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[Music]

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[Applause]

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hi thank you I'm Elizabeth cada mio I'm

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a postdoc at the Jet Propulsion lab and

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today we'll be talking about a method

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I've been developing for the analysis of

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organic and inorganic ions using

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capillary electrophoresis with

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capacitively couple of contactless

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conductivity detection or CEC 4d for

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short so the target for this method is

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really analysis of the samples from this

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ocean world I don't have to explain to

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this group why this world these worlds

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are important or interesting so I'm not

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going to waste my time doing that but

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I'm going to say that one of the targets

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one of the things I would like to

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possibly use this for is incorporation

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into a potential larger mission for life

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detection and there was this wonderful

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paper that was published that sort of

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delineates how you might search for life

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and detect life on other worlds and the

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method that I'm developing would sort of

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fall on the rung that describes

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molecules and structures conferring

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function so I would be looking for

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patterns within classes of molecules

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that are not random and therefore might

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be indicative of life and the classes of

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molecules that I'd be looking at are

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things like amino acids or carboxylic

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acids because these are things that we

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find for carboxylic acids they're

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important intermediates and metabolic

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processes and they can sort of be

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component of cell membranes and things

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like that I'm also interested in this

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method and it probably has its strengths

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mainly and have ability assessment and I

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want to take a moment to make sure that

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we are clear about the difference

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between life detection and habitability

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so my Cliff Notes version of this is for

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life detection you're looking for

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evidence of some sort of metabolism

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habitability is just asking whether or

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not that environment could support a

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metabolism and it's much more

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complicated than that but those are the

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cliffnotes for now and so the basis for

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looking an habitable environment would

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be looking for something like water

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organic molecules and energy sources and

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having these are not simply enough you

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need to have them sort of co-located so

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any organism that might be present could

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actually use them for metabolism so to

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do this we're going to first search for

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these organic molecules and energy

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sources in the soluble chemistry of the

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environment and that will ensure not

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only that they are co-located but also

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that you

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a solvent that is able to participate

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and support the electron transport

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processes that are essential for

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metabolism so there are different

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organic molecules that we will be

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looking for and different energy sources

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that we will be looking for the specific

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targets that I've sort of used to

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develop my method for inorganic ions we

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have over here we based it off of

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seawater for a lot of them because if

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we're looking at ocean world's our ocean

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is sort of a decent model for and also

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we suspect there will be a lot of

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chlorine there will be a chloride there

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will be sulfate so we're looking for a

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bromide chloride carbonate sulfate and

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we're also looking for metabolic energy

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sources so microbial life on earth can

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use oxyanion species like sulfate and

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nitrate and perchlorate in order to

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metabolize I also want to take a moment

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to specifically focus on perchlorate

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just briefly if we are expecting these

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ocean worlds to have a lot of chloride

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and we think they might be highly

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oxidizing I think it is important that

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we consider the presence of perchlorate

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as a potential ion that could either

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interfere or support metabolic life in

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these oceans and so I wanted to make

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sure that that is part of our our search

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in these inorganic sources we're also

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looking for carboxylic acids so I sort

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of split them into these short chain

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carboxylic acids and potentially longer

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chain ones so here we have a sort of

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loose selection of ones that are found

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in meteorites that are shorter chain

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their mono carboxylic acids their branch

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they have hydroxy groups attached to

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them and then we have these which are

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sort of Diane tri carboxylic acids which

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are products and intermediates in

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metabolism we're also interested in some

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mono carboxylic acids that are longer

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chain lengths that are derived from

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lipids and could be potential components

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of cell walls

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so the technique we're using seee is

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ideal for this sort of analysis because

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it inherently separates charged species

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in an aqueous solution which is exactly

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what we're looking for

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so to do a cex parent you have a thin

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capillary about 50 microns in our case

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we fill it with a background electrolyte

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we inject sample into one end and then

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we apply a voltage and what happens is

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under the effect of that voltage the

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species of the charged species in the

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sample will separate based on their mass

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and there are there's their size and

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their charge and so at the other end you

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can place a detector and you can detect

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the different species that are already

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separated the detector detection method

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that we have chosen is a form of

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conductivity detection and we chose this

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because it will detect any charged

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species so we don't actually need to

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know what the charged species is so long

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as we can detect that that it is

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different from our background

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electrolyte it doesn't require any

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labeling or derivatives ation so we

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could use this in conjunction with other

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detection methods so we could put

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perhaps the c4d detector in line with a

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mass spec and get even more information

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out of it in the end so if you noticed

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when I talked about the targets that I

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was interested in I talked mostly about

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anions and the reason for that is that I

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have a lot of lab mates who have done a

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lot of work on the cation side of this

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here we have a paper that was published

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last year by a Philip O stock motto for

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da Santos and I encourage you if you're

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interested to look up that paper and he

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what he did was he used the same CEC 4d

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technology in order to separate

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inorganic cations and amino acids

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simultaneously and the background

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electrolyte he used for that was acetic

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acid and then we have another method

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that was developed by Jessica Kramer

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who's giving a talk later today that I

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encourage you all to go to that is done

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she's got excellent work developing

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chiral separations of amino acids with

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very low detection detection limits and

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the electrolyte that she's been using

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so as I go forward to develop my method

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I'm looking for using it in these in

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situ environments where we're really

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trying to minimize the complexity of our

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instruments and our methods because we

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don't have a lot of space we don't have

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a lot of power and so we want to have it

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be as simple as possible but we're not

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really willing to give up any of the

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science that we're looking for and so my

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approach to doing this is to take those

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cation methods that my lab mates have

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developed and develop an anion version

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that is compatible so it should be run

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on the same capillary with the same

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equipment and ideally use as many of the

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so this is exactly what I did so method

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1 I basically took the acetic acid based

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method that was developed for the

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simultaneous separation of inorganic

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cations and amino acids and I titrated

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it with sodium tetraborate until it

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reached around the pKa of acetic acid

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and I used that to separate whoops

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that's the wrong button use that to

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separate the the inorganic ions that I

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was interested and then the small very

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highly charged oxalic acid and then I

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basically did the inverse in order to

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develop the carboxylic acid version of

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this so I took the sodium tetraborate

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that's being used for the chiral amino

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acids and I titrated that with acetic

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acid to about the pKa of sodium

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tetraborate and I use that to separate

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out these carboxylic acids and this is

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very interesting because we can get it's

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very convenient I don't know if it's

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interesting because we have basically we

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have this division here and at this

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point everything below that are these

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sort of mono carboxylic acids these

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hydroxy acids these things which are

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typically found in meteorites and and

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the like and on the other side we have

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the highly charged ion tri carboxylic

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acids and things that are typically

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found as metabolic intermediates it

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might be more interesting in that

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respect we were also interested in these

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longer chain carboxylic acids so these

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are the potential components of cell

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membranes and things like that and so we

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find that earlier on even earlier than

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many of the hydroxy and branched mono

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carboxylic acids we get this

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sweet of them that come out I ran this

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between a 3-carbon long mono carboxylic

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acid all the way up to a 20 and we were

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able to get up through 14 car back

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carbon long mono carboxylic acids which

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starts to approach your cell ability

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solubility limit for these so I think

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this is pretty indicative that if we're

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looking in the soluble chemistry we can

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get most of them using this method and

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we can get them in chunks along the way

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so we also felt it was important to test

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this in a natural sample because

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standards are great because standards

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are great so I found a pond that was

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certainly full of life it had a lot of

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green in it and a lot of turtles and so

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we analyzed the sample from there we did

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not dilute it the only pretreatment that

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we did was we hydrolyze the cells by

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heating them up to 180 degrees for about

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30 minutes which would hopefully break

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apart the membranes and get anything

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that was inside of them out and when we

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ran that we found that we found the

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chloride and saw the sulphate which are

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the dominant anions in organic anions in

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that solution and then we also found

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citric acid which we would expect

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because if you have a lot of metabolism

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in a very large life filled pool you

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would expect citric acid from the citric

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acid cycle to be there

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we found carbonate and we found tiny bit

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of mono carboxylic acids and then we

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found this little peak down here which

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turns out to be indicative of

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potentially a lot of amino acid so we

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can't separate any amino acids with this

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method but what we do get when we spike

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a solution with amino acids is this

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immediate large dip that looks exactly

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like this which may be indicative of the

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amino acids great so I am ahead of time

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so moving forward on this method there

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are things we want to do so right now

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we've tested in this very friendly life

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filled pool but we want to be able to

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make sure that we can use this on things

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that are a little more analogous to the

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ocean world's that we're hoping to use

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it on in the future so we did some field

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work and we went and collected samples

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from Owens Lake and Mono Lake which are

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too hyper saline alkaline lake

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environments which could be analogous to

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these and so we're hoping to use this

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method to analyze those samples and then

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we're also beginning to develop a

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simultaneous method because well it's

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beautiful and simple to say we're just

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going to use the reagents from these

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cation methods and use the capillary and

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use all the equipment and there's a

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simplicity that makes that that really

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exciting it still requires two analyses

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and so if time is your issue then it's

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actually not as simple as we had hoped

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and so I'm attempting to develop a new

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method that will do this all at once and

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we're still using the same acetic acid

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base but we're adding triethylamine

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instead of boring as to adjust the pH of

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this and using this we can separate all

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of the primary inorganic ions with the

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exception of the chlorate and

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perchlorates to both oxychloride species

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seem to collude and we can actually get

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all of the carboxylic it's acids that we

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were interested in all the way up to c10

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so it kind of crashes out at that point

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but we're able to get all of the other

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ones including citric acid and we're

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able to take carbonate and things like

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that

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shameless plug for the rest of my lab

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mates so we're part of the chemical

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analysis and life detection group at JPL

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and a lot of us are giving talks and

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posters this week so this is a Nate and

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Constantin have posters tonight and

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they're talking about developing the

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hardware and qualifying the hardware for

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the instruments that might use these

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kinds of methods they've done a lot of

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terrific work so I suggest you go by and

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talk to them I already talked about

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Jess's talk for chiral amino acids

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fernanda is giving a talk on her

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chemical laptop which is a portable

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microchip device it's automated and

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she's done a lot of X

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with that so you can see that tomorrow

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and Florian is giving a talk about his

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liquid extractors so even if we had a

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soil sample or something that wasn't

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necessarily already in our perfect

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liquid form he's developed a tool that

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will help us get it into the form we

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need for this and then on Thursday Peter

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our group supervisor is going to give an

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excellent overview of all of the work

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that we've been doing developing these

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types of techniques for for these

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environments and then Aaron is going to

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be talking later about a microfluidic

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ion analyzer that we've been developing

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as well which is complemented with all

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of this so with that I will take any

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questions we've got plenty of time for

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questions hi Lawrence Tyler from hooey

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um so there's a pretty big difference

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between a small amino acid like glycine

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and like a really huge one like prolene

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or tryptophan um do you have any idea

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why this technique can't tell the

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difference between those things yes so

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it has to do with basically the speed

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with which they move so the method that

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detects them but doesn't separate them

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is because the Y actually I'm not sure

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it would even measure glycine they all

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just move so fast under that because in

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addition to separating based on our

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charge in size when you apply the

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electric field you also get what's

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called an electrostatic flow so the

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solution the background electrolyte gets

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pulled along with these ions toward the

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negative end and so if that's happening

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it can if it's too fast it can sometimes

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squish together those small highly

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charged positive ions and so the reason

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the negative ones separate is because

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they have a draw backwards that the

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positive ones don't have so you really

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just get them all at once and you don't

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get any separation because they move

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yes okay um yeah so she was asking about

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the size of the peaks given that the

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abundance of life we'd expect in that

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pond right okay

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yes also it can be quantitative

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I didn't quantify that because I'm still

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working on really making that a little

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more I guess robust would be the phrase

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but especially with the mono carboxylic

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acids probably what's present there just

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isn't soluble in the water they're

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probably still longer chains that we

385
00:14:32,860 --> 00:14:31,490
just can't get to in terms of the citric

386
00:14:35,260 --> 00:14:32,870
acid I'm not sure how much I would

387
00:14:37,389 --> 00:14:35,270
expect to see to be honest I haven't

388
00:14:41,410 --> 00:14:37,399
quite finished doing that it was our

389
00:14:44,680 --> 00:14:41,420
sort of first attempt at a at an

390
00:14:46,870 --> 00:14:44,690
analysis like this so it could just be

391
00:14:49,569 --> 00:14:46,880
that that when you break apart a Cell

392
00:14:51,040 --> 00:14:49,579
right maybe I'm not sure how much citric

393
00:14:52,900 --> 00:14:51,050
acid you expect to get out with that

394
00:14:54,370 --> 00:14:52,910
metabolism but it's something that we're

395
00:14:56,980 --> 00:14:54,380
kind of looking at and trying to figure

396
00:14:58,630 --> 00:14:56,990
out how much we expect sort of per cell

397
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and how many cells we might need to say

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00:15:04,990 --> 00:15:01,310
that we found something and iterative we

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00:15:10,620 --> 00:15:09,139
so let's thank our speaker again we've

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got our meal