for the most current version of this manual
with up-to-date procedures and best practices for operating your SC-50.
http://www.vinmetrica.com/FAQ
Table of Contents:
Materials Provided in the Kit............................................................................................................... 2
Things you will need............................................................................................................................ 2
Why Test for Malic Acid?.................................................................................................................... 3
Theory of Operation............................................................................................................................. 3
Setup.................................................................................................................................................... 4
Setting up the MLF Analyzer for the first time........................................................................ 4
Procedures.......................................................................................................................................... 5
Measuring Malic Acid............................................................................................................ 5
Confirming Procedure………………… .................................................................................. 7
Concentration Determination…………………………………………………………...….….8
Data Interpretation..…………………………………………………………………………...9
Finishing Up……………………………………………………………………………………...….10
Representative Data ……………………………………………………………………..…….........10
Warranties, Liabilities & Hazards.....................................................................................................11
Appendix A –Troubleshooting and FAQs ……………………....................................................12
*US Patent pending

2
SC-50 Manual v3.1
Materials provided in the kit:
1. Vinmetrica MLF analyzer unit
2. Malic Reagent set:
Boost juice™ (Part Number: SC-50-4)
***Keep in Refrigerator***
Malic Acid (1M) concentrate (Part Number: SC-50-5)
***Keep in Freezer***
Malic Acid Standard Solution (0.40 g/L) (PN: SC-50-11)
***Keep in Freezer***
Malic Acid Standard Solution (0.10 g/L) (PN: SC-50-12)
***Keep in Freezer***
3. Biopressure Agent (PN: SC-50-3-10 or -20)
***Keep in Freezer***
4. Reaction Vials (5)
5. 1 ‘check’ vial
6. 10 ml serological pipette
7. 15 ml conical tube
8. Plastic bulb pipette (2)
9. 100 mL polypropylene beaker
10. Plastic Scoop
Things you will need:
1. Vinmetrica SC-100A or SC-300.
2. One 9 volt battery.
3. Distilled water, which usually can be found at your local grocery store. It’s handy to have a wash
bottle for rinsing. Rinse bottle available from Vinmetrica (Part number SC-100-17)
4. Microwave oven, hot plate or stovetop (for boiling wine samples)
5. (Optional) Additional Biopressure agent for running more than 5 samples, as provided in the kit.
Comes in sets of 10 (SC-50-3-10) or 20 (SC-50-3-20).
6. (Optional) BNC to RCA adapter - See under “Setup”, Step 4, for more information.
Figure 1. The SC-50 MLF
Analyzer with vial insertion
assembly

3
SC-50 Manual v3.1
Why Test for MalicAcid?
In winemaking, malolactic fermentation (MLF) converts malic acid to lactic acid, with CO2
being the byproduct of the reaction. MLF, which is typically carried out in most red wines and in some
white varietals, plays an important role in the finished wine’s feel and taste. MLF reduces titratable
acidity, increases pH, and produces flavors often characterized as “soft” or “buttery”.
Sulfite, as free SO2, inhibits the bacteria that carry out MLF. Therefore free SO2levels must be
kept low during MLF, carrying risks that the wine is left unprotected against oxidation and microbial
contamination. As soon as MLF is done, then, SO2should be raised to appropriate levels for protection
of the wine. Thus it is important to know when MLF is done, and the best way to do this is to measure
malic acid levels in the wine.
Theory of operation:
The SC-50 MLFAnalyzer, with the SC-300 or -100A, can be used to determine malic acid
concentration in wine. It relies on the biochemical MLF reaction caused by enzymes found in certain
bacteria, including lactobacilli and oenococcus strains, and in the Biopressure agent component of the
kit. These bacteria live on a variety of nutrients, but their production of CO2results almost entirely
from the enzymatic transformation of malic acid to lactic acid:
Malic Acid Lactic Acid
HOOC–CH2–CH(OH)–COOH CH3–CH(OH)–COOH + CO2
The CO2creates pressure, which is converted by the MLF Analyzer to a signal for the SC-300 (or
SC-100 or -100A). The CO2pressure signal is directly proportional to the amount of malic acid in the
sample. The device is set so that when malic acid levels exceed 0.1 g/L1(suggesting MLF is
incomplete), the user is alerted to this state by audible and visible indicators. The level of Malic acid
can be calculated from the digital readout by a calibrator of Malic acid provided with the kit. Detection
limit is below 0.04 g/L. The assay takes 30 minutes, or up to 60 minutes if the Confirmation Procedure
is performed (Page 8).
1On the original SC-100 model, the device will signal at a Malic Acid level of about 0.5 g/L. However, the sensitivity and
accuracy of malic acid values lower than this are comparable to those determined with the SC-100A and SC-300.

4
SC-50 Manual v3.1
Setup
Setting up the MLFAnalyzer for the first time:
1. The MLF Analyzer runs on one 9 volt battery. To insert the battery, slide open the battery
housing on the bottom of the back of the unit by gently pushing the lid.
2. Low Battery Detection: If the battery is good, the POWER LED will stay illuminated when the
switch is pressed ON. When the battery level is too low, the LED on the POWER switch will
flash on briefly and then shut off. Replace the battery.
3. Before connecting the SC-50, turn on your SC-100A or 300 (on the SC-300, select SO2mode,
press Enter) and wait for the initialization and green ‘Proceed’ light.
4. Now attach the MLF Analyzer to your SC-100A or 300 via the SO2electrode connector
protruding from the top. Be sure to secure the electrode plug to the connector to ensure proper
function. Note: on earlier SC-100A or -300 models, there is just one BNC-style connector for
this purpose - see Figures 2 –4. On newer models (June 2014) there is a separate phono- style
SO2connector that will require an adapter for SC-50 models that have a BNC plug (Figure 5)–
contact us if you need one.
5. Prepare all Reaction Vials:
a. Remove Biopressure Agent from freezer and allow to come to room temperature.
b. Using the small plastic spoon provided, measure out two (2)level scoops (approx. 35-40
mg) of Biopressure Agent into each Reaction Vial.
a. Due to the consistency of the Biopressure Agent, we recommend rinsing and
completely drying the plastic spoon after filling approximately 5 vials.
Figure 2.Attach the SC-50’s
cable to the BNC or RCA
connector on the SC-300 (or
100)
Figure 3. Be sure the cable
plug is screwed into place on
the BNC connector or firmly
inserted into the RCA plug.
Figure 4. Recommended
arrangement of the
equipment
Figure 5. The adapter for
the SC-50.

5
SC-50 Manual v3.1
c. Cap all Reaction Vials.
d. Keep out all Reaction Vials needed for MLF assay.
a. For Quick Check Mode, you will need one (1) reaction vial per wine sample.
b. For Quantify Mode you will need at least two (2) reaction vials, one (1) for the
Malic Acid Standard and one (1) per wine sample. (See Quantify Mode, pg 8, for
further information).
e. Place all remaining reaction vials in freezer until use.
f. Thoroughly rinse plastic measuring spoon with DI water.
Procedures
Biopressure Assay method:
There are two modes for using your SC-50 to assess MLF status by Biopressure assay. Both modes use
steps 1 through 18 below.
In the Quick-check Mode, you set the instrument such that if your sample has 0.1 g/L or more
of malic acid, you will see the STOP LED on the SC100/300 illuminate and the tone will sound,
alerting you that MLF is not complete. There is an optional confirmation step (see
Confirmation Procedure, page 8) that you can run after the test to verify the validity of a
result.
In the Quantify Mode, you set the instrument to higher sensitivity and run the assay with one or
more malic acid standards provided in the kit. This allows you to determine the actual malic
acid concentration in your wine samples. (See Quantify Mode Procedure on page 8).
We recommend running the Quantify Mode until you are familiar with the workings of the instrument.
Note: the following procedure is designed to be run at room temperature, between 66 and 73°F
(19 - 23°C). Results may differ if temperatures are outside this range. SeeAppendix for more
information.
1. Take out all Reaction vials needed for assay from freezer. One Reaction vial per sample is
necessary. If you will run the assay in Quantify Mode you will need at least one additional
reaction vial (See Quantify Mode Procedure on page 8). Allow Reaction vials to come to
room temperature before initiating assay. Note that you can run multiple wine samples at one
time.
2. Using the 10 mL serological pipette, measure a 10 mL wine sample and dispense into the 100
mL plastic beaker or other suitable container. (Serological pipette can be rinsed with DI water
for re-use; remove cotton plug if necessary)
3. Place wine sample in microwave and boil for 90 seconds at medium power. Actual time
depends on your microwave oven’s power; the object is to get the wine sample to a gentle boil
for 60 seconds without having the wine splash out of beaker. If you are unsure about the power
of your microwave, we suggest you experiment with a couple of wine samples beforehand to
determine what the best settings are for your microwave oven. You may also bring wine to a
gentle boil on a stove top for 60 seconds.

6
SC-50 Manual v3.1
4. Let wine sample cool to room temperature, then pour the sample into the 15 mL conical tube. If
the sample has lost volume due to evaporation, add DI H2O to restore the volume to 10.0 mL,
then pour the sample back into 100mL beaker.
5. Gently mix the bottle of Boost Juice. Using a plastic bulb pipette, add 5 drops (150 µL) of the
Boost juice into the 10 mL boiled wine sample. Mix wine gently. Caution: Use care when
handling. Boost Juice contains materials that may be a skin irritant. Do not ingest. If
there is contact with skin, rinse with plenty of water.
6. Disconnect the vial insertion assembly from the SC-50 by unscrewing the luer lock connector
on the bottom of the instrument. (Figure 6.) The disconnected vial insertion assembly will be
used to zero the pressure immediately after capping the sample. Note: The luer lock connector
should be reattached tightly to the SC-50 (step 8) before taking a measurement in step 15.
7. Using the 10mL serological pipette, transfer exactly 3.0 mL of the wine sample from step 5 into
the reaction vial with the Biopressure agent. Immediately cap the vial tightly, then quickly stand
the vial upright and position the disconnected vial insertion assembly such that the needle
punctures the septum of the reaction vial (see figures 7 and 8, but note that in this case the SC-
50 is NOT connected to the vial insertion assembly). Keep the vial insertion assembly with its
needle inserted through the septum for about 3 seconds. This step equilibrates the vial’s internal
pressure to ambient pressure, i.e., it “zeroes” the initial pressure.
8. Remove the vial from the insertion assembly and repeat step 7 for each sample to be analyzed.
When done, be sure to reconnect the vial insertion assembly to the SC-50 by firmly screwing its
luer lock fitting back on to the SC-50.
9. Shake all vials gently but thoroughly for approximately 10 seconds.
10. Allow sample to incubate at room temperature for 30 minutes. While incubation is occurring,
gently shake the vial at the 10 minute and 20 minute mark.
11. Make sure the SC-50 is attached as described in step 4 of the Setup section. Turn on both
instruments. If using an SC-300, select SO2mode and press the ENTER button.
Figure 6. Unscrew the Luer Lock
connector.

7
SC-50 Manual v3.1
12. Turn on the SC-50 by pushing the green button. The green LED should illuminate. If it
illuminates briefly and then goes out, replace the battery.
13. Initialize the SC-50 by turning the black LEVEL knob until non-zero numbers appear on the
Vinmetrica SO2Meter. You are setting the baseline value for ambient air pressure. Adjust until
the SC-100A or SC-300 meter reads the appropriate level: 20 ± 1 for the Quick-check Mode, or
45 ± 1 for the Quantify Mode (Note: every time device is turned on, it should be initialized)
14. Immediately following the 30 minute incubation period, shake the vial once more for 10
seconds and then proceed to the next step immediately. It is best to shake the vial by holding the
cap, as holding the glass vial may change its temperature, resulting in a less accurate pressure
reading. DO NOT remove the cap of the vial.
15. This is the CO2pressure reading step. [Note: Make sure all the parts of the vial insertion
assembly are secured tightly before proceeding.] Place reaction vial upright on your work
surface. Position the opening of the vial insertion assembly over the vial. Gently push the
assembly down onto the vial as far as it will go and hold it in place (i.e. so that the assembly’s
rubber-sheathed needle is inserted into the vial’s septum); see figures 7 and 8.
16. Record the highest value that appears on the meter right after insertion of the needle. This
process should take no longer than 5 seconds. For calculation purposes, call this value “a”.
[Note: If values tend to drift upwards, hold the insertion assembly at the very top or by the
flanges at the bottom to minimize heat transfer from your hand to the vial.]
17. Release the vial by letting go of the assembly. If you are not performing multiple reactions,
continue to the next step. When performing multiple reactions, once the vial is removed, the
analyzer should stabilize back to the original initialization value. If needed, re-initialize the
instrument by repeating step 13.
Figure 7. Position the opening of the vial
insertion assembly over the vial.
Figure 8. Gently but firmly push the assembly
down onto the vial as far as it will go.

8
SC-50 Manual v3.1
18. For the Quick-check procedure, if Red LED light appears and tone is audible, MLF is not
complete. If Green LED stays on, MLF is near or at completion. See “Quick-check Data
Interpretation”section below.
Confirmation Procedure:
To ensure your Quick-check assay is working correctly, you can confirm the validity of the assay result
using the 1M malic acid concentrate with a “just-completed” wine sample.
1. Add Malic Acid concentrate: After recording the CO2 response in step 16 above, open up the
Reaction vial and, using the plastic bulb pipette, add 1 drop (30 uL) of 1M Malic Acid to the
sample. This is equivalent to 1.3 g/L malic acid in the 3mL wine sample.
2. Immediately re-cap the reaction vial and equilibrate it with the disconnected insertion assembly
as in step 7 above. Mix the vial thoroughly.
3. Repeat steps 10-15 from above, letting the sample incubate again for 30 minutes and
performing the CO2pressure reading step. Record the data. [If your instrument has turned off, it
will need to be reinitialized.] To determine what these numbers mean and how to use them, please
see the data interpretation step below.
Quantify Mode Procedure:
The Biopressure Assay Procedure above will allow you to quantitatively assign a value (in g/L) for
malic acid in your wine samples by including the steps below when you run the assay.
1. You will run at least one “Standard Vial” at the same time as your wine samples. This will re-
quire one additional Reaction Vial.
2. It is advised to run all your samples within a few minutes of each other. Be prepared to begin
the standard vial right after adding wine samples to the Reaction Vials (Step 7 under Procedures,
above).
3. Take a new Reaction Vial (already at room temperature) and, using a clean 10 mL serological
pipette, add 3.0 milliliters of the 0.1g/L Malic Acid Standard Solution to the Reaction Vial.
This is now your Standard Vial. Cap the Standard Vial tightly. You may also run a 0.4 g/L
standard or any higher concentration if you want to make one; however concentrations above
about 2 g/L will give results that exceed the measurement limits of the device.
4. Complete the assay following steps 8-17 of the Biopressure Assay Procedure (note that you ini-
tialize the SC-50 at 45 for this mode). Make sure to record the highest value that appears on the
meter for the Standard Vial. Call this value “c”
5. Measure the blank response.
a. Add 3.0 mL DI water to the check vial, capping tightly.

9
SC-50 Manual v3.1
b. Read the vial as mentioned in Steps 15 & 16 in the section above, “Measuring Malic
Acid by Biopressure Assay”. Record this value. Call this value “b”
c. If you are running multiple wine samples at one time, you only need to take the “blank”
reading once. If the instrument turns off, the “blank” will need to be re-done.
Calculation
1. You should now have three values a,b, and c, in order to calculate the concentration of malic
acid:
a= Wine sample result
b= Blank Vial result
c= Standard Vial result for standard concentration S (e.g., 0.10 g/L)
2. The malic acid (MA) content, in grams per liter, is given by
MA, g/L = S *(a - b)/(c - b)
(i.e. subtract bfrom a, subtract bfrom c, divide the first difference by the second, then
multiply that result by S, the concentration of your standard (usually 0.10 g/L )
Example: in a Quantify mode assay with an SC-300, awas 75.0, bwas 59.2, and cwas 129 for a
0.10 g/L standard, (i.e., S was 0.10). So the malic acid concentration was
0.10 x (75.0 - 59.2)/(129 - 59.2) = 0.02 g/L malic acid
Having determined the concentration in g/L, you should use this table as a rough guideline for
status of MLF in your wine
MalicAcid Concentration, g/L
MLF status
Above 1
Not started or just started
0.4 –1.0
Incomplete, probably started
0.1 –0.4
Progressing well
0.05 –0.1
Nearly complete, probably OK
Below 0.05
Complete
Quick-check Data Interpretation
The SC-50 is designed to give a readout of approximately 50 nA (nanoAmps) current on the SC-100A
and SC-300 when the level of malic acid in the wine is at 0.1 g/L. At this level of the current, the red
LED and the audible tone of the SO2meter come on in the customary manner for SO2titrations. This
signals that malic acid is at or above 0.1 g/L and therefore MLF is not complete. If the malic acid level
is below 0.04 g/L, a signal less than 50 nAwill appear, the green PROCEED light will stay on, and no

10
SC-50 Manual v3.1
tone will be heard. At this point you will probably want to verify this result by following the
confirmation procedure above. At the end of the confirmation procedure the reading should increase by
several hundred nA for the SC-100A and SC-300. This confirms that when malic acid is present, the
system responds appropriately.
Finishing Up:
1. Turn off both devices and detach the SC-50 MLF analyzer from the BNC connector on the top
of your other SC series device. You can remove the battery if desired.
2. If you have unused reaction vials, place these back in the freezer as well as the 1M Malic Acid
and the 0.4 and 0.1 g/L Malic Acid Standard Solution. Place the Boost Juice™ back in the
refrigerator.
3. Rinse all plastic ware with DI water and store the unit and plastic accessories in a safe place.
4. Vinmetrica is considering enacting a recycling program for all used reaction vials. If you are
interested in participating, please hold on to them until further notice. We thank you in advance
for your participation.
5. Rinse out all used reaction vials with DI (distilled) water two to three times and air dry with lid
removed. Reaction vials can be re-used. Save all reaction vials and caps.
Representative Data:
Analyses performed at Vinmetrica Labs on an SC-50 with an SC-300. This table is for reference
purposes only; your data may be different. The Red Wine had completed MLF; the White wine had not.
Representative Data
Sample
Sample result
(a)
Standard result ©
(c, S = 0.4)
Check Vial
Response
(b)
Malic Acid
Concentration
(g/L)
White wine
702
---
59
0.92
Red Wine
63
---
59
0.006
0.4 g/L malic acid
349
59
---
Technical assistance: [email protected] tel. [phone]