better performance for DPPH assay

Today, was thought to be lazy. But then…ah, skip all those craps…lol

I remembered boss wanted me to run the standard antioxidant compound, BHA (butylated hydroxyanisole) as reference. But KC was not here today. So asked help from another senior, Lisa to call KC to get it. Lastly, I got the BHA, and get the amount I wanted. Wow, the smell…ehem…@@, my lab coat now add on another smell…=.=

Boss insisted me to run 2 compounds today, which were BHA and another compound with functional group of O-methyl group. When the time I got the BHA, was already near noon. So went to have lunch then come back to do. Prepare the stock solutions of the 2 compounds. I was “praying” that, no more errors, please. At the meanwhile, I was very careful in every steps. I don’t want boss to show me the “black face” again. Ok, when start to do the DPPH assay, everything is rushing. Rushing with time. I was competing with the time intervals.

After I prepared the stock solutions, I ran to downstairs to initialize the UV-VIS spec. Then when was near the 30th minute, got everything ready. My precaution steps were good enough, the BHA standard’s readings were nice. Can see the obvious and distince colour change of DPPH solution from purple to light purple or from purple to light yellow.

When running the sample that needed to be tested, my control’s reading is lower than the other concentration’s…what’s wrong. Damn it, I have to cheat a bit…(don’t imitate me). However, if excluded that, my result was nice. Hooray!!!

So damn busy and rush today…but I think I am calm enough…^^ If not, I can’t get the results that supposed to be.

Felt happy for myself today… because boss smiled and explained to me the results that I got.

rush with time

Today, run DPPH assay on one sample alone (of course alone), but this is the second time I do it, so I was hoping I am not nervous and mess around that time. This time I know I need to be very very careful. Boss also there to look around and clear some things, but I am not that nervous anymore, in fact, I am calm that time…haha ^^ Oh yeah!

Make my pace as fast as possible to dry all the glasswares needed. Wow, for just to test one compound, I need more than 10 of 10mL volumetric flask, sample tubes, sample vials, a lot of micropipette tips, a lot of beakers to stabilize the flasks n tubes, and my energy to shake vigorously to mix the compounds in the flask.

Ok, use samples prepared to make up initial concentration: 500ppm. Then the next is stock solutions preparation which are 250, 100 (x2), 50 (x2), 40, 20, 10, 5, 2.5 and 0 ppm in concentration. 0 ppm is just consists of DMSO and DPPH. Well, DMSO is the solvent used to dissolve the sample. DPPH is the free radical. Firstly, 5mg or 0.005g of sample is dissolved in DMSO and top up with DMSO to 10mL in volumetric flask. Then, start the using of sample tubes…from 250 ppm to 2.5 ppm. All has to be precise…if not I will get scolded by boss…lolz. Ok, prepare stock solution. It took me a lot of time…!!! After prepare that, it is almost 1pm. Went to have lunch, to save time, I only get 100 plus.

Next is the step of preparing DPPH solution. Got the reagent from the refrigerator, take a sample vial, a beaker and one small spatula to get DPPH measured. Dissolve the 1.97 mg of DPPH powder in methanol then transfer to 50mL volumetric flask then top up with methanol then shake vigorously to mix. Cover the whole flask with aluminium foil as stated in manual and keep in dark. In the meanwhile, prepare the samples for DPPH assay. 1mL of sample solution plus 4mL of DPPH solution in sample vials. Well, DPPH solution is added in time intervals of 3 minutes between each concentration. Aiks, why I am not thinking of this, while prepare stock solution, I can prepare the solution in sample vials to avoid from using so many sample tubes (because end up I need to wash a lot of glasswares…lol).

DPPH assay can be run when I am ready. Get a stopwatch, prepare sample tubes to allow easy filling of DPPH solution to sample vials by micropipette. Get set, ready! First 4mL of DPPH solution is added to first concentration, 2.5ppm. This is the challenging part. Within time intervals I need to fill the particular amount of DPPH solution. After I fill the last sample vial of 250ppm concentration, I need to get the UV-VIS spectrophotometer set up, initialization, the cuvettes, distilled water, tissue paper, all get ready within 15 minutes. 5 minutes before the 30th minute, I need to get ready to get all my things to downstairs to the UV-VIS spec there. When it is the 30th minute, the 3 minutes time intervals is rushing me. Get the readings in triplicate. Wash the cuvette for the next concentration. Blank for this test is pure DMSO.

When I measure the first concentration, which is 0ppm, it showed 1.254 for absorbance, I was shocked there, because so far I do the spectrophotometric assay, the absorbance seldom reach that high. But this one, is high until second last concentration. I was a bit panic that time. Thought I was wrong. Then I asked a senior, luckily she done DPPH assay before, she said my result was ok. Fuh, relieved. It would get that high absorbance, as long as not reach 2.xxx for absorbance then is ok.

Ended up the absorbance of 5 and 10ppm got problem. Oh gosh!!! I already very careful. Maybe I shall be even even more careful next time. Tomorrow will be run with a standard, which I need to do 2 times of absorbance reading, standard and sample’s readings. Will make it better and more efficient for next round! I can do it!!!

random

Woke up around 7am, was very reluctant to wake up…feel like I just sleep for not long… >.<

Around 8 something, prepare myself to go to church.

Then is events in church…

Back around 11:35am, rest a while, then clean my room. Then after that (actually I cleaned partially -> is scheduled), then I napped not long after that. Maybe 1pm something like that, woke up by a call at 2pm? I think so. Then another call at 3:26pm, I woke up and went to collect my clothes and hanged it back to place, then I continue nap. Woke up by a call at 5:17pm, prepare myself then went to dinner.

Round and round at new town, damn…nothing to eat… Ended up went to old town, eat some light ones, kuey teow soup, both my friend and me with a lot of fish balls…lol. Then chatted about our stuffs. Round again to search for shop mending shoes then went to Watson.

Back home. Feel direction-less. I felt so tired, but I scared to lie on bed, scared I sleep like how I did in the afternoon. I see myself at the mirror, my eye bags… @@ and the swollen eyes… What happened to me? I thought I rested enough…@@

>.<

Today, after the high blood pressure and nervous and tension and whatever by my boss, I felt very hungry at lunch time.

Then I met my friend. She said long time didn’t see me around then said I looked nicer (变美了) –> 我差点“吐血”. I am not 暗爽 here. NO!!! Because I don’t agree… >.< But her first response seeing me is: wah! 做么这样瘦了? My response was: Huh? 有瘦到咩? I think shirt that I wore today is quite big…or if I really looked thinner, that means…?!

For being asked on why getting thinner, really no comment. I can only say that I got 暗爽 and also thinking of other thing…which won’t be stated here…

nice day

Reached campus around 9:45am. Then was thinking what to do next. Today sure need to perform DPPH assay, and need to wait for my boss in order to start it. Not long later, saw my boss arrived campus. Then later, he came to me and said the best UV-VIS spectrophotometer is in malfunction state…>.< and asked me to search for lab officer to look for other UV-VIS spectrophotometer (UV-VIS spec.) that is available. Then start my walking and searching for lab officer…a lot of searching and finding in between; cut it short, found other UV-VIS spec that is available.

Start work around 11am, I supposed. Then cut it short the story, finally we (my partner and me) made my boss mad…lolz. But still I don’t consider it as scold…(face too thick), I consider it as…hmm, suddenly I can’t find a word…lol. Mistakes in between. Ok, I know I shall not make mistakes, but this can’t be avoided. I will explain the reason here. Well, almost everybody is scared of my boss, everybody will be nervous when see my boss is around the lab. We are not used to his style, and, and his look and whatever it is, is giving us the force, tension, suddenly both of us nervous like hell. Our hands were shivering when we were doing our work. Blood pressure suddenly spike high, is true, oh gosh, blood circulation would be very good in condition like that. Work that time was preparing stock solution for DPPH assay, so steps: pipette…pipette…pipette…fingers were exercised that time…lol

Then, while my partner was preparing the stock solution for second compound, I was asked to prepare DPPH solution. Dissolve the DPPH solid in methanol and top up to 100mL with methanol in volumetric flask. Fuh, this one easy, but still he is watching…tension…@@ Even I top up the flask with methanol, my hands were shivering…=.=

Ok, the reading by UV-VIS spec. At first the result was weird. Then only we realized the mistake…@@ Nearly got scold again… Never mind. Learn from mistake. Then my job is to deal with the machine and my partner was dealing with jotting down of result (like that the work is finished faster, for that time). Open -> close -> click “auto zero” -> open -> put in blank/sample -> close…these steps repeated for at least 18 times…my hands… And it seems gonna rain that time, and those steps were driving me mad…@@

Then the result looked ok. He smiled when he saw the result. Finally boss smiled. Yeah! Next week, no more trial and error, means we really need to do it in real and no mistakes is allowed. We have “assignment” to do, which is plot the graph and determine the IC (underscore 50) and need to show to my boss. So, assignment ahead!

By the way, actually under his supervision, although is tension and causing high blood pressure, yet can make us produce good work and promote efficiency as boss is a demanding person. His research assistant, KC was kinda like telling and advising me how to do and what to react next time. At least this is a good experience, if without boss’s supervision, we might not produce high efficiency work. Come on, keep it up!!! ^^

today…

Today, I found my boss. Ok, start work on Thursday. Boss really damn busy… ><

My lunch today is one cup of 龟苓膏 and one cup of ice lemon tea. See? Those cooling stuff, to cool my body that is getting very hot.

Generated conversation…

Me : 我吃这些就够了…

Friend : 要死啊? 已经这样瘦瘦干干，还不吃?! 等下变排骨…

Me : 吃了我会很热，虽然我很饿，但我可以忍… 最多我晚餐多吃…

Friend : 唉…

Dinner time. I ate 2 rounds. (Even though I am hungry after heavy dinner MOST of the time –> a big big plateful of mixed rice, I won’t proceed to second round) But today, aiya, pamper myself a bit… One plate of mixed rice and one bowl of 板面… Some friends saw that, then they said that I shall eat that way… (If I always eat like that, I will be bankrupt VERY SOON)… >.<

Lame-ing here, people normally look at me with weird sight with the amount of food I eat… >.<

today is Sunday

Woke up early in the morning, then prepare myself to go to church for worship. p/s: I am not a Christian. Just join to know other things. So far I attended a few worships, I learned a lot. But too bad, before I went to church, on the way to wait for bus, I felt headache. (I think I had not get enough sleep)

The worship started. My head is damn heavy. Instead I looked up to the pastor, my head look down as though I was sleeping…== With that pose, it eased my headache a little better… Then journey back. Sat in front of my laptop a while, then I headed to nap. Again, slept until sweating…ISH!!! 3 hours of nap, my headache gone…^^

Dinner time. Eat and chat with my friend. From still 天亮 (nearly 6) talked until 天黑 (after 7pm)…lolz. We talked about our FYP stuffs. My friend talked A LOT. Well, I am not complaining, I am happy, at least the atmosphere during dinner is not dull…most importantly, is full with laughter, because of some craps we cracked…LOLZ… I did enjoyed the dinner…^^

Continue chat, with another friend. Talked about plans for birthday for a friend. Talking craps again, craps is common among us, if not, is so not like us…lol

Watching photos uploaded by friends on FB. Feel great too because most of them smile. At the same time, multitasking in searching journals. Journals…@@

Overall, today I enjoyed the happy moments although is just simple…^^ Thanks…

-random-

Am thinking of getting myself another belt. Not fancy belt, but FORMAL style belt. Well, I had one, however, suddenly feel like I don’t really keen of that fashion. I prefer those guys style ones…lolz! I mean it, at least I think those guy style ones looked more formal than the one I had now.

Ya, maybe I will consider to get one…lolz

new stuffs to be learned

My FYP title, the previous one (because boss changed the previous title, now doing a new title), was dealing with dihydropyridines, which I still have not much idea on what are they actually, until now. Now, indoles. I only know about Indole Test in Microbiology. What we need to do for new title is synthesize indoles derivatives and use them to test antioxidant and antimicrobial assay. Boss keeps on asks us (my partner and me) to search for journals. The problem now is I don’t even know what is indole, how much can I find?

Ok, I am talking craps. I am not asked to be spoon fed. Ok, in these few days (when I really in mood and trying to bear the hell weather), I get myself the direction to get more info and widen my search. So, it is like studying Organic Chemistry now…haha! Higher degree of Organic Chemistry, as this topic can’t be found in the Organic Chemistry “text books” that we used to refer to. Means, I need help from Google. Hope I get to learn and understand as much as I can for my own sake.

By the way, is fun and challenging! From what you don’t know, self-learn until you know about it. Fun is because this topic sounds interesting (lolz), challenging is because need to study when this kind of moment is supposed to be semester break (LOLZ) Anyhow, is good to make my brain active rather than let it becomes blunt in the hell long break.

Indoles, Fischer indole synthesis, indole derivatives, 2,3-benzopyrrole and 1-benzazole (it’s other names), its antimicrobial activity, its DPPH antioxidant activity, etc… I read some of the journals, the product name of indoles is damn hell long…@@ lolz…

Ok la, I shall ganbatte for the new stuffs!!! Ciaoz…

is kinda satisfaction

Boss this whole week is SUPER DUPER BUSY!!! So, consider a break for his supervisees. Today still going to campus because made appointment yesterday with Chai to meet him for explanation for certain questions of his paper. Well, he explained to me and my response is O.o… finally I got the answer that I was not sure with. Am actually glad and happy because finally my biology subject got at least B. I don’t expect much because I know my limitation (the memorizing power). Then chatted a while before I left his room.

Headed to Dr. Teh’s room for explanation of Metabolism II final paper. Ok, this subject is taken in Year 2 Semester 2. So why now only…? Reasons: the time matter and the reason to approach her. Finally during this short semester, I met her and told her my purpose. She “praised” for my 执着…lolz. Yup, indeed, because I expected higher for this subject but…and, and I like her. That’s why…

Lastly, my questions were solved!!! Feel satisfied. My brain needs to be trained more…lolz