This problem also affects 512e Advanced format HDD's. I have not found any official information either. Everyone (HDD manufacturers, Tech sites etc.) pretends 'partition alignment' can solve all problems and that's it.
But you can find many forums where users experienced slowdowns with aligned partitions but FAT formatted media. I have referenced another link above:
https://msfn.org/board/topic/151798-doe ... -clusters/
Unfortunately I do not know any DOS software that can do this. But you can use RMPrepUSB on Windows to get the necessary info. Here's an example of my Advanced Format 1GB Western Digital with 'perfectly 4K aligned' 98GB FAT32 volume shown by RMPrepUSB:
FAT32
000B Bytes Per Sector = 512 (0200h)
000D Sectors Per Cluster = 64 (40h)
000E Reserved Sectors before first FAT = 32 (0020h)
0010 Number of FATs = 2 (02h)
0011 Root Entries = 0 (0000h)
0013 Total Log Sectors (small) = 0 (0000h)
0015 Media Descriptor = 248 (F8h) HDD
0016 Sectors per FAT table = 0 (0000h)
0018 Sectors per Track = 63 (003Fh)
001A Number of Heads per Cylinder = 255 (00FFh)
001C Hidden Sectors preceding Partition = 2048 (00000800h)
0020 Total Log. Sectors (big) = 204797952 (0C34F800h)
0024 Log. Sectors per FAT = 25000 (000061A8h)
0028 Mirroring Flags = 0 (0000h)
002A Version No. = 0 (0000h)
002C Cluster No. of Root Dir Start = 2 (00000002h)
0030 Log. Sector No. of FS Info Sector = 1 (0001h)
0032 First logical sector number of a copy of the three FAT32 boot sectors, typically 6 = 6 (0006h)
0040 Physical Drive Number = 128 (80h) First Fixed Disk
0042 Extended Boot Signature = 41 (29h)
0043 Volume ID Serial No. = 3256109152 (C2144860h)
0047 Volume Label = WDC_FAT
0052 FileSystem Type = FAT32
First FAT begins at LBA 2080
Second FAT begins at LBA 27080
Root Directory begins at LBA 52080
First file data (cluster 0) begins at LBA 52144
Drive 0 WDC WD10EZEX-00BN5A0 F/W Rev.=01.0 Serial No.= [ bytes = 00 00 00 00 00 00 00 00 ]
Reported size 1,000,200,658,432 bytes (931.5094GiB) Last LBA 1,953,516,910
RMPrepUSB Max 1,000,194,048,000 bytes (931.5032GiB) Last LBA 1,953,503,999
@Edit:
In case of Advanced Format 4K sector HDD's it's enough if you can get FAT/Clusters start at an LBA address divisible by 8 (8 * 512 = 4096). But for flash media more strict alignment can be necessary.