Tikhonov, 40, last played for Spartak in a Champions League match against Real Madrid in September 2000, before being unceremoniously dropped by club boss Oleg Romantsev days later.
He supposedly retired at the end of last year before joining Spartak as assistant coach for the start of the new season but registering as a player.
The former Russia winger was one of the most active players on the Luzhniki pitch on Wednesday before being substituted to a standing ovation in the 53rd minute.
Spartak scored twice in three minutes through Argentine defender Marcos Rojo and Brazilian striker Welliton midway through the second half.
Krasnodar striker Spartak Gogniyev pulled one back for the Premier League newcomers late in the game but the home side held on to record their first victory in any competition for over a month.
Head coach Valery Karpin resigned on Monday after seeing Spartak lose four out of their last five matches and drop to the bottom of the table.
Tikhonov was pleased with his and the team's efforts but did not say if he would play again this season.
"I'm happy to last 50 minutes and help my team," he said.
"I'm also happy to see so many fans in such difficult times for our team. We can play better than we did tonight and we'll try to improve little by little."
ZENIT LOSE AGAIN
CSKA Moscow got the better of Zenit St Petersburg for the second time in 10 days, this time on the pitch when they won 2-0 to reach the semi-finals where they face city rivals Spartak.
Last week, CSKA were awarded a 3-0 "technical" win over Zenit in a league match, which ended in a 1-1 draw, after the Russian champions did not include any home-grown players in their lineup.
On Wednesday, goals from Ivory Coast striker Seydou Doumbia and Russian international Sergei Ignashevich either side of the interval gave CSKA a well-deserved victory in St Petersburg.
Elsewhere, Dynamo Moscow suffered a 2-1 upset by unfashionable Rostov, putting more pressure on their embattled Montenegrin coach Miodrag Bozovic.
Rostov will face second division side Alania Vladikavkaz, who reached the last four without kicking a ball after their quarter-final opponents Saturn Ramenskoye were liquidated with debts of more than $26 million at the end of last year.
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