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Those are generally speaking the bosses I consider to be hardest ones for me in the game, DLCs included, but obviously a lot of it depends on the difficulty setting and the way you play the game. For example in a 0% playthrough someone like Vanilla can be quite a challenge compared to for example Miriam who doesn't do a lot in a 0% playthrough.
On the other hand on a regular playthrough on higher difficulties such as hell or BEX, Cocoa 2 and Ashuri 2 can be a good road block if you attempt to try them really early in the game.
Only boss who almost made me rage quit my BEX playthrough was Noah 2 (yes, the blue floating phase), aside from that I never felt rage quitting the game but the Pillar fight from the Order DLC made me take about a year break from my BEX playthrough until I cleared that DLC first on hell difficulty, got better at the game and was able to continue my BEX playthrough after that.
So, I don't think there is an easy answer. Every time you defeat a boss that you thought was the hardest one in the game there might be something even harder, and at the same time the one you defeated feels just a little bit easier by comparison.
I remember that when I fought Irisu for the first time on my very first playthrough on the normal difficulty, I thought it was the hardest thing I had ever done and there's no way the game could get harder than that. Oh, how naive I was back then. I mean, come on. Irisu on normal difficulty is something you can almost do blindfolded.
But yeah, SP Miru truly is a nightmare to see and SP Irisu really showed what a final boss should look like. I have nothing but praise for those two in that regard.
In all seriousness, if you know me at all you probably already know my answer.
Irisu SP is the hardest boss fight in the entire game, by a mile. Not even a discussion honestly.
But even then there'll always be other roadblocks too, depending on how you play.
AFD, for example, makes you face Ashuri 2 very early without many upgrades at all, turning an already hard earlygame fight into an absolute nightmare. And I still haven't beaten that one boss before Chapter 7 due to the nature of how AFD handles buffs. But that's a silly mode anyway. Almost every boss was "the hardest boss" in that (with very few exceptions).
If you do a 0% run... I'd say it's probably Seana, due to the fight taking place underwater. I know she always took a lot of tries for me in the two 0% runs I've done. And they were both on Casual!
Lets see... The hardest boss that took me the longest to beat on my playthroughs of this game so far.
BEX (All clear, including BNA DLC) Irisu SP.
Right Path took about 15 hours of attempts. +5 additional hours for Extreme+ path, so you could say total is 20 hours on this boss. Its simply the most brutal fight in the game and has some of the nastiest patterns as well.
The scariest thing? I cant even beat her consistently. If you told me to go beat any other boss in the game. I think I could do it without major issues, I feel confident and could probably win 8 times out of 10.
But NOT Irisu SP. In her case is more like 4 times out of 10. Maybe less. Theres just too many trouble spots where the fight can go downhill FAST, its extremely punishing due to its nature!
Impossible (All clear, does NOT include BNA dlc):
Fairy Duo is the hardest one I feel. Its very hard, and Zero Offense (which activates randomly) can very easily screw you over. Patterns are quite hard, as well, honestly they are arguably harder than most of Forgotten Maiden's attacks.
AFD+BEX (All clear, incuding BNA DLC) Irisu SP.
Yeah, she returns. And this time its even worse...took almost 20 hours of attempts to beat her Extreme+ version. And if you think about it, I already had 20 hours of experience on vanilla BEX so holy crap, this many hours and the freaking boss is still difficult. If that doesnt convince you the fight is the spawn of satan, nothing will.
I would like to see someone beating this boss even with Consumables or Buffs. Is probably still very hard. But I hardly find videos that cover AFD mode (a shame, its a cool game mode and I believe it does deserve more attention)
BEX 0% Item run- I havent even got to Noah yet but just thought I would mention it anyway:
Alius III is the hardest boss so far. Actually, the whole segment is very crazy; but the 3rd amulet charge is worth the pain...
Also Ashuri 2 on BEX 0% item was even harder than its AFD-BEX version. I still dislike her Cut-In attack. I die to it almost every attempt lol.
There wasnt any boss or particular spot that was much harder than anything else on Casual 0% run... Plurkwood and Keke were really annoying, and so was getting to Aruraune in the dark, but thats about it. 0% Casual is a rather chill experience IMO.
As you can see, assuming the gamemode/run can do BNA Bosses and you are wiling to go that far, I am pretty sure Irisu SP is ultimately the absolute hardest thing in the game. If you ask me at least, its a complete no-brainer.
Rita SP on Extreme+ is very luck based, if you have bad luck then that may be even harder. But you can just reset; its not consistently difficult like Irisu SP.
2nd hardest fight in the game overall is Miru SP, and possibly Forgotten Maiden if you dont use checkpoints.
Also Syaro SP IMO is quite weak on BEX. Almost feels like a normal post-game boss tier. Her attacks are actually quite tricky, but they hit for rather low damage so its relatively easy to survive..
So yeah thats my take on this. I think Irisu SP being the hardest was probably intended as well.