 One for nephilometry. Depending on how you, basta, something that will help you differentiate the two, okay? Moving forward, let's go to osmometry. Osmometry sounds familiar, okay? Sounds easy. Because osmometry is the measurement of osmolality. Osmolality in where? Osmolality in your acquiesolution such as your serum, your plasma, and your urine, okay? And usually, okay, we have osmotically active components or substances such as what? These are your glucose, your urea, and your sodium, okay? Glucose, urea, and sodium are usually when added to the solution, they increases, okay? They increases the osmolality of your solution, okay? It increases the osmolality of your solution. Having mentioned that now, okay? Having mentioned that now, sir, how do we measure osmolality? We measure osmolality how? By measuring your colegative properties. And what are these colegative properties? We have your osmotic pressure. We have your boiling point, your freezing point, and your vapor pressure. So collectively, they are called your colegative properties, okay? And these colegative properties are related to one another, but much more related to your osmolality, okay? It is related to your osmolality. So in short, we are indirectly measuring osmolality using your colegative properties. And what is the effect of osmolality to these colegative properties? So when osmolality in the solution increases, what happened? Osmotic pressure and boiling point both increases. The freezing point and the vapor pressure, this is vapor pressure, are both depressed or are both decreased. Are you getting me? So if you understand me pa ano na lang sa chat box, okay? So osmolality again, if the osmolality increases, okay? If the osmolality increases, osmotic pressure, your boiling point are affected, how? It increases. What about sir, your ano? What about my freezing point and your vapor pressure? Those two are all, what happens to them is that they are decreased. Are we clear? Clear po tayo, can you answer in the chat? Yes. So in osmometry, okay, osmometry is based on measuring changes on the colegative property of a particular solution, okay? So once that there are increase of this particular particle, take for example, there are a lot of glucose, a lot of glucose, a lot of BUN or a lot of sodium, it will now affect your osmolality, okay? It will affect your osmolality. And take note, we were mentioning a while back that we measure osmolality by measuring its colegative properties. And among those four colegative properties, osmotic pressure, boiling point, freezing point and vapor pressure, the most commonly used method for measuring your osmolality and measuring changes in colegative properties of a solution is what? Your freezing point depression, okay? Your freezing point depression, okay? Here's the thing, take for example, your water. What is the freezing point of water? Zero degree Celsius po. Zero degree Celsius, correct? Okay, so here's the thing, okay? We started out friends, Char. So the thing is, okay, take for example, you have your water. We added glucose, we added BUN, okay? We added also sodium, which are osmotically active components. What will happen? It will increase its osmolality, okay? And you would also realize that water with those impurity would take longer time or higher temperature before it boils and lower temperature, lower than zero degree Celsius before it freezes. So naging gets yung point, the more that there are those particles already, the higher the osmolality will be. And so the changes in the colegative property will also happen, okay? So is that clear, chat box? Is that clear? Are we clear with osmometer? Okay, thank you so much, okay? Okay, moving forward, let's go now to your electrochemistry. Funds familiar, okay? Sounds familiar, maybe some of you are having a deja vu, okay? Yes, these are the things that you summarized in koale before. Now we are explaining it and maybe just to remind everyone, why am I focusing now on terbidimetry down to mass spectrometry? The thing is, most of the time, they are the topics that are neglected because we are focused on spectro, flame, fluoro, chemi, and even AAS. AAS should be there, okay? AAS should be included there, okay? Included yung AAS hat. Most of the time, sa sobrang haban ng mga topics na yun and it would really take time for students to grasp the topics. It would actually cover the entire time that should be allotted for terbidimetry down, okay? For this topic. So kung mapapansinin ninyo, sabi ko I opted to really cover everything so since we're ending the semester by December, to cover everything, na ayoko naman na naloloka kayo. That's why I allotted your laboratory time for the topics number one to electrophoresis plus AAS, okay? So let's move forward to your electrochemistry, okay? For electrochemistry, electrochemistry involves now the measurement of your current or your voltage that are generated because of the activity of your specific ions. So specific ions give of specific current or voltage depending on the analytic technique that we are using. So right now, we will be differentiating four, actually plus one, plus conductance. We will be differentiating five electrochemistry techniques. One is potensiometry, colometry, voltammetry, amperometry. Do not be afraid, okay? Do not be afraid because most of these are actually terms. That's why I need you guys to read and study. And just a matter of your midterm exam will be the last week of October. So we need to really be ready. We really need to study harder, okay? It's midterms. It's time to polarography not kasama. The one that are here, Angel. So moving forward, okay? So analytes undergo electrochemical changes, electrochemical reactions specifically, oxidation and reduction. So when a particular ion reduce or oxidize, they will now affect the electrode in our instruments. And in measuring your current and your voltage, these are actually generated by the activities of specific ions. Yung ngayong sinasabi ko kanina. So specific ions have their own potential energy, the current and the voltage that are being generated. So let us go now to the first one which is your potensiometry. In potensiometry, okay? In potensiometry, the measurement of potential or the voltage between two electrodes in the solution is the basis for the variety of procedures for measuring analyte concentration. So meaning to say, we have two electrodes. In what are those electrodes? We have your reference electrode and we also have your indicator or your analytical or your measuring electrode. We'll get to that later on. But please write this down, everybody, that in potensiometry, we are able to measure the concentration of a particular substance by measuring the difference between the two electrodes. The reference and the indicator, the difference between the two that will now reflect the concentration of the ion that you are measuring. Are we clear? So let us go now to the two electrodes. In potensiometry, later naman, lalabas pauletyo. Okay, let us discuss the two electrodes. We have your reference electrode and we also have later on your measuring electrode. So your reference electrode is, of course, an electrode. An electrode that is constant and completely insensitive to the composition of the solution under study. So what are we trying to say here? That when it comes to reference electrode, the reference electrode should not be affected by the one you are measuring. So meaning to say, if I put in electrical current in the system, if I put in an electrical current in the system, the reference electrode will not get affected by whatever substance I am measuring. For potensiometry, please write it down. In potensiometry, we are usually using this in the measurement of your pH. In the measurement of your pH or, more specifically, the measurement of your hydrogen ion. The measurement of your hydrogen ion meaning to say, since we are talking about pH, the reference electrode should not be affected by your hydrogen ion. Are we clear? The reference electrode should not be affected by your hydrogen ion. So what should be the ideal characteristic of your reference electrode? Number one, it should conform to the NERS equation. Nakikita nyo nga ba kung paano ko tatanongin sa quiz? All of the following are true about a reference electrode except, okay? So dapat kilalan nyo nako by this time when it comes to the quizzes. So next, it should exhibit a potential that is constant with time, okay? Number one, reference electrode should be constant, okay? Because if there is fluctuation with the potential or the voltage within your reference electrode, it will affect your measurement, okay? It will also affect our measurement. So having said that, your reference electrode should also be able to return in its original potential after being subjected to small current, okay? So dapat nakakabalik siya agad. And of course, last but definitely not the least, there should be, it should only exhibit little hysteresis or lags with temperature cycling. What do you mean by temperature cycling? Your reference electrodes because your electrodes will be subjected into different current, electrical current, different level, so to speak. And different level of electrical current also mean different temperature. So meaning to say, magiiba-ibayong temperature niya and that reference electrode should be able to cope up to that or adapt to that fast, which just little lag, okay? Little, small lang yung lags nila. Are we clear with the reference electrode? So that is the reference electrode now. So in the laboratory, we usually use two major reference electrodes. One is the mercury-mercurious chloride and the silver-silver chloride reference electrode. So let us differentiate one from the other. So mercury-mercurious chloride is a frequently used reference electrode, although the disadvantage here is that it is low to reach a new stable voltage, okay? So meaning to say, yung hysteresis na pinag-uusapan natin, it would take time for your mercury-mercurious chloride to move into one voltage to the other, okay? One voltage to the other, okay? It would take time for it to adjust. Aside from that, okay? Aside from that, not only does we have issue when it comes to it changing in a new voltage, your mercury-mercurious chloride is also unstable at 80 degree centigrade. Wala bang magtatanong sakin? So probably you haven't did your advanced reading. So here, okay? The thing here is that in bishop, okay? Those who are using bishop, this is the one written, okay? But when you go to Henry's, Henry summarized it na, okay? Because ang nakalagay kay Henry's is that greater than 60 degrees hindi na kaya ni mercury-mercurious chloride. Let me just say it again. If it is greater than 60 degrees, higher than 60 degrees, mercury-mercurious chloride will already become unstable. Sir, what are we going to follow? We're going to follow what? We're going to follow what reference? We're going to follow the Henry 60. Well, eventually ganan din naman yan, di ba, above 80, ganan na rin yan. 80 degrees, okay? Higher than that, ang stable nasi mercury-mercurious chloride. On the other hand, we also have another electrode which is your silver-silver chloride. Your silver-silver chloride is another reference electrode that is being used. But this time, here's the thing with silver-silver chloride. It has, it can be used at higher temperatures, okay? Because it can go up greater than 60 degrees and even greater than until 275 degrees centigrade, okay? And aside from that, your silver-silver chloride are also compact electrode comparing it to your mercury. Am I clear? Clear po tayo. So in electrochemistry, ganito yung mga yari. Electrochemistry, you will try to differentiate potensiometry from columnetry, from ISE ang pyrometry, voltametry. And within potensiometry, punta lang mo na tayo po eq potensiometry, you should be able to differentiate what reference electrode is and what a measuring electrode is. And in the reference electrode, there are two, okay? Your mercury-mercurious chloride and your silver-silver chloride, okay? And by this time, I just want to congratulate everyone because you made this far and alam ko, there are a lot of things that you're studying right now and looking back on my third year, nakaka, ano, nakaka, nakaka, ano, nakaka, nakaka mabombard ng maraming information and I know that you're going through that right now, but I know you will, ano, you will get through this, okay? So, um, let me just clarify, mercury is unstable above 60 degrees, but silver-silver chloride is stable, stable, stable above 60 degrees, okay? Stable until 60 degrees, all right? So, I hope that's clear. Okay, let's move forward. We have your reference electrode. Now we have your measuring electrode, also known as your indicator electrode, also known as your analytical electrode, okay? Analytical electrode. Sir, can we, sir, pag sinagot ko ba indicator? Sinagot ko analytical, sinagot ko measuring, yes, kahit anong isagot mo kapatid, tama yan, okay? So, measuring electrode, sir, it was mentioned here that it should be sensitive to hydrogen ion, because in this case, what we are measuring is what again, your pH, pH or your hydrogen ion concentration. So, meaning to say, opposing now, okay, comparing it now to your reference electrode, your measuring electrode should be sensitive to the analyte that you want to measure. Your, um, your reference should be insensitive. It should not be affected. It should not be affected by the analyte that you are measuring. So, here's the thing now. In potentiometry, the concentration of your ions in the solution can be calculated how from the measured potential difference, the measured potential difference between your reference electrode and your measuring electrode. And the measured cell potential, okay? The measured cell potential is related to the molar concentration by nurse equation. So, what, sir? Medyo na lost ako dun, okay? Can you please repeat that? Ito lang yung sinabi niya. The difference or the measured potential, the measured potential that we came up with the difference between reference and the measuring electrode is equal to the concentration of your ion, okay? And if you want to, um, how is that depicted that is depicted by your nurse, uh, nurse equation. Are we clear, chat box? Okay. So, ayun ha. So we're moving now, okay? Potentiometry, okay? Potentiometry, difference ng reference electrode and ng, um, measuring electrode is equal to cell potential which is directly proportional to your molar concentration. Let's move forward to your ayun selective electrode. Ayun selective electrodes, okay? Is an electrochemical transducer capable of responding to one specific ayun. One specific ayun, sir? Yes, one specific ayun. So, meaning to say, we have one ISE only dedicated committed to your pH, one for your sodium, one for your calcium, and one for your potassium, okay? So this ayun selective electrodes, okay? This ayun selective electrodes are very sensitive and selective on the ayun that they are measuring. So meaning to say, they only want to measure what they are made to measure, okay? Hindi sila yung flexible adap kung ano yung binigay ano yung unang instruction yun na yun, okay? So there are different membranes, okay? Your ayun selective electrode are made up of different membrane and those different membrane set apart one ayun selective electrode from the other. So meaning to say, aglass, ayun selective electrode are specific and sensitive measurement of hydrogen ayun, okay? Sir, baka may nalili ito sa inyo, no? Bakit tinasin nga binis, sir? Hydrogen ayun tapos bag pH? Guys, pH is the pH measurement is equal to hydrogen ayun. Nagigets nyo ko. Yung hydrogen ayun natin, yung hydrogen ayun concentration is expressed by pH. Clear? Chat box? Okay, wala pang sumagot. Okay? I'm just making sure baka may nalulos sa atin. So glass, ayun selective electrode are dedicated for your pH. Malaya, ah, different na to. We're already on the ayun selective electrode na. Ayun selective electrode. Another, okay. So ayun selective electrode. So where was I? Okay, okay. Here I am. In the ayun selective electrode, in the ayun selective electrode, we have now the glass aluminum silicate. The glass aluminum silicate is dedicated for your sodium. Your ayun selective electrode, dioptylphenylphosphonate is for your calcium. In your ayun selective electrode that has a valinomycin membrane is specific for potassium. Okay? Guys, spent time to write this for because not only that it is in your book, lumabas na po ito sa board exam. So meron ka nang 4 points, ka patiid. You are 4 points ahead of being a top-nature on your September on the, ano, the 2022 board exam. Oh, yes naman, di ba? So moving forward now for after your ayun selective electrode, we go now to your electro chemistry, which is your columnetry. In your columnetry, we measure the quantity of electricity expressed in columns that is needed to convert an analyte to a different oxidation state. Different oxidation state. And usually in columnetry we are measuring your chloride ayun in your serum, in your plasma, in your CSF, and even in your sweat. Even your sweat sample. Yes, literally all fluid that you can imagine that is coming out in the human's body is being measured by a medical technology. Sir, even my tears, can be a sample for your bacteriology if you may want to add your tears. So moving forward now, sir, bakit po na ka bold, bakit po na ka underlying sa sweat test, what condition is it selected? Why do we measure chloride in sweat? What are we trying to diagnose or what are we trying to what disease are we looking for? Anyone from the chat box? Anyone? Okay, very good. Okay. The correct answer is what? The correct answer is okay, the correct answer is cystic fibrosis. Cystic fibrosis. CSF is not a disease. CSF is cerebrospinal fluid. Cerebrospinal fluid. So in sweat test, we are measuring chloride in your sweat to diagnose or to determine your cystic fibrosis also known as your myocovisidosis. Myocovisidosis. Sir, spelling, cystic fibrosis. Guys, eto ha, once that you hear whenever you hear whenever you hear chloride sweat test lagging cystic fibrosis. Okay, cystic fibrosis or your myocovisidosis. Myocovisidosis. Cystic fibrosis. Cystic fibrosis. Cystic fibrosis. Okay, it's on the chat box already. Myocovisidosis. So moving forward now, that is for your that is for your column 3. So let's try to compare potensiometry difference between your reference and your measuring electrode. The difference between the potential on your column 3 the amount of electricity that you need to oxidize a particular sample. So in this case, for example, for chloride it will now it will now combine with your silver ion. So the amount of the amount of silver ions released by ionization is directly proportional to the amount of chloride in the ion. So kung gaano ka daming silver yung kailangan mong i-release for it to couple with your chloride to become silver chloride, that is also the amount of your chloride, obviously. So that is for your column 3. Let's go now to your amperometry. What is amperometry? Amperometry this time is the measurement of the current flow produced by an oxidation reduction reaction. Are we clear? Let me just try to differentiate column 3 from amperometry. Amperometry. So here, here you applied electricity to oxidize so what you are measuring is how much should I need to apply for it to oxidize. In amperometry there is oxidation there is reduction in the system the thing that you are going to measure now is the current flow from the oxidation reduction. Nakikitaing difference can you see the difference guys in the column 3 ikaw nagbigay ng electricity for it to oxidize. In amperometry, there is already oxidation, there is reduction in the system. What you want to measure now is the current flow produced by the reaction and now the change in the current is measured through the cell and the change is directly proportional to the partial pressure of oxygen present in the specimen which lead me to my next point that amperometry is usually being used in the measurement of your partial oxygen. Are we clear? Hello? Are we still clear chat box? Okay. So that is for amperometry that is for amperometry. For example there is already a constant constant current flow in the system but after oxidation and reduction, there will be changes and that is what you want to measure that is what you want to measure again directly proportional to the oxygen level. Now we go to voltometry. No, we go to voltometry. At voltometry, it is a method now in which a potential is applied in an electrochemical cell and the resulting current is measured. The resulting current is being measured. The advantages guys of voltometry is that it's very sensitive and are capable of multi-element measurement. Unlike your ISE, one is to one specific is to specific. This one, your voltometry own is very sensitive and can measure multi-element can do multi-element measurement. And aside from that, it also consumes only a minimal amount of analyte. So, kunting sample ang kanyang consume. At voltometry, it is used in the measurement of heavy metals such as your lead which is your anodic stripping voltometry. Anodic stripping voltometry. I hope you guys are copying the power point and also reading your book side by side because I did technically, my power point is summarized in yung book. Nandyan po, chapter 4 and page 46 of your Henrys. So, let's go now to the last one. That is voltometry. Let's go to the last which is your conductance. Your conductance is an electrolytic conductivity is a measure of the ability of a particular solution to carry an electrical current. Conductance is the opposite of resistivity or resistance. Activity is the opposite of resistivity. So, in this particular in this particular technique, conductance is used in what? In measuring water impurity. So, the appear water would be free from ions, would be free from, they would be free from other elements. Meaning to say, it will not become a conduct. Yung hindi siya, it will not become a conductor. So, it can also be used in measuring your urea. And these are being used as detectors for your high performance liquid chromatography which will be discussed even next meeting. And also in cell counters and capillary electrophoresis. So, that is for your conductance. So, to wrap it up in electrochemistry we have five, your potentiometry we have your we have your potentiometry, kolometry, amperometry, voltametry and conductance. Okay? Voltametry and your conductance. Let me just correct myself. Okay? Let me just correct myself. Milaya, okay? Hi-O-1 Hi-O-1 pala ito. Hi-O-2-O-3. So, Milaya have a question. As or under, yung measuring electrode, yung measuring electrode natin is for your potentiometry. Potentiometry siya. Okay? So, I'll entertain your questions later. So, let's move forward to your chromatography. Let's move forward to your chromatography. And by the way, chromatography, have you done chromatography before in your clinical chem? Okay, may isa paulit question nakakatawa. Ibig sabihin nandong kayo sa Henry's kasi nakita yung I opted not to include impedance because that will be discussed in the automation and hematology. And even flow cytometry. Are we clear? So, impedance and flow cytometry are actually more specific for your more specific for your clinical hematology. The five are your ano? Potentiometry, kolometry, amperometry, voltametry and your conductors. One, two, three, four, five. Okay, let's go now to your let's go now to your chromatography. So, chromatography is a technique that separates mixture into individual components, okay? And it is used to separate complex mixtures on the basis of different physical interactions between the individual compound and the stationary phase of the system. So, later on, we'll be discussing basic components of chromatography. But let me just let me just say what chromatography does first. So, chromatography, take for example I have a water, I have a solution rather, I have a solution that contains sodium chloride and sodium potassium in chloride. Or take for example merong ano siyang merong siyang different components like drugs or different therapeutic drugs. And I want to measure each one of them, okay? I want to measure each one of them. I can perform chromatography. So, number one, chromatography separates your mixture. Take for example you have a homogenous mixture. You are able to separate each component from the other using chromatography. But not only that, okay? Not only that you separate the mixture according to its component but using chromatography you are also able to identify what specific compound that is, okay? What specific compound that is which will be discussed in matter of three slides from now. And aside from that, in chromatography okay, not only are you able to separate them not only are you able to identify them but eventually you are also able to measure them, okay? You're also able to measure them, okay? So, that is what chromatography is all about. So, let us go to the basic components first, okay? The basic components of your chromatography are first your mobile phase. Your mobile phase is a solvent or a mixture whereby your sample is added, okay? Your mobile phase. Take for example ito yung ano ito yung, take for example I am the sample. The mobile phase is obviously the mobile, okay, yung mga sasakyan that would and the stationary phase is where the mobile phase flows. So, meaning to say ito yung kalsada, okay? So, take for example, okay? We want to we want to identify one from the other we want to identify take for example, there are three students in a mobile phase, okay? There are three students I am making an analogy para may imagine ninyo. There are three students okay, there are three students hindi natin alam kung saang school sila hindi natin alam kung saang school sila tagaroon, okay? But because of but because of the chromatography, we will be able to identify them. So, all of them are on the same mobile phase and then all of them will be passing through the same stationary phase, okay? Take for example, do lang tayo sa u-belt para tabi-tabi yung school okay, you would be able to identify them because one will get off spanya, U.S.T., one will get off at gastambide, your U.E., and one will get off moraita in F.E.U. So, in that case now, you're able to identify among those students U.S.T., Tomasyan, Tamarau and U.E. Redwar yours. So, parang ganun din sa mga components natin. You're able to identify which one is cocaine, you're able to identify which is which one is degoxine, which one is ampetamin, which one is tetrahydrocannabinol because of their appearance in the chromatography. Are we still clear? So, that's separated now after from the mixture once that they are separated they will now be called LOA. So, let me just show you a picture of an LOA. Okay? This is an example. So, as you can see all of these things are belonging to the same same solution tanina. But now, we are able to separate them. So, this B1 here, B2 here, the C1 the E1 here, all of them are signifying an individual analyte. Are we clear? Chat box? Are we clear? So, all of those components okay, all of those components are separated. Okay, ay walang sumasago sa chat box. Okay? So, that is for your chromatography. Okay? That is for your chromatography. So, moving forward now moving forward how are these components separated from one another? One big factor there is because of their polarity. Okay? One reason there is their polarity. The attractive forces that encompasses the total interaction of the solvent molecule meaning to say the mobile phase with the sample molecules and also the attractive forces between your sample and the stationary phase. So, meaning to say the solution that passes through your mobile phase there is an effect causing its separation already and as they pass through the mobile phase okay, as they pass through the mobile phase they are also separated on how they interact with the mobile phase. Okay? How they interact with the mobile phase. That is how they are separated from one another because of their different polarities. Okay? Because of their different polarities. Sir, now that they are separated, I want to identify them. How can I identify them? You can identify them by using their, by identifying their retention time. Retention time, you have a formula there. Retention time is the time it takes for a compound to elute. Okay? For a compound to elute or for them to separate. Okay? So you will now ang inay naman, sorry. Okay, you will now identify okay, you will now identify a particular compound because of their retention time. So, the value is a characteristic of a compound and it's related to the strength of its interaction with the stationary phase and that mobile phase. So, meaning to say, the retention time, therefore can be used to determine the compound's identity. Are we clear? So, when I ask, how can I identify a compound's identity using chromatography? The answer is because of its retention time. Are we clear? Sir, may job pa ulit po, sir. Yung kasi may jila wala po ako. Okay, so again, okay? So, we are able to identify the identity of a compound because or due to their retention time. Okay? Due to their retention time. Moving forward now, I have only like a couple of four slides and this will be the final 10 minutes and the final coverage for the quiz on Monday. So, we have four modes of operation. Okay? We have four sir, ano po yung mobile phase ng mga analyte? The mobile phase of an analyte are the take for example, it can be usually solid siya. Eto later. Merong paper, merong merong paper, merong gel, but usually in general most of the mobile phase are solid. I want you to look, I want you to imagine electrophoresis. Electrophoresis are somehow similar with chromatography. It's just that electrophoresis has current chromatography you have the mobile phase that drives the component into the mobile, into the stationary phase. Okay? So, ayan. So later on the different chromatographic techniques actually differ on the mobile phase rather than the stationary phase. Merong tayong gas chromatography, merong tayong liquid chromatography. So, I'll explain that in the succeeding topics. Let me just finish this first. So, in the chromatography we have mode of separation that is the adsorption. First is the adsorption the partition, steric exclusion, and ayon exchange. So, let me go first to adsorption. Okay? Adsorption. So, adsorption is based on competition. So, competition between the sample and the mobile phase. They compete in the absorptive sites of the stationary phase. So, take for example, it is your Wattman paper. Okay? Your Wattman paper. Take for example, you have here your alcohol and your water. You want to separate one from the other. So, they would compete from one another. So, the molecules that is most soluble again ha, most soluble, ibig sabihin it really did again talk about the interaction of your molecule and your mobile phase and the interaction as well of your sample or your analytes to the stationary phase. So, nakatepende the solubility of your molecules okay will determine who would move faster in the in the solution. So, what do i mean by moving faster? So, as they move faster parang ito they would now be separated from one another. This is your B2, this is your B1. This one here are, we can say that this was absorbed easily because it has higher solubility or it is the most soluble between the two compounds. That's why it is the one that is up here. And moving forward usually usually your adsorption usually your adsorption is seen in liquid solid chromatography. Liquid solid chromatography. Moving forward now, let's go to your partition. Your partition, let me just clear the air immediately. The partition is usually done or usually the separation technique in your liquid-liquid chromatography. Sir, liquid-liquid chromatography, what do you mean by that? Meaning to say liquid's immobile phase liquid then see stationary phase. Liquid-liquid. So let us go here. So separation is based on relative solubility again solubility again of your compound inorganic and acueyo solvent. Inorganic and acueyo solvent. So what happened here is that you will be adding an immisable solvent. Usually an organic solvent. So meaning to say nonpolar. What will happen is that after adding that, sabi natin immisable, it's like water and oil. They will not mix. But you are pretty much aware that in that acueyo solution there are some organic or some nonpolar compounds. And what you want to do is to extract that which leads me now into saying that partition is also extraction. Partition and extraction are one in the same. So what happen is that molecules containing a polar in a nonpolar group in an acueyo solution when an immisable organic solvent is added and then what will happen what we will do is to shake it. You will shake it and then depending ang dinagdag mo is an organic solvent, the organic compounds in the solution will move into the organic solvent. The one that you added. Take for example, nag-add ka ng immisable organic compound. Yung organic compound na nang dun sa solution mo will be migrating to the immisable. When it gets back, it will now move it will now extract the organic from the solution. So that is the partition. So usually liquid-liquid chromatography liquid-liquid chromatography. So let us move forward. We only have four minutes. So we also have here your steric exclusion. Steric exclusion is similar to your adsorption. What is the difference? Not only is it affected by its solubility but also affected we are trying to differ. Sir Panapag, they have the same solubility. How can I differentiate them? The answer is steric exclusion because it is now based on their size and their shape. So it is a variation of a liquid-solid chromatography. Usually the solid stationary phase is a gel. So it is all known as a size exclusion or molecular exclusion or molecular sieve chromatography. Yung ano niya. Either size exclusion chromatography, molecular exclusion chromatography or molecular sieve chromatography. How does it look like? Ganyta siya. This is your size exclusion chromatography. Can you see it? Chat box. Please confirm. Take for example this ions here, the one in blue and the one in red are of the same solubility. So you would have a hard time differentiating or separating the two but it was answered by size exclusion chromatography. Your steric exclusion whereby smaller compounds will migrate faster into the stationary phase. And the larger the larger molecules would take longer time would take longer time for them to migrate into the mobile phase. Meaning to say, again you're able to identify them because of the difference now of their retention time. Are we clear with that? So ganan siya ha, it is composed of gel. The last but not the least is an ion exchange chromatography. Ion exchange chromatography solute mixtures are separated by magnitude and charge of their ionic species. So stationary phase are your resins. Your resins can be cut ion or an ion resin. So let me just try to differentiate the four ways to separate using your chromatography. We had your adsorption we had your partition we have your steric exclusion and then we have your ion exchange. Here's the thing. So for adsorption chromatography we separate them according to their solubility. How fast are they absorbed from the mobile phase. Your partition on the other hand is based on their relative solubility. We have two phases one that is the solvent that you are adding is imesable to the other. So meaning to say hindi sila nagmimix. What you will do there is to separate the organic from the inorganic from one another and that is done through partition chromatography. On the other hand, we also have your size exclusion chromatography or your steric exclusion whereby you separate molecules not only based on their polarity or solubility but also based on their size and their shape. San Thomas helpful. It is very much helpful if both of your sample has the same absorption had the same solubility and now you want you know that there are two different components in there and you want to separate that what you're going to do is to separate them based on their size and their shape using size exclusion chromatography. And last but not the least is your ion exchange chromatography whereby you are trying to separate your molecules you separate your compounds based on what? Based on their ionic charges. So if you are having an ionic if you are having an ionic resin, you will now be attracting of course the positively sign the cut ion if you have the cut ion you will be attracting now in your will be separating now the anion. So that is for the methods of separation. The rest of the chromatography will be discussed on Monday so your quiz will be from spectrophotometry up until ion exchange chromatography. So that would be all for today. Thank you so much. So I'm stopping the recording now.