#Chloroformed: (5/10)

[gallifreyanhotfive]

Random Doctor Who Facts You Might Not Know, Part 21

Missy was one of Amelia Pond's childhood therapists. She was looking for the Doctor but came too early.

River Song knows Venusian aikido.

Raj Kahnu, one of the Rani's favorite experiments, considered the Rani to be his mother. The Rani never acknowledged this.

The Master once tried to combat the Third Doctor's Venusian aikido with Martian kendo but was defeated when the Doctor switched to Mercurian kung fu.

The Battle of the Bands Beyond the Stars was an intergalactic televised music competition where the losers get incinerated by a laser cannon. Clara and the Twelfth Doctor were forced to compete after Clara accidentally insulted the monarch. During their episode, a band composed of five different versions of the Master also played (Missy, Crispy Master, Bruce Master, Tremas Master, and Saxon Master), trying to hypnotize the viewers, but they were disqualified after they started fighting each other.

The Twelfth Doctor thinks of his Tenth and Eleventh selves as "Manic Pixie Dream Doctors."

The Tenth Doctor, meanwhile, is very concerned about where the Twelfth Doctor got extra regenerations from and believed he might be the Valeyard.

In his Masterplan Journal, the Saxon Master admitted that living as all the female Masters in woman's clothes felt "strangely liberating" and that he should get more in touch with his feminine side.

You can listen to the Fifth Doctor speak Gallifreyan in the audio Cold Fusion.

The Eighth Doctor meant to visit the opening night of the Braxiatel Collection but was prevented from doing so by the Kotturuh Crisis.

The Doctor spent his 1000th birthday with two broken ribs on board a spaceship.

Chloroform is effective on Time Lords.

The Seventh Doctor once broke the galactic record for continuous spoon playing at 67 hours.

The Doctor once took seven people who were so wealthy that they were bored of their lives to a place he called "Purgatoria." He brought each of them one by one into a separate room where he brought them to their breaking points and let them experience a story from his life first hand.

Galileo Galilei discovered what he thought was a planet between the sun and Mercury called Phaiton. It was actually a stellar manipulator that had been forgotten about by the Time Lords and lost until the Eleven found it.

Gender is subject to fashion trends on Gallifrey. For example, during some eras of Gallifrey, the female gender went out of fashion to such an extent that women were completely absent.

The Doctor used to sing songs about windmills to his daughter Miranda.

Part 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28

[suiana]

dear psycho, what do i do when my crush doesn't like me back? (yandere! psycho x gn! reader) (angstober day 4)

does your crush have a clear disinterest in you? have they repeatedly told you that they want nothing to do with you? are you looking for a way to get them to love you? here's what you should do!

step 1: meet up with them. make sure that they're alone with you! it's the first step to making sure that they fall for you! remember to dress good too! we need to make a good first impression :)

step 2: bring them over to your house. make sure you bait them with a good reason or they won't follow! (tried and tested)

step 3: knock them out. you can use any method but i prefer to use chloroform. I don't like seeing my darling in pain :( and when their eyes roll back as rhey collpase into my arms oh god i swear- (redacted)

step 4: tie them up and watch them for hours as you wait for them to wake up. just... observe them. stare at their pretty eyes, cute lips... just take in every little thing that they do. you're meant to worship your darling after all... hah...

step 5: surprise them with your love! tell them how much you love them, kiss them and drown them in your affection! it helps to prep them for the next step :D

step 6: promise an eternity with them. you deseve to be with them forever. so end their life and preserve their body so that no one else but you gets to be with them. ah... their lifeless body looks so delicate like this... unmoving and lifeless....

step 7: regret everything you've done and hope for a miracle. fuck i miss them i shouldnt have done that. im such an idiot im sorry im sorryimsorryimsorryimsorryIMSORRY

step 8: PLEASE COME BACK I NEED YOU I CAN'T BREATHE I MADE A MISTAKE

step 9: ...

....

.....

kill yourself so you can be with them forever. you deserve it. a happy ever after with your darling in the afterlife :) ... what do you mean no? aren't you glad? this is what you're waiting for, yes? after all, you're a fucking psycho. no one else but darling will love you. so do it, end your miserable life already. darling is waiting for you! regrets mean nothing when darling is there :)

step 10: have a happy life with your lovely darling♡

[staydandy]

The Lost Tomb (2015) - 盗墓笔记 - Whump List

List by StayDandy Synopsis : Wu Xie is an antique shop owner who comes from a family of tomb raiders. As he continues the family trade with his team of tomb raiders, he finds lost treasures of the Warring States as well as the answers to the tragedies of his family’s past. With the help of his grandfather’s notes and his team – his experienced Uncle Wu Sanxing, San Xing’s loyal helper Pan Zi, the quiet Zhang Qiling, and the resourceful Pang Zi – Wu Xie sets out to find the lost treasures as well as the people responsible for the massacre of his family. (MDL) AKA : Grave Robbers’ Chronicles | Grave Robbery Note

Whumpees : Wu Xie played by Li Yi Feng (center forefront) • Zhang Qi Ling played by Yang Yang (2nd from left) • High Shao [Wu Xie's friend] played by Leon Lee (not pictured) • Pan Zi played by Wei Wei (3rd from right)

Country : ���🇳 China Genres : Action, Adventure, Mystery, Supernatural, Bromance

Notes : This is a Full Whump List • Adapted from novel 1 of "The Grave Robbers' Chronicles" (盗墓笔记) by Kennedy Xu (南派三叔) • My favorite episode is pink • Suggested watch order of series (not including movies & spin-offs) : -- 1. Mystic Nine (2016) >> 2. The Lost Tomb (2015) -- 3. The Lost Tomb 2: Wrath of the Sea (2019) -- 4. The Lost Tomb 2: Explore with the Note (2021) -- 5. The Lost Tomb 3: Ultimate Note (2020) -- 6. Adventure Behind the Bronze Door (2024) -- 7. Tomb of the Sea (2018) -- 8. Reunion: Sound of the Providence (2020) -- 9. Reunion: Sound of the Providence 2 (2020)

Related Lists : The Lost Tomb 2: Wrath of the Sea (2019) - Full List • The Lost Tomb 2: Explore With the Note (2021) - Full List • The Lost Tomb 3: Ultimate Note (2020) - Full List • Adventure Behind the Bronze Door (2024) - Full List

Episodes on List : 8 Total Episodes : 10

*Spoilers below*

01 : Wu Xie shot in the arm … punched in the stomach, collapses … High Shao tied up & mouth taped

02 : Wu Xie & High Shao head pain from sharp noise, Wu Xie entranced, kicked into water … Pan Zi attacked & bitten by flesh-eating bug, bug flung onto Wu Xie, bitten … High Shao pukes … Zhang Qi Ling cuts his own hand, collapses … Wu Xie hit in the head & knocked out, nightmare.. Zhang Qi Ling exhausted from blood loss, sleeping.. carried/helped to walk

04 : Pan Zi attacked by corpse-eating bugs, heavily bitten up, helped to walk, bug dug out of a wound (ewww) … Wu Xie attacked & bit (by human)

05 : Dizzy, passes out, was poisoned by bite, given temporary fix … High Shao kicked to the ground … Wu Xie unsteady, helped to walk … dizzy, passes out & falls from a great height, caught … punched, choked, coughing, neck bruised, unconscious

07 : (near end) Head pain, falls

08 : … continued from previous ep. ... Falls, wind knocked out of him by the landing, Zhang Qi Ling cuts his own hand … High Shao tied up … Wu Xie dizzy, passes out

09 : (near end) Tripped

10 : High Shao hit with batons … knocked out with a injection, Wu Xie knocked out with chloroform.. Wu Xie & High Shao kidnapped & tied up.. mouths taped

More Whump Lists for this show: love-me-a-lotta-whump

[ky-randomshitposting]

MacGyver 2025 Writing Promts!

Not all are MacGyver based so if you're not part of that fandom feel free to still use these!

Go wild, combine days, skip days, use these for art instead and whatever you want! Feel free to tag me in any fics or art you make (Even if I'm not in the fandom!)

January 1 • New Years Confessions 2 • Mission gone poorly 3 • Nightmares 4 • Hurt/Comfort 5 • 5-1 6 • Sick Day 7 • Team Meeting Under Different Circumstances 8 • Diner Au 9 • Codex 10 • Whump 11 • The Organization Causes Problems 12 • James Macgyver Being A Shitty Father 13 • Matilda Rescuing The Team 14 • Disavowed 15 • Peaceful Day At The Park 16 • Day Off 17 • Mac + Gun 18 • Working With The Enemy 19 • Dealing With Grief 20 • Movie Theater Au 21 • Insomnia 22 • Double Drabble 23 • Cuddling 24 • Car Accident 25 • Murdoc Returns 26 • Mac Gets The Team Out Of Trouble 27 • Riley Finds Something She Can't Hack 28 • Love Confessions 29 • Kidnapping 30 • Wilderness Survival 31 • Bomb Disarming

February 1 • One Bed Au 2 • Comfort 3 • Alone 4 • Corruption Arc 5 • Fake Relationship 6 • Oversight And Mac Never Meet 7 • Fake Death 8 • Chloroform 9 • Gunshot Wound 10 • Afghanistan Days 11 • Cairo 12 • Jack Being The Team's Dad 13 • Therapy 14 • Poisoning 15 • Swiss Army Knife 16 • Protecting Cassian 17 • Murdoc + Handcuffs 18 • Supermax 19 • Mission Gone Right 20 • Physical Exhaustion 21 • Working Out/Training 22 • Assassination 23 • In Love With Someone Else 24 • Secret Relationship 25 • Unable To Save Everyone 26 • Torture 27 • Self Doubt 28 • Marriage Vows

March 1 • Mac + Chemicals 2 • A Deal With The Devil 3 • Casino 4 • Jack Returns From The Kovac Mission 5 • Mac Asks For Help 6 • Bozer's Cosmetics 7 • Murdoc Saves The Team (Possibly Accidently) 8 • Black Pepper + Wound 9 • Trapped 10 • Under Water/Downing 11 • OneShot 12 • Celebration 13 • Making Bets 14 • Birthday Celebration 15 • Running Away 16 • Forest Fire Verses Team 17 • Song Fic 18 • Bioweapon 19 • Nikki Lies 20 • Thornton Gets Caught 21 • Trying Something New 22 • Watching The Sunset/Sunrise 23 • Jack Saves Mac/Riley 24 • Loud Fight 25 • Nearly Avoided Plane Crash 26 • Bounty Hunters 27 • Amnesia 28 • Doctor's Visit 29 • Tramatic Injury 30 • Corn Maze + Lost + Murdoc 31 • Paranoia

April 1 • Terrible No Good Day 2 • Stalking 3 • Carnival 4 • Hunter/Hunted 5 • Stranded 6 • Heist 7 • Mac Leaves The Team 8 • Break Up 9 • First Kiss 10 • Prison Escape 11 • Black Site 12 • Safehouses 13 • Flash Backs 14 • Slowly Dying On The Phone/Last Phone Call 15 • Funeral 16 • Medical Leave 17 • Mac Breaks Another Phone 18 • Secret Diary 19 • Party 20 • Unexpected Relationship 21 • Coming Out 22 • Canon Typical Violence 23 • Magic Au 24 • Library Au 25 • Hurt/No Comfort 26 • Drunk 27 • Pen Pals 28 • Arachnophobia 29 • Self Care 30 • Terrorist Organization

May 1 • Acrophobia 2 • Ducktape + Panic + Mac 3 • Love Triangle 4 • Fire + Extinguisher 5 • Matty Being Strict/Concerned 6 • Hidden Injury 7 • Rare Pair 8 • Stargazing 9 • Explosion 10 • The Ghost 11 • Jill Morgan 12 • Samantha Cage Joins 13 • Secrets 14 • Undercover Op 15 • Role Reversal Au 16 • Prison Breakout 17 • Bozer + Sticky Notes 18 • Riley + Trapped 19 • Matty Goes Missing 20 • Sandbox Days 21 • Hidden Injury 22 • Mac + Shoelaces 23 • Murdoc On Comms 24 • Experimenting Gone Wrong 25 • Cartel 26 • Corrupt C.I.A 27 • Amber Causing Problems 28 • Hostage Situation/Negotiation 29 • Handling Depression 30 • Trauma Response/Flinching 31 • Day At The Beach

June 1 • Haircut 2 • Dying Saving Someone 3 • Choose A Random Item And Find A Way For Mac To Use It 4 • Jumping Out A Window 5 • Stealing A Painting 6 • Safe Cracking 7 • Losing Hope 8 • Cassian's Summer Break 9 • Nicholas Helman 10 • Jonah Walsh 11 • Flower Picking 12 • James Macgyver Dies 13 • Paperclips 14 • Lost Swiss Army Knife 15 • Riley Or Bozer Tries To Quit Or Quits The Team 16 • Mac Goes With On The Kovac Mission 17 • Collapse Building 18 • Chemical/Oil Spill 19 • Chernobyl 20 • War Injury 21 • Cruel Choices 22 • Leaving Someone Behind (And Coming Back For Them Later) 23 • Riley Hacks The Government 24 • Jill Morgan Lives 25 • Cassian Finds Mac Cool 26 • School Reunion Science Fair 27 • Video Game Night 28 • Breaking And Entering 29 • Jack + Captured 30 • Hidden Fears

July 1 • Jigsaw Scenario 2 • Mac Pretending To Be Murdoc 3 • Riley's Undercover Op 4 • Write A Story With A Word Count Outside Of Your Usual 5 • Bar Night 6 • Talking To The Moon 7 • Late Night Run 8 • Bones (Broken, Found, Fossils ect) 9 • Jack's Medical Leave 10 • Panic Attack 11 • A Favor 12 • Aromantic Or Asexual Character 13 • Nyctophobia 14 • Murdoc's Hit List 15 • The Organization's Assassination Attempts 16 • Mac + Gun Power 17 • Medical Emergency 18 • Late Night Phone Call 19 • Angst 20 • Fake Sleeping 21 • Drugged 22 • Brain Fog 23 • Poor Cooking 24 • The Team Saves Mac 25 • Soul Mate Au 26 • Blizzard 27 • Stressful Drive To The ER 28 • Adopting A Pet 29 • Stabbed 30 • Haunted 31 • Orginal Character

August 1 • Area 51 2 • The Team Finds Matty's Files 3 • Mac Has Trust Issues 4 • Jack & Mac's Co Dependency 5 • Ticking Noise (Could Be A Bomb, A Clock, Who Knows) 6 • Mac Hates Medical Leave 7 • Bakery Au 8 • Murder Mystery 9 • Jack + Bomb + Mac 10 • Happy Ending 11 • Sleepless Nights 12 • Accidental Murder 13 • Matty Retires 14 • Samantha Doesn't Return To Australia 15 • Taking Down A Cult 16 • Alternative Au Where James Never Left Mac 17 • Bozer Finds Out About The Phoenix 18 • Riley Joining The Team 19 • Amber Returns After Murdoc Had Warned Her Not To 20 • A Phoenix Day Off (Probably Interrupted) 21 • Meaningful Coversations (At The Worst Time Possible) 22 • Mac And Riley Fall Asleep On Jack 23 • Internal Bleeding 24 • Wishing Upon A Star 25 • Father/Son Bonding (Jack & Mac or Murdoc & Cassian) 26 • Babysitting 27 • Lack Of Taking Care Of Oneself (Noticed By Another Team Member) 28 • Mac Being Reckless 29 • The Team Travels Somewhere New 30 • Ethan Reigns & Matty 31 • Fainting

September 1 • Breakfast In Bed 2 • Horror Movies 3 • Celebrating A Holiday 4 • Crutches 5 • Suffocation 6 • Bed Rest 7 • Airplane Travel 8 • Hiding In Plain Sight 9 • Crossover Au 10 • Unhealthy Habits 11 • Hand To Hand Combat 12 • Winter Time 13 • Running Out Of Time 14 • A Miracle 15 • Murdoc Leaves A 'Gift' 16 • Secret Service 17 • Night Light 18 • Lying 19 • Feigning Innocence 20 • Leaving Clues 21 • Dramatic Murdoc 22 • Vacation + Cocktails 23 • Anxiety 24 • Heat Exhaustion 25 • Long Term Undercover Op 26 • Lone Survivor/Survivor Guilt 27 • Broken Glass 28 • Ambulance Ride 29 • Flame Thrower 30 • Improper Use Of Chemicals

October 1 • Gasoline 2 • Bazooka 3 • Fireman's Carry 4 • Paperclip Collection 5 • Fascination/Obsession 6 • Taking Apart Electronics And Making Something New 7 • Multiple Love Interests 8 • Ufo 9 • Visiting A Pumpkin Patch 10 • Bozer Accidently Starts A Fire 11 • Nikki Is Not Who She Claims (Again) 12 • The President Is In Danger 13 • Welcome Party 14 • A Letter Goodbye 15 • Missed Calls 16 • Pain Relief 17 • Zombie Apocalypse 18 • Vampires 19 • Reckless Driving 20 • Transgender Character 21 • Moving Away 22 • Teacher Au 23 • Nightshade 24 • Hypothermia 25 • Helicopter 26 • Smoke 27 • Camping 28 • Light bulb + Lightning 29 • Push Pins 30 • Mac + Skyscraper 31 • Halloween

November 1 • Quarantine 2 • Acid 3 • Tar Pit 4 • Murdoc + Sniper Rifle 5 • Abandoned Warehouse 6 • Ice Skating 7 • Exchanging Letters 8 • Doing Drag For A Mission 9 • Insanity 10 • Dealing With Psychopaths 11 • Long Term Memory Loss 12 • Dazed 13 • Loss Of A Friend 14 • Forgetting Your Own Name 15 • Mission Gone Poorly (But Ends Up Fine) 16 • Rescuing People From A Cave 17 • Jill Catches Murdoc 18 • Bomb Hidden In Package 19 • Mixed Feelings 20 • Evacuation 21 • Bookcase Door 22 • Old Photo Albums 23 • Jack's Dog Tags 24 • Facing Their Biggest Fear 25 • Volcano Eruption 26 • Team Evaluation 27 • Held Captive Long Term 28 • Murdoc's Father 29 • Prom Night 30 • Injured Jack

December 1 • Drinking Hot Cocoa 2 • Electrical Fire 3 • Fixing The Toaster 4 • Movie Night (Die Hard Perhaps) 5 • A Letter Containing A Threat 6 • Home On The Range 7 • Jet Engine 8 • Fighting (Verbally Or Physically) 9 • Injured Matty 10 • Flower Picking 11 • Derealization 12 • Hallucinations 13 • Almost Happy Ending (Something Goes Wrong At The End) 14 • Snow Skiing 15 • Caroling 16 • Desi + Mac + Codex 17 • Saying 'I Love You' Too Late 18 • Bonfire 19 • Snow Day 20 • Team Meeting 21 • Ice Cubes + Mac 22 • Chemical Burns 23 • Cassian + Murdoc + Holidays 24 • Holiday Baking 25 • Christmas 26 • Mac Making A Tazer Out Of What He Has In His Pockets 27 • Fire Escape (Comes In Handy) 28 • Mistletoe 29 • Movie Theater Au 30 • Night Vision 31 • Sharing A Kiss On New Years Eve

(Some Days May Be Similar Or The Same Accidently)

[holmesillustrations]

Vote for your favourite, the top 9 will proceed in the bracket. Since theyre all different shapes and sizes, make sure to click into the full views!

Paget Eliminations // Other Artist Eliminations

Full captions and details for each illustration below the cut:

"The old woman faced round and looked keenly at him" Charles Doyle, Study in Scarlet (1888 Ward, Lock, & Co. Novel) Characters: ‘Mrs Sawyer’, Holmes, Watson

[Holmes entering Watson’s consulting-room] Harry C. Edwards, Final Problem (McClure’s) Characters: Holmes, Watson

"Miss Violet Smith, Teacher of Music." FD Steele, Solitary Cyclist (Collier’s) Characters: Violet Smith

Collier’s Cover FD Steele, Missing Three-quarter (Collier’s) Characters: Holmes, Pompey

"She fought her way out again." Arthur Twidle, Wisteria Lodge (The Strand) Characters: Warner, Murillo, Lopez, Signora Duranda

"The fellow gave a bellow of anger and sprang upon me like a tiger." Alec Ball, Lady Frances Carfax (The Strand) Characters: Hon. Phillip Green, Watson, Holmes

"Well, Holmes," I murmured, "Have you found out anything?" Frank Wiles, Valley of Fear (The Strand) Characters: Watson, Holmes

"He was gripped at the back of his neck by a grasp of iron, and a chloroformed sponge was held in front of his writhing face." Alfred Gilbert, His Last Bow (The Strand) Characters: Von Bork, Holmes

"A nick in the parapet, fifteen feet from the body, interested Holmes strangely." GP Nelson, Thor Bridge (Hearst’s International) Characters: Holmes

"Here you are!" he cried, waving a paper over his head." HK Elcock, Three Garridebs (The Strand) Characters: Evans, Nathan Garrideb, Holmes, Watson

"I extend the same warning to you... take your reputed talents to some other field." FD Steele, Blanched Soldier (Liberty) Characters: Holmes, Col. Emsworth, Ralph (Butler), James Dodd

"One night my cries brought Leonardo to the door of our van." FD Steele, Veiled Lodger (Liberty) Characters: Leonardo, Griggs (clown), Mr Ronder and Eugenia

[proxylynn]

Can Peter use chloroform on me to help me sleep if I ask?

Peter: I mean...You do know it doesn't work as fast as movies and things show, right? You have to inhale it for like a good 5 to 10 minutes.

Lynn: Sweets...You tested this, didn't you?

Peter: *sweating nervously* M-Maybe...?

[jamietarttsnorthernattitude]

Answer the questions and tag five fanfiction authors you know!

Tagged by the talented @kvetchinglyneurotic

1. How many fandoms have you written in?

One! Two if you count the truly bad LOTR fic I wrote in high school.

2. How many years have you been writing fanfiction?

I wrote a story or two in high school and then nothing for a long time. I started really writing almost a year ago (just looked published my first fic on 5/21)

3. Do you read or write more fanfiction?

Writing because I basically only read Ted Lasso fan fiction recently and it’s just slowed its output.

4. What is one way you've improved as a writer?

I’ve learned to outline a bit more than just winging it like I did for so long. I’m not sure if there’s a specific thing I’ve improved on, just generally I think (I hope).

5. What's the weirdest topic you researched for a writing project?

How to make chloroform lol but mostly just injury related things, spleen removal, stuff like that. Total normal whump writer things.

6. What's your favourite type of comment to receive on your work?

I truly love any and all comments, I’d be lying if I didn’t say I loved the long, detailed type with quotes, etc but I also love the ones that are just a sentence of reaction like one I got that just said “good fucking fic” and I LOVED that.

7. What's the most fringe trope/topic you write about?

My online circle contains a lot of whump-enthusiasts but probably whump although gen stories sometimes feel very fringe.

8. What is the hardest type of story for you to write?

All of them? I can’t do anything short, which is why whumptober was so hard to get though, I want the whump but then I want the comfort and I just don’t know how to write something concise that is satisfying to me.

9. What is the easiest type?

I don’t know if I find anything about writing easy. Different parts of the story are easier than others but I haven’t found a type that is easy.

10. Where do you do your writing? What platform? When?

Wherever I can, but usually from my couch. I use google docs but have wanted to look into other platforms but none seem to have exactly what I want. Whenever my brain tells me to write.

11. What is something you've been too nervous/intimidated to write, but would love to write one day?

Hockey AU for sure, along with an idea I have for a post-series fic where Michelle and Henry move to London and Ted stays.

12. What made you choose your username?

I was looking for a new one that I could use here and on ao3. When I mentioned this to the brilliant @jamiesfootball they suggested something to incorporate my love of Noah Kahan (many many of my titles are from his songs) so jamietarttsnorthernattitude was born. I’d like to think Jamie and I share a Northern Attitude.

Trying to tag who hasn’t been tagged but these things spread quickly and I might have missed people so trying @fanficfanattic @jamiepoptart @nativestarwrites @lunar-years @providing-leverage and anyone else who would like to play

[gerardway-is-my-babygirl]

Okay, now I need your Renfield headcanons!! And I need to know if you agree/disagree with mine 😄

I hope you know I started stimming so hard when I got this notif and I'm still actively stimming now AGHJ

I absolutely agree he wouldn't keep up with "popular" music now, he's definitely still listening to the stuff he somehow got exposed to when he was younger and hes just chilling with not knowing who *Insert current pop artist here* Is

I also absolutely love him using Dracula's punishments as a way to punish himself!! Also him reading poetry... I would pay good money to hear nick hoult read me literally anything. Even the bee movie script. Also between you and me he DEFINITELY ruins his underwear at least 5 times a week

Time for my hcs!! This Is gonna be a long post

-I Imagine he has never had a phone before and he has no Idea how they work. Yes I know he has a laptop In the original script but that man Is 126 years old he would not know how to work modern technology. Rebecca gives him money to buy a phone and he buys a shitty nokia from 10 years ago because he doesn't like that the keys on a new phone are flat

-He's autistic!!!! He struggles with social cues and loud/crowded spaces, which Is why he likes his job so much. It gives him a peaceful atmosphere to work In and thats why he uses the chloroform to silence his victims Instead of letting them scream. Some stims I think he'd do are: rocking back and forth, dancing, fidgeting with things In his hands, chewing on things and doing his little giggle

-His main love language Is touch. I Imagine he loves feeling Dracula's face and teeth, he'd spend hours running his fingers across Dracula's cheeks If he could. He also enjoys cuddling and holding hands with Rebecca(platonically) because he never got any affection from Dracula.

-His favourite colours are pink and orange(which Is funny considering they are my least favourites) and he owns so much pink clothing. He's always struggled with being traditionally masculine and now that he lives In a day and age where gender expression Is more fluid and colours are less gendered he Is free to wear pink shoes and pink shirts!!

-He struggled with his sexuality for a long time before and even for quite a while after meeting Dracula. He never really loved his wife romantically and It didn't help that they were forced Into their marriage by their parents and societal expectations. They were very distant but Renfield just thought that maybe she wasn't the right one, but there wasn't anything he could do so he just put up with It and repressed his feelings. Then he met Dracula and finally realised hes gay, though he kept trying to hide It from himself until eventually his wife found out about the letters he had been sending to Dracula where the vampire flirted with him. She got angry and that was the last straw for Renfield that made him finally decide to go to Transylvania

-I think he'd really like rats and maybe even get one as a pet after leaving Dracula. He'd probably name It something cute like snuffles or mr whiskers. Maybe even Remy because It sounds like Renny(which I Imagine Rebecca calls him)

-He's VERY possessive over Dracula, to the point where he felt jealousy when he found out about Drac's wives and sort of lost himself a bit which contributed to his short descent Into madness when he was locked up In the asylum

-He loves birdwatching and just zoning out while watching them fly around. He can't do It as much now because he lives In the city but when he and Drac lived In Europe they often had houses on the outskirts of forests so he could do It a lot more back then

-I think after he and Rebecca defeated Dracula he definitely asked to "borrow" a pair of handcuffs and she looked at him suspiciously before hesitantly agreeing. Then he proceeded to get himself stuck In them a few weeks later and had to call her to help because he lost the key under his bed

-I think he'd like baking pastries and lots of sweet tasting things since they remind him of Lillian and how much she loved sweet treats, plus It helps him forget the bitter taste of Dracula's blood. He's very bad at cooking though, like absolutely atrocious. Its like when you make a sim cook something like eggs and toast and they burn the oven to a crisp

-He does crocheting and crosstitching!! He loves making gifts for Rebecca and the DRAAG support group

-Hes a very sentimental person and ends up holding onto things he doesn't really need but cant let go of. He still has one of Lillian's old teddy bears from when she was a baby and he also kept a lot of Dracula's clothes and rings along with his coffin and cane

I'm gonna stop there because this Is far too long but thank you so much for asking AGHHHH the autism cannot be contained

[playstation2horrorgame]

shuffle your on repeatr playlist and tag 10 people

i was tagged by @houndsofdoom

starkiller-bear ghost

2. assimilate-skinny puppy

3. key entry extraction 2: holly wood the cracked- cooheed and cambria

4. jenny, you're barely alive-rilo kiley

5. pet- a perfect circle

6. harvey-her's

7. cop car-mitski

8. said the spider to the fly-the paper chase

9. chloroform girl-polkadot cadaver

10. the small of your back the nape of your neck (the flood)-the paper chase

i tag: @weenhands @pomesa @faerieyuri whoever wants to do this idc

[mmmph82]

Part 5- Kidnapped in a BMW

Abducted by @tyler-lawson

What seems like hours later, I am slowly awakened to being unchained from the cage. His forehead presses onto my gas mask looking directly into my eyes as his breath fogs up the lens of the mask.

“Bad news. I am afraid that your flight as been cancelled…. And you will be staying with me for a while longer.”

The smile on his face means he cancelled it himself, but my cock trying to swell into the locked chastity device is certainly not complaining.

He drags the black leather sleepsack into the closet and begins to unwrap the chains from my legs. I shake my head several times and try to push the gag out of my mouth but it’s locked and molded inside and impossible to move. Once he unwraps the chains from my legs they immediately get pushed down into the sleepsack. I rattle the chains from my wrist in protest.

His head once again leads forward into my eyes of the gas mask.

“Don’t worry those chains will come off in a few minutes. Now we can play the easy way or the hard way. Are you going to be good when I remove the chains from your hands?”

I try to give a very straight face but he knows I’m nothing but trouble. He gets more chloroform ready to add to the rag attached to the hose of the gas mask. I shake my head moaning and pleading through the gag that I would be good.

He puts the bottle down but keeps it nearby. He begins to unchain my hands and slowly and unwraps all of the chain from around my torso and body.

“Give me your hands, nice and slow and do not make any fast moves or you will be sleeping no again in just a few minutes”

I slowly move my arms forward from behind. He turns around to grab some restraints and I know I only have two seconds. I take my padded hands and quickly put them over his head and twist his arms. After a few minutes of scuffling on the floor, I am able to pull my legs from out of the sleeping bag and roll over and shut the closet door and dart into one of the bedrooms where there is a fire door. My padded hands try to open the door when the alarm goes off. It is obnoxious and loud. I realize I may not be able to climb the ladder with these mitts locked on my hands and run back down the hallway about the same time as the closet door swung open. He tries to grab my leg but barely misses. I begin farting up the stairs and running for the main door but it’s locked. I get jiggling it with my padded hands and messing with the lock but it won’t open.

“MPPHHFUCK,” I attempt to say through the locked, molded gag inside my mouth. I am able to pull the gas mask off of my head and throw it on the floor.

Suddenly the alarm turns off but there is no sound from Tyler. I know I am locked inside this house and he could sneak up on me at any second. Either way, I am screwed. I slowly sneak up another flight of stairs and hide in one of the bedroom closets. I try to stand flat inside the closet against the wall and the door. I try to stay as still as possible as five minutes goes by… 10…15. There is still no sign of Tyler.

I slowly emerge from the closet and Tyler is sitting on the bed with the bedroom door shut and locked, and with an evil look on his face. I come over slowly and kneel in front of him and put my padded hands behind my back.

His gloved hands rub my beard and the side of my face as he smiles at me.

“That was … cute. But, did you really think you were going to escape from this house? You know how much I love security and locks.”

He grabs me with his arms and holds me into a cuddle. His gloved hands rub my beard again and the molded gag locked inside my mouth, admiring how I cannot make a sound.

He then quickly takes his gloved hands and pulls my padded hands above my head as he drags me further up the bed and locks them to a chain that is already secured to the bed frame. This was not shocking as I have been locked to this bed before many times and know all about his restraints onto the bed. He then unlocks the molded gag from my head and the large gag slowly pulls out of my mouth. Before I could say anything, a thick leather hood with a muzzle is pulled over my face. He quickly ties the strings in the back and then adjusts all the buckles around the head and underneath the jaw to make sure I cannot make a sound. I hear the dangling of more chains as they are added to each ankle and locked to each corner of the bed frame. I try moving my hands and legs but I know it’s useless.

His gloved hands then rub my hooded face and his fingers slide all the way down to my locked cock as he playfully slaps my balls and wiggles the chastity device, reminding me that I am his prisoner. The door shuts and locks behind him as I am in the dark trying to move around in the chains. They are much more restrictive than any time we have played, but I realize this is punishment.

[jimmyandthegiraffes]

Tintin in the Land of the Soviets (1929-30)

Vehicles

Train - 3

Car - 6 (5 times as driver, once as passenger)

Boat - 2

Plane - 2 (once as pilot, once as passenger)

Motorbike - 1

Total: 14 vehicles, 6 crashes

Health

Slept - 3 (snored - 3)

Ate - 2 (one bowl of cabbage soup)

Sneezed - 2

Injuries - received 10 bruises, 13 head injuries (mostly minor), was shot at 3 times and hit once, was knocked unconscious twice. Was chloroformed once but wasn't rendered unconscious.

Frozen - 1

Drunk - 1

Physical Activity

Enacted violence against another person - 12

Trees climbed - 1

Swam - 1

Horses ridden - 1 (badly)

Le Petit Vingtième

Articles written - 1

Emotions

Cried - 2 (once when leaving Belgium and once when he thought Snowy had been killed)

Angry - 3 (twice at Snowy for disobedience)

Danced - 4

Scared - 1 (in a haunted house)

The Law

Arrested - 4

Sentenced to death - 3

Imprisoned - N/A bc i forgot to count

Joined the army - 1

QUIFF DOWN - 4

[geneticteacher123]

Preparation Histology Sections: 1- Fixing (Fix samples one day with 10% Formalin or 75% ethanol alcohol or Carnoy’s fluid to prevent autolysis for possible living state to give suitable texture for staining) , 2- Dehydration (With series dilutions of ethanol alcohol (70%, 80%, 95% and 100%) to pull water from samples)) and Cleaning (Using Xylene or Chloroform or Benzene), 3- Embedding (By Pour fresh melted wax 60 degree Celsius in sample then allow wax settle and cool and solidify surrounding sample), 4- Sectioning (Using knife in microtome machine to cut (4-5) micron thick section to mount in glass microscope slide), 5- Staining (by remove paraffin from sample by dissolving with xylene the rehydrate section in descending series dilutions of ethanol (100% to 96% to 80%) for 1 minute next remove excess dye by tap water after that Dehydrate section in ascending series dilutions of ethanol (80% to 96% to 100%) for 1 minute and finally clear slide with xylene and enjoy examining) #geneticteacher

[lobotomized-angel]

1 — do you have a darling ? or multiple ? if not , what is your ideal darling like ?

~~~ I do,, I have a couple <3 I have 5 different ones,, 2 I talk about in this account

2 — how much would you change about yourself for your darling / yandere ?

~~~ Everything. Whatever makes them like me more. From appearance to personality to speech patterns to my eating habits to sleeping habits to my alcohol intake to my bad habits to things that will ruin me and my life as I know it,, etc. Anything and everything I will change for them.

3 — who is someone you couldn’t live without ( platonically ) ?

~~~ Uhm...... I don't know,, I'm not sure...... I don't really have friends or close friends....... I don't really have anyone like that,, unless family counts and that would be my brother and my dad

4 — would you be okay being locked up

~~~ Very much so,, Just make sure I have books,, notebooks, writing utensils,, food and something I can listen to music on and I will be the happiest I ever could be <3 (sex is also a need for my survival...... to high of a libido mixed with hypersexuality and it legit makes it hard to live.)

5 — would you want to lock up your darling ?

~~~ I would <3 They don't need to do anything themselves,, I can do it. I need them safe and well cared for and I can take care of them better than what anyone else can and could and will.

6 — how much of someone’s life do you want to control ? what aspects of it ?

~~~ Mmm...... I'm not too sure,, Like I'm not much of something of controlling their life but I would want to know who they're talking to 24/7 and what they're doing and where they're at 24/7. They will mostly be controlling me and what I do whether they realize it or not...

7 — how much of your life are you willing for someone else to control?

~~~ All of it. I need it to survive and function,, I have zero thinking abilities for myself and can barely function without someone telling me what to do even though I despise it so much. I hate it when I already know what I'm doing or what I need to do or what I'm going to do but when I don't,, I need the direction and pointers...... I need the dominance and demanding stuff.

8 — would you ever let your darling go , if you had kidnapped them ?

~~~ No. I kidnapped them for a reason,, The only way they're "leaving" me is if they're dead and even then they will not be escaping me because they will be mine for eternity. The only way I would let them go is if they promised to stay with me and be with me and love me,, essentially stockholm syndrome <3

9 — what kidnapping method do you think about the most ?

~~~ Drugging <3 Chloroform,,

10 — what is your yandere to darling ratio ? ( how much are you of each ? )

~~~ I think I'm 100 percent of each,, I'm not quite sure,,

11 — can you handle long distance relationships ? for how long ?

~~~ Yes and no,, I need communication constantly and if I'm long distance we need to be talking all day every day. If it's long distance we will need to meet at some point if it's truly serious and will last otherwise no. I will never ever do nor handle long distance because I cannot physically see them and it makes my mental health worse.

12 — what’s an object you can’t live without ?

~~~ Books <3

13 — what would be one object you’d most likely steal from your darling ?

~~~ Their clothes,, maybe their cologne. Their pillow maybe even,,

14 — consider this a free pass to ramble about your darling or partner :)

~~~ I don't know which one to ramble about and most of my thoughts are NSFW at the moment,, I can't find the right words to do so,, so I'm going to pass on this one......

15 — would you ever hurt your darling / partner ? how ?

~~~ I don't ever want to but if I did physically. I love hurting people,, making them bleed...... I just want their okay before I do it. I don't want them to hate me

16 — how would you celebrate your darling’s birthday ?

~~~ EEEEEE THIS IS A PERFECT QUESTION RIGHT NOW,, SO,, My Blaze's birthday is next week on Monday,, I would love to spoil him to halcyon and back,, I would love to wake him up with a nice yummy breakfast,, spoil him with gifts and take him out to lunch and dinner,, buy him anything he wants,, go see a movie,, take him to a concert of one of his favorite bands if I could,, hell even get a private meet and greet with one of his favorite bands that sadly no longer tours,, like for example a band like Pink Floyd. I would love to take him to a Weezer concert. We both share a love and passion for music and love to create our own so it's truly something so intimate and precious,, for dinner maybe get something on the way home for us to eat from the concert and when we get home,, we cuddle up in bed and share the most sensual, romantic, maybe a little rough, intimate party beneath the sheets <3 I of course would make him a homemade card,, and I would make him treats,, cookies,, brownies,, cupcakes,, cake pops,, anything,, This is about My Blaze,,

17 — what would you say to your darling / partner right now if you could ?

~~~ That I love him and I want to be with him. This goes for my Blaze and my Blondie Delight

18 — how much are you willing to support your partner no matter what ?

~~~ They could be worse than Richard Ramirez, John Wayne Gacey, Ted Bundy, Albert Fish, etc of nasty disgusting predators and I would support them through all of it. I will always be by their side and their number one supporter. When I love,, I love deep and nothing will ruin that love.

19 — if anything , what would be most likely to bring you to k!ll your darling ?

~~~ Probably jealousy or fear of them leaving me. A BPD episode,, something of that sort,,

20 — is there anything you dislike about your darling ? or : what’s the least desirable trait in a partner to you ? 

~~~ For my Blaze it's probably the fact he's Christian,, I don't like the religion much but,, knowing he doesn't mind me being into crystals and stuff makes me feel a little better about it. For my Blondie Delight,, the fact he has a girlfriend and that he flirted with me still. He lead me to believe he was single and I embarrassed myself and hurt myself very bad by giving him a note,, all I wanted was to be his friend and get to know him and he turned me down because of this supposed girlfriend. I fucking hate her.

I hope you enjoy reading this,,

[nothumerus]

when you get this, list 5 songs you like to listen to, publish. then, send this to 10 of your favourite followers <3

1. You're one of them, aren't you? - The Paper Chase

2. Chloroform Girl - Polkadot Cadaver

3. The road giveth - Rent Strike

4. From here to utopia - Ramshackle Glory

5. Date rape - Sublime

[helen0086]

 Coenzyme Q10 Detection Technology

In 1957, Prof. Grane of the Institute of Enzyme Research of the University of Wisconsin isolated a new quinone compound from the lipid extract of bovine heart mitochondria [1]. The compound is an orange-yellow crystal with a melting point of 48-49 ℃, capable of reversible oxygenation and reduction, and mainly involved in mitochondrial electron transfer. The compound is coded as Q-275 (Q is the initials of quinone, and 275 is the maximum absorption at 275 nm).

In 1958, American scholar Folkers and his team synthesized a series of coenzyme Q compounds, confirmed the structure of Q-275 and named it coenzyme Q10 [2]. In 1961, Mitchell, a British chemist, proposed the theory of "chemotaxis" in the study of energy conversion in living organisms and revealed the role of coenzyme Q10 in the energy conversion system of mitochondria [3], and was awarded the Nobel Prize in Chemistry in 1978. Since then, people have gradually recognized coenzyme Q10, and its applications have been widely and deeply studied.

Coenzyme Q10 (CoQ10), also known as ubiquinone, is chemically known as 2,3-dimethoxy-5-methyl 6-deca-isopentadienylbenzoquinone and consists of a benzoquinone ring and polyisoprene side chains. The number of isoprene units in the coenzyme Q series varies by species, with humans having 10 units. Coenzyme Q10 is available in both oxidized (ubiquinone, CoQ10, Ubiquinone) and reduced (ubiquinol, CoQ10H2, Ubiquinol) forms, and its chemical structure is shown in Figure 1.

Fig. 1 Chemical structures of oxidized (a) and reduced (b) forms of coenzyme Q10

Coenzyme Q10 is an important component of the mitochondrial respiratory chain, where it acts as an electron carrier and participates in electron transfer and ATP production. Furthermore, the cellular functions of coenzyme Q10 are multifaceted: it is present in all cell membranes, it limits the toxic effects of free radicals, it is a component of low-density lipoprotein (LDL), and it is involved in the aging process. Its deficiency is associated with a variety of diseases, such as mitochondrial disease, cardiovascular disease, age-related diseases, tumors, liver disease, kidney disease, etc. Panthenol is also a powerful antioxidant. Panthenol is also a powerful antioxidant, preventing lipid peroxidation in biological membranes [4].

With the deepening of the research on coenzyme Q10, the application of coenzyme Q10 is becoming more and more extensive. In addition to its use as a drug, it also has many applications in nutraceuticals, cosmetics and dietary supplements. Coenzyme Q10 is an endogenous substance, but its concentration in living organisms is very low. The analysis and determination of CoQ10 is important for the clinical diagnosis of diseases and the quality control of drugs and health products. In recent years, many analytical methods for the determination of coenzyme Q10 have been developed, which are summarized and discussed in this paper.

1 Coenzyme Q10 extraction and sample preparation

Coenzyme Q10 is insoluble in water and methanol at room temperature, slightly soluble in ethanol, soluble in acetone, 1-propanol, and soluble in organic solvents such as hexane and chloroform. Pharmaceuticals and dietary supplements such as tablets, capsules and softgels can be dissolved in ethanol, 1-propanol and other solvents, and analyzed by ultrasonication or filtration.

The isolation and enrichment of coenzyme Q10 from complex biological matrices is a laborious process.

Conventional liquid-liquid extraction is the most commonly used extraction method. This method is simple and has a large processing capacity, but has the disadvantage of high solvent consumption and some solvents can interfere with subsequent detection. Often the solvent is evaporated under N2 protection after extraction and redissolved in a mobile phase or other solvent. Whole blood samples were immediately dosed with the anticoagulants heparin or EDTA, and the plasma was centrifuged at low temperature and stored at -80°C. The plasma was then analyzed for the presence of coenzyme Q10 in the plasma. Coenzyme Q10 was extracted from plasma as follows [5]: Methanol was added to the plasma to precipitate the proteins, and the plasma was extracted with hexane. The mixture was rotated and shaken for 15 min, then centrifuged for 5 min, and the supernatant was extracted and the solvent was evaporated. The supernatant was dissolved in acetonitrile before analysis.

Coenzyme Q10 was extracted from animal heart tissue [6]. The extraction of coenzyme Q10 from animal heart tissue [6] was performed by precise weighing, transferring to homogenization tubes containing lysis medium A (containing garnet and zirconia beads), adding 1-propanol and the antioxidant 2,6-di-tert-butyl-4-methylphenol (BHT), shaking, centrifugation, and collection of the supernatant, which was analyzed immediately. The extraction of coenzyme Q10 from muscle tissue is most often done directly using muscle homogenate, or sometimes mitochondria are extracted from the tissue under ice-cold conditions, and then the mitochondrial suspension is diluted with 1-propanol, centrifuged, and the organic layer is extracted with ethanol and hexane [7]. The one-step extraction method is to use a suitable organic solvent to extract coenzyme Q10 while precipitating proteins. Yang et al. [8] studied the one-step precipitation of plasma proteins with different organic solvents (methanol, ethanol, acetonitrile, and acetone), and found that acetone was the best precipitant, and the extraction yield ranged from 71.00% to 93.07%, and was simpler than the operation of liquid-liquid extraction.

The solid phase extraction (SPE) technique can also be used for the extraction of coenzyme Q10. On-line SPE techniques are less time-consuming, less expensive, and reduce sample loss and contamination problems. The technique is usually automated using a programmable on/off valve [9]. However, protein precipitation is required prior to extraction.

Molecularly imprinted polymers (MIPs) are specialized molecular recognition techniques that have been developed in recent years. Molecularly imprinted polymers (MIPs) are formed by mixing template molecules with functional monomers, cross-linkers and initiators. After polymerization, the template molecules are removed and binding sites and cavities complementary to the templates in size, shape and function are formed [10], allowing selective recognition and adsorption of molecules structurally similar to the templates.

Contin et al. [10] synthesized MIP using coenzyme Q0 as the template, methacrylic acid as the functional monomer, acetonitrile as the pore-forming agent, ethylene glycol dimethylacrylate as the cross-linking agent, and benzoyl peroxide as the initiator. MIP was used as an adsorbent for solid-phase extraction of coenzyme Q10 from liver samples using dispersive solid-phase extraction. In addition, MIP synthesized in the same way could be used as the filling adsorbent for solid-phase extraction of coenzyme Q10 in urine. In addition, the MIP synthesized by the same method can also be used as the filling adsorbent of polypropylene columns for solid-phase extraction of coenzyme Q10 in urine, and the columns can be reused four times [11]. Compared with the traditional solid-phase extraction, MIP as a polymer adsorbent for solid-phase extraction has the advantages of simple synthesis, low cost, good stability, porous, and high selectivity for target molecules [11].

Sometimes it is necessary to maintain the original oxygenated and reduced state of coenzyme Q10 in the samples during the extraction process, which causes great difficulties due to the oxidizability of CoQ10H2. In this case, the temperature can be controlled at a low temperature of 4 ℃ during the extraction process [6,12], shortening the extraction time and using anhydrous extract will increase the stability of CoQ10H2 [13], and the use of HCl-acidified ethanol as a diluent can also prolong the stability of CoQ10H2 and prevent the auto-oxidation of CoQ10H2 [12]. BHT is an antioxidant often added in the extraction of plasma and tissue samples, which can prevent the oxidation of CoQ10H2 [6,12,14]. However, the addition of BHT to CoQ10H2 extracts from dietary supplements and pharmaceuticals was found to increase the oxidation of CoQ10H2 [13,15]. The difference in matrix composition between plasma samples and dietary supplements may be the main reason for the loss of antioxidant capacity of BHT [13].

Biological samples for coenzyme Q10 extraction include plasma, leukocytes or platelets, muscle, fibroblasts and urine [16]. Muscle biopsy is the best choice for studying coenzyme Q10 status in mitochondrial diseases, but it is very invasive; the correlation between the levels of coenzyme Q10 and tissues in plasma, blood cells and urine has been controversial, but the determination of coenzyme Q10 in these samples has an important role in therapeutic monitoring [16]. However, the determination of coenzyme Q10 in these samples is important for therapeutic monitoring [16].

The methods used to extract coenzyme Q10 from biological samples are summarized in Table 1.

Table 1 Extraction methods of Coenzyme Q10

Simple operation, large processing capacity, high solvent consumption, some solvents may interfere with the subsequent detection.

Plasma, animal heart, muscle homogenate, mitochondria

Online Solid Phase Extraction

Less time-consuming and costly, reducing sample loss and contamination.

plasma (medicine)

Molecular Blotting Techniques

Low cost, good stability, high selectivity for target molecules, and can be combined with solid phase extraction.

Animal liver, urine

2 The main assay for Coenzyme Q10

2.1 High Performance Liquid Chromatography (HPLC)

HPLC is currently the main analytical method for analyzing coenzyme Q10 in various matrices. The main detectors coupled with HPLC are ultraviolet (UV), tandem mass spectrometry (MS/MS), electrochemistry (ECD), fluorescence (FL), chemiluminescence (CL), etc. The separation effect of HPLC is good, and each detector has its own characteristics.

2.1.1 HPLC-UV

HPLC-UV is the most commonly used method for the determination of coenzyme Q10, and has become the national standard for drugs and health foods [17, 18]. It has been widely used for the determination of coenzyme Q10 in pharmaceuticals [15, 19-22], health foods or dietary supplements [15, 20], plasma [14, 23] and tissues [10]. Conventional C18 or C8 reversed-phase chromatographic columns can separate either one form of coenzyme Q10 (usually oxidized) or both oxidized and reduced forms.

Liposomes are a new type of pharmaceutical dosage form formed by the self-assembly of lipids (mainly phospholipids and cholesterol) with a bilayer structure similar to that of a cell membrane, which can encapsulate hydrophilic or hydrophobic drugs. Ruiz-Garcia et al. [21] prepared a small monolayer of liposomes encapsulating coenzyme Q10, phosphatidylserine, and fat-soluble vitamin C (6-o-palmitoyl-L-ascorbic acid) by thin-film hydration. The prepared samples were freeze-dried, solubilized in chloroform and determined by HPLC-DAD at two analytical wavelengths.

Clementino et al. [22] prepared lecithin/chitosan nanoparticles encapsulating simvastatin and coenzyme Q10. The chitosan-modified liposomes showed higher stability and narrower particle size distribution. The content of simvastatin, simvastatin hydroxylate and coenzyme Q10 was quantified by reversed-phase HPLC-UV method to account for possible degradation products. The encapsulation rate was determined and the in vitro release of the drugs was studied. According to the study, the serious side effects of statins, such as rhabdomyolysis, were associated with the decrease of coenzyme Q10, so the co-encapsulation of these two drugs is of great significance.

Coenzyme Q10, as a fat-soluble vitamin coenzyme, is often measured in conjunction with other fat-soluble vitamins. Franke et al. [14] analyzed 25 substances including 25-OH-vitamin D3, 25-OH-vitamin D2, retinol, tocopherols, carotenoids (including their stirrup isomers), and oxidized and reduced coenzyme Q10 in plasma on a fusion-nucleated 2.6 μm particle size C18 column in tandem with a C30 column, which is good at separating carotenoid isomers, and in conjunction with a six-pass valve. D2, retinol, tocopherols, carotenoids (including their stirrup isomers), and oxidized and reduced coenzyme Q10 in plasma. The switching of the six-way valve allows coenzyme Q10 to flow from the C18 column to the detector while the carotenoid isomers are eluted on the C30 column, avoiding the difficulty of separating these two substances on the same column. In addition, if a pressure-resistant UV-Vis detector is added between the C18 and C30 columns, it is possible to separate all substances without switching the six-way valve, but special software is required to control the two detectors and to acquire and process the data. It has also been reported that retinol, six carotenoids, two tocopherols, and coenzyme Q10 (10 fat-soluble vitamins) can be measured in human plasma using a MYC30 column, and the total amount of the oxidized form of coenzyme Q10 was measured by oxidizing coenzyme Q10H2 first with FeCl3 [23].

The HPLC-UV method is highly accurate and reproducible, with LOD generally on the order of μg-mL-1 and sometimes on the order of ng-mL-1 with highly sensitive detectors [14].

2.1.2 HPLC-MS/MS: HPLC-MS/MS has been developed rapidly and applied more and more widely. This method utilizes the high separation efficiency of HPLC for complex samples combined with the high sensitivity and high selectivity of mass spectrometry, which can detect low content samples under the background of complex matrix, and is widely used in the analysis and determination of target compounds in biological samples.

The main types of tandem mass spectrometry are triple quadrupole mass spectrometry [5,7,11,25,26], quadruple linear ion trap mass spectrometry [13,24], and hybrid quadruple orbit trap mass spectrometry [12], etc. Most of them use electrospray ionization, multiple reaction monitoring (MRM), and positive ionization mode. Due to the low sensitivity of [M + H]+ analysis of coenzyme Q10, ammonium adducts, i.e., [M + NH4]+, are often used to improve the sensitivity of the mass spectrometric response. By adding a certain amount of ammonium acetate to the mobile phase, [NH4]+ forms a stable five-membered chelated ammonium cation with coenzyme Q10 [8]. The formation of Li adducts has also been reported to greatly increase the sensitivity [24].

The electrostatic field orbitrap mass spectrometry (Otbitrap) is a new type of high-resolution mass spectrometry, which has the advantages of high resolution, high mass accuracy, and wide dynamic range, etc. Pandey et al. [12] applied HPLC-hybrid quadruple orbitrap mass spectrometry (Q-Orbitrap) to rapidly determine the redox state of coenzyme Q9 and coenzyme Q10. Two scanning modes, full MS/AIF and tSIM/data-dependent secondary scanning (tSIM/ddMS/MS), were compared, and it was found that full MS/AIF had higher signal sensitivity and good peak shape. During sample preparation, coenzyme Q9 and coenzyme Q10 were extracted with BHT-containing hexane to limit the oxidation of the reduced form, and the Kinetex C18 column, with fused-core SiO2 packing and smaller particle size (2.6 μm), was found to have higher column efficiency, better resolution, and good peak shape. Oxidized and reduced forms of coenzyme Q9 and coenzyme Q10 were analyzed in brain, heart, liver, adipose tissue, and muscle of healthy mice with a small amount of sample (<5 mg) and a very short analysis time (4 min). the LOD ranged from 0.01 to 0.49 ng mL-1 .

Due to the complexity of the biological sample matrix and the low concentration of coenzyme Q10, sample pretreatment is very important. Becerra et al. [11] analyzed coenzyme Q10 in human urine by molecularly imprinted polymer solid-phase extraction (MIP-SPE) coupled with HPLC-MS/MS. The pretreatment process concentrates the coenzyme Q10 by at least 5-fold. The high degree of sample purification reduces the ion suppression caused by the matrix effect of mass spectrometry. The analytical system does not interfere with protein or white blood cell elevations, which is important in cases of coenzyme Q10 deficiency with renal impairment.

The HPLC-MS/MS method uses a lot of internal standards, and the selection of suitable internal standards is also an effective way to eliminate matrix effects. Commonly used internal standards include coenzyme Q9 [5, 11, 25], coenzyme Q4 [12], and the isotopes of coenzyme Q10, coenzyme Q10-2 H6 [7] and coenzyme Q10-2 H9 [26], which are structurally similar to coenzyme Q10. Structural analogs of coenzyme Q10, such as coenzyme Q4 and coenzyme Q9, have many advantages. They are also endogenous ubiquinones and are present in human plasma at very low concentrations, or at least at levels that do not interfere with their use in analytes at the concentrations required for analysis, and therefore do not interfere with the quantification of analytes. In addition, it separates well from coenzyme Q10 [5]. A potential source of error in mass spectrometry is ion suppression, especially in electrospray ionization mass spectrometry, where the response signal of the analyte is altered and often suppressed if an interfering substance interferes with the ionization of the analyte on the surface of the droplet, or if there is competition. The use of an isotope internal standard is a good solution to the problem of ion suppression. By co-eluting the isotope internal standard with the analyte, the effects of various effects can cancel each other out, and the matrix effect can be minimized and the sample recovery can be better [7].

2.1.3 HPLC-ECD

Electrochemical detectors (ECDs) are widely used because of their high sensitivity, good selectivity and low price. Coenzyme Q10 can undergo a reversible redox reaction and can be detected by an ECD.

The commonly used detection methods are coulometric or voltammetric analysis. Different voltages are set according to the redox potentials of the substances to be measured. For oxidized coenzyme Q10, it is usually reduced to its reduced form first, and then oxidized as the original reduced coenzyme Q10 in the sample. This method can measure both oxidized and reduced coenzyme Q10 simultaneously.

Yubero et al. [27] used HPLC-ECD to determine coenzyme Q10 in urine and gave reference values for the pediatric population. An ESA Coulochem II electrochemical detector was used, and the cell voltages were -600 mV and +600 mV. The amount of coenzyme Q10 in urine fluctuated greatly at different times of the day, and the morning urine with the smallest fluctuation was chosen as the sample. The results were expressed as the amount of coenzyme Q10 per gram of particulate protein. The reference standards for children are: 2-10 years old: 24-109 nmol; 11-17 years old: 43-139 nmol. This assay provides a noninvasive method for assessing renal coenzyme Q10 status in patients with renal disease, but it is not currently available and requires up to 30 mL of urine per sample.

Schou-Pedersen et al. [6] determined reduced and oxidized coenzyme Q10 in canine plasma and cardiac tissue by HPLC-ECD and compared it with HPLC-MS/MS. The ECD was performed by fluid dynamic voltammetry using an RS6011 ultra-analytical cell at a voltage setting of 500 mV. A guard cell at -600 mV was used prior to the analytical cell to reduce oxidized coenzyme Q10 eluting from the column. Mass spectrometry was performed using a Waters Micromass Quattro micro API triple quadrupole mass spectrometer with multiple reaction monitoring (MRM) and the internal standard CoQ10-2 H9. Both methods used the same column with slightly different mobile phase ratios and additives. The results showed that CoQ10H2 was approximately 30% lower in the HPLC-MS/MS method than in the HPLC-ECD method, which may be due to differences in the calibration stock solutions or to accelerated oxidation during storage or analysis in the LC-MS/MS system. Therefore, the two methods are not interchangeable. In terms of sensitivity, the sensitivity of the two methods was comparable for coenzyme Q10H2, whereas the sensitivity of the HPLC-ECD method was higher for coenzyme Q10.

2.1.4 HPLC-FL and HPLC-CL

HPLC with a fluorescence (FL) detector is widely used for the determination of various substances in biological samples due to its high selectivity and sensitivity. Coenzyme Q10 is not a fluorescent substance and needs to be derivatized before determination. Nohara et al. [28] measured CoQ10 and CoQ10H2 in blood by post-column derivatization using HPLC using 2-cyanoacetamide and CoQ10 and CoQ10H2 heated under alkaline conditions to produce fluorescent products. The fluorescence emission and excitation wavelengths were 442 nm and 549 nm, respectively.

HPLC coupled with a chemiluminescence (CL) detector has also been reported for the determination of coenzyme Q10.Kishikawa et al. [29] used dithiothreitol (DTT) as a reductant to reduce quinone to semiquinone radicals, and semiquinone radicals converted dissolved oxygen to superoxide anion, which reacted with luminal to form CL.Accordingly, coenzyme Q10 was determined in plasma by HPLC-CL, and other components in plasma were not interfered with. Coenzyme Q10 in plasma was determined by HPLC-CL, and other components of plasma were not interfered.

Both methods require a reaction coil between the column and the detector, and require two or three pumps to mix the various reaction reagents with the coenzyme Q10-containing eluent after the column and then into the reaction coil, which is a cumbersome operation. In recent years, the literature in this area is relatively scarce.

2.2 Spectrophotometric and fluorescent methods    

The Enzyme Labeler, also known as Microplate Reader, is an instrument for reading and analyzing the results of Enzyme Linked Immunosorbent Assay (ELISA) experiments. The basic principle of ELISA is similar to that of spectrophotometer or photoelectric colorimeter, using plastic microplates instead of cuvettes, usually 48-well, 96-well, or larger, with low reagent consumption, high speed, and good repeatability. Multifunctional enzyme labeling instrument often has a variety of detection functions such as absorbance, fluorescence, chemiluminescence, etc., in the medical and health inspection has been widely used.

Fukuda et al. [30] developed a rapid microtiter plate method for the determination of coenzyme Q10 using the redox cycle of quinone. Coenzyme Q10 was reduced to ubiquinone radical by NaBH4, and then the ubiquinone radical was oxidized to ubiquinone and superoxide anion radical by dissolved oxygen. The superoxide anion radical converts 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride (INT) into a pink methanol dye. It has a strong absorbance at 510 nm, which increases with increasing concentrations of coenzyme Q10. The absorbance of Mazanine dye was measured quickly and easily by a microplate reader. As an application of this method, the content of coenzyme Q10 in cosmetics was successfully determined with an LOD of 14.8 nmol-L-1 . The proposed method can be used for the rapid high-throughput analysis of ubiquinone-containing products.

Fei et al. [31] developed a new method for the determination of coenzyme Q10 in serum and urine of Alzheimer's disease patients by fluorescence spectrophotometry, also using a microplate reader. The method is based on the fact that the chemical derivative between ethyl cyanoacetate (ECA) and coenzyme Q10 is fluorescent and can be detected by fluorescence spectrophotometer (FS-ECA) at λex/em = 450/515 nm. The results showed that serum and urine levels of coenzyme Q10 were significantly lower in Alzheimer's patients than in age-matched controls. This method has similar LOD and LOQ as the HPLC-UV method.

The FS-ECA method has some advantages over the HPLC-UV method, such as easier sample preparation, faster detection speed, and similar accuracy and specificity [31].

The important role of liposomes as a new drug dosage form for co-administration and targeted delivery was described in the literature [21,22], and liposomes can also be used as micro-containers to protect and concentrate reagents.Román-Pizarro et al. [32] prepared a new type of magnetic liposomes (MLs) containing hydrophobic magnetic gold nanoparticles (Fe3 O4 @ AuNPs) and the long-wavelength fluorophore cresyl violet for the determination of coenzyme Q10 in foodstuffs. AuNPs) and a long-wavelength fluorophore, cresyl violet, were used for the determination of coenzyme Q10 in food. First, the MLs were introduced into the flow-through system using a flow injection device and retained in front of the detector for 300 s by means of a solenoid device to achieve preconcentration. Then, a coenzyme Q10 solution containing the surfactant Triton X-100 was injected into the flow-through system. The surfactant caused the solubilization of the MLs and the release of cresyl violet, which was oxidized by coenzyme Q10, resulting in a decrease in the fluorescence signal. The concentration of coenzyme Q10 is directly proportional to the decrease in fluorescence signal. The LOD of this method is lower than that of the LC-UV method, but the equipment required is simpler and less expensive.

2.3 Electrochemical analysis    

The redox properties of CoQ10/CoQ10H2 allow the determination of coenzyme Q10 by electrochemical analysis. The methods are generally voltammetric, such as cyclic voltammetry (CV) [33], differential pulse voltammetry (DPV) [34], square wave voltammetry (SWV) [35], etc. The samples can be pharmaceuticals, dietary supplements, animal and plant tissues, etc. The samples can also be used for the determination of CoQ10/CoQ10H2. Samples can be pharmaceuticals, dietary supplements, plant and animal tissues, etc.

Li et al. [34] investigated the electrochemical reduction mechanism of coenzyme Q10 at a silver electrode and developed a DPV method for the direct determination of coenzyme Q10 in plant and animal tissues. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) revealed that the reduction of coenzyme Q10 under anoxic conditions is a reversible one-electron, one-proton reduction and forms a stable semiquinone radical (coenzyme Q10H-), which is quenched by oxygen in an oxygen-filled environment. This is the reason why coenzyme Q10H2 is able to scavenge oxygen radicals due to its antioxidant function. Under the optimized experimental conditions, the DPV method can be used to determine coenzyme Q10 in complex samples, and it is rapid, sensitive, and highly selective, with an LOD of 3.33 × 10 -8 mol-L-1 .

Graphene, a single atom thick planar sheet composed of carbon atoms heterogeneously linked by sp2 in a honeycomb lattice, is a new type of sensor material [35]. Screen printing is a practical technique for manufacturing low-cost disposable sensors [35].

The new graphene sensor developed by this technology can be used for the determination of coenzyme Q10 and α-lipoic acid. The MnO2-modified screen-printed graphene electrode (MnO2/SPGE) has a larger capacitance and electrically active surface area, which facilitates electron transfer and significantly improves the oxidation performance of coenzyme Q10 and α-lipoic acid. The MnO2-modified screen-printed graphene electrode coupled with square wave dissolution voltammetry (SWV) can be used for the simultaneous determination of coenzyme Q10 and α-lipoic acid in dietary supplements with high sensitivity and practicality.

The electrochemical mechanism of coenzyme Q10 is complicated by the different electrodes and media. In a 1.1:1 methanol-ethanol solution, the electrochemical reaction of coenzyme Q10 at the glassy carbon electrode was controlled by adsorption, and the sensitivity of the determination could be improved by pre-enrichment [33]. In anaerobic ethanol solution, the cathodic process of coenzyme Q10 at the silver electrode was one-electron-single proton reduction [34], while the oxidation on MnO2/SPGE showed two-electron-single proton transfer [35]. Michalkiewicz [36] investigated the anodic oxidation of oxidized coenzyme Q10 in acetic acid solution using a glassy-carbon electrode and a carbon-fiber microelectrode coupled with voltammetry, respectively. The oxidized coenzyme Q10 in acetic acid solution was studied by

The results show clear oxidation peaks or waves in the potential range above 1.5 V. The presence of these signals cannot be linked to the well-known redox pair CoQ10/CoQ10H2, but may be attributed to the irreversible and diffusion-controlled two-electron oxidation of methoxy in coenzyme Q10 (formation of two additional quinone groups at the 2 and 3 positions of the ring). the total number of electrons involved in the CoQ10 anodic oxidation is much greater than two, suggesting that the oxidation also takes place in the unsaturated isopentadienyl side chain. The total number of electrons involved in CoQ10 anodic oxidation is much higher than two, suggesting that oxidation also occurs in the unsaturated isoprene side chain. The oxidation of the oxidized coenzyme CoQ10 has been rarely reported, is much less readily accessible than that of coenzyme Q10H2, and the mechanism of oxidation has yet to be demonstrated.

2.4 Other analytical methods   

Supercritical fluids are substances whose temperature and pressure exceed the critical point, in a state of gas-liquid indistinguishability, with a density close to that of liquids, and a viscosity close to that of gases, and a high diffusion coefficient. Supercritical fluids with high diffusivity and low viscosity, very suitable for mobile phase. The supercritical fluid with more research and application is supercritical CO2. Supercritical fluid chromatography-tandem mass spectrometry (SFC-Ms/Ms) with supercritical CO2 as mobile phase can be used for the determination of coenzyme CoQ10 in rat plasma [8]. The method uses one-step acetone method to precipitate the protein and extract CoQ10 from the sample. Due to the low sensitivity of [M + H]+ of CoQ10 in mass spectrometry, methanol containing ammonium acetate was used as a post-column compensating solvent to provide [M + NH4]+ and improve the sensitivity. Supercritical CO2 is non-toxic, non-flammable, and relatively inexpensive, so it is widely used. Due to its non-polar nature, it is well suited for the analysis of fat-soluble compounds and can greatly reduce the use of organic solvents.

Other analytical methods include: high performance thin layer chromatography (HPTLC) [37], Fourier transform near infrared spectroscopy (FT-NIR) [38], nuclear magnetic resonance hydrogen spectroscopy (1 H- NMR) [39], etc. HPTLC is simple and rapid, but the sensitivity is not very high, and can be used for the analysis of coenzyme Q10 in raw materials and pharmaceutical preparations. FT-NIR does not require complex sample pretreatment, but requires a certain number of samples to establish a calibration model and obtain results through complex statistical analysis, and is generally used for the initial screening of the target analyte. 1 H- NMR also does not require complex sample pretreatment, can be both calibrated models, and obtained through complex statistical analysis, and is generally used for the initial screening of target analytes. FT-NIR does not require complex sample pretreatment, but requires a certain number of samples to establish a calibration model and obtain the results through complex statistical analysis, and is generally used for the initial screening of target analytes.1 H-NMR also does not require complex sample pretreatment, and can be used both qualitatively and quantitatively, with low sensitivity, and can be used for routine analysis of preparations. These methods can be used as a useful supplement to the quantitative analysis of coenzyme Q10.

3 Simultaneous determination of oxidized and reduced coenzyme Q10

In many methods, the total amount of coenzyme Q10 is determined by adding oxidizing agents such as FeCl3 to oxidize coenzyme Q10H2 to the oxidized form, and then the total amount of the oxidized form is determined. However, coenzyme Q10 coexists in both oxidized and reduced forms in biological matrices, and sometimes it is necessary to determine the two forms of coenzyme Q10 in biological samples, drugs and supplements separately. Commonly used methods include HPLC-UV [14, 15], HPLC-MS/MS [12, 13, 24, 25, 40], HPLC-ECD [6, 41], HPLC-FL [28], and so on.

Coenzyme Q10H2 standards are sometimes not readily available and can be obtained by reducing oxidized coenzyme Q10 with reducing agents such as NaBH4 [12-14,24,25,28,40,41]. The reduction of coenzyme Q10 at low concentrations may be incomplete, even if the amount of NaBH4 is 8800-fold excess [13], so the reduction process needs to be controlled at a certain concentration range. The reaction is usually carried out at low temperature and in the dark, and sometimes ED-TA is added to the solution after the reaction [12,13,15,25,41], which mainly binds to the metal ions that catalyze the oxidation process and acts as an antioxidant. Even if a standard of coenzyme Q10H2 is available, it may be partially oxidized and needs to be reduced before use [15], or the absorbance of the stock solution (ε = 4010) can be measured spectrophotometrically at 290 nm to determine the exact concentration [6].

Whether the quantitative results were expressed as the total amount of coenzyme Q10 or as the oxidized and reduced amounts, respectively, could affect the sample preparation. In order to maintain the reduced state of coenzyme Q10H2, in addition to the rapid operation at low temperature and the addition of antioxidants such as BHT during the preparation process, researchers have different opinions on whether the extracted solution should be re-dissolved by evaporating the solvent in the presence of N2. Some of them evaporate the solvent and re-dissolve it during sample pretreatment [12,14,15,24], while some scholars believe that there will be significant oxygenation of Coenzyme Q10H2 during solvent evaporation, so the extracted solution should be immediately dissolved in the presence of N2 [12,14,15,24].

Determination [6 ,13 ,25 ,28 ,40 ,41].

In fact, coenzyme Q10H2 is highly unstable during extraction and determination.Yamashita et al.[41] found that coenzyme Q10H2 was stable only at -78 °C, and the rate of oxidation of coenzyme Q10H2 to coenzyme Q10 increased with increasing storage time and temperature.Claessens et al.[25] found that significant oxidation of coenzyme Q10H2 to coenzyme Q10 occurred after 2 h in 1-propanol extracts of human plasma, so routine analysis was limited to 12 samples per batch in order to keep the total run time within 2 h. In addition, coenzyme Q10H2 is not stable at the same temperature as coenzyme Q1010. Claessens et al. [25] found that in 1-propanol extracts of human plasma, significant oxidation of coenzyme Q10H2 to coenzyme Q10 occurred after 2 h. Therefore, routine analyses were limited to 12 samples per batch in order to keep the total run time within 2 h. The results of this study are summarized below.

Due to the uncontrolled nature of oxidation, it has been suggested that almost all mitochondrial coenzyme Q10H2 is oxidized during sample pretreatment, and therefore quantification of total coenzyme Q10 in isolated mitochondria does not require ubiquinol oxidation [7]. Nevertheless, attempts have been made to control the rate of oxidation of coenzyme Q10H2 during analysis or to know the extent of oxidation. The choice of internal standards has helped to realize this desire. Structural analogs of coenzyme Q10 and coenzyme Q10H2, such as diethyl- or dibutyl-coenzyme Q10 [14] and dipropoxy-coenzyme Q10 [40], are sometimes used as internal standards in the analysis of coenzyme Q10 and coenzyme Q10H2. They are structurally very similar to CoQ10 and CoQ10H2 and exhibit the same chemical behavior as the analytes, especially with respect to artificial oxidation, which makes it possible to accurately back-calculate the original CoQ10H2/CoQ10 ratio [14].

The CoQ10H2/CoQ10 ratios in biological tissues varied, and Claessens et al. [25] showed that the plasma CoQ10H2/CoQ10 ratios in healthy volunteers without nutrient supplementation ranged from 22.3 to 64.4, with an average of 41.7, whereas Yamashita et al. [41] showed that the plasma CoQ10H2/CoQ10 ratio was about 95/5, suggesting that plasma coenzyme Q10 is mainly in the reduced form. These results indicate that plasma coenzyme Q10 exists mainly in the reduced form.

Changes in the CoQ10H2/CoQ10 ratio have important physiological significance and are associated with many functional disorders and diseases. Measurement of the CoQ10H2/CoQ10 ratio is useful in exploring the mechanisms of many diseases. Oxidative stress has been defined as a disturbance of the pro-oxidant-antioxidant balance in favor of the former, and is considered to be a causative factor in aging and degenerative diseases such as cardiac diseases, diabetes mellitus and cancer [41]. There is a consensus that the CoQ10H2/CoQ10 ratio is an important parameter in the assessment of oxygenation stress [24, 25, 41].

Another study showed that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TC-DD) damaged mouse liver in a dose-dependent manner. Tang et al. [40] investigated the mechanism, and found that exposure of mouse liver samples to TCDD resulted in a decrease in the total amount of coenzyme Q10, a decrease in the level of coenzyme Q10H2, and an increase in the CoQ10H2/total CoQ10 ratio. This may be due to the inhibition of succinate dehydrogenase in the electron transport chain. In addition, the decrease in the total amount of CoQ10 implies that CoQ10 is degraded by external environmental influences, which was confirmed by Temova Rakua et al [42]. They found that oxidized coenzyme Q10 was degraded during storage of dietary supplements and drugs containing coenzyme Q10, and that oxidized coenzyme Q10 was converted to reduced coenzyme Q10H2, especially in the presence of antioxidants such as vitamin C.

4 Summary

Coenzyme Q10 is an important electron carrier and antioxidant component of the mitochondrial respiratory chain and is widely found in human cells. Coenzyme Q10 deficiency may be associated with a variety of diseases. Although it is an endogenous substance, it can be used as a drug or dietary supplement to treat or ameliorate certain related diseases. Therefore, the selection of efficient isolation and analytical methods is of physiological and clinical importance. Liquid-liquid extraction is the most common method for the extraction of coenzyme Q10, while the extraction of reduced coenzyme Q10H2 requires temperature control and the addition of antioxidants. Solid-phase extraction and molecular blotting techniques have also been applied in the extraction of coenzyme Q10 from biological samples, which have greatly improved the extraction efficiency and detection sensitivity.

Coenzyme Q10 can be detected by a variety of methods, and currently the most commonly used method is HPLC. In clinical and pharmaceutical analysis, miniaturization of instrumentation by reducing column diameter and length and particle size is one of the major trends in improving separations [43]. HPLC-UV is easy to use as a standard method, has good stability, is not very sensitive, but is generally sensitive enough to meet the requirements for the simultaneous analysis of a variety of components.

HPLC-MS/MS has high sensitivity and good selectivity, and has unique advantages for the analysis of coenzyme Q10 in complex matrices, such as biological samples, but the operation of the instrument is complicated and the price is expensive. The electrochemical analysis method is simple, fast and sensitive, and has certain applications in the analysis of coenzyme Q10. The HPLC-ECD method is convenient for the simultaneous determination of oxidized and reduced Coenzyme Q10. Coenzyme Q10 co-exists in both oxidized and reduced forms in almost any sample. Some analytical methods are capable of determining both the total amount of coenzyme Q10 and the oxidized and reduced forms, while others can only determine the total amount, depending on the HPLC separation. The characteristics of the various methods, their determination formats and their applications in samples are shown in Table 2.

Table 2 Comparison of different analytical methods for Coenzyme Q10

Easy and fast to use, sometimes requires color development or derivatization, matrix may be interfering

Coenzyme Q10 is mainly present in reduced form in organisms, and the ratio of CoQ10H2/CoQ10 is clinically important, with greater bioavailability of CoQ10H2 in drugs and dietary supplements. Therefore, the simultaneous determination of oxidized and reduced coenzyme Q10 is a future development. The distribution of the two forms, their interconversion and their biological significance will be a focus of future research, which also brings opportunities and challenges to the study of extraction and analytical methods for both forms of coenzyme Q10.

In order to meet the clinical needs, coenzyme Q10 can be prepared by microbial fermentation [44] or chemical synthesis in addition to extraction from biological samples. Chemical synthesis is divided into total synthesis [45] and semi-synthesis [46], and the intermediate of semi-synthesis is ganiol. Microbial fermentation can be used for large-scale industrial production.

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#coenzymecoq10 #Coenzyme Q10 #Q10 #coq10

[wyrmfedgrave]

Pics:

1 shows another side to the Lord of the Earth. But, it's probably just a nightmare...

2 reveals that Little Cthulhu was happiest as a studious & precocious Old One...

3 adds a magic rite cookbook to our dream library of Old Masters...

4 soothes the all too active brain down just enough to open the dream gates to a painless desolution of will...

Addendum: A Good Anaesthetic.

Jellygraphed in 1899(?), this zine covered several anesthetics & how to make them.

(Not recommended as these gasses are all highly reactive & explosive.)

Of ammonium nitrate, HPL stated that breathing it for a moment causes 1 to become intoxicated. Then, you fall asleep & end up insensible to pain.

(I think Lovecraft meant after you turn the ammonium nitrate into nitrous oxide - laughing gas! - then you can use it as an anesthetic...)

Then, we move along to ether, which HPL notes is somewhat poisonous but, not fiery. Because of this, it's used much more extensively.

Next up is chloroetherene (the ether/chloroform mix?), which came from Dutch chemists.

Lovecraft mentions the apparatus needed for experiments of this type.

And then come the ads for (HPL's) "Good Books":

1. Chemistry 1 & 2 - 5¢ each.

2. Simple Chemistry - 10¢.

3. Chemical Magic - 10¢.

4. Art of Fusion - 5¢.

5. Lovecraft's Manual of Explosives - 10¢!!

(Lovecraft!! We really don't know you, do we?!)

Definitions:

1. Ammonium nitrate is a white to brown crystalline salt compound that's a highly flammable explosive.

It has many other uses, among which are: fertilizers, explosives, rocket fuel, meat curing, antibiotics & making matches.

2. Nitrous oxide, which HPL identified as nitrogen protoxide. It's a colorless, sweet smelling & tasting gas that dentists once used. Nowadays, a local anesthetic is injected right at the point of worst pain.

3. Ether is another colorless, pleasant smelling & partly poisonous gas that's also used as an industrial solvent!

During its early use, it caused several deaths!!

4. Chloroetherene (vinyl chloride) is a sweet smelling & colorless gas that's also highly flammable.

Extra Warning: This chemical burns quite easily!! It should be handled with Extreme Care...