The only way to be 100% certain about sex, is when a female becomes gravid. There is no other definitive method that I'm aware of, unless you want to attempt to 'milk' suspect males. As anyone who has kept clown loaches in a group setting in an aquarium knows, there is a definite hierarchy among the group. I suspect that only the larger more dominate females will become gravid in each group, possibly in some groups only one dominant female. I have a mix of sex, with a female that is currently gravid, and she's been this way for months. I can only take an educated guess as to what sex the others are.